HIPPOCRATIC JOURNAL OF UNANI MEDICINE
Volume 10, Number 1, January - March 2015 Hippocratic J. Unani Med. 10(1): 1 - 136, 2015
CENTRAL COUNCIL FOR RESEARCH IN UNANI MEDICINE Ministry of Ayurveda, Yoga & Naturopathy, Unani, Siddha and Homoeopathy (AYUSH) Government of India Hippocratic Journal of Unani Medicine
Chief Patron Minister of AYUSH, Government of India
Patron Secretary, Ministry of AYUSH, Government of India
International Advisory Board Prof. G.N. Qazi, New Delhi, INDIA Prof. Talat Ahmad, New Delhi, INDIA Prof. Ranjit Roy Chaudhury, Delhi, INDIA Hakim Syed Khaleefathullah, Chennai, INDIA Dr. Fabrizio Speziale, Paris, France Dr. Suraiya H. Hussein, Kuala Lumpur, MALAYSIA Mrs. Sadia Rashid, Karachi, PAKISTAN Prof. Allauddin Ahmad, Patna, INDIA Prof. Ikhlas A. Khan, USA Dr. Maarten Bode, Amsterdam, THE NETHERLANDS Prof. Abdul Hannan, Karachi, PAKISTAN Prof. Usmanghani Khan, Karachi, PAKISTAN Prof. Rashid Bhikha, Industria, SOUTH AFRICA Dr. S.S. Handa, Haryana, INDIA Prof. Ram Vishwakarma, Jammu, INDIA Prof. Irfan Ali Khan, Hyderabad, INDIA
Editorial Board Prof. Wazahat Husain, Aligarh Prof. V.H. Talib, Dehradun Dr. (Mrs.) Nandini Kumar, New Delhi Prof. K.M.Y. Amin, Aligarh Dr. O.P. Agarawal, Delhi, INDIA Dr. A.B. Khan, Aligarh Prof. Y.K. Gupta, New Delhi Dr. (Mrs.) Neena Khanna, New Delhi Prof. A. Ray, Delhi, INDIA Dr. (Mrs.) Yasmeen Shamsi, New Delhi Dr. S. Asad Pasha, New Delhi Dr. Mohammad Khalid Siddiqui, Faridabad Prof. S. Shakir Jamil, New Dlehi Dr. Ghufran Ahmed, Aligarh Prof. Mansoor Ahmad Siddiqui, Bengaluru Dr. M.A. Waheed, Hyderabad
Editor-in-Chief Prof. Rais-ur-Rahman Director General Central Council for Research in Unani Medicine (CCRUM)
Associate Editor Dr. Khalid M. Siddiqui, Deputy Director General, CCRUM
Assistant Editors Dr. Wasim Ahmed Azmi, Deputy Director, CRIUM, Lucknow Dr. Munawwar Hussain Kazmi, Deputy Director, CRIUM, Hyderabad Dr. Shariq Ali Khan, Research Officer, RRIUM, Aligarh Mr. Aminuddin, Research Officer (Botany), CCRUM Mr. Shamsul Arfin, Research Officer (Chemistry), CCRUM Mr. Mohammad Niyaz Ahmad, Research Officer (Publication), CCRUM
Managing Editor Dr. V.K. Singh, Consultant (Botany), CCRUM
Editorial Office CENTRAL COUNCIL FOR RESEARCH IN UNANI MEDICINE Ministry of Ayurveda, Yoga & Naturopathy, Unani, Siddha and Homoeopathy (AYUSH), Government of India 61 - 65, Institutional Area, Janakpuri, New Delhi – 110 058, India Telephone: +91-11-28521981, 28525982, 28525983, 28525831/52/62/83/97, 28520501, 28522524 Fax: +91-11-28522965 • Email: [email protected] • Website: www.ccrum.net
Annual Subscription: ` 300/- (India) US $ 100/- (Other Countries) Single Issue: ` 150/- (India) US$ 50/- (Other Countries) Payments in respect of subscription may be sent by bank draft marked payable to Director General, CCRUM, New Delhi.
On behalf of Central Council for Research in Unani Medicine (CCRUM) published and printed by Prof. Rais-ur-Rehman Director General, CCRUM at CCRUM headquarters, 61-65 Institutional Area (Opposite ‘D’ Block), Janakpuri, New Delhi – 110058 and printed at Rakmo Press Pvt. Ltd., C-59; Okhla Industrial Area (Phase I), New Delhi - 110020 Contents
1. Therapeutic Evaluation of Unani Coded Drug UNIM-104 in Cases of Non-Alcoholic Fatty ...... 1 Liver Disease (NAFLD) – A Preliminary Clinical Trial Hafiz C. Mohammad Aslam, Zaheer Ahmed, Shahida Begum, Aijaz Ahmad, K. Kabiruddin, Shaikh Nikhat, Javed Ali, Abdul Kareem and Syed Jameeluddin
2. Exploration of Mechanism of Antiinflammatory Action of Habbe-Gule-Aakh ...... 9 Musarrat Nafees, N.A. Khan, K.M.Y. Amin and Ghufran Ahmad
3. Toxicity Study of Qurs-e-Hudar in Experimental Animals ...... 15 Mohd. Nadeem, Mohd. Urooj, Habibur Rahman and Shariq Ali Khan
4. Comparative Analytical Study of the Efficacy of Unani Formulations with Reference to Oligospermia 23 M. Anas and M.M.H. Siddiqui
5. Re-designing of the Unani Preparation From Powder Form into Tablet and its Standardization ...... 31 Along with HPLC Profile Aziz ur Rahman and Tajuddin
6. Development of Standard Operative Procedure (SOP) of a Toothpaste Prepared with the ...... 43 Ingredients of Sunun Poste Mughilan Mohammad Rashid, Shariq Shamsi and Roohi Zaman
7. A Review on Phytochemistry, Pharmacological Properties and Biotechnological Studies in...... 53 Valeriana officinalis L., An Important Medicinal Herb Anwar Shahzad and Taiba Saeed
8. Some Depletive Plants of Kumaon Himalaya Used in Unani Medicine for Treating ...... 73 Non-Communicable Diseases and Their Conservation Strategies Zaheer Anwar Ali, Sarfraz Ahmad and Shariq Ali Khan
9. Quality Evaluation of Jawarish-e-Ood Kibreet...... 85 Rampratap Meena, D. Ramasamy, S. Mageswari, P. Meera Devi Sri, Shamsul Arfin and Aminuddin
10. Standardization of Unani Drug Post-e-Anar (Fruit rind of Punica granatum L.) ...... 95 Nazish Siddiqui, M. Masihuzzaman Ansari, Alvia Khan, Mohd. Bilal Tafseer, Saba Viquar and Abdul Haleem
11. A Study on Diversity of Unani Medicinal Plants Used for Non-Communicable Diseases in ...... 105 Southern Western Ghats of Tamil Nadu R. Murugeswaran, K. Venkatesan, Aijaz Ahmed and Aminuddin
12. Pharmacognostical Standardization of Flowers of Althaea officinalis Linn. (Gul Khatami) ...... 117 Nitin Rai, Rampratap Meena, S. Mageswari, Shamsul Arfin, Aminuddin and Rajeev Kr. Sharma
13. Development of Standards of Sapistan (Cordia dichotoma Forst. f.) ...... 129 Abdul Haleem, Abdul Latif, Abdur Rauf, Nazish Siddiqui and Sumbul Rehman
• Instructions to Contributors
Editorial
The world-wide interest in traditional medicine has led to renewed efforts to study their medical efficacy and safety on scientific lines. This is particularly true since the dangers and shortcomings of modern medicines have started getting more apparent. Currently there had been many advances in the strategies for discovery and evaluation of drugs, particularly plant based. The input of bio-chemistry to pharmacology followed by clinical trials puts more emphasis on the mode of action of drugs, albeit it becomes clear that the activities of most drugs are not confined to one single mode of action. This has led to generating lot of new researches and there is, therefore, an enormous need for exchange of this vital information for wider use by scientific community across the world engaged in the search of new drugs of plant origin. Although the Unani medicines have been in use for centuries and are known for their therapeutic efficacies, there is a need to scientifically establish their efficacy and safety in order to achieve global acceptance. Organized research work in this system is, therefore, a need of the hour. In this direction, Central Council for Research in Unani Medicine, through its clinical, drug research, literary research, survey & cultivation of medicinal plants programme is contributing significantly for last three decades. Vitiligo, sinusitis, filariasis, eczema, malaria, infective hepatitis, asthma are some of the conditions where Unani therapies have earned recognition after scientific validation.
The Council has been publishing the peer reviewed Hippocratic Journal of Unani Medicine (HJUM), mainly to bring out fundamental and applied aspects of Unani Medicine. The journal also publishes recent advances in other related sciences and traditional medicines as well as different streams of medical sciences, which have bearing on validation and scientific interpretation of various concpts and strengths of Unani medicine.
In view of an overwhelming response, the journal earlier published twice a year, its periodicity had been changed to quarterly w.e.f. January 2008 to accommodate more articles for quick dissemination of research data among scientific community. The journal has sufficient room for invited articles from luminaries of modern medicine and sciences as well as scholars of Unani medicine. The broad areas being covered include clinical research on single and compound Unani drugs, validation of regimental therapy, Clinical and experimental pharmacological studies, standardization of single and compound drugs, development of standard operating procedures, ethnobotanical studies, experimental studies on medicinal plants and development of agro-techniques thereof, and literary research on classics of Unani medicine. The journal is also open for studies on safety evaluation of Unani and other herbo-mineral drugs, nutraceuticals, cosmotherapeutics, aromatics, oral health, life style disorders, sports medicine etc. and such other newer areas which are the outcome of modern day living.
The current issue of this journal provides 13 original and review papers in the areas of clinical research, drug standardization, pharmacology, ethnobotanical surveys and allied disciplines contributed by eminent scholars in their respective fields. It is hoped that data presented will contribute significantly in R&D sector of traditional drugs and prove to be an excellent exposition of current research efforts of scientists in this direction. Council acknowledges the authors for their contributions included in this issue and hope for their continued support in this endeavor. We wish to ensure the readers to bring out the future issues of the journal on time.
We at the CCRUM have been constantly striving to reach to higher standards and make HJUM the leading journal of Unani medicine and related sciences. In this context, we thank our learned reviewers for their invaluable inputs in improving the manuscripts. We sincerely hope and trust that the mission can be accomplished with active partnership of quality-conscious individuals and institutions. Through these lines we seek your cooperation and support in materializing our dreams about the HJUM. In this regard, we request you for your as well as your colleagues’ contributions for publication in and subscription to the journal. Further, we will appreciate if the journal is introduced far and wide. We would also welcome esteemed suggestions for achieving the highest standards of quality for the journal.
April 22, 2015
(Prof. Rais-ur-Rahman) Editor-in-Chief
Therapeutic ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract Evaluation of atty liver disease is caused by an accumulation of fat in the Unani Coded hepatocytes. Non Alcoholic Fatty Liver Disease (NAFLD) has been reported worldwide, estimated to be around 10–24% in various populations. There is no Drug UNIM-104 Fstandard medical treatment specifically for “Non-alcoholic, non-Diabetic and non- in Cases of obese Fatty liver Disease”, hence a preliminary clinical study has been conducted at RRIUM Chennai, to evaluate the safety and efficacy of the Unani coded drug Non-Alcoholic UNIM-104 in the treatment of NAFLD. It was observed that there was reduction in Fatty Liver the sign and symptoms at the end of trial with statistical improvement (p< 0.05) in Disease Hounz field (HF) Units value on CT scan of liver. (NAFLD) – A Keywords: Non-Alcoholic Fatty Liver Disease (NAFLD), Fatty liver, Tashhamul Preliminary kabid, Unani Medicine, UNIM 104 coded drug. Clinical Trial Introduction
*Hafiz C. Mohammad Aslam, Non-alcoholic fatty liver disease (NAFLD) is a common clinic-pathological condition Zaheer Ahmed, characterized by significant lipid deposition in the hepatocytes of the liver Shahida Begum, Aijaz Ahmad, parenchyma. The pathological picture bears a striking resemblance to that of K. Kabiruddin, alcohol-induced liver injury, but it occurs in individuals who deny a significant Shaikh Nikhat, history of alcohol ingestion. It comprises of a wide spectrum of liver damage, Javed Ali, ranging from simple macrovesicular steatosis to steatohepatitis, advanced fibrosis, Abdul Kareem and cirrhosis (Angulo, 2002; Angulo et al., 2003). Subsets of NAFLD which and progress to cirrhosis are being increasingly recognized as a major cause of liver- Syed Jameeluddin related morbidity and mortality with the potential to progress to liver failure (David, Regional Research Institute of 2005). The term non-alcoholic steatohepatitis, (NASH) and several other terms Unani Medicine, have been used to refer to this entity, including pseudo alcoholic liver disease, 1 West Mada, Church Street, Royapuram, Chennai-600013 alcohol-like hepatitis, diabetic hepatitis, non-alcoholic Laennec’s disease, and ○○○○○○○○○○○○○○○○○○○○○○○○○○○ steatonecrosis (Ludwig et al., 1980). However, seeing that the disease represents a spectrum of pathology, the umbrella term “NAFLD,” was first introduced in 1986, became the preferred one (Sheth et al., 1997). The spectrum ranges from simple benign fatty liver (hepatic steatosis) to NASH, characterized by fatty change with lobular inflammation, hepatocellular injury, and Mallory hyaline, progressive fibrosis, and cirrhosis, and has been defined both histologically and clinically (Tominaga et al., 1995).
NAFLD has been reported worldwide but geographic variations in prevalence are evident. Although the worldwide prevalence has not yet been determined, it has been quoted as 10–24% in various populations (Angulo, 2002; Angulo et al., 2003) and it is estimated to be the most common liver disease in the western world, and
*Author for correspondence
Hippocratic Journal of Unani Medicine 1 January - March 2015, Vol. 10 No. 1, Pages 1-7 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ its prevalence is likely to increase (Tominaga et al., 1995). It affects all racial and ethnic groups and has no age or sex predilection.
According to Unani system of medicine, the Kabid (liver) is the largest glandular organ in the body, It breakdown the nutrient materials, participate in the growth and development of human body and repair the wear and tear of the body (Badal- e-Ma-Yatahallal). According to Ibn-e-Sina, Kabid (liver) is considered the seat or centre of Quwa-Al-Tabi’yah (Natural Faculty) (Ibn-e-Sina 1906). It is responsible for the absorption, digestion, retention, metabolism and assimilation of food and excretion of Fudhlat (waste products) (Abusahal Masihi 810-895 A.D). In Unani classical text, Su-e-Mizaj-e-Kabid (alteration in normal temperament of liver) particularly due to kasrate buroodat (excessive coldness) somewhat resembles to NAFLD, in which liver is loaded with Fudhlat (waste products) and toxins which ultimately help in deposition of Raddi Shaham (abnormal excessive fats) (Ibn-e- Rushed, 1126-1198).
Till now there is no standard medical treatment for fatty liver diseases although several possible treatments are under investigation including Vitamin E and C, Ursodiol and other medications.9 In contrast Unani literature is replete with drugs administered for fatty liver like conditions which has the potential to detoxify the liver. However, considering necessity of novel medical intervention, the Central Council for Research in Unani Medicine, Ministry of Ayush, Govt. of India, had initiated a preliminary clinical study to test the safety and efficacy of the Unani coded drug UNIM-104 in the treatment of non-alcoholic, non-obese and non- diabetic fatty liver disease.
Materials and Methods
The study was a single blind, prospective clinical trial approved by the Institutional Ethical Committee of the Institute. Altogether 35 cases from GOPD of RRIUM, ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Chennai were enrolled in the study during the period 2010-2012 after getting voluntary informed consent from the participants. However, 5 patients were lost to follow up due to personal reasons. Subjects of either gender in the age group of 40-60 years, non-alcoholic, non-diabetic & non-obese, with or without sign & symptoms like fatigue, loss of appetite, malaise, dull ache in upper right abdomen, nausea, hepatomegaly and BMI ≤ 29.9 were included in the study. Diagnosis and grading of fatty liver was done on the basis of Ultra sonogram and CT Scan of upper abdomen. Subjects with pregnancy or lactation, those unwilling to give consent to abide by the protocol, systemic illness (cardiac, gastric, hepatic, renal, neurological disorders etc) were excluded from the study.
Clinical assessment and laboratory investigations were carried out at baseline and at 30 days interval and CT scan of upper abdomen for measuring liver Hounz
Hippocratic Journal of Unani Medicine 2 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ field (HF) unit at base line and end of the study to monitor the prognosis. The Unani coded drug UNIM-104 was given in form of ‘Majoon’ in a dosage of 5 gm twice a day with water before food for a period of 90 days. Patients were advised strictly to refrain from consumption of alcohol, high carbohydrate and high fat content diet.
Following investigations were carried out using standard laboratory techniques.
Blood: Heamogram with ESR, Fasting Blood Sugar, S. Bilirubin, SGOT, SGPT, S.Alkaline Phosphatase, S. Urea, S. Creatinine, S.Cholestrol, S.Triglycerides, HDL, LDL.
Urine: Routine and Microscopical examination.
Statistical analysis of the data was carried out using Student t test (paired). p value < 0.05 was considered to be significant.
Results and Discussion
NAFLD is an increasingly important chronic liver disease with a wide spectrum of histopathology, ranging from bland steatosis to cirrhosis. It is often asymptomatic and discovered incidentally on routine laboratory screening. In the absence of established therapies, treatment is generally directed at optimizing body weight and controlling risk factors.
In the present study it was observed that the prevalence is more in low socio economical class in age group between 40-45 years which might be due to patients attending RRIUM, Chennai from lower strata of society. It was also observed that 53.33% patients were of Balghami (phlegmatic temperament) and 36.66% were of Damavi Mizaj (sanguine temperament) suggesting that those with colder temperament and robust built are more susceptible to fatty liver thereby
○○○○○○○○○○○○○○○○○○○○○○○○○○○ substantiating the claim made by Unani physicians (Ibn-e-Sina -19.6), Ibn-e- Rushad, 1980). Majority of the patients were overweight (BMI ≥ 25), demonstrating that obesity is a predisposing risk factor for the development of NAFLD (Table-1).
Though majority of the trial participants were asymptomatic which corroborate various study findings, however dull ache in right hypochondrium was present in 69% of the cases Improvement in most of the sign and symptoms like nausea, anorexia and reduction in dull ache in right hypochondrium was recorded at the end of trial (Table 2) indicating the Mushtahi, Muhallil, Musakhkhin, and Muqawwi kabid properties of polyherbal coded research formulation (UNIM 104).
No any appreciable improvement was observed in liver function tests and lipid profile as these parameters were within normal range during the course of trial. (Table 3 & 4) There might be no biochemical evidence of liver disease as stated
Hippocratic Journal of Unani Medicine 3 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 1: Demographic data
S.No. Demographic data Unit Mean ± SD P value
1 Age Mean ± SD 44.16 ± 7.29
2 Gender Male 16 (53.33%) Female 14 (46.67%)
3 Duration of disease Mean ± SD 9.66 ± 10.57 (months)
4 BMI (Kg/m2) Base line 28.05 ± 10.17 After treatment 27.1 ± 1.68 > 0.028
5. Body weight (kg) Base line 73.8 ± 10.17 After treatment 71.26 ± 10.11 > 0.42
6. Mizaj Damavi (sanguine) 11 (36.66%) Balghami (phlegmatic) 16 (53.33%) Saudavi (melancholic) 2 (6.66%) Safravi (choleric) 1 (3.33%)
Table 2: Improvement in Sign & Symptom in NAFLD cases treated with UNIM-104
Status No. of Fatigue Malaise Nausea Loss of Hepato- Dull ache Cases Appetite megaly in RUQ
Baseline 30 12 9 6 10 7 21 (100%) (40%) (30%) (20%) (33.3%) (23.3%) (70%)
After 30 2 Nil Nil Nil Nil 6 Treatment (100%) (6.6%) (0%) (0%) (0%) (0%) (20%)
Table 3: Biochemical parameters in NAFLD cases treated with UNIM-104 ○○○○○○○○○○○○○○○○○○○○○○○○○○○ S.No. Parameters Base Line After 90 days P-Value
1 AST (U/Lt) 20.72 ± 2.58 19.93 ± 2.29 >0.21
2 ALT (U/Lt) 24.41± 2.88 24.62 ± 3.29 > 0.79
3 Alkaline phosphatise 6.27 ± 1.06 7.08 ± 1.53 > 0.20 (K.A Units)
4 Urea (mg/dl) 19.75 ± 5 21.85 ± 4.73 > 0.10
5 Creatinine (mg/dl) 0.76 ± 0.07 0.78 ± 0.05 > 0.20
by Kumar et al. (2005). Laboratory values may be normal in up to 80% of persons with fatty liver according to Tierney et al. (2010).
Hippocratic Journal of Unani Medicine 4 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 4: Lipid profile in NAFLD cases treated with UNIM-104
S. Parameters Base After After After P No. Line 30 days 60 days 90 days value
1 Serum 221.82 211.93 205.89 211.41 > 0.34 Cholesterol ±±±± (mg/dl) 45.83 40.73 37.80 38.21
2 Serum 136.79 127.58 125.20 131.96 > 0.47 Triglycerides ±±±± (mg/dl) 43.46 37.42 49.21 43.72
3 HDL (mg/dl) 42.55 - - 42.17 > 0.47 ±± 1.86 2.21
4 LDL (mg/dl) 152.17 - - 142.93 > 0.31 ±± 38.31 32.74
Computed tomography is the most reliable imaging modality in the diagnosis of fatty liver because there is a close correlation between the CT attenuation value and the degree of fat deposition in the liver parenchyma. Non-contrast CT attenuation value of the normal liver are on average 8 Hounsfield units (HU) higher than that of the spleen (Lavine et al., 2000). Normally the attenuation of the hepatic parenchyma is 50 to 75 HU in a non-contrast CT scan and declines by 1.6 HU for each milligram of triglyceride accumulated per gram of liver tissue (Subbarao et al., 2003, Crowley et al., 2007 & Goldman et al., 2008). The study revealed significant improvement (p< 0.01) in terms of H.U. unit values which enhanced from 36.69 to 46.95 (Mean) indicating that test drug (UNIM 104) significantly
○○○○○○○○○○○○○○○○○○○○○○○○○○○ decreased the accumulated fat from liver due to its Muhallil (Anti-inflammatory), Musakhkhin (calorifaecient) and Muqawwi kabid (liver tonic) properties. The haar temperament of UNIM 104 may be responsible in mobilization and redistribution of Radi Shaham, (morbid fat) which eventually decreases the deposition (Fudhlat or waste products like alcohol, fats, toxins) of liver (Table 5 & Figure 1).
Table 5: Improvement in C.T Scan H.F Unit values in NAFLD cases treated with UNIM-104
CT Scan finding Base Line After 90 days P-Value
Liver Hounz field 36.69 ± 10.33 46.95 ± 9.58 <0.01** (HF) unit
Hippocratic Journal of Unani Medicine 5 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○
Figure 1: Improvement in C.T Scan H.F Unit values in NAFLD cases treated with UNIM-104
No adverse effects were observed during and after the trial with the test drug. The maintenance of laboratory investigations throughout the course of study attests that the test drug is safe for the patients (Table 3).
Conclusion
It may be concluded that coded Unani formulation UNIM-104 is safe and effective in the management of Non Alcoholic Fatty liver. It has encouraging potential for mobilization and redistribution of deposited Raddi Shaham (abnormal excessive fats) from the liver with no major adverse effects. However, large, long-term, controlled studies may throw more information about its efficacy.
Acknowledgment
The authors are thankful to the Director General and Dy. Director General, Central
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Council for Research in Unani Medicine, New Delhi, for their valuable suggestions, timely guidance and providing technical facilities in conducting the trial at RRIUM, Chennai.
References
Angulo, P., 2002. Non-alcoholic fatty liver disease. N. Engl. J. Med. 346 (16):1221– 1231.
Angulo, P., Lindor, K.D., 2002. Non-alcoholic fatty liver disease. J. Gastroenterol. Hepatol, 17 (Suppl):S186–S190.
Crowley, L.V., 2007. An introduction to human disease – pathology and pathophysiology correlations. 7th ed. Jones and Bartlett Publishers, U.S.A., pp. 569-578.
Hippocratic Journal of Unani Medicine 6 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ David, A. Sass, 2005. Non-alcoholic Fatty Liver Disease: A Clinical Review. Digestive Diseases and Sciences 50(1):171–180.
Goldman, L., Ausiello, D., 2008. Cecil medicine. 24th ed. Saunders Elsevier, Philadelphia, pp. 996-998.
Ibn-e-Rushd Abdul Walee Mohammad (1126-1198), 1980. Kitab-ul-Kulliyat. Published and Translated by the CCRUM, New Delhi, pp 182-289.
Ibn-e-Sina, 1906. Al-Qanoon Fit-Tib (Urdu Translation by G.H. Kantoori), Vol. 2. Nawal Kishore Press, Lucknow, p. 20.
Kumar, V., Abbas, A.K., Fausto, N., 2005. Robbins and Cotran Pathologic basis of disease, 7thed. Saunders Elsevier, New Delhi, pp. 35-36, 904-907.
Lavine, J.E., 2000. Vitamin E treatment of non-alcoholic steatohepatitis in children: a pilot study. J. Pediatr 136(6):734–738.
Ludwig, J., Viggiano, T.R., McGill, D.B., Oh, B.J., 1980. Non-alcoholic steatohepatitis : Mayo Clinic experiences with a hitherto unnamed disease. Mayo Clinic Proc. 55(7):434-438.
Sheth, S.G., Gordon, F.D., Chopra, S., 1997. Non-alcoholic steatohepatitis. Ann. Intern. Med. 126(2):137–145.
Subbarao, K., Banerjee, S., Aggarwal, S.K., Bhargava, S.K., 2003. Diagnostic radiology and Imaging. 2nded. Jaypee brother’s Medical publisher’s Ltd, New Delhi, pp. 486-495.
Tierney, L.M., McPhee, S.J., Maxine, A., 2010. Current Medical Diagnosis & Treatment. 49th ed., McGraw Hill, U.S.A., pp. 614-617.
Tominaga, K., Kurata, J.H., Chen, Y.K., 1995. Prevalence of fatty liver in Japanese children and relationship to obesity: An epidemiological ultrasonographic ○○○○○○○○○○○○○○○○○○○○○○○○○○○ survey. Dig. Dis. Sci. 40(9):2002–2009.
Hippocratic Journal of Unani Medicine 7 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 8 Exploration of ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract Mechanism of abbe-Gul-e-Aakh (HGA) is an important pharmacopoeal drug of Unani medicine commonly used in a number of inflammatory diseases Anti- such as arthritis and gout etc. It contains four ingredients of plant origin drugs. All inflammatory the ingredients have been evaluated individually and shown to possess analgesic, Hanti-inflammatory and related effects. HGA as such has also been studied on Action of experimental models of acute, sub-acute and chronic inflammation and reported Habbe-Gule- to be safe and effective anti-inflammatory agent. However, its mechanism of anti- Aakh inflammatory effect has not been studied so far. Therefore, in the present study HGA was evaluated experimentally to detect its possible mechanism of action. 1*Musarrat Nafees, 2N.A. Khan, The study was carried out by inoculating the potent chemical mediators like 2K.M.Y. Amin Serotonin (5-HT), Bradykinin, Histamine and PGE1, directly in albino rats by and subplantar injection to cause edema. The animals were treated with 1000 mg/kg 2Ghufran Ahmad of 50% hydroalcoholic extract of HGA one hour before injecting the edemagen. HGA produced 28.0% (p<0.02) inhibition against 5-HT and 41.01% (P=0.001) 1Dept. of Ilmul Advia, Z.V.M. Unani Medical College & against PGE1. Although HGA inhibit the Histamine and Bradykinin induced Hospital, K.B.Hidayatullah Road, inflammation also but the inhibition was found non-significant (16.30% and 3.30% Azam Campus, Camp, respectively). It may be concluded therefore that HGA mainly acts by its anti-5HT Pune-411001 and anti-PGE1 activity. 2Dept. of Ilmul Advia, Ajmal Khan Tibbiya College, Keywords: Unani system of medicine, Habb-e-Gul-e-Aakh, Anti-inflammatory, Aligarh Muslim University, Aligarh-202002 Mechanism of action
Introduction
Habbe-Gule-Aakh (HGA) is an anti-arthritic polyherbal formulation of Unani medicine. It is commonly used in the treatment of inflammatory diseases, mainly
○○○○○○○○○○○○○○○○○○○○○○○○○○○ in arthritis and other diseases of joints (Lubhaya, 1979; Khan, ynm).
It is a solid dosage preparation available in a pill form of a gram size containing four ingredients wiz. Gule-Aakh (Calotropis procera), Zanjabil (Zingiber officinale), Filfil siyah (Piper nigrum) and Barg-e-Bans (Bambusa arundinacea Retz.) in equal proportion (Lubhaya, 1979, Khan, ynm). All the four ingredients of HGA are described to possess anti inflammatory activity, in Unani literature (Attar, 1988; Ibn Baitar, 1999; Ibn Sina, 1887). Their anti inflammatory activity has also been proved in various scientific studies (Mascolo et al., 1988; Vendruscolo et al., 2006; Bang et al., 2009; Muniappan et al., 2003).
Inflammation is a series of pathological changes associated with local vascular reaction and cellular response of the living tissue to a sub-lethal injury. All
*Author for correspondence
Hippocratic Journal of Unani Medicine 9 January - March 2015, Vol. 10 No. 1, Pages 9-14 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ inflammatory reactions are constructed from the same few elements but the mediators that affect the elements are different. A number of chemicals play major roles to carry out the process of inflammation (Harsh Mohan, 2000). As chemical mediators influence inflammatory response, in the very same manner drugs play roles to influence different mediators in order to show their effects on inflammation. This influence of drug on chemical mediators is referred as the mechanism of action of that particular drug. HGA has been described in Unani literature vividly and is widely prescribed by the physicians in the management of chronic inflammatory diseases such as arthritis, gout and lumbago etc. Further it has also been studied scientifically for anti-inflammatory and antiarthritic effect (Nafees et al., 2012) and shown to possess significant ameliorating property but its mechanism has still not been elucidated. Therefore present study was undertaken to explore its mechanism of antiinflammatory action in experimental animals. The efficacy of HGA was determined against four important inflammatory mediators
viz. Serotonin (5-HT), Bradykinin, Histamine and PGE1. Since these mediators intervene at different stages of inflammation therefore the exploration of mechanism will also indicate appropriate therapeutic application of HGA in different acute, sub acute and chronic forms of inflammation.
Materials and Methods
Barge-Bans (Bambusa arundinacea Retz.) was supplied by Forest Research Institute (FRI), Dehradun while the remaining three drugs were purchased from Dawakhana Tibbiya College, Aligarh Muslim University, Aligarh, which has valid drug license issued by Govt. of India. Botanists at department of Ilmul Advvia and department of Botany, AMU, Aligarh identified the drug samples. A voucher specimen has been deposited in the department of Ilmul Advia, AMU, Aligarh, for future reference. ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Preparation of test drug
All the ingredients were dried under shade and the powder was made from each drug separately. Then the equiproportional mixture of the four powders was extracted in 50% alcohol with the help of Soxhlet’s apparatus for a period of 6 hours. The extract was filtered and evaporated on a hot plate till it dried. The yield percentage of the extract was calculated with reference to the crude drug and it was found to be 40.23%. The dried extract however was reconstituted at the time of administration to animals in to a fresh suspension by adding distilled water to it. The suspension was administered orally with the help of a gastric cannula.
Hippocratic Journal of Unani Medicine 10 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Test for mechanism of anti-inflammatory activity
The method described by Parmar et al. (1978) was employed to detect the possible mechanism of antiinflammatory effect of HGA. Albino rats of either sex, weighing 100-150 gm were divided into 8 groups of 6 animals each. Four groups served as control and remaining four were used as test groups for each edemagen. Right hind paw edema was induced by subplantar injection of 0.1 ml. of chemical mediators viz. 5-HT (1gm/ml) to Group I and II, Histamine (1mg/ml) to Group III
and IV, Bradykinin (20μg/ml) to Group V and VI, PGE1 (1μg/ml) to Group VII and VIII one hour after the HGA treatment. The volume of the hind paw was measured by plethysmometer (UGO Basile7041) at 0 hour and at predetermined intervals
of 30 minutes after administration of 5-HT, Bradykinin and PGE1 and 1 hour after histamine injection. Each group received 1000 mg/kg of 50% ethanolic extract of HGA while the animals in Control Group received the vehicle 1 hour before the edemagen administration. Results were statistically analysed using Student’s t- test. A P-value less than 0.05 was considered significant.
Results and Observation
The mean increase in the paw volume produced by 5-HT was found to be 0.308
ml in control group and 0.218 ml. in test group. PGE1 produced edema of 0.206 ml. in control group and 0.11 ml. in test group. Histamine produced a mean increase of 0.235 ml. in control group and 0.195 ml. in test group while Bradykinin produced edema of 0.305 ml. and 0.300 ml. in the control and test group, respectively (Table 1, Fig 1).
Table 1: Effect of HGA on inflammation induced by different endemagen
Groups Paw volume (ml.) Mean Percentage ‘P’ mean ± SE increase value
○○○○○○○○○○○○○○○○○○○○○○○○○○○ in Paw of After treat- volume inhibition ment (ml.) 5-HT Control 1.52±0.15 1.83±0.17 0.308 28.0 <0.02 Treated 1.86±0.13 2.08±0.13 0.218
PGE1 Control 1.89±0.11 2.10±0.10 0.207 41.01 0.001 Treated 1.62±0.13 1.74±0.12 0.110 Histamine Control 1.74±0.12 1.97±0.03 0.235 16.30 N.S. Treated 2.00±0.04 2.15±0.02 0.195 Bradykinin Control 1.84±0.22 2.14±0.24 0.305 3.30 N.S. Treated 1.51±0.06 1.81±0.12 0.300
Hippocratic Journal of Unani Medicine 11 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 1: Effect of HGA on inflammation induced by different endemogen
Thus, HGA produced 28.0% (p<0.02) inhibition against 5-HT and 41.01%
(P=0.001) against PGE1. Although HGA inhibit the Histamine and Bradykinin induced inflammation also but the inhibition (16.30% and 3.30%, respectively) was non-significant.
Discussion
HGA produced significant effect against 5-HT and PGE1 induced edema suggesting that its action is mediated through the inhibition of these two edemagens. As far as the remaining two chemical are concerned HGA produced little response that did not amount to be significant statistically. The inflammatory process is mediated by many chemicals. Vasodilatation is caused by Histamine,
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Kinins and particularly by Prostaglandin E and Prostacyclin. Many factors that are able to increase vascular permeability have also been identified. There are two or more phases of increased permeability governed by different mediators. An initial phase begins within the few minutes of injury and lasts for half an hour. It is usually caused by vasoactive amines like Histamine and Serotonin (Rang et al, 2003, Ritchie, 1990). The delayed phase which reaches a maximum in few hours and persists as long as the inflammation lasts is mediated by prostaglandin E. In the present study HGA was observed to produce significant antiinflammatory
effect against PGE1 and 5-HT induced inflammation. The maximum effect however
was found against PGE1 induced inflammation while the effect against 5-HT induced inflammation was although significant statistically but was comparatively of lesser degree. Therefore, it can be inferred that HGA is less effective in early phase of inflammation. This is so, because 5-HT, Bradykinin and Histamine are
Hippocratic Journal of Unani Medicine 12 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ the chemicals which mediate the inflammatory reactions in early phase and the test drug found to be less effective against these mediators. On the other hand
HGA was able to reduce the inflammation mediated through PGE1 enormously therefore it may be deduced that the test drug is more effective in delayed phase of inflammation and may be therapeutically useful in chronic inflammatory conditions. This finding is in conformity with our previous report where it has been shown that HGA has very striking antiarthritic effect (Nafees et al., 2012). A drug specially that having antiinflammatory activity such as HGA if used over a long period of time, is liable to induce side effects particularly the gastritis and gastric ulcer etc. However, one of the ingredients of HGA viz. Zanjabil (Zingiber officinale) possesses comparatively weak anti-inflammatory effect but has been reported to protect the gastric mucosa significantly (Nanjundaiah et al., 2011). Inclusion of Zanjabil in the compound is probably meant to minimize the side effect of HGA which is likely to be used for long period of time in chronic inflammatory condition.
It indicates that HGA is effective and safe PGE1 inhibitor.
Conclusion
It can be concluded therefore that HGA produces anti-inflammatory activity mainly
through inhibiting PGE1. Since this mediator is responsible for delayed phase of increased permeability and inflammatory response therefore it is likely that the test drug will be more useful in the management of chronic inflammatory conditions.
References
Attar, H.Z., 1988. Ikhtiyarat-e-Badiyee (Kanpur). Matba Munshi Naval Kishor, Lucknow, p. 393. Bang, J.S., Oh, D.H., Choi, H.M., Sur, B., Lim, S., Kim, J.Y., et.al., 2009. Anti-
○○○○○○○○○○○○○○○○○○○○○○○○○○○ inflammatory and antiarthritic effects of piperine in human interleukin 1β-stimulated fibroblast-like synoviocytes and in rat arthritis models. Arthritis Res Ther. 11(2): 49. Harsh, Mohan, 2000. Text book of pathology, 4th ed. Jaypee Brothers Medical Publishers (P) Ltd., New Delhi, pp. 114,121-122. Ibn-e-Baitar, 1999. Al-Jamiul Mufradat-al-Advia Wal Aghzia, Vol III & IV. Central Council for Research in Unani Medicine, New Delhi, pp. 274, 276, 377, 378, 380. Ibn-e-Sina, 1887. Tarjuma-e-Qanoon. (Translated by Moulvi H. S. Ghulam Husnain), Vol II. Matba Munshi Naval Kishor, Kanpur, pp. 93,184,186,187,197. Khan, A.G., ynm. Maseehul Mulk ke Murakkabat. Vol.II. Urdu Dawakhana, pp. 77-78.
Hippocratic Journal of Unani Medicine 13 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Labhaya, R., 1979. Delhi ke Muntakhab Murakkabat, Goswami Kutubkhana, Delhi, p. 89.
Mascolo, N., Sharma, R., Jain, S.C. and Capasso, F., 1988. Ethnopharmacologic investigation of Calotropis procera flowers. J. Ethnopharmacol. 22(2): 211-221.
Muniappan, M. and Sundraraj, T., 2003. Anti-inflammatory and anti-ulcer activities of Bambusa arundinacea. J. Ethnopharmacol. 88(2-3): 161-167.
Nafees, M., Khan, N.A., Amin, K.M.Y., Ahmad, G., 2012. Evaluation of Anti- inflammatory activity of Habbe-Gule-Aakh in albino Rats. Hippocratic Journal of Unani Medicine 7(4): 1-9.
Parmar, N.S., Ghosh, ,M.N., 1978. Anti-inflammatory activity of gossypin isolated from HVLinn. Indian Journal of Pharmacology 10:277-93.
Rang, H.P., Dale, M.M., Ritter, J.M., Moore, P.K., 2003. Pharmacology, 5th ed., Churchill Livingstone, New York, p. 219.
Ritchie, A.C., 1990. Boyd’s Text Book of Pathology, Vol.I, 9thed., Lea and Fabiger, Chesterfield, pp. 61,62,73,74.
Siddaraju, M., Nanjundaiah, Harish Nayaka, Mysore, Annaiah, and Shylaja M., Dharmesh, 2011. Gastroprotective Effect of Ginger Rhizome (Zingiber officinale) Extract: Role of Gallic Acid and Cinnamic Acid in H+, K+-ATPase/ H. pylori Inhibition and Anti-Oxidative Mechanism. Evidence-Based Complementary and Alternative Medicine. Article ID 249487.
Vendruscolo, A., Takaki, I., Bersani-Amado, L.E., Dantas, J.A., Bersani- Amado, C.A., Cuman, R.K.N., 2006. Anti-inflammatory and antinociceptive activities of Zingiber officinale Rosc. essential oil in experimental animal models. Indian J. Pharmacol. 38(1): 58-59. ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 14 Toxicity Study ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract of Qurs-e-Hudar he present study was carried out to investigate the safety of Qurs- in Experimental e-Hudar by conducting acute and sub-acute toxicity in Swiss albino mice & rats respectively. Acute toxicity was determined by administering aqueous extract of Animals TQurs-e-Hudar orally to two groups of mice of six each at a dose 1gm/kg and 2gm/ kg body weight. The animals were observed for gross behaviour and mortality for *Mohd Nadeem, 24 hours after drug administration. The formulation was well tolerated by the Mohd Urooj, animals and no abnormality was observed in the general behaviour of the animals Habibur Rahman and and no overnight mortality was recorded. Similarly, Sub-acute toxicity was Shariq Ali Khan determined in albino rats by orally administration of aqueous extract to three groups of seven animals each at a dose ranges from 1 gm/kg and 3 gm/kg body weight Pharmacology Research Unit for 28 days. The results of haematology and biochemistry profile done on 29th Regional Research Institute of Unani Medicine, day were found to be normal and no changes were observed in organ to body wt Post Box 70, Aligarh-202002 ratio of liver, heart, kidney and spleen.
Keywords: Acute toxicity, Sub acute toxicity, Qurs-e-Hudar, Zingiber officinalis, Strychnos nux-vomica, Detoxification
Introduction
Some drugs of the plant, animal and mineral origin used in Unani system of medicine have components that are naturally toxic in their properties. Such drugs, are therefore, detoxified in order to reduce/ overcome their toxicity. In Oriental medicine Strychnos nux-vomica L. (Loganiaceae) seeds have been used in activating the channels, alleviating pain and reducing swelling (Bensky and Gamble, 1989). The pharmacological effects of this plant have also been known to increase spinal reflexes and stimulate respiratory and sensory centres of the cerebral cortex (Chung and Shin, 1989). Azaraqi is a deadly poison and is detoxified ○○○○○○○○○○○○○○○○○○○○○○○○○○○ before its use for the preparation of drug. Its main constituent is alkaloid strychnine with human LD50 as 0.7-2.1 mg/kg (Jackson and Marsh, 1997). In the present study the drug has been evaluated for its toxicity in Swiss albino mice and rats of either sex.
Determination of efficacy and safety of herbal remedies is necessary because many people using these agents as self medication. Since there is limited data available about the safety of the commonly used herbal remedies, therefore, effort to elucidate health benefits and risk of herbal medicine should be intensified (Anonymous, 2011). Toxicity testing in animal is typically the initial steps to determine the effect profile of test substance and potential hazards which occurs due to short term exposure of test substance. A toxicity study provides information
*Author for correspondence
Hippocratic Journal of Unani Medicine 15 January - March 2015, Vol. 10 No. 1, Pages 15-22 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ on the hazardous properties and allows the substance to be ranked and classified according to the Globally Harmonized System (GHS) for the classification of chemicals which cause acute toxicity (OECD 2000).
Formulation
Qurs-e-Hudar is a poly herbal formulation containing below mentioned constituents in equal parts
S.No. Common Name Botanical Name
1 Azaraqi Strychnos nux-vomica
2 Zanjabeel Khushk Zingiber officinalis
3 Filfil Siyah Piper nigrum
Methodology
This study was carried out in Pharmacology Research Unit, Regional Research Institute of Unani Medicine, Aligarh, and conducted in accordance with the protocol approved by Institutional Ethics Committee of RRIUM Aligarh.
Procurement of Drug:
The Qurs-e-Hudar formulation was procured in the form of Tablet from Central Research Institute of Unani Medicine, Hyderabad.
Detoxification of Azaraqi (Nux Vomica) Seeds:
Azaraqi was detoxified by pharmacy section of Central Research Institute of Unani Medicine, Hyderabad, as per the method given in National Formulary of Unani ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Medicine (Anonymous, 1981b). 70 grams of Azaraqi seeds were buried in Peeli Matti (yellow clay) for 10 days. The clay was kept moist throughout. The seeds were then removed and washed. The seed coat was peeled off with the help of a sharp knife and the cotyledons were separated after removing the embryo. The healthy cotyledons were then washed with hot water. The cotyledons in a cloth bag were boiled in 2 lit of milk till it evaporated (the care was taken that the cloth bag should not touch the bottom of the container).The water washed cotyledons were then used subsequently.
Animals
The study was carried out in Swiss albino mice (20-25 g) and rats (100-150g) of either sex, for acute & sub-acute toxicity determination respectively. The animals
Hippocratic Journal of Unani Medicine 16 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ were procured from M/S Rahat Hussain Enterprises Biological Suppliers, Babri Mandi, Aligarh. They were acclimatized to the conditions for one week before experimental study. The animals were maintained in a standard environmental condition at a room temperature of (25±2 degree Celsius) with 12 Hrs light/Dark cycles, humidity (50-55%), and had free access to food pellets. The study was conducted after approval of protocol from Institutional Ethics Committee of RRIUM, Aligarh.
Preparation of Drug Extract
The tablets of the drugs were crushed into fine powder and a weighed quantity was steeped in acidulated distilled water. The water soaked mass of the drug was warmed over water bath and kept for 24 hours at room temperature. During this period it was occasionally stirred. After 24 hours it was filtered through a filter paper and filtrate was dried over water bath. The aqueous extracts of the drug thus obtained was used in different doses selected according to OECD guidelines for safety evaluations.
Acute Toxicity Study
The current study was carried out in accordance to Organization of Economic Co-operation and Development (OECD) guideline for testing of chemicals. Swiss Albino mice of either sex weighing 20-25 g were randomly selected and divide into two groups of six mice each. Mice were kept fasted overnight (12hrs) with free access to water prior to administration of dose ranging 1gm/kg body weight and 2 gm/kg body weight as per limit test of OECD guideline. The aqueous extract of the drugs was administered orally. The animals were kept in polypropylene cages after drug administration and were observed for behaviour pattern (salivation, lacrymation, lethargy, sleep, clonic convulsions, vigorous rolling and ○○○○○○○○○○○○○○○○○○○○○○○○○○○ head tilt) & mortality at 1 hour, 2 hour, 3 hour, 4 hour, 5 hour, 6 hour, 12 hour, 24 hour and thereafter once every day up to 14 days after drug administration.
Sub-Acute Toxicity Study
Swiss Albino rats of either sex weighing between 100-150 g were randomly selected and divided into three groups of seven animals each. Rats were kept fasted overnight (12hrs) with free access to water prior to administration of dose ranging 1gm/kg and 3gm/kg body weight for 28 days as per limit test of OECD guideline. Group I was kept as normal control which received distilled water for 28 days, while in the IInd and IIIrd groups aqueous extract of the drug was administered orally at a dose of 1gm/kg and 3gm/kg body weight for 28 days. The animals were kept in polypropylene cages after drug administration and were
Hippocratic Journal of Unani Medicine 17 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ observed for Gross behavior (salivation, lacrymation, lethargy, sleep, clonic convulsions, vigorous rolling and head tilt) & mortality at 1 hour, 2 hour, 3 hour, 4 hour, 5 hour, 6 hour, 12 hour, 24 hour and thereafter once every day up to 28 days after drug administration. On 29th day blood was collected of all the three groups of rats through retro-orbital plexus for estimation of SGOT,SGPT and Serum alkaline Phosphatase, Serum urea and Serum Creatinine, Serum cholesterol, Serum triglyceride and Serum HDL, Percentage hemoglobin, ESR, Total leukocyte count and Differential leukocyte count. After collection of blood the animals in all the three groups were sacrificed and liver, heart, kidney and spleen were excised out for determination of organ to body weight ratio. SGOT, SGPT were estimated by Reitman’s and Frankel, 1957 method (Reitman et al., 1957). Alkaline Phosphatase was estimated by Bessey and Brock, 1946 method (Bessay et al., 1946). Serum urea was estimated by GLDH, Ureas method given by (Tiffany et al., 1972), while Serum Creatinine was estimated by Jaffe’s method given by (Bower’s et al., 1980). Serum HDL was estimated by Phosphotungstic Acid method given by (Burstein et al.,. 1970), while Serum cholesterol and Triglyceride were estimated by CHOD-PAP method given by (Roeschlau et al., 1974) and GPO- Trindev method given by (Mcgowan et al., 1983). ESR and DLC were estimated by Westergreen and Leishman stain methods given in Medical Laboratory Technology (Mukherjee, 1990). TLC was estimated by Hemoaltometry method given by (Plum, 1936). Percent hemoglobin was estimated by Sahli’s Acid Haematin method given by (Newcomer, 1919).
Statistical Analysis
Statistical analysis was performed by using unpaired t test calculating p value at 5 % level. All values are expressed as Mean±SEM (standard error of mean). p value less than 0.05 found to be considered statistically significant ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Results and Discussion
Acute Toxicity Study
The effect of oral administration of single dose of aqueous extract of Qurs-e- Hudar in Swiss albino mice shows that the formulation was well tolerated by the animals and no abnormality was observed in the general behaviour of the animals and no overnight mortality was recorded. Herbs and supplements can be toxic when used for inappropriate indication, or prepared inappropriately, or used in large excessive dosages or for a prolonged duration of time. Since it is polyherbal formulation other ingredients present in formulation helps in reducing the toxic effect of active component.
Hippocratic Journal of Unani Medicine 18 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Sub-Acute Toxicity Study
The values of all the parameters including liver functions, renal functions hematology and organ body weight ratio found to be normal as compared to control group. The effects of the studied drug on organ body weight ratio in control and treated animals are presented in Figure 5. There were no significant changes observed in organ body weight ratio of the control and the animals treated with various doses. Figure 1, 2, and 3 is graphical representation for the effects of
Fig. 1: Effect of Qurs-e-Hudar on Liver Function Test ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 2: Effect of Qurs-e-Hudar on Renal Function Test
Hippocratic Journal of Unani Medicine 19 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ drug on the biochemical parameters. There were no significant changes in AST and ALT levels in all the treated animals compared with the control. Similarly lipid profile of treated animal was found to be normal as compared to control group as shown in Figure 3. The values for hematological parameters of treated group as compared to control showed no significant changes in Hb, TLC, ESR, and % lymphocyte as shown in figure 4. It can be concluded on the basis of above observation that drug is quite safe.
Fig. 3: Effect of Qurs-e-Hudar on Lipid Profile ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 4: Effect of Qurs-e-Hudar on Hematological Parameters
Hippocratic Journal of Unani Medicine 20 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 5: Effect of Qurs-e-Hudar on Organ to the Body Weight Ratio
Conclusion
The formulation ‘Qurs-e-Hudar’ was well tolerated by the animals and no abnormality was observed in the general behavior of the animals and no overnight mortality was recorded. There were no findings of any organ toxicity and hematological changes as laboratory findings were normal. It can be concluded on the basis of above observations that drug studied is quite safe in animals.
Acknowledgement
The authors would like to record their gratitude to Director General, Central Council for Research in Unani Medicine, New Delhi, for providing an academic and
○○○○○○○○○○○○○○○○○○○○○○○○○○○ research environment at the institute to work with excellence and meticulousness. The authors would also like to thank Dr. R.S Verma, Research Officer (Biochemistry) for his technical support and co-operation in carrying out the biochemical investigations in biochemistry laboratory of RRIUM, Aligarh.
References
Anonymous, 1981b. (reprinted in 2006). National Formulary of Unani Medicine, Ist edition. Part 1, GOI, Ministry of H&FW, New Delhi. p. 319.
Anonymous, 2011. Toxicity testing overview. Toxicity Testing Resource Centre (TTRC), pp 2-25.
Bensky, D. and Gamble, A., 1986. Chinese Herbal Medicine, Eastland press, Sealtle, p. 646.
Hippocratic Journal of Unani Medicine 21 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Bessay, O.A., Lowry, O.H., Brock, M.J., 1946. A method for the rapid determination of alkaline phosphates with five cubic millimeters of serum. Journal of Biological Chemistry 164:321-329.
Bowers, L.D., Wong, E.T., 1980. Kinetic serum creatinine assay I. The role of various factors in determining specificity. Clin.Chem 26 (5):551-554.
Burstain, M., Scholnic, H.R., Morphin, R., 1970. Rapid method for the isolation of lipoprotein from human serum by precipitation with polyanions J. Lipid. Res 11:583-595.
Chung, B. and Shin, M.K., 1989. Dictionary of Folk Medicine. Young Lin Press, Seoul. p. 972.
Jackson, T.A. and Marsh, F.P., 1997. Test methods for vertebrate pest control and management material.American Society for testing and materials, Philadelphia, pp. 1-4.
Macgowan, M.W., Artiss, J.D., Strangberg, D.R., Zak, B., 1983. A peroxidase- coupled method for the colorimetric determination of serum triglycerides. Clin Chem. 29 (3):538-542.
Mukherjee, K.L., 1990. Medical Laboratory Technology, Vol. 1, 3rd Edition. Tata, McGraw-Hill Publishing Company Limited, New Delhi, pp. 228-30.
Newcomer, H.S., 1919. Absorption Spectra of Acid Hematin, Oxyhemoglobin and carbon monoxide hemoglobin. A new hemoglobinometer J .Biol.chem. 37:465- 496.
OECD, 2000. Guidance Document on the Recognition, Assessment and Use of Clinical Signs as Humane Endpoints for Experimental Animals Used in Safety Evaluation. Environmental Health and Safety Monograph Series on Testing and Assesment No 19.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Plum, P., 1936. Accuracy of hematological counting method. Acta. Med. Scandinav 90: 342-364.
Reitman, S., and Frankel, S., 1957. A colorimetric method for determination of serum glutamic oxaloacetic and glutamic pyruvic transaminases.Am.J.Clin.Pathol.28:56-63
Roeschlau, P., Bernt, E., Gruber, W., 1974. Enzymatic determination of total cholesterol in serum. Z Klin. Chem, Klin Biochem. 12 (5):226.
Tiffany, T.O., Jansen, J., Burtis, C.A., Overton, J.B., Scott, C.D., 1972. Enzymatic kinetic rate and end- point analyses of substrate by use of a GeMSAEC fast analyzer. Clin. Chem. 18: 829-840.
Hippocratic Journal of Unani Medicine 22 Comparative ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract Analytical retrospective study was conducted to elaborate the Study of the successful treatment for oligospermia. For comparison, four studies with six common parameters were taken into account involving one hundred twenty six Efficacy of Adiagnosed patients with idiopathic oligospermia. The study was divided into four Unani groups namely A, B, C and D and it was observed that among the six common parameters, the semen volume was improved in C and B group, semen liquefaction Formulations time was raised in A, B and C groups, sperm count was maximally raised in D with Reference group, sperm motility was improved in A group, and sperm morphology was almost to Oligospermia equal in all groups and pH of semen has shown no change in all groups. Keywords: Oligospermia, Unani treatment, Herbal Medicine, Qillat-e-Huwaniya. *M. Anas and Introduction M.M.H. Siddiqui As per the WHO guidelines, a sperm count of <20 millions/ml is considered as Department of Ilaj-bit-Tadbeer, oligospermia (Dhaliwal et al., 2011). It is one of the main factors of infertility in A.K. Tibbiya College, Aligarh Muslim University, 60% of couples. Among them male infertility is involved in 40% while 20% of the Aligarh-202002 cases there is a combination of both male and female factors. The factors responsible for male infertility are low sperm count, abnormal sperm morphology and impaired sperm transport. As a remedy, a number of nutritional supplements are used to improve sperm count and sperm motility including Amino acids like carnitine, arginine, minerals like zinc, selenium, numerous vitamins and antioxidants like vitamin B12, vitamin C, vitamin E and glutathione and co-enzymes Q10 in treating male infertility (Mukherjee et al., 2003). Male infertility also involves harmful environmental and occupational risk factors along with underlying nutritional imbalances therefore; to overcome these factors including correction of nutritional imbalances to produce optimum sperm count and their functions, a multifaceted therapeutic approach can be a management tool to improve male ○○○○○○○○○○○○○○○○○○○○○○○○○○○ infertility (Prdani, 1976; Solepure et al., 1979; Sengupta, 1982).
The Unani medical literature regarding male sexual disorders is enriched with its contents and therapeutics but the terminologies of various disorders are ill defined that may be due to primitive knowledge of Physiology of the ancient and traditional Unani scholars. But the contents of therapeutics of ancient Unani physicians clearly indicate that they were well aware about oligospermia. Unani Medical literature has several authentic manuscripts that are rich and full of knowledge of sexual disorders. To explore the things on scientific parameters to validate the ancient knowledge before the present medical fraternity and public domain has encouraged us to analyzed oligozoospermia which is a major challenge before sexual internist and general physician as well.
*Author for correspondence
Hippocratic Journal of Unani Medicine 23 January - March 2015, Vol. 10 No. 1, Pages 23-29 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Purpose of the study
Extensively and critically reviewing and analyzing already carried out four studies which have similar methodology using different drugs so as to put forth the most promising drug/drug combination that can be reassessed for its efficacy by using more comprehensive and standard protocols.
Material and Methods
This retrospective study was carried out to elaborate the promising drug combination studied in the past for the treatment of Qillat-e-huwaniya-e-maniyat (Oligospermia) on one hundred Twenty Six patients pre-diagnosed with idiopathic oligozoospermia. The study was divided into four groups namely A, B, C and D. In Group A the test drug “Safoof-e-Saalab” (a combination of Moosli Sainbhal (Bombax malabaricum DC), Saalab Panja (Orchis latifolia Linn) and Satawar (Asparagus racemosus Wild), in Group B the test drug was a powdered combination of “Saalab Misri (Orchis latifolia Linn), Khoolanjan (Alpinia galanga Wild) and Koonch (Mucuna pruriens Bak)”, in Group C the test drug was Laboob- e-Kabir Khaas (a Unani polyherbal pharmacopieal preparation) and in Group D the test drug was a powdered combination of Saalab Gatta (Orchis laxiflora Linn), Salaajeet (Styrax officinale Linn), Moosli Safaid (Asparagus adscendens Roxb), Moosli Sainbhal (Bombax malabaricum Linn) and Sataawar (Asparagus racemosus Wild). All the drugs have six gram dose in each case orally twice a day for a period of sixty days.
Results and Discussion
In the study the average semen volume before treatment was observed 3.5ml, 2.1 ml, 2.9ml, and 3.4ml in groups A, B, C and D respectively. After treatment it
○○○○○○○○○○○○○○○○○○○○○○○○○○○ was improved to 4.7ml, 3.8ml, 4.6ml and 4.4ml in the same sequence. The average improvement in semen volume was observed as 1.2ml, 1.7ml, 1.7ml and 1.0 ml in groups A, B, C, and D respectively. The maximum improvement was noted in group B and C cases, this improvement of the semen volume was may be due to drugs such as Orchis latifolia and Asparagus racemosus which possess tonic and spermatagogue activities (Alok et al., 2013)
In the study, the sperm count improvement was observed as 2.8 million sperm count in group A, 6.5 million sperm count in group B, 6.4 million sperm count in group C and 8.2 million sperm count in Group D. This effect of the formulations may be due to the drugs like Asparagus adscendens, Bombax malabaricum, which enhances the production of sperm (Sinha and Jagdale, 2013) and the drug like Orchis latifolia which correct the sluggishness of sperms and helps in better
Hippocratic Journal of Unani Medicine 24 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 1: Effect of test samples on volume of semen
Before treatment After treatment
Group Volume of No.of %age Average No.of %age Average Improve- semen patients volume patients volume ment in ml in ml in ml in ml A 2.0-3.9 14 66.7 3.5 06 28.5 4.7 1.2 4.0-7.0 07 33.3 15 71.5 B 2.0-3.9 25 83.3 2.1 25 83.3 3.8 1.7 4.0-7.0 05 16.7 05 16.7 C 2.0-3.9 17 68 2.9 01 04.0 4.6 1.7 4.0-7.0 08 03 24 96.0 D 2.0-3.9 39 78 3.4 16 32.0 4.4 1.0 4.0-7.0 11 22 34 68.0
(Siddiqui, 1991; Qasmi, 1994; Khan, 1996; Ahmad, 2011)
penetration. The formulation ingredients like Styrax officinale is suppose to be a Muqawwi-e-Aza-e-Raisa wa Umoomi (General tonic) and Muqawwi-e-Baah (Aphrodiasic) leads to increase the capacity of testes to produce more sperms.
Table 2: Effect of test samples on mean sperm count in millions
Group Mean sperm count Mean sperm count Mean improvement before treatment after treatment in sperm count A 9.9 12.7 2.8 B 14.3 20.8 6.5 C 13.3 19.7 6.4 ○○○○○○○○○○○○○○○○○○○○○○○○○○○ D 14.6 22.8 8.2
(Siddiqui, 1991; Qasmi, 1994; Khan, 1996; Ahmad, 2011)
Table 3: Effect of test samples on sperm motility in percent
Group Mean sperm motility Mean sperm motility Mean Improvement before treatment after treatment in sperm motility A 35.0 50.0 15.0 B 42.6 53.9 11.3 C 41.4 54.5 13.1 D 37.0 47.0 10.0
(Siddiqui, 1991; Qasmi, 1994; Khan, 1996; Ahmad, 2011)
Hippocratic Journal of Unani Medicine 25 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ The increase in sperm count may also be attribute to the drugs like Asparagus adscendens and Orchis latifolia which contain Protein Saponins, micronutrients like Iron, Selenium and Zinc (Ainslie, 1984; Kirtikar and Basu, 1996)
Similar to above, it was observed that group A has the maximum mean improvement in sperm motility followed by group C, group B and lastly group D.
Table 4: Effect of test samples on sperm morphology
Group Mean normal sperm Mean normal sperm Mean before treatment after treatment Improvement A 37.0 47.0 10.0 B 39.4 50.0 10.6 C 42.0 52.8 10.8 D 37.0 44.8 07.8
(Siddiqui, 1991; Qasmi, 1994; Khan, 1996; Ahmad, 2011)
Table 5: Effect of test samples on Liquefaction time
Before treatment After treatment
Group Lique- No.of % Avg. No.of % Avg. Improve- faction patients in pts in ment time min min in min (in min) A 06-10 14 66.7 11 00 00 18 07 11-15 04 19.0 13 62.0 16-20 03 14.3 08 38.0
○○○○○○○○○○○○○○○○○○○○○○○○○○○ B 06-10 19 63.3 10 01 3.3 15 05 11-15 08 26.6 16 53.3 16-20 03 10.1 13 43.3 C 06-10 15 60 9.4 01 04 14.9 5.2 11-15 09 36 13 52 16-20 01 04 11 44 D ≥ 25 05 10 25 12 24 17 -08 25-30 15 30 10 20 30-35 20 40 16 32 35-40 10 20 12 24
(Siddiqui, 1991; Qasmi, 1994; Khan, 1996; Ahmad, 2011)
Hippocratic Journal of Unani Medicine 26 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ The motility of the sperms increased may due to the combined and stimulant effect of the herbs. This activity is attributed to the drugs like Orchis latifolia, and Styrax officinale that improve the strength by virtue of general tonic as well as aphrodisiac properties (Ainslie,1984; Khan, 1885; Ghani, ynm; Majoosi, 1993)
In the study the effect of test samples on sperm morphology showed that trialed combinations have restored normal morphology of the sperms to a significant level. All the groups showed equal mean improvement but with a little difference i.e 10.8 and 10.6 in the Group A and B followed by 10.0 in Group C and 7.8 in Group D. The improvement in sperm morphology may possible due to actions of the drugs like Asparagus adscendens, Bombax malabaricum, and Asparagus racemosus (Kirtikar and Basu, 1996; Ghani, ynm; Majoosi,1993) The possible explanation to improve the morphology of the sperm may due to the strengthening of the Quwwat–e–Tanasuliya (reproductive power) of the body which in turn strengthens its Quwwat–e–Musawwira (Formative power) that is responsible for the formation and maintenance of the shape of sperm.
In the parameter ‘liquefaction time’ of semen, it was observed that in group A, B and C liquefaction time was increased by 07, 05, and 5.2 minutes respectively and this effect was attributed to the action of drugs like Asparagus adscendens, Bombax malabaricum, and Asparagus racemosus which possess Mughalliz and Moallid Mani effect claimed by various writers (Khan, 1885; Ibne sina, 1906; Majoosi, 1933;and Rabban, 1996)
Table 6: Effect of test samples on pH of semen
Before treatment After treatment
Group pH of semen No. of pts Percentage No. of pts Percentage A 7.0-7.9 05 23.8 05 23.8 ≤ 8 16 67.2 16 67.2 ○○○○○○○○○○○○○○○○○○○○○○○○○○○ B 7.0-7.9 10 33.3 10 33.3 ≤ 8 20 66.6 20 66.6 C 7.0-7.9 06 24.0 06 24.0 ≤ 8 19 76.0 19 76.0 D 7.0-7.9 20 40.0 20 40.0 ≤ 8 30 60.0 30 60.0
(Siddiqui, 1991; Qasmi, 1994; Khan, 1996; Ahmad, 2011)
Finally, it was observed that there was no change in the pH of semen before and after the treatment in all the groups. This means that all the formulations experimented do not interfere with pH of the semen.
Hippocratic Journal of Unani Medicine 27 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Conclusion
Over all, it was concluded that all the four combinations studied possess aphrodisiac/ rejuvenative, spermetogenesis, spermetagogue, antidepressive an anxiolytic activity. They increase the level of testosterone, libido. These action made the formulations therapeutically active to treat male oligospermia.
Although all the four groups have nearly similar therapeutic uses and are trialed for the same conditions but as per observations, semen volume was increased more in group C and B cases, semen liquefaction time is increased in group A, B, and C cases, sperm count was increased more in group D cases as compared to other groups, sperm motility is increased in group A as compared to rest of the groups, as far as sperm morphology is concerned it increases in all groups. Therefore, to manage oligospermia the formulation of all four groups may be specifically used as per the individual case requirement in respect to the parameters taken into account.
References
Ahmad, B., 2011, “Qillat Haiwaniyat-e-Manviyah (Oligospermia) ka Tahqeeqi mutala aur uskey Ilaj me Salab Gattah, Salajeet, Moosli Safaid, Moosli Sainbhal, Satawar ki Ifadiat ka Jaizah”, M.D (Moalejat) Thesis, Faculty of Unani Medicine, A. M. U. Aligarh.
Ainslie, W., 1984. Materia Medica, Neeraj Publication House, Delhi, pp. 24-25 and 368-369
Alok, S., Jain, S.K., Verma, A., Kumar, M., Mahor, A., and Sabharwal, M., 2013. Plant profile, phytochemistry and pharmacology of Asparagus racemosus (Shatavari): A review. Asian Pacific Journal of Tropical Disease 3(3): 242– 251. ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Dhaliwal, L.K., Gupta, K.R. and Majumdar, S., 2011. Treatment of oligospermia with Speman: A formulation of plant origin. Indian Medical Gazette, pp. 375- 379.
Ghani, N., ynm. Khazeenul Advia, , Idara Kitab ul Shifa, Delhi, pp. 542,543,788- 789,820,821,1271,1272
Ibne Sina, Abdullah Ibne Hassan, 1906. Al Qanoon Fit Tib, Vol 1st and 2nd . Munshi Naval Kishore Press, Lucknow, pp. 67-68
Jagdale, S.P., Shimpi, S., Chachad, D., 2009. Pharmacological Studies Of ‘Salep’. Journal of Herbal Medicine and Toxicology 3 (1) 153-156
Khan, A., 1885. Moheet-e- Azam, Vol 2 and 4. Matba Nizami, Kanpur, pp. 11,47 and 113-114
Hippocratic Journal of Unani Medicine 28 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Khan, B., 1996. Qillat haiwaniyat (Oligospermia) per Laboob Kabeer Khas ki Ifadiat ka Jaizah, M.D (Moalejat) Thesis, Faculty of Unani Medicine, A. M. U. Aligarh.
Kirtikar, K.R. and Basu, 1996. Indian Medicinal Plants, Vol 4, 2nd edition. International book distributors, Rajpur, Dehradun, pp. 2412, 2413, 2499-2501
Majoosi, A., 1933. Kamil-us-Sana, Umada-al-Matab , Lucknow, pp. 39-40
Mukherjee, K., Tripathi, A. and Kulkarni, K.S., 2003. Evaluation of the efficacy of Speman in the management of male subfertility. Indian J. Clinical Practice 13(11): 29-31.
Pardanani, D.S., Delima, R.J., Rao, R.V., Vaze, A.Y., Sheth, A.R. and Jayatilak, P.G., 1976. Effect of an indigenous drug (Speman) on human accessory reproductive function. Indian J. Surgery 38(1): 34-39.
Qasmi, M.A., 1994. Qillat wa Zoaf Haiwaniyat Manviyah (Oligospermia) wa Zoaf Baah (Sexual Weakness) me Salab Misri, Khoolanjan aur Tukhm Koonch ki Ifadiat ka Jaizah. M.D (Moalejat) Thesis, Faculty of Unani Medicine, A.M.U., Aligarh.
Rabban, Tabri, 1996. Firdaus Al Hikma Fit Tib. Hamdard Foundation Press, Karachi, pp. 706-715
Sengupta, S. 1982, A Clinico-pathological study on the effect of Speman on spermatogenesis in cases of oligospermia. Probe XXI (4): 275-276
Siddiqui, M.S.H., 1991. Qillat wa Zoaf-e-Haiwaniyat Manviyah (Oligospermia) ka Tahqeeqi mutala aur uske ilaj me Safoof Salab ki Ifadiat ka Jaizah. M.D (Moalejat) Thesis, Faculty of Unani Medicine, A.M.U., Aligarh.
Sinha, B.N., Pandey A., Diwan A., Saini, 2013. Bombax malabaricum DC: A Salutary Boon. International Journal Pharmaceutical Innovations 3(2):17-19.
Solepure, A.B., Nirmala M. Joshi, Deshkar B.V. and Muzumdar, S.R., 1979. Effect ○○○○○○○○○○○○○○○○○○○○○○○○○○○ of Speman on quality of semen in relation to magnesium concentration. The Indian Practitioner, pp. 663-665.
Hippocratic Journal of Unani Medicine 29 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 30 Redesigning of ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract the Unani ablet is one of the most suitable and preferred solid dosage Preparation form used all over the world. Almost all drug molecules can be formulated as a tablet and the process of manufacturing of tablets is very simple, and flexible. from Powder TOne can administer 0.01 mg to 1 gm of a drug as a tablet by the oral route. Form into Therefore, in the present study an anti-arthritic pharmacopoeial preparation in powder form having ingredients Suranjan Talkh(Colchicum luteum), Zanjabeel Tablet and its Khushk (Zingiber officinale) and Elwa (Aloe vera) were re-designed and modified Standardization for use in the form of Tablet (Qurs) and standardized. Organoleptic characters of tablet, tests for weight variation, uniformity of diameter and thickness, hardness, Along with disintegration time and friability of tablets were conducted for standardization and HPLC Profile the values obtained indicated the compliance with the pharmaceutical standards. HPLC profile of tablet and qualitative analysis of chemical constituents present in *Aziz ur Rahman the tablet were also determined. Furthermore, tablets were also tested for the and presence of pesticidal residue by comparing HPLC profile of pesticides and tablet Tajuddin in identical conditions and the result shown absence of pesticides in the formulation.
Department of Saidla, These tablets can be used as an alternative of powder form of the given formulation A.K. Tibbiya College, and the findings will help to set the standards for future reference. Aligarh Muslim University, Aligarh-202002 Keywords: Re-design, Colchicum luteum, Zingiber officinale, Aloe vera, Tablet, and HPLC
Introduction
The Unani System of Medicine and its pharmaceutics are most suitable to the people of the Indian subcontinent as it was not only the part of culture and heritage but has a long cherished history during the ages of different dynasties and reigns. But in this scientific era fast paced life style emerged with exploration of various ○○○○○○○○○○○○○○○○○○○○○○○○○○○ scientific inventions in the biomedical fields and pharmaceutical sciences in accordance with the need and requirement of the modern age. The pharmaceutical industries of traditional and herbal medicine came up with the invention of convenient dosage form development in accordance with the demand of biomedical requirements as well as to provide better compliance to the patient.
Tablet is one of the most convenient and appropriate solid dosage form provided an accurately measured dose in a convenient portable package; and can be designed to protect unstable medications or disguise unpalatable ingredients. Tablets are most stable of all oral dosage forms and easy to swallow. They are very economical also (http://tabletsdosageform).
*Author for correspondence
Hippocratic Journal of Unani Medicine 31 January - March 2015, Vol. 10 No. 1, Pages 31-42 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Therefore, in view of above and in order to achieve global acceptance, a modification has been made to pharmacopoeial Unani formulation mentioned in “Ilaj al Amaraz” (Khan, 1870) i.e. from powder form in to the tablet as most of the dosage forms falls in this category, by adding Gum Acacia (S. d. Fine Chemical Ltd.) as an excipient along with its standardization which can also provide the better compliance to the patient. Further, one of the ingredients of the formulation that is Aloe is extremely bitter in taste and the consumption of the drug per oral in powder form would be more distasteful to the patient which can be easily used in the form of tablet. Parameters for tablet standerdisation include organoleptic characters, physical parameters, qualitative analysis and HPLC profile of tablet. In the form of tablet the formulation seems convenient to take and easy to handle by the practitioners and patients. The dosage of the formulation becomes more specific and accurate in the form of the tablet.
Ingredients of formulation
(1) Sonth (Zingiber officinale Linn. – Dried Rhizome) 3.5 g (2) Suranjan talkh (Colchicum luteum Baker – Dried Corm) 3.5 g (3) Elwa (Aloe vera Linn. – Dried Exudate) 7.0 g
Material and Methods
Crude drug collection and authentication
The raw materials were procured from the local market of Aligarh and the samples were authenticated in pharmacognosy section of the department of Ilmul Advia, Faculty of Unani Medicine by Prof. S. H. Afaq and found within range of standards as mentioned in Ayurvedic and Unani Pharmacopoeia of India (Anonymous, 1999, 2001 and 2007). ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Preparation of tablet
The two ingredients of formulation ‘Sonth’ and ‘Suranjan talkh’ were powdered in an electric grinder and Elwa was made Mushawwa (broiled or roasted) by keeping in an Apple, covered it by the process of Kapoorti, and heated in an oven, till it was brown. Now, Elwa was taken out of apple and dissolved in distilled water as required, after few minutes it was filtered and then concentrated. All the three ingredients were mixed together in order to make Lubdi (dough). The mixture was dried in shed and then powdered in a mortar. This powder was mixed with a suitable inert substance viz. powder of Gum Acacia (S.d. Fine Chemical Ltd.) as excipient. The material in the requisite degree of fineness mixed and damped with a moistening agent (purified water). The moistened material was made into granules by passing through a sieve. The granules were dried in shed and again
Hippocratic Journal of Unani Medicine 32 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ passed through a sieve for uniformity of size. Tablets of 500 mg were made by Automatic tablet making machine in Dawakhana Tibbiya College, AMU, Aligarh (Anonymous, 1968; 1970).
Standardization of the tablet dosage form
The tablets were evaluated for various quality control parameters like organoleptic characters (appearance, color, smell, taste and texture), weight variation, diameter, thickness, hardness, friability and disintegration time. Qualitative analysis and HPLC profile of tablet was also done. Tablets were also tested for the presence of pesticides by comparing the HPLC profile of tablet with the HPLC of standard samples of pesticides.
Organoleptic characters
Twenty tablets were taken to check for appearance and any discoloration or degradation, if any, by visual method (Patel et al., 2010). Tablets were also tested for their smell, taste and texture. All the data obtained were based on random selection and multiple observations.
Weight variation of tablet
Ten tablets were selected randomly and weighed individually by an Electronic weighing machine to check for weight variation and was expressed in mg. Weight ± 5% deviation is acceptable for the tablets / pills having average weight 250mg or more, hence considered as appropriate (Subrahmanyam et al., 2006; Dandagi et al., 2006).
Diameter of tablet
Uniformity of Diameter was assessed by picking three tablets randomly and the
○○○○○○○○○○○○○○○○○○○○○○○○○○○ diameter was measured individually by using a Digital Vernier Caliper and expressed in mm. (Subrahmanyam et al., 2006; Dandagi et al., 2006).
Thickness of tablet
Uniformity of Thickness was assessed by picking three tablets randomly and the Thickness was measured individually by using a Digital Vernier Caliper and expressed in mm. (Subrahmanyam et al., 2006; Dandagi et al., 2006).
Hardness of tablet
Hardness or Tablet crushing strength (the force required to break a tablet in a diametric compression) was measured using Monsanto Tablet Hardness Tester and expressed in kg/cm2 (Subrahmanyam et al., 2006; Vijaya and Mishra, 2006).
Hippocratic Journal of Unani Medicine 33 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Determination of disintegrating time
The rate of disintegration was measured by a Disintegration-Testing Apparatus using the two media, the aqueous and the acidic and expressed in minutes. For measurement in aqueous medium double distilled water was taken and for measuring in acidic medium simulated gastric fluid (pH about 1.2) was prepared without enzyme by dissolving 1gm of NaCl in 500 ml of distilled water, adding 7 ml of concentrated HCl, and diluting the volume to 1000 ml with water (Subrahmanyam et al., 2006; Anonymous, 1989) and modified by Afaq et al. (1994).
Friability test
Friability is the loss of weight of tablet in the container / package, due to removal of fine particles from the surface. Friability of tablets was determined by using Roche Friabilator and expressed in percentage (%). This device subjects the tablets to the combined effect of abrasions and shock in a plastic chamber revolving at 25 rpm and dropping the tablet at a height of 6 inches in each revolution. Pre weighed sample of tablets were placed in the friabilator and were subjected to 100 revolutions. Tablets were de-dusted using a soft muslin cloth and re-weighed. The friability (f) is calculated by the following formula.
⎛ W0 ⎞ f = ⎜1– — ⎟ x100 ⎝ W ⎠
Where, W0 is the weight of the tablets after the test and W is the weight of the tablets before the test (Subrahmanyam et al., 2006; Vijaya and Mishra, 2006).
Qualitative analysis of chemical constituents
The qualitative analysis of different chemical constituents, present in the tablet
○○○○○○○○○○○○○○○○○○○○○○○○○○○ was carried out according to the scheme proposed by Bhattacharjee and Das (1969). This scheme has been given in the form of Flow Chart (Fig. 1).
The powder of compound formulation was extracted with petroleum ether. The petroleum ether extract (I) was tested for free phenols, alkaloids and sterols/ terpenes. A part of this extract was saponified and sap portion (II) was tested for fatty acids, whereas, unsaponified portion (III) was tested again for alkaloids, phenols, and sterols/terpenes for confirmation. The defatted marc was divided into two portions. One portion was extracted with hot water and the other with ethanol (70%). The aqueous (IV) and ethanolic (V) extracts were tested for alkaloids, flavonoids, saponins, sugars and tannins. Aqueous extract was extracted with ether and ether soluble portion (VI) was tested again for alkaloids, sterols/ terpenes, whereas, water-soluble portion (VII) was tested for glycosides. The water- soluble portion was again hydrolyzed with 5% hydrochloric acid and extracted
Hippocratic Journal of Unani Medicine 34 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 1: Scheme for qualitative analysis of chemical constituents in the tablet
with chloroform. The aglycone portion (VIII) was tested for insoluble hydrochloride of alkaloid. Chloroform soluble portion (IX) was tested for alkaloids and sterols/ terpenes, whereas; water-soluble fraction (X) was tested for alkaloids. One part of this water-soluble portion was basified with any alkali (ammonia) and extracted with immiscible solvent (ether). The solvent soluble part (XI) was again tested for alkaloids.
Hippocratic Journal of Unani Medicine 35 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Qualitative chemical analysis of the tablet for above mentioned compounds were carried out by different tests and methods for the determination of the presence of active compounds (Overtone, 1963; Harborne, 1973 and Afaq et al., 1994).
HPLC profile of the tablet
General HPLC profile of the hydroalcoholic extract of the tablet was done by a Cyber lab’s HPLC system equipped with a single pump and porous silica with 5 ìm diameter, C 18 4.6×250 mm column and software driven peaks were obtained. The pressure, temperature and flow rate was 7 Pa, 250C and 1.0 ml/min, respectively. Detector for HPLC was UV and the wave length was 254 nm. Mobile phase for HPLC profile of extract consisted of acetonitrile: water (55:45). The hydroalcoholic extract of coarsely powdered drug was determined with the help of Soxhlet’s apparatus for 6 hours, extract was filtered and allowed to evaporate on water bath. 20mg of hydroalcoholic extract was dissolved in 1ml of HPLC grade distilled water and used for study.
Determination of Pesticidal residue
Common pesticides (Chloropyriphos, DDT, Parathion, Malathion and Endosulphan) were obtained from Sigma-Aldrich, mixed properly and dissolved in Acitonitrile. This standard was injected in the C18 column of HPLC instrument (Cyber lab, USA) considering the retention time in the identical conditions as in the HPLC profile of the test drug and software driven peaks were obtained. These peaks were compared with the peaks of tablet.
Results and Discussion
Organoleptic characters are the first step to identify the drugs and also indicate
○○○○○○○○○○○○○○○○○○○○○○○○○○○ the status and condition of the drug. If any discoloration or black spots appears, it shows the degradation or decomposition of the in the tablet (Patel et al, 2010). Organoleptic properties like colour, taste, smell, appearance and texture were noted and depicted in Table 1.
Table 1: Organoleptic description of formulation tablet
Appearance Tablet Colour Dark Brown Smell Agreeable Taste Pungent Texture Hard
Hippocratic Journal of Unani Medicine 36 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ A good quality tablet should be accurate and uniform in weight, diameter and thickness. But diameter and thickness of a tablet can vary without any change in its weight. Tablet’s diameter and thickness are controlled by the machine tooling, includes a die and punch. This is called a station of tooling. The thickness is determined by the amount of tablet material and the position of the punches in relation to each other during compression (http://en.wikipedia.org/wiki/Tablet). It should be controlled within a ±5% variation of standard value (http:// www.pharmainfo.net/namanm/evaluation-tablets). The uniformity of weight, diameter and thickness was found to be within the prescribed limit and mean values are shown in Table 2.
Tablet requires a certain amount of strength or hardness and resistance to friability to withstand mechanical shakes during manufacture, packaging, shipping and handling by the pharmacist and patient. The resistance of the tablet to chipping, abrasion or breakage under above mentioned condition before usage depends on its hardness and friability. Hardness and Friability loss indicate that tablets have good mechanical resistance and have ability to with stand abrasions. A loss less than 1% is considered acceptable by industrial standard and all the tablets were found well within approved range to give good handling properties without breakage or excessive friability problems.
Tablet should be hard enough to withstand manufacturing, packaging, and transportation process. However they cannot be too hard since that may alter disintegration or release of the drug product. Disintegration roughly indicates the possible pattern of dissolution of active substance. Hence, the experimental conditions closely mimic the situations that a tablet encounters in gastrointestinal tract, in terms of temperature, pH and mechanics. Physical parameters such as area, hardness, surface characteristics and size can significantly affect the rate of disintegration of drugs contained in a complex system. After administration, ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 2 : Parameters for standardization of tablet
S.No. Parameters Result (Mean±SE) 1. Weight variation of Formulation Tablet (g) 0.525 ±0.01 2. Uniformity of Diameter (mm) 13.54 ± 0.00 3. Uniformity of Thickness (mm) 4.77 ± 0.01 4. Hardness (kg/cm2) 2.67 ± 0.17 5. Friability (%) 0.47 ± 0.01 6. Disintegration time in the water (minutes) 12.17 ± 0.39 7. Disintegration time in the simulated gastric 10.14± 0.52 fluid (minutes)
Hippocratic Journal of Unani Medicine 37 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ the tablet should disintegrate readily for quick absorption in gastro-intestinal tract; therefore, the tablets were also subjected for the evaluation of Disintegration time. According to the test the tablet must disintegrate and all particles must pass through the 10 mesh screen in the time specified. If any residue remains, it must have a soft mass. It is mentioned that uncoated tablets must disintegrate in not more than 45 minutes (Anonymous, 1989). Results for hardness, friability loss and disintegration time illustrated in Table 2.
The therapeutic properties of the crude drugs are mainly due to presence of physiologically active chemical constituents present in the drugs. The presence and absence of active compounds are shown in Table 3.
HPLC Analysis
The HPLC profile of the formulation was recorded as the obtained graph can be compared with the batches in future. The HPLC pattern shows 15 peaks and the peak no. 1 is major peak having concentration 84.632% and retention time was found to be 1.98 min. The details are depicted in Fig. 2 and Table 4. HPLC is an important qualitative and quantitative analytical procedure. By doing HPLC we can also determine the impurities in the formulation. If there is any change in no. of peaks or retention time or area of peaks from standard it indicates adulteration or deterioration in the drug. The HPLC profile of the mixture of different pesticides
Table 3: Qualitative analysis of chemical constituents
S.No. Tested for Result 1. Alkaloids + ve 2. Amino acids - ve 3. Fixed oil / Volatile Oil + ve ○○○○○○○○○○○○○○○○○○○○○○○○○○○ 4. Flavonoids + ve 5. Glycosides + ve 6. Phenols + ve 7. Proteins - ve 8. Resins - ve 9. Saponins + ve 10. Sterols / Terpenes + ve 11. Sugar (Reducing) - ve 12. Sugars (Non-reducing) - ve 13. Tannins + ve
Hippocratic Journal of Unani Medicine 38 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig 2: HPLC Profile of the Tablet
Table 4: HPLC Profile of the Tablet
ID Name Retain.T Height Area Conc Tail. Theo. Factor Plate 1 1.908 22622 185305.8 84,632 1.43 1165 2 2.509 678 3404.8 2.536 1.00 4974 3 2.600 782 4709.3 2.926 2.00 3716 4 2.758 334 2141.5 1.250 1.89 3689 5 2.950 679 4883.2 2.540 1.00 3354 6 3.092 332 5293.5 1.242 4.54 749 ○○○○○○○○○○○○○○○○○○○○○○○○○○○ 7 3.512 167 2262.3 0.625 1.50 1339 8 3,853 145 1907.9 0.542 1.62 1709 9 4.236 245 3758.5 0.917 0.90 1520 10 4.473 138 1264.5 0.516 1.38 4750 11 4,773 177 3669.7 0.662 1.28 1056 12 5,407 234 5956.6 0.875 1.64 890 13 6,302 14 127.0 0.052 1.35 9616 14 7,472 171 1821.3 0.640 1.41 9808 15 8.025 12 93.6 0.045 1.07 21100 Sum: 26730 226599.8 100.0000
Hippocratic Journal of Unani Medicine 39 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ in Acetonitrile depicted in Fig. 3 and table 5, shows there is no any peak corresponds to the HPLC profile of tablet indicating absence of pesticides in the test drug. A comparison of both of them is also shown in Fig. 4.
Fig 3: HPLC Profile of the mixed pesticides
Table 5: HPLC Profile of the Tablet
ID Name Retain.T Height Area Conc Tail. Theo. Factor Plate ○○○○○○○○○○○○○○○○○○○○○○○○○○○ 1 1.092 23 82.9 1.197 1.19 1830
2 1,768 261 6405.7 13.580 1.24 103
3 2.268 54 378.2 2.810 2.25 2091
4 2.912 210 2042.2 10.926 1.12 1787
5 3,203 1009 11936.6 52.497 2.02 1461
6 4.030 21 294.6 1.093 1.22 1645
7 5.665 199 2523.1 10.354 1.27 3979
8 6.058 145 1701.5 7.544 1.73 5313
Sum: 1922 25364.7 100.0000
Hippocratic Journal of Unani Medicine 40 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig 4: Comparison of HPLC profile of tablet and mixture of pesticides (Peaks with sky blue and red colour indicates HPLC profile of tablet and pesticides respectively)
Conclusion
This study assumes great significance as it will provide a key of diagnostic characters specially HPLC profile which serves as an important tool in laying down the standards for quality assurance. The parameters applied for standardization of formulation viz. organoleptic characters, friability and HPLC etc. may be taken as standard parameters for future reference.
The study also offers an improvement in Unani health care system by redesigning ○○○○○○○○○○○○○○○○○○○○○○○○○○○ of powder (Safoof) form which is less convenient to use into the more convenient tablet form.
Acknowledgment
The authors are grateful to the, Dept. of Ilmul Advia, F/o Unani Medicine, A.K. Tibbiya College, Aligarh Muslim University, Aligarh, for providing significant assistance to carry out this work.
References
Afaq, S.H., Tajuddin., Siddiqui, M.M.H., 1994. Standardization of Herbal Drugs. Publication Division, AMU, Aligarh, pp. 33-34, 41-42, 100, 143-146.
Hippocratic Journal of Unani Medicine 41 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Anonymous, 1968. British Pharmacopoea, General Medicine Council. Pharmaceutical Press, Bloomsbury square, London, pp. 27-28(A.V.), 1276- 1277, 1286-1288, 982-985 (Tablet). Anonymous, 1970. Pharmacopoeia of India, 2nd ed. Ministry of Health & F.W., Manager of Publications, Delhi 238-239(A.A.), pp. 496-497. Anonymous, 1989. Food and Drug Regulations. Ministry of Health, USA, Section C. 01.015. Anonymous, 1999. The Ayurvedic Pharmacopoeia of India (Edn. 1st) Ministry of Health and Family Welfare, Govt. of India, New Delhi, Part-I, Vol.2; 12-14(Z.O.) Anonymous, 2001. The Ayurvedic Pharmacopoeia of India, (Edn.1st), Ministry of Health and Family Welfare, Govt. of India, New Delhi, Part-I, Vol.1: 62(A.V.); pp. 103-104(Z.O.). Anonymous, 2007. The Unani Pharmacopoeia of India, Ministry of Health and Family Welfare, Deptt. of AYUSH, Govt. of India, New Delhi, Part-I, Vol. 1: 82- 83(A.V.), 88-89(Z.O.). Anonymous, 2007. The Unani Pharmacopoeia of India, Ministry of Health and Family Welfare, Deptt. of AYUSH, Govt. of India, New Delhi, Part-I, Vol. 4: 7- 8(Z.O.). Bhattacharjee, A.K. and Das, A.K., 1969. Phyto-chemical screening of Some Indian Plants. Quart. Jour. Crude Drug. Res. 9. Dandagi, P.M., Halakatti, P.K., Mastiholimath, V.M., Patil, M.B., Manvi, F.V., 2006. Rapidly disintegrating Domperidone tablets. Indian Drugs 43 (7): 594-597. Harborne, J. B., 1973.. Phytochemical methods, Chapman and Hall, London, p. 70. http://en.wikipedia.org/wiki/Tablet. http://tabletsdosageform.blogspot.com/2008/07/tablet-dosage-form.html. http://www.pharmainfo.net/namanm/evaluation-tablets. ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Khan, S., 1870. Ilaj-ul-Amraz. Matba Munshi Naval Kishore, Lucknow, p. 365. Overtone K.H., 1963. Isolation, Purification and preliminary observation in elucidation of structures by physical and chemical methods. Bentley Interscience Pub., New York, p. 34. Patel S.R., Patel P. R., Vora C.N., Patel N.D. and Patel J.K., 2010. Formulation, process parameters optimization and evaluation of delayed release tablets of rabeprazole sodium. International journal of pharmacy and pharmaceutical science 2 (3): 144-156. Subrahmanyam, C.V.S., Setty, J.T., Mutta, S.K., Swamy, S.M.V., 2006. Laboratory Manual of Industrial Pharmacy, Edn. 1st. Vallabh Publication, Delhi, pp. 29-39. Vijaya, K.S.G., Mishra, D.N., 2006. Rapidly disintegration oral tablets of Meloxicam. Indian Drugs 43(2): 117-121.
Hippocratic Journal of Unani Medicine 42 Development of ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract Standard he present study was designed to convert a classical Unani Operative pharmacopoeial preparation ‘Sunun Poste Mughilan’ (a tooth powder) into toothpaste form with the same ingredients as contained in powder form. This Procedure Twork was undertaken with the objective of developing the Standard Operating (SOP) of a Procedure (SOP) for manufacturing process of toothpaste. The legitimacy of SOP was determined by assessing the quality of product in three different batches. Toothpaste Each batch was assessed three times for spreadability, foam formation and Prepared with dispersion time in water. The group which was in range with standard limits was selected as standard batch. The procedures adopted for the preparation of the Ingredients standard batch was taken as the SOP. of Sunun Poste Mughilan Keywords: SOP, Sunun, Toothpaste, Spreadability, Tooth powder.
*Mohammad Rashid, Introduction Shariq Shamsi and In Unani system of medicine, toothpowders are commonly known as Sunun which Roohi Zaman contain finely powdered drugs. Toothpowders are prepared both with herbal and
Department of Ilmul Saidla, mineral drugs but more often by combining the two. These powders usually contain National Institute of Unani Medicine, more than one ingredient. Apart from the maintenance of oro-dental hygiene Kottigepalaya, Magadi Main Road, toothpowders are also described in Unani medicine to be used in various Bangalore-560091 pathological conditions of tooth, gum, throat and buccal cavity. Although toothpowders are in use since ancient periods but due to rapid changes in modern cult, use of toothpowder is declining day by day even in rural population. Further toothpastes also have definite advantages over toothpowders which also pave the way for its wider acceptability. Toothpowders bear less spreadibility than toothpaste and usually contain coarse particles, hence chances of abrasions,
○○○○○○○○○○○○○○○○○○○○○○○○○○○ friction to enamel, dentin and soft tissue ablations are much high, leading to various periodontal diseases like dental caries, hypersensitivity of teeth, pyorrhoea, toothache, bleeding gums, gingivitis and recession of gums etc. Secondly, it is very difficult to protect powders usually containing hygroscopic, deliquescent (tending to melt or dissolve in humid environment) and aromatic materials from decomposition. Thirdly, the toothpaste has largest share of dental cleansing and care preparations. Further, chances of least wastage at use, attractive consistency, and easy distribution in mouth, and over all convenient and soft use further add to their acceptability.
In recent years acceptability and demand for natural products has increased many fold mainly because of the safety concern which is almost unavoidable with most
*Author for correspondence
Hippocratic Journal of Unani Medicine 43 January - March 2015, Vol. 10 No. 1, Pages 43-51 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ of the products prepared with synthetic raw materials. In Unani system of medicine a number of formulations intended to be used to maintain the oral health and hygiene and to ameliorate various diseases of tooth, gum, mucus membrane and throat etc are available but most of them are available in powder form such as Sunun, Zaroor, Qula and Mazmaza etc. The age-old Pharmacopoeial formulation Sunun Poste Mughilan (SPM) is used to strengthen the dental roots and impart the sparkle to the teeth (Said, 1997; Kabiruddin, ynm) and used commonly by the patients and healthy person alike. However, the change of its form is desired in view of the advantage of paste over powder and also the convenience of use of paste form. Therefore an attempt has been made to convert SPM into toothpaste. The Standardization of its procedure and product has also been undertaken to set the standard of a genuine sample and establish the SOP. Three variables i.e. spreadibility, foaming power and dispersion time were taken as the attributes for its standardization and quality determination. During the conversion of SPM into the toothpaste, different batches (F-1, F-2, and F-3) were prepared and assessed on three variables as mentioned above.
Materials and Methods
Procurement of Raw drugs
All the ingredients of SPM were procured from the raw drug dealer, except the bark of Acacia arabica which was collected from the campus of National Institute of Unani Medicine (NIUM), Bangalore. Dr. R. Sumathi, Botanist at RMR, FRLHT- IAM, Bangalore authenticated the plant drugs. The voucher specimens (Areca catechu-2741, Acacia catechu-2742, Zingiber officinalis-2743, Piper nigrum-2744, Acacia arabica-2745) have been deposited in the museum of the Institute of Ayurveda and Integrative Medicine, Bangalore. Other constituents needed to
○○○○○○○○○○○○○○○○○○○○○○○○○○○ prepare the toothpaste were procured from the Srinivasa Products, 5th Block, Rajaji Nagar, Bangalore.
Preparation of extracts
The crude drugs (Table 1) were dried in oven at 350C and coarse powder was prepared after grinding each drug separately. Then each powder was extracted separately by continuous hot extraction (Soxhlet extractor) with double distilled water at 1000C. The extracts were dried on water bath.
All the ingredients of SPM were used in the ratio of 4:1:1:1:0.1:0.1 as mentioned in Hamdard Pharmacopoeia (Table 2). Toothpaste was prepared by taking extract of all the plant drugs while the mineral drug (Silicate of magnesia) was taken as such and mixed with the extracts. Ten percent of total amount of extract of all the
Hippocratic Journal of Unani Medicine 44 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 1 : Ingredients of Sunun Poste Mughilan
Unani Name Scientific Name Part Used
1 Post Kikar Acacia arabica Bark 400gms
2 Burnt Supari Areca catechu Nut 100gms
3 Sange Jarahat Silicate of magnesia Stone 100gms
4 Kath Safaid Acacia catechu Extract 100gms
5 Zanjabeel Zingiber officinalis Dried Rhizome 10gms
6 Filfil Siyah Piper nigrum Seed 10gms
Table 2 : Composition of extracts of Toothpaste
Sl.No. Ingredients Gm
1 Acacia Arabica 5.56
2 Areca catechu 1.39
3 Silicate of magnesia 1.39
4 Acacia catechu 1.39
5 Zingiber officinalis 0.14
6 Piper nigrum 0.14
Total 10
plant drugs and Sange Jarahat was taken for the formulation of toothpaste. Other ingredients required to give the form of paste were also used (Table 3).
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Preparation of Toothpaste
Heated Liquid-Phase Process
Ten gm of extract of all plant drugs and Sange Jarahat was weighed first in the ratio of 4:1:1:1:0.1:0.1. All the ingredients were mixed with Sorbitol, Water and Saccharin sodium dehydrate in container No. 1. The solution was then heated with continuous stirring for 15 minutes at 40 0C.
In another container (No. 2), Calcium Carbonate, Carboxy Methyl Cellulose and Preservatives (Methyl Paraben and Propyl Paraben) were taken and mixed properly. A hot solution of humectants, water, extracts and sweetener which was prepared after proper heating and stirring was then, slowly added with mixing to the powder which was prepared in No. 2 container. The resulting mass was mixed
Hippocratic Journal of Unani Medicine 45 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 3 : Composition of Toothpaste
Sl. Ingredients Formulations Property No. Qty. Used (%) (% w/w) F1 F2 F3
1. Extracts 10 10 10 Active Ingredient
2. Calcium carbonate 35 45 25 Abrasive
3. Sodium lauryl sulphate 1.5 2.5 1.5 Surfactant
4. Sorbitol 30 20 20 Humectant
5. Sodium Carboxy 1 1 1 Binding agent Methyl Cellulose (CMC)
6. Sodium saccharine 0.3 0.3 0.3 Sweetener
7. Methyl paraben 0.1 0.1 0.1 Preservative
8. Propyl paraben 0.02 0.02 0.02 Preservative
9. Titanium dioxide 0.5 0.5 0.5 Opacifier
10. Peppermint oil 1 1 1 Flavouring agent
11. DD Water 20.58 19.75 40.58
well to get thick paste. Finally the Sodium Lauryl Sulphate, peppermint oil, Titanium dioxide were added and mixed properly.
Determination of spreadibility, foaming and dispersion time
Three different batches of toothpaste were prepared with different percentage of ingredients. Each batch was assessed three times for spreadability, foam
○○○○○○○○○○○○○○○○○○○○○○○○○○○ formation, and dispersion time in water.
Determination of Spreadibility
One gm of paste was placed at the centre of the glass plate (10X10 cm) and another glass plate was placed over it carefully. Above the glass plates 2 kilogram weight was placed at the centre of the plate. Sliding of the plate was avoided. The area (cm) of spread of paste was measured. The experiment was repeated three times and the average was recorded (Akelesh et al., 2010; Pael and Kamani, 2009).
Determination of Foaming Power
Five gram of paste was placed in a 100 ml glass beaker. To this 10 ml of water was added and the beaker was covered with a watch glass and allowed to stand
Hippocratic Journal of Unani Medicine 46 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ for 30 minutes, this operation was carried out to disperse the toothpaste in water. The contents of the beaker were stirred with a glass rod and the slurry was transferred to a 250 ml graduated measuring cylinder. During this transfer it was ensured that no foam was produced and no lump paste went into the measuring cylinder. The residue left in the beaker was transferred with further portion of 5-6 ml of water to the cylinder. The content of cylinder was adjusted to 50 ml by adding sufficient water and the content was maintained at 30°C. The contents of the cylinder were stirred with a glass rod to ensure a uniform suspension. Stirring was stopped when the temperature of the content reached to 30 0C and then 12 complete shakes were given by hand. The cylinder was then allowed to stand for 5 minutes and the volume of foam with water and the volume of water only was noted.
Foaming power was determined by the following formula (Akelesh et al., 2010):
Foaming power = V1 – V2
V1 - Volume in ml of foam with water
V2 - Volume in ml of water only
Determination of Dispersion Time in Water
The USP tablet disintegration test unit was used. 150 ml of distilled water at 370C+1 was placed in the cylinders of the apparatus. One gram of the toothpaste was placed, cautiously, onto the gauze of the basket. The movement of the basket was maintained at the normal speed of the apparatus and the temperature was kept at 370C+1. The time elapsed till no paste was present on the gauze was determined and was taken as a measure of dispensability of the paste in water. A time ranging from 10-30 minutes is considered satisfactory (Pander, 1993). ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Results
The quality of the toothpaste was determined by assessing each batch three times for spreadability, foam formation and dispersion time in water and the mean was regarded as standard parameter value with an aim that if the paste will qualify the criteria of quality then it will be assumed that the procedure adopted to prepare the paste is also standard. The results of all batches are given in Table 4 and Fig. 1. The batch F3 with abrasive and humectant shows spreadability, foam formation and dispersion time in water as 7.47 ± 0.09 cm, 122.33 ± 1.20 ml and 26 ± 0.58 min, respectively. Most appropriate result among three batches was found in this group. The values of all the above parameters were compared with the standard values given by Bureau of Indian Standards and with other published work (Table 5) (Fig 2). The findings on comparison were found to be within the normal range.
Hippocratic Journal of Unani Medicine 47 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 4 : Results of Formulations Batches
Sl. Formulations Spreadability Foam formation Dispersion time No. (cm) (ml) (min) 1 F1 6.00 ± 0.12 97 ± 1.2 45 ± 0.63 2 F2 4.2 ± 0.12 11 ± 0.58 55.89 ± 0.35 3 F3 7.47 ± 0.09 122.33 ± 1.20 26 ± 0.58 ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 1
Table 5: Comparison of F3 with the optimum values mentioned in Bureau of Indian Standard
Values Spreadability Foam Formation Dispersion time (cm) (ml) in water Toothpaste value 7.47 122 26 Standard value (Max.) 8.5 Min 50 10-30 min
Hippocratic Journal of Unani Medicine 48 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 2
Discussion
An important Unani powdery dentifrice SPM was successfully converted into paste form using the standard procedures and approved excipents while retaining the ingredients of the formulation. The quality standards of the newly converted formulation were tested on some important variables and it was found that the formulation qualify the criteria necessary to establish its standard and thereby the genuineness. The study demonstrated that the paste attained almost same standard as that mentioned in Bureau of Indian Standard suggesting that the ○○○○○○○○○○○○○○○○○○○○○○○○○○○ procedure adopted to prepare the paste was also genuine and standard and may be taken as the standard operating procedure.
One of the criteria for paste to meet quality standard is that it should possess good spreadability. The spreadability is used to denote the extent of area to which a paste readily spreads on application to the skin or affected part, as the therapeutic efficacy of a formulation also depends upon its spreading ability (Patel, 2012). Spreadability is imperative characteristic of toothpaste and shows the consistency of paste. The toothpastes are homogenous in nature and it should not separate into liquid and solid ingredients. The large spread area as seen in case of the paste of SPM demonstrated that it possesses good consistency (Das et al., 2013). Thickness or the consistency felt in the mouth upon using the toothpaste is important because it is directly related to the usage of the dentifrice. If toothpaste
Hippocratic Journal of Unani Medicine 49 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ is too thick, it may be difficult to spread around the teeth and it may not be used properly. If toothpaste is too thin, it may not be too effective in cleaning. Additionally, an adequate amount of thickness in paste is required to keep the abrasives properly dispersed throughout the toothpaste (Mason Stephen, 2000). This depends heavily on the amounts of gelling agents in the toothpaste, as well as on the fraction of the toothpaste which is solid. Hence, determination of spreadability is very important in evaluating paste characteristics. The mean value of spreadability of toothpaste was found to be 7.47 ± 0.09 cm which is almost equal to that of the standard value of 8.5.
A controlled level of the foam in toothpaste is necessary for suspending and removing the debris, rinsing the mouth and giving an intense perception of cleaning action. This is due to the surface tension reducing properties of detergents (Nanda et al., 2009). The quality of the foam, i.e. volume and stability or lifetime, depends upon many factors such as, the amounts and properties of humectants, polymers, and abrasives used (Pader Morton, 1997). The foam volume depends mainly on the amount of surfactant. The ratio of amount of surfactant and foam volume can be best fit line for experimental data. The foaming character was studied for nature, stability and washability of toothpaste (Mithal and Saha, 2010). The mean value of foaming power of toothpaste was found to be 122.33 ± 1.20 ml which is significantly higher than the minimum required value of 50.
Dispersion time is used to measure dispersion power of toothpaste in water. A time ranging from 10-30 minutes is considered satisfactory. The mean value of dispersion time in water was found to be 26 ± 0.58 min which was found to be within the normal range of 10-30. Thus the all three parameters the paste form of the preparation was tested for were found to be commensurate with the standard values.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Conclusion
The study demonstrated that the conversion of ‘Sunun Poste Mughilon’ (SPM) into paste form was successful as the finished product was found to qualify all the criteria laid down for toothpastes of standard quality. It was concluded therefore that the procedure used to prepare the toothpaste is also standard and it may be used as the SOP for preparation of other herbal and herbo-mineral toothpastes.
Acknowledgement
The authors are grateful to Prof. Mansoor Ahmad Siddiqui, Director, National Institute of Unani Medicine, Bangalore, for providing the financial and logistic support.
Hippocratic Journal of Unani Medicine 50 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ References
Akelesh,T., Kumar, R.S., Jothi, R.P.V.R., Raj, A., Venkatnarayan, R., 2010. Evaluation of Standards of Some Selected Cosmetic Preparations. JPRHC 2(4): 302-306.
Das, Ishita, Suki Roy, Shreta Chandni, Karthik, L., Gaurav Kumar and Bhaskar Rao, 2013. Biosurfactant from marine actinobacteria and its application in cosmetic formulation of toothpaste. Der Pharmacia Lettre 5 (5):1-6.
Kabiruddin, M., Al-Qarabadin Faisalabad, ynm. Kashmir Book Deport, Faisalabad, p. 568.
Mason, Stephan, 2000. Dental Hygiene, Poucher’s Perfumes, Cosmetics and Soaps, pp. 217-253.
Mithal, B.M., Saha, R.N., 2010. A Handbook of Cosmetics. Vallabh Prakashan, Delhi.
Nanda, S., Nanda, A., Khar, R.K., 2009. Cosmetic Technology. Birla Publication Pvt. Ltd.
Pader, Morton, 1997. Surfactants in Dental Products. Surfactants in Cosmetics. (Eds. Martin M. Rieger and Linda D. Rhein). CRC Press.
Pader, M., 1993. Dentrifice Rheology. In: Rheological properties of cosmetics and toiletries. Marcel Dekker Inc., New York.
Patel, R., Kamani, R., 2009. Formulation Optimization and Evaluation of Mometazone Furate Cream. Journal of Pharmacy Research 2(10:1565-1569.
Patel, H., Panchal, M.S., Shah, S., Vadalia, K.R., 2012. Formulation and Evaluation of Transdermal Gel of Sildenafil Citrate. IJPRAS 1(3):103-118.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Rashid, M., 2013. Formulation and Evaluation of Unani Toothpaste Composed of Ingredients of Sunun Post Mughilan: A Unani Tooth Powder. Journal of Research in Unani Medicine 2(2): 1-9.
Said, H.M., 1997. Hamdard Pharmacopeia of Eastern Medicine. Sri Satguru Publication, A Division of Indian Books Centre, Delhi, pp. 166-167.
Hippocratic Journal of Unani Medicine 51 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 52 A Review on ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract Phytochemistry, edicinal plants are serving mankind to cure ailments Pharmacological since time immemorial and today they are equally important even with a higher potential, because of the understanding of the mechanism of action with the advent Properties and Mof modern technique and extensive research. The present review looks into the Biotechnological medicinal potentials of Valeriana officinalis. To encompass the major activities of this plant, information is collected pertaining to phytochemistry, pharmacological Studies in properties and biotechnological investigation carried out in various laboratories Valeriana world-wide. Valeriana officinalis, known as Sumbu-ul-teeb in Unani system of medicine, is an important source of various bioactive compound viz. valerenic officinalis L., acid, valepotriates, alkaloids, flavonoids, lignans etc. Pharmaceutical application An Important of valerian is due to its sedative, anticonvulsant, antidepressant, antihypertensive, Medicinal Herb hypnotic, antispasmodic, antidysmenorrheal and anxiolytic activity. The literature reviewed on biotechnological approaches include in vitro protocols for large scale production of valerian, genetic transformation and elicitation studies undertaken *Anwar Shahzad and for enhanced production of valerenic acid. Taiba Saeed Keywords: Valeriana officinalis L., Tagar, Valerenic Acid, Valepotriates, Sedative Plant Biotechnology Laboratory, Department of Botany, Aligarh Muslim University, Introduction Aligarh-202002 In recent times an upsurge is witnessed worldwide towards the use of medicinal plants as a whole or in the form of crude drug in traditional systems of medicine such as Ayurveda, Sidha and Unani, which draw the attention of modern science researchers to understand the isolation, application of bioactive compounds and mechanism of action to cure diseases in a more precise way. Now a huge amount of research outcomes are available to be utilised in a judicious way. The present review deals with the application of Valeriana officinalis an important medicinal ○○○○○○○○○○○○○○○○○○○○○○○○○○○ plant of traditional system of medicine to cure various diseases as well as the use of biotechnological tools to conserve its fast depleting population because of indiscriminate collection, and the production of bioactive compounds through hairy root culture, elicitation, and genetic transformation for the overexpression of gene related to desired metabolites.
Botanical Name : Valeriana officinalis L.
Common name : All-heal, Garden heliotrope, Great wild Valerian, Amantilla, Phu, Nard, Theriacaria, Marinella, Genicularis and Terdina.
Unani name : Tagar
*Author for correspondence
Hippocratic Journal of Unani Medicine 53 January - March 2015, Vol. 10 No. 1, Pages 53-71 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ The genus Valeriana, with about 200 species, belongs to the Valerianaceae family and grows throughout the world (Piccinelli et al., 2004). Valeriana officinalis L. one of the medicinally important species of Valeriana, is a perennial flowering herb commonly known by various names such as all-heal, garden heliotrope, great wild Valerian etc (Cunha et al., 2003). Two other ancient names are “nard” and “phu”. “Nard” is derived from a Sanskrit word meaning “strong smell” and “phu” or “fu” refers to the usual exclamation of disgust that attends the experience of smelling the dried root. The word valeriana might have been derived from the Roman province of Valeria, or from Valerianus, a Roman emperor, or from a certain Valerius who first used the herb as medicine, while other writers believe it came from the Latin word valere (to be in health). The plant is native to Europe and Asia and has naturalized in eastern North America. It appears in moist places with mild climates, mainly in forests and river margins. It is cultivated in low lying, damp sandy humus with lime fertilizer (Cunha, 2005). Other important species of Valeriana includes V. fauriei used in Traditional Chinese Medicine and Japanese Medicine, V. capensis used in African traditional medicine, V. edulis used in Mexico and V. wallichii is used in India (Hikino et al., 1975; Iwu, 1993; Huang, 1999). The collective species V. officinalis L. is very polymorphic, with a number of naturally occurring subspecies that differ from each other by their degree of ploidy (Bos et al., 1997). This plant grows from a short rhizome to approximately 1.5 m in height, it flowers, and then dies back again in the winter. The rhizome is light greyish brown, about the size of a finger joint, bearing many rootlets. V. officinalis L. has pinnately-divided leaves, typically with 6 -10 pairs of lanceshaped leaflets, and bears many small white or pink hermaphroditic flowers in a dense head of several stalked clusters. These heads bear small (5 mm) tapered seeds (Andrews and Basu, 2005). The fresh root has no odor, while the dried root smells distinctly unpleasant, akin to old gym socks, due to isovaleric acid (Bisset, 1994; Flemming, 1998).
○○○○○○○○○○○○○○○○○○○○○○○○○○○ V. officinalis (Tagar) is one of the important constituents of many Unani formulations (Anonymous, 1969) which have been reported to possess some interesting pharmacological effects (Ahmad et al., 2012).
Phytochemistry
This plant contains over 150-200 chemical constituents, including the essential oils and their sesquiterpenoid derivatives (valerenic acids), epoxy iridoid esters (valepotriates), alkaloids, flavonoids and lignans which are mainly produced in the root and rhizome (Cramer et al., 2006). Among these constituents, valerenic acids and valepotriates are often regarded as active principles in valerian commercial and medicinal preparations and its crude extract is widely used in many countries (Houghton, 1999).
Hippocratic Journal of Unani Medicine 54 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Essential Oil
The essential oil contains about 150 constituents which includes monoterpenes, sesquiterpenes, less volatile sesquiterpenic carboxylic acids. Borneol (Carretero, 2000; Cunha, 2005) mainly in the form of ester viz. bornyl acetate and bornyl isovalerate , camphene, á and â-pinene are the major monoterpenes found in essential oils of V. officinalis (Carretero, 2000; Barnes et al., 2002; Heinrich et al., 2004). Sesquiterpenes include valeranone (Dewick, 2002; Heinrich et al., 2004), valerenal (Carretero, 2000; Dewick, 2002; Heinrich et al., 2004), valerenol (Barnes et al., 2002; Heinrich et al., 2004), valerianol (Carretero, 2000; Heinrich et al., 2004), bisabolene, caryophyllene, (Heinrich et al., 2004) pacifigorgiol and ledol (Barnes et al., 2002). Valerenic acids, acetoxyvalerenic, hydroxyvalerenic are less volatile sesquiterpenic carboxylic acids present in essential oil (Carretero, 2000; Dewick, 2002; Heinrich et al., 2004; Cunha, 2005). Isovaleric and hydroxyisovaleric acids are liberated when drying and storing the plant which gives the valerian its characteristic smell (an unpleasant aroma) which is not noted in the fresh plant (Weiss, 1988; Dewick, 2002).
Valepotriates
The valepotriates are epoxy iridoid esters (Dewick, 2002) that contain valtrate (Carretero, 2000; Heinrich et al., 2004; Cunha, 2005) which is about 80% of the valepotriates, acevaltrate (Cunha, 2005), dihydrovaltrate (Carretero, 2000; Heinrich et al., 2004; Cunha, 2005) isovaltrate (Carretero, 2000; Dewick, 2002; Heinrich et al., 2004) isovaleroxy-hydroxydihydrovaltrate (Caron and Reidlinger, 1999; Carretero, 2000). The valepotriates are very unstable molecules as they can be easily decomposed by heat, humidity or even pH variations (www.naturaldatabase.com). Therefore, they could only be present in fresh plants. However, small quantities of these compounds exist when the plant material is made to dry at a temperature lower than 40ºC (Carretero 2000). Being insoluble ○○○○○○○○○○○○○○○○○○○○○○○○○○○ in water valepotriates could be obtained only in non-aqueous preparations. Although valepotriates were once thought to be the active ingredients, these compounds being chemically unstable degrade readily, poorly absorbed and are not found in infusions and tinctures (Foerster et al., 1984; Houghton, 1988; Lin et al., 1991; Reichert, 1998). Instead, their degradation products, baldrinals which are unsaturated aldehydes, are found in such preparations, and may account for much of valerian’s sedative effect. The baldrinal and homobaldrinal are the degradation products of valtrate and isovaltrate respectively (von der Hude, 1986; Barnes, 2002; Heinrich, 2004).
Other Constituents
Apart from essential oils and valepotriates, the plant contains other bioactive compounds including amino acids, alkaloids, flavonoids, lignans etc. Arginine,
Hippocratic Journal of Unani Medicine 55 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ glutamine, tyrosine, ã-aminobutyric acid (GABA) are the important amino acid constituents. The major flavonoids present in V. officinalis are linarin (Fernandez et al., 2004), methylapigenin, hesperidin (Marder et al., 2003; Fernandez et al., 2004). Alkaloidal constituents include chantinine, valerine, valerianine, actinidine, methyl-2-pyrrole ketone (Torssell and Wahlberg, 1967; Franck et al., 1970; Janot et al., 1979; Duke, 1985). Hydroxypinoresinol is an important lignan found in this plant (Houghton, 1999). Other valerian constituents are caffeic and clorogenic acids, â-sitosterol, tannins, choline etc (Barnes et al., 2002)
Pharmacological Properties
Valerian has been used as a medicinal herb since at least the time of ancient Greece and Rome. Hippocrates described its properties, and Galen later prescribed it as a remedy for insomnia. Pharmaceutical application of valerian is due to its sedative, anticonvulsant, antidepressant, antihypertensive, hypnotic effects, antispasmodic and anxiolytic activity. The pharmacological effects of valerian have primarily been attributed to the valepotriates (iridoid esters), volatile oils, monoterpenes, and sesquiterpenes constituents (Wang et al., 2010; Ansari et al., 2013). Lately, the potential cytoprotective effect of aqueous extract of V. officinalis on human neuroblastoma cells has also been demonstrated (Madureira de Oliveria et al., 2009). In 1998, valerian was the tenth most popular herbal cure sold in the United States. Now, valerian extracts are available as dietary supplements, which are primarily composed of dried root or extracts from the root, formulated into tablets or soft gelatin capsules are extensively used with an estimated 210 million doses sold annually in the United States and 125 million doses sold annually in Europe (Patocka and Jakl, 2010).
Cardiovascular Activity
○○○○○○○○○○○○○○○○○○○○○○○○○○○ The coronary dilating and antiarrhythmic effects of valerian extract has been demonstrated in rabbits, mice and cats. Petkov (1979) reported that valepotriates an important component of Valerian, prevented the appearance of acute coronary insufficiency as well as vasopressin-induced arrhythmia, provoked a short-lived increase of coronary blood flow, and had moderate positive inotropic and negative chronotropic effects. In mice, valeranone another bioactive compound found in small quantities in valerian and in larger amounts in its relative, Nardostachys jatamansii, exerted weak hypotensive effects (Morazzoni and Bombardelli, 1995). A significant increase in coronary blood flow, a transient fall in blood pressure and a decrease in heart rate was noticed when cats were intravenously injected with valerian extracts (Zhang et al., 1982). Valerian is included in a German heart tonic to maintain neuro-cardiac stability (Mowrey, 1986). In an open, multicenter trial of 2,243 patients with a variety of functional cardiac disorders, an herbal
Hippocratic Journal of Unani Medicine 56 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ combination (valerian, hawthorn, cereus and camphor) was associated with improvement in 84% of patients (Busanny-Caspari 1986). Circosta et al. (2007) reported significant anticoronaryspastic, antibronchospastic, antihypertensive activities from ethanolic and aqueous extracts of V. officinalis L. roots in anaesthetized guinea-pigs and were found similar to those exhibited by nifedipine.
Anxiolytic Activity
Hattesohl et al. (2008) evaluated the pharmacological profile of different extracts derived from V. officinalis L. including two commercially available extracts and the newly developed preparations VAL SE 35E and phytofin Valerian 368. Therefore tests for sedative, anxiolytic, antidepressant as well as for myorelaxant properties were conducted in rodents. The results revealed that none of the valerian extracts displayed sedative or myorelaxant effects when used up to maximum dosages of 500 or 1000 mg/kg body weight. However pronounced anxiolytic of the 45% methanolic and 35% ethanolic extract as well as of phyotofin Valerian 368 was noticed in the elevated plus maze test in a dose range of 100–500 mg/kg body weight. Additionally and different from its primary extract (35% ethanolic extract) phytofin Valerian 368 showed antidepressant activity in the forced swimming test after subacute treatment. They concluded that anxiolytic and antidepressant activity may contribute to the sleep-enhancing properties of valerian. Further, Murphy et al. (2010) reported primary anxiolytic activity of valerian due to the presence of valerenic acid and the enhanced anxiolytic effect of valerenic acid in the presence of GABA. During their experiment rats were administered either ethanol (1ml/kg), diazepam (1mg/kg), valerian root extract (3ml/kg), valerenic acid (3mg/kg), oral solution of valerenic acid and exogenous GABA (75 mg/kg and 3.6 mg/kg, respectively) and assessed for the number of entries and time spent on the open arms of an elevated plus maze. Results showed that there was a significant reduction in anxious behaviour when valerian extract or valerenic ○○○○○○○○○○○○○○○○○○○○○○○○○○○ acid exposed subjects were compared to the ethanol control group.
Sedative/anticonvulsant Activity
Rosecrans et al. (1961) evaluated the biological activity of the different root extracts of Valeriana officinalis wherein two fractions (V103 and VI15) were found to potentiate the pentobarbital sleeping time in rats. In mice, intraperitoneal injections of valerenic acid, valerenal and whole herb extracts produced significant sedation, ataxia and anticonvulsant effects (Veith et al., 1986). Intraperitoneal injections of 100 mg/kg had sedative effects as strong as barbiturates; doses of 400 mg/kg led to death. In comparison with diazepam and chlorpromazine, valerian extract had weak anticonvulsive properties. Valerian root extract (Valdispert) reduced motility and increased thiopental and pentobarbital-induced sleeping time. Even the aroma
Hippocratic Journal of Unani Medicine 57 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ of valerian root exerted sedative effects in mice (Buchbauer et al., 1992). Marder et al. (2003) reported the sedative and sleep enhancing property of hesperidin, a compound isolated from V. officinalis. They also demonstrated the ability of 6- Methylepigenin, another compound from V. officinalis, to potentiate the sleep enhancing property of hesperidin. Fernandez et al. (2004) further reported the presence of the flavone glycoside linarin and its sedative and sleep-enhancing properties that are potentiated by simultaneous administration of valerenic acid. An intraperitoneal coadministration of linarin (4 mg/kg) and valerenic acid (5 mg/ kg) had sedative and sleep-enhancing effects as evidenced by the remarkable reduction in the exploration of holes, the time mice spent head dipping and the number of their rearings as assayed in the holeboard test and also produced a striking increase in the sleeping time induced by sodium thiopental whereas the administration of linarin and valerenic acid independently at the above mentioned doses did not increase the sleeping time induced by sodium thiopental.
Gastrointestinal Activity
Valerian is traditionally used in the treatment of intestinal spasms, colic, and “nervous stomach”. Valerian has a bitter flavor, and bitters have historically been used to enhance appetite and digestion. Valerenic acid, valtrate and valeranone exert spasmolytic effects in guinea pig ileum through direct effects on smooth muscle (Hazelhoff et al., 1982; Busanny-Caspari, 1986).
Antidysmenorrheal activity
Valerian seems to be an effective treatment for dysmenorrhea, probably because of its antispasmodic effects. In a double-blind, randomized, placebo-controlled trial, 100 students were randomly assigned to receive valerian (dose 255 mg) 3 times daily for 3 days beginning at the onset of menstruation, for 2 consecutive
○○○○○○○○○○○○○○○○○○○○○○○○○○○ menstrual cycles. At baseline and during the intervention cycles, the pain severity was evaluated with a visual analog scale and the systemic manifestations were assessed using a multidimensional verbal scale. The pain severity at baseline did not differ significantly between the groups. After the intervention, the pain severity was significantly reduced in both groups, but the extent of the reduction was larger in the valerian group, with the difference between the 2 groups being statistically significant. The total scores of the systemic manifestations associated with dysmenorrhea decreased after the intervention, but there was no significant difference between the groups, with the exception for syncope (Mirabi et al., 2011).
Attention enhancing activity
In Germany valerian is sometimes used to treat attention deficit hyperactivity disorder (ADHD) in children (Cavazzuti, 1969). German studies during 1960’s
Hippocratic Journal of Unani Medicine 58 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ reported that valerian could antagonize the hypnotic effects of alcohol, enhancing concentration and coordination (Mowrey, 1986). In a randomized, placebo- controlled, double-blind study, valepotriates demonstrated a dose-dependent increase in concentration abilities in 24 healthy volunteers. Also, valeropotriates when given in combination with alcohol did not affect blood alcohol levels, sedative effects or effects on driving performance (Mayer and Springer, 1974). There are no controlled trials evaluating valerian’s use in treating attention deficit hyperactivity disorder (ADHD) (Mowrey, 1986).
Other Neurological Activity
Unlike diazepam, valerian did not affect spontaneous ambulation and rearing or approach-avoidance conflict in mice in a water-lick conflict test. On the other hand, valerian and imipramine significantly inhibited immobility induced by a forced swimming test in rats and significantly reversed reserpine-induced hypothermia in mice, leading researchers to conclude that valerian may be a useful antidepressant (Morazzoni and Bombardelli, 1995). Among 80 hospitalized geriatric patients enrolled in a placebo controlled trial for 14 days, those assigned to an aqueous valerian extract had significant improvements in mood and behavioral disturbances as well as sleep (Kam-kohl et al., 1984). Among 121 patients with sleep disturbances enrolled in a controlled trial, those assigned to an alcoholic extract of valerian (600 mg daily for 28 days) had a significant improvement in depression, mood and global functioning as well as sleep; no significant side effects were reported (Vorbach et al., 1996).
Propagation
Conventional Methods
○○○○○○○○○○○○○○○○○○○○○○○○○○○ The plant is conventionally propagated through seeds as well as from root cuttings. Although Valerian can grow in any ordinary soil, but prefer moist humus rich loam having pH of 6-7. It is often found to flourish in damp and shady places, though some drought resistant forms also occur in chalk and limestone hills. The plant may easily be propagated by portions of the old rootstocks either in the autumn months or in spring. The divisions of the rootstock are spaced 30 cm × 60-90 cm. A good crop of the drug may be harvested in the autumn season if the rootstock divisions are planted very early in autumn in time to become well established before the onset of frost. The summer cultivation requires the weeding of beds and cutting off all floral stalks to avoid exhaustion and promote the formation of larger rootstocks. Soil should be rich in phosphorous (or supply a fertilizer) as the leaves turn purple when the plant is grown in phosphorous-deficit soil. Seeds are directly sown by simply pushing them into the soil during spring season. Avoid
Hippocratic Journal of Unani Medicine 59 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ covering them as they require light and a temperature of 68 to 70°F to germinate. Their germination takes one to two weeks. The nursery raised seedlings need consistent moisture, grow slowly, need protection from faster growing weed and are first transplanted at a spacing of 18-20 cm in rows 30 cm apart. The seedlings are then re-transplanted to their permanent sites at the same spacing as used for root stock divisions. Application of farmyard manure with proper mixing into the soil before the transplanting of seedlings favours the good growth of the plant. The formation of large root stock is promoted by raising a low ridge of soil along the bases of plant. For the better development of the rhizome flowering tops must be cut off as they appear. Many of the young plants do not flower in the first year, but produce a luxuriant crop of leaves, and yield rhizome of good quality in the autumn. The rhizomes and roots of the plants, propagated from the divisions of the rootstock may be harvested in the autumn of the first year’s growth, though the yield is generally low. The bud-raised plants do not attain a size suitable for harvesting before the end of second growing season (Anonymous, 2003).
In Vitro Methods
Abdi and Khosh-khui (2007) established a highly reproducible micropropagation protocol through adventitious shoot bud regeneration from leaf explants. They reported influential role of size as well as orientation of leaves, type of plant growth regulators on shoot bud induction. Maximum shoot regeneration (74 %) was observed from abaxial side of leaves with the production of about 7.4 mean number of shoot on MS (Murashige and Skoog, 1962) supplemented with 1 mg/L 6-Benzyl adenine (BA). Multiplication of shoots was further achieved by transferring shoots regenerated from leaves on MS medium supplemented with Isopentenyl Adenine (2-ip) and indole-3-acetic acid (IAA) alone or in combination. The highest multiplication rate (93.3 %) was observed on combination of 2-iP (6 mg/L) and IAA (0.3 mg/L) which induce the formation of about 13.7 mean shoot number. á- ○○○○○○○○○○○○○○○○○○○○○○○○○○○ naphthalene acetic acid (NAA) at 1 mg/L was the optimal treatment for in vitro root induction in their study.
Abdi et al. (2008) further evaluated the potential of silver nanoparticle (35 nm average size) for removal of bacterial contaminants from in vitro cultures of V. officinalis. Their experiment involved the treatment of explants with different concentration (25, 50 and 100 mg/L) of silver nanoparticle before and after surface sterilization for three different time periods (30, 60 and 180 min). Explants were cultured on MS medium supplemented with 5 mg/L KN and 0.1 mg/L NAA. Their results revealed the highest percentage (89 %) of disinfected explants by using 100 mg/L of silver nanoparticle solution for 180 min after surface sterilization while same amount of silver nanoparticle solution used for the similar duration resulted in only 32 % disinfected explants when the nano silver solution was used
Hippocratic Journal of Unani Medicine 60 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Figure 1: Steps involved in micropropagation protocol of Valeriana officinalis L.
Step A. In vitro induction of shoots from nodal explant in culture media in V. officinalis; Step B. Multiplication and proliferation of cultures for enhanced production of V. officinalis; Step C. In vitro rooting in tissue culture raised plantlets; Step D. Acclimatised in vitro raised plantlet ready to transfer in field
Hippocratic Journal of Unani Medicine 61 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ before surface sterilization. However, no affect on contamination was observed when silver nanoparticle solution was used without surface sterilization of explants. However, the authors did not notice any affect of nano silver solution on the characters measured. On the basis the above mentioned experiment, it was concluded that NS had a good potential for removing of the bacterial contaminants in plant tissue culture procedures.
Muntean (2008) reported somaclonal variation among the valerian (V. officinalis), micropropagated plantlets. During their experiment optimum callus induction was observed on Murashige and Skoog (MS) medium supplemented with 10 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) while maximum regeneration was achieved upon transferring the callus to MS medium containing combination of 2.5 mg/L BA and 0.5 mg/L IAA. In vitro rooting of regenerated shoots was obtained on MS medium containing 1 mg/L indole-3-butyric acid (IBA). About 19 plantlets regenerated from the callus were successfully hardened followed by their transfer to the greenhouse. The genetic analysis through RAPD marker was further performed during their study in order to investigate any detectable variation among the parent plant and the regenerants. They found four somaclonal variants out of 19 regenerants and the obtained somaclones were different from the parental plants by at least one polymorphic amplification product. However, the remaining regenerants were found to be genetically stable as compared to the elite donor plants. The seven primers generated 13 new RAPD markers in the somaclonal variants that were not found in the donor plant fingerprints. Moreover, two out of four somaclonal variants were polymorphic, compared with the donor plant, with two or more primers.
Reza et al. (2009) reported indirect shoot regeneration from leaf and petiole derived callus. In their study callus was developed on MS medium supplemented with different combinations of different auxins (2,4-D, NAA, picloram) and N6-
○○○○○○○○○○○○○○○○○○○○○○○○○○○ furfuryladenine (KN) from petiole and leaf segments wherein highest callus induction frequency was obtained on 1.0 mg/L 2,4-D for petiole (98 %) and leaf (100 %) explants. For shoot regeneration the calli were transferred to a medium supplemented with various concentrations of BA or KN and they observed maximum shoot regeneration frequency on 0.2 mg/L BA or KN with the production of about 13 adventitious shoots per callus segments derived from leaves. MS medium supplemented with 1 mg/L NAA was reported to be the optimal rooting media during their investigation on which 100 % regenerated shoots developed roots with an average of 10.7 roots per shoot within 3 weeks. The in vitro raised plantlets were further successfully hardened and transferred to greenhouse with 80 % survival rate.
Zebarjadi et al. (2011) developed in vitro regeneration protocol via indirect organogenesis from leaves, stems and roots explants excised from 35 day old in
Hippocratic Journal of Unani Medicine 62 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ vitro germinated seedlings. The highest callus formation frequency was achieved on MS medium augmented with 0.1 mg/L KN and 2 mg/L 2,4-D for leaf and root explants (100 %) while MS medium containing 0.1 mg/L KN and 10 mg/L 2,4-D induce maximum callus (100 %) from stem explants. Their results further revealed maximum shoot regeneration frequency of 60 % on MS medium containing 0.5 mg/L NAA and 2 mg/L BA in leaf-derived calli. Regenerated shoots were subsequently transferred to medium lacking growth regulators and then MS medium supplemented with 2 mg/L IBA for in vitro rooting.
Tansaz et al. (2014) investigated the effects of three different culture media formulations viz. MS, B5 (Gamborg et al. 1968) and MB (new combined medium containing half MS and B5 salts and vitamins, 10 ml/L Fe EDTA) and various combinations of plant growth regulators (BA and NAA) on in vitro regeneration and growth parameters of V. officinalis. Excised crown with 2-3 segments having one bud was used as explants. Among the different media tried during their study, MB medium exhibited highest regeneration frequency alongwith maximum induction of mean number of microshoots. However, highest percentage of regenerated plantlet (80 %) with maximum mean shoot number (9.43) and mean shoot length (7.04 cm) was obtained on MB medium supplemented with BAP (0.5 mg/L) in combination with NAA (0.25 mg/L). B5 medium without any PGR exhibited the lowest regeneration frequency of 40 % which increased to 75 % on supplementation with 1.5 mg/L BA and 0.25 mg/L NAA.
Ghadheri and Jafari (2014) for the first time reported efficient and rapid propagation protocol of V. officinalis for large-scale production of uniform raw materials for future pharmaceutical compound extraction as well as the quantification of the two industrially important bioactive compounds (i.e. valerenic acid and valtrate) accumulated in tissue culture-raised plants. Petiole and leaf explants were used for the callus induction and the three types of callus could be recognized by color
○○○○○○○○○○○○○○○○○○○○○○○○○○○ (creamy green, light or dark green) and texture (compact, nodular and friable) depending on the PGR concentrations/combinations used. Compact, nodular, yellowish green callus usually formed through leaf and petiole explants on medium containing 4.65 or 9.30 ìM KN in combination of 2.26 ìM 2,4-D showed the highest regenerative capacity with the formation of maximum shoot buds in both types of explants. IBA at a concentration of 2.46 ìM was found to be the best rooting treatment, which promoted the highest frequency of rooting (98 %). Successful acclimatization of rooted plantlets was achieved in potting medium containing 5:1 mixture of peat and perlite with 95.34 % survival rate. The contents of valtrate and valerenic acid in biomass extracts from petiole-derived plants were significantly higher than leaf-derived plants as well as the seed-raised field-grown plants. Maximum valtrate (6.98 mg g”1 DW) and valerenic acids (3.02 mg g”1 DW) contents were quantified in root tissue of petiole derived plants raised on medium
Hippocratic Journal of Unani Medicine 63 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ with higher concentration of Kn (9.30 ìM) in combination with 2.26 ìM 2,4-D. Further they also conducted RAPD analysis to screen tissue culture induced genetic variations in high level valerenic acid-accumulated plants using 15 RAPD primers (OPA 1-15) wherein only 4 primers viz. OPA-3, 5, 8, and 12 produced reproducible and scorable bands ranging from 3 to 8, with the generation of 104 amplification products of 200 to 1400 bp. Their results showed no differences between the plants raised from in vitro cultures but a slight variance in only one electrophoresis band, produced by OPA-8 primer, with seed-raised field grown plant was reported by them which demonstrates significant difference of these plants with seed raised plants from the metabolite content point of view. They lastly concluded that tissue culture-raised plants had higher genetic stability than did the seed-propagated plants.
Bhat and Sharma (2015) evaluated the effect of light and low temperatures as
well as pretreatment of seeds with GA3 on seed germination along with the development of in vitro propagation protocol through shoot tip explants. Their results revealed increased germination rate (48 %) in unchilled seeds pretreated
with GA3 (200ppm) for 24 hr in dark as compared to non pretreated seeds with only 10 % germination under dark conditions. Further maximum germination rate of 60 % was obtained through pre-chilling treatment of seeds for 72 hr and keeping them in light conditions during their study. MS medium supplemented with BA (1mg/L) and IAA (0.5 mg/L) was found to be the optimal treatment for shoot multiplication and elongation with 50 % response.
Genetic transformation and Elicitation studies for enhanced metabolites production
Gränicher et al. (1992) established hairy root cultures in V. officinalis var. sambucifolia by infection of sterile plantlets with Agrobacterium ○○○○○○○○○○○○○○○○○○○○○○○○○○○ rhizogenes strain R1601. Different hormone-free liquid media were used for the growth of transformed roots and the yield of different valepotriates viz. isovaltrate, valtrate, didrovaltrate, isovaleroxyhydroxydidrovaltrate content was quantified through HPLC. The highest overall valepotriate content (10.3 % dry wt) which was about 4 times the amount found in the roots of 9-month-old nontransformed plants, was observed in half strength Gamborg B5 medium supplemented with 2 % sucrose. They further demonstrated the production of about 44 mg/g dry wt valepotriates from hairy roots cultured in MS liquid medium supplemented with 2 % sucrose for 50 days.
Zebarjadi et al. (2011) carried out genetic transformation studies in V. officinalis under in vitro conditions by co-cultivating leaf, stem and root derived callus with two different strains of A. rhizogenes viz. AR15834 and LBA 9402 for the
Hippocratic Journal of Unani Medicine 64 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ induction of hairy root cultures. During their experiment, leaf, stem segments 2 (1 cm ) and calli were immersed in a suspension of A. rhizogenes (OD600 = 0.6-1) for 5 to 7 min followed by subsequent co-cultivation on solidified MS medium without any hormone and MS medium containing (0.1 mg/L NAA+1 mg/L BAP), (0.5 mg/L BAP+0.5 mg/L NAA+0.5 mg/L 2,4-D) for 2 days in darkness. Explants were then transferred onto selective medium containing 250 mg/L cefotaxime and 20 mg/L rifampicin, and further subcultured onto fresh selective medium every 15 days. Un-inoculated explants were used as a control and after 28 days of culture, the number of produced roots on selective medium to the number of cultured explants was determined. The results obtained demonstrated LBA9402 strain to be better than AR15834 strain in terms of both mean number of hairy root formation (68.62%) and production of the respective hairy root (90%) while leaf and root calli were found to be the best explants for transformation with both the strains. The optimal growth of the hairy roots occurred on selective MS medium containing 20 mg/L rifampicin and 250 mg/L cefotaxime without any plant growth regulators. Genomic DNA of putative transgenic and non-transgenic (control) regenerated hairy roots were analyzed by PCR for presence of rol B gene, using specific primers.
Torkamani and Samadi (2014) studied the influence of four different levels of Calcium and Potassium (half, full, two and four fold concentrations) on enhanced accumulation of valerenic acid in hairy root cultures of V. officinalis. Hairy root cultures were established via A. rhizogenes A13 strain using various explants derived from 42-day-old sterile seedlings and the transgenic status of the roots were confirmed by the presence of rol B and rol D amplified products during Polymerase chain reaction (PCR) analysis. The induced hairy root cultures of V. officinalis were maintained in the media and after 35 days were used for investigations of biomass accumulation and valerenic acid production through ○○○○○○○○○○○○○○○○○○○○○○○○○○○ high-performance liquid chromatography (HPLC). The results showed highest valerenic acid accumulation of about 0.69±0.03 mg/g DW from hairy roots cultured on media with two-fold calcium, which was 1.92 times higher than normal culture (0.36± 0.01 mg/g DW). The study also demonstrated the positive effect of double concentration (880 mg/L) of calcium in media on growth of transformed hairy roots. However, they further reported decrease biomass accumulation but 1.2 fold increase in valerenic acid production than control roots on the media with 3.32 mg/l (four times of control concentration) potassium concentration. Their results revealed the positive role of calcium in intensification of the valerenic acid production in V. officinalis hairy roots cultures.
Tokramani et al. (2014a) also studied the effect of exposure time (3 and 7 days) of different calcium and magnesium concentrations (2 to 6 fold than basal MS
Hippocratic Journal of Unani Medicine 65 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ medium) on the growth of hairy roots and valerinic acid production in hairy root cultures. They used hairy root cultures grown in MS liquid medium without elicitors for 8 weeks for the quantification of valerinic acid through HPLC. Their results revealed highest production of valerenic acid (1.83 ± 0.06 mg/g DW) in hairy root cultures exposed to 6-fold calcium for 7 days which was 7.9 times higher than that of control culture (0.23 ± 0.01 mg/g DW) while (6-fold) of magnesium at exposure time of 7 days resulted in the accumulation of valerenic acid at (1.11 ± 0.03 mg/g) DW which is 4.2-fold higher than that of non-elicited control.
Torkamani et al. (2014b) further increased valerenic acid production in the selected hairy root line ‘LeVa-C4’ through the use of three different elicitors’ viz. Fusarium graminearum extract (FE), methyl jasmonate (MJ) and salicylic acid (SA). They use wild-type strain ‘A13’ of A. rhizogenes for the induction of hairy roots and 23- day-old hairy root cultures were further inoculated into 25 ml MS liquid medium supplemented with various concentrations of MJ (50, 100, 200 ìM), SA (50, 100, 150 ìM) or FE (0.5, 1, 2% v/v) for different exposure times (3 and 7 days). They found 1% FE and 100 ìM L-1 MJ highly efficient for increased levels of valerenic acid after 7 days of elicitation which were 12.31 (3.02 mg/g DW) and 6 times (2.296 mg/g DW) higher than that of non-elicited control (0.24 mg/g DW) respectively. They further demonstrated that FE did not exert any negative effects on biomass yield of hairy root and no significant increase in valerenic acid content was noticed through the use of salicylic acid as elicitor.
Parizi et al. (2014) established hairy root cultures through A. rhizogenes (strain ATCC 15834) mediated genetic transformation and further determine the effect of different media viz. MS, B5 media (full and half strength), N6 medium and a modified MS medium without phytohormones, on the growth parameter of hairy + - roots. They also studied the effect of different NH4 /NO3 ratios (00:20, 10:20, 20:20, 20:10, 20:00, 20:40) in MS medium on hairy root growth and the effect of
○○○○○○○○○○○○○○○○○○○○○○○○○○○ these treatments were evaluated after 21 days of culture in relation to hairy root growth. Their results revealed B5 and ½ B5 media to be the best basal medium with the production of maximum dry weight of hairy roots i.e. 3.85 and 3.67 g/L respectively while N6 medium exhibited lowest hairy root growth with the production + - of only 0.86 g/L dry weight after 21 days. Further among the different NH4 /NO3 ratios in MS media tested, MS medium supplemented with a 20:20 mM and 20:40 + - mM ratio of NH4 to NO3 produced the maximum biomass of 1.80 and 1.62 g/L respectively after 21 days.
Acknowledgement
Ms. Taiba Saeed acknowledges UGC, for providing financial assistance under the scheme SRF-Maulana Azad National Fellowship (Award No. MUS-UTT-2624).
Hippocratic Journal of Unani Medicine 66 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ References
Abdi, G. and Khosh-khui, M., 2007. Shoot regeneration via direct organogenesis from leaf segments of Valerian ( Valeriana officinalis L.). Internat. J. Agricult. Res. 2: 877-882.
Abdi, G., Salehi, H., Khosh-Khui, M., 2008. Nano silver: a novel nanomaterial for removal of bacterial contaminants in valerian (Valeriana officinalis L.) tissue culture. Acta Physiol. Plant. 30: 709–714.
Ahmad, F., Rashid, H., Bhatia, K., Rehman, H., Kaur, M., Anjum, S., Ansari, R.A., Raisuddin, S., 2012. Modulatory Effect of a Unani Formulation (Jawarish amla sada) on Cyclophosphamide induced Toxicity in Tumour Bearing Mice. British J. Med. Res. 2: 454-468.
Andrews, M.J., Basu, A., 2005. Process for the extraction of Valerian roots. US Patent 6,913,770 B2.
Anonymous, 2003. The Wealth of India. A Dictionary of Indian Raw Materials and Industrial Products. Vol. 10. CSIR, New Delhi, p. 426.
Ansari, Dugaheh, M., Meisami, F., Torabian, Z. and SharifiFar, F., 2013. Antioxidant effect and study of bioactive components of Valeriana sisymbriifolia and Nardostachys jatamansii in comparison to Valeriana officinalis. Pak. J. Pharm. Sci. 26: 53 58.
Barnes, J., Anderson, L.A., Phillipson, J.D., 2002. Herbal Medicines. A guide for healthcare professional, 2nd ed. The Pharmaceutical Press, London, pp. 468-476.
Bhat, B. and Sharma, V.D., 2015. In vitro propagation of the Garden Heliotrope, Valeriana officinalis L. : Influence of pre-chilling and light on seed germination. Ind. J. of Exp. Biol.53: 184-188. ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Bissett, N.G., 1994. Herbal Drugs and Phytopharmaceuticals. MedPharm CRC Press, Stuttgart, p 566.
Bos, R., Woerdenbag, H.J., Hendriks, H., Scheffer, J.J.C., 1997. Composition of the essential oils from underground parts of Valeriana officinalis L. s.l. and several closely related taxa. Flav. Frag. J. 12: 359-370.
Buchbauer, G., Jager, W., Jirovetz, L., Meyer, F., Dietrich, H., 1992. Effects of valerian root oil, borneol, isoborneol, bornyl acetate and isobornyl acetate on the motility of laboratory animals (mice) after inhalation. Pharmazie 47: 620-622.
Busanny-Caspari, E., 1986. Indikationen: Funktionelle Herzbeschwerden, Hypotonie und Wetterfuhligkeit. Therapiewoche 36: 2545-2550.
Hippocratic Journal of Unani Medicine 67 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Caron, M.F., Riedlinger, J.E., 1999. Valerian: a practical review for clinicians. Nutri. in Clin. Care 2: 250-257.
Carretero, M.E.A., 2000. Terpenos II: iridóides y sesquiterpenos. Pan Actual Med. 24: 1115-1119.
Cavazzuti, G.B., 1969. Study of the clinical pharmacology of an association of gamma-amino- beta-hydroxybutyric acid and plant extracts used in therapy of children with behavior disorders and sleep disorders. Clin. Ter. 51:15-29.
Circosta, C., Pasquale, R.D., Samperi, S., Pino, A., Occhiuto, F., 2007. Biological and analytical characterization of two extracts from Valeriana officinalis. J. of Ethnopharmacol. 112: 361-367.
Cramer, K., Charrois, T., Vohra, S., 2006. Practical management of adverse effects and drug interactions. Clinical Brief 139: 39-41.
Cunha, A.P., 2005. Farmacognosia e fitoquímica. Fundação Calouste Gulbenkian, LIsboa, pp. 410-411.
Cunha, A.P., Silva, A.P., Roque, O.R., 2003. Plantas e produtos vegetais em fitoterapia. Fundação Calouste Gulbenkian, Lisboa, pp. 634-635.
Dewick, P.M., 2002. Medicinal natural products: a biosynthetic approach. 2nd ed. John Wiley and Sons Ltd, UK, pp. 190-191.
Duke, J.A., 1985. Handbook of Medicinal Herbs. CRC Press, Boca Raton.
Fernandez, S., Wasowski, C., Paladini, A.C., Marder, M., 2004. Sedative and sleep-enhancing properties of linarin, a flavonoid-isolated from Valeriana officinalis. Pharmacol. Biochem. Behavior 77: 399-404.
Fleming, T., 1998. PDR for herbal medicines. Medical Economics Company, Inc. Montvale, NJ. ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Foerster, W., Becker, H., Rodriguez, E., 1984. HPLC analysis of valepotriates in the North American genera Plectritis and Valeriana. Planta Med. 50:7-9.
Franck, B., Petersen, U., Huper, F., 1970. Valerianie, a tertiary monoterpene alkaloid from valerian (1). Angew Chem. Int. Ed. Engl. 9:891.
Ghaderi, N. and Jafari, M., 2014. Efficient plant regeneration, genetic fidelity and high-level accumulationof two pharmaceutical compounds in regenerated plants of Valeriana officinalis L. South Afr. J. Bot. 92: 19-27.
Gränicher, F., Christen, P., Kapetanidis, I., 1992. High-yield production of valepotriates by hairy root cultures of Valeriana officnalis L. var. sambucifolia Mikan. Plant Cell Rep. 11: 339-342.
Hippocratic Journal of Unani Medicine 68 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Anonymous, 1969. Hamdard Pharmacopeia of Eastern Medicine. Hamdard National Foundation, Pakistan.
Hattesohl, M., Feistel, B., Sievers, H., Lehnfeld, R., Hegger, M., Winterhoff, H., 2008. Extracts of Valeriana officinalis L. s.l. show anxiolytic and antidepressant effects but neither sedative nor myorelaxant properties. Phytomed. 15: 2-15.
Hazelhoff, B., Malingre, T.M., Meijer, D.K., 1982. Antispasmodic effects of valeriana compounds: an in-vivo and invitro study on the guinea-pig ileum. Arch. Int. Pharmacodyn. Ther. 257: 274-287.
Heinrich, M., Barnes, J., Gibbons, S., Williamson, E.M., 2004. Fundamentals of pharmacognosy and phytotherapy. Churchill Livingstone, London, pp. 234- 245.
Hikino, H., Kato, T., Takemoto, T., 1975. Constituents of wild Japanese valerian roots (4). Yakugaku Zasshi 95: 243-245.
Houghton, P.J., 1988. The biological activity of Valerian and related plants. J. Ethnopharmacol. 22: 121-42.
Houghton, P.J., 1999. The scientific basis for the reputed activity of valerian. J . Pharmacy & Pharmacol. 51: 505-512.
Huang, K.C., 1999. The pharmacology of Chinese herbs. CRC Press, Boca Raton.
Iwu, M.M., 1993. Handbook of African medicinal plants. CRC Press, Boca Raton.
Janot, M.M., Guilhem, J., Contz, O., Venera, G., Cionga, E., 1979. Contribution to the study of valerian alkaloids (Valeriana officinalis, L.): actinidine and naphthyridylmethylketone, a new alkaloid (author’s transl). Ann Pharm. Fr. 37:413-20.
Kamm-Kohl, A., Jansen, W., Brockmann, P., 1984. Moderne Baldriantherapie gegen nervose Storungen im Senium. Med Welt. 35: 1450-1454. ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Lin, L.J., Cordell, G.A., Balandrin, M.F., 1991. Valerian-derived sedative agents I. On the structure and spectral assignment of the constituents of valmane using the selective INEPT nuclear magnetic resonance technique. Pharm Res. 8: 1094-1102.
Madureira de Oliveria, D., Barreto, G., Valverde, G., De Andrade, D., Saraceno, E., Bertolino, L.A., Capani, F., Dos Santos, El Bacha, R., Giraldez, L.D., 2009. Cytoprotective effect of Valeriana officinalis extract on an in vitro experimental model of Parkinson disease. Neurochem. Res. 34: 215-220.
Marder, M., Viola, H., Wasowski, C., Fernandez, S., Medina, J.H., Paladini, A.C., 2003. 6-Methylapigenin and hesperidin: new valeriana flavonoids with activity on the CNS. Pharmacol. Biochem. Behavior 75: 537-545.
Hippocratic Journal of Unani Medicine 69 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Mayer, B., Springer, E., 1974. Psychoexperimental studies on the effect of a valepotriate combination as well as the combined effects of valtratum and alcohol. Arzneimittel-Forschung 24: 2066-2070.
Mirabi, P., Dolatian, M., Mojab, F., Majid, H.M., 2011. Effects of valerian on the severity and systemic manifestations of dysmenorrhea. Internat. J. of Gynecol. & Obstetrics 115: 28-288.
Morazzoni, P., Bombardelli, E., 1995. Valeriana officinalis: traditional use and recent evaluations of activity. Fitoterapia 66: 99-112.
Mowrey, D.B., 1986. The scientific validation of herbal medicine. Keats Pub., New Canaan, Conn, p. 316.
Muntean, L., 2008. Assessment of somaclonal variation in valerian (Valeriana officinalis L.) by RAPD fingerprinting analyses. Proceedings of the Fifth Conference on Medicinal and Aromatic Plants of Southeast European Countries, (5th CMAPSEEC), Brno, Czech Republic, 2-5 September, p. 107.
Murphy, K., Kubin, Z.J., Shepherd, J.N., Ettinger, R.H., 2010. Valeriana officinalis root extracts have potent anxiolytic effects in laboratory rats. Phytomed. 17: 674-678
Parizi, A.P., Farsi, M., Nematzadeh, G.A., Mirshamsi, A., 2014. Impact of different culture media on hairy roots growth of Valeriana officinalis L. Acta Agricult. Sloven.103: 299- 305
Patocka, J. and Jakl, J., 2010. Biomedically relevant chemical constituents of Valeriana officinalis. J. Appl. Biomed. 8: 11 18.
Petkov, V., 1979. Plants and hypotensive, antiatheromatous and coronarodilatating action. Am. J. Chin. Med. 7: 197-236.
Piccinelli, A.L., Arana, S., Caceres, A., Bianca, R.E.V., Sorrentino, R., Rastrelli,
○○○○○○○○○○○○○○○○○○○○○○○○○○○ L., 2004. New lignans from the roots of Valeriana prionophylla with antioxidative and vasorelaxant activities. J. Natural Prod. 67: 1135-1140
Reichert, R., 1998. Valerian clinical monograph. Quarterly review of natural medicine; Fall, pp. 207-215.
Reza, A.G., Morteza, K.K., Akhtar, S., 2009. Rapid Micropropagation through Shoot Regeneration of Valeriana officinalis L. Horticult. Environ. Biotechnol. 50: 467-471.
Tansaz, M., Zamani, A., Otroshy, M., 2014. Rapid in vitro shoot regeneration of Valeriana officinalis. Plant Tissue Cult. Biotech. 24: 263 271
Torkamani, M.R.D. and Samadi, A., 2014. Influence of minerals on valerenic acid accumulation in hairy root cultures of Valeriana officinalis L. Iranian J. of Plant Physiol. 5: 1187-1193.
Hippocratic Journal of Unani Medicine 70 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Torkamani, M.R.D., Abbaspour, N., Jafari, M., Samadi, A., 2014a. Elicitation of Valerenic Acid in the Hairy Root Cultures of Valeriana officinalis L (Valerianaceae). Trop. J. Pharm. Res. 13: 943-949
Torkamani, M.R.D., Jafari, M., Abbaspour, N., Heidary, R., Safaie, N., 2014b. Enhanced production of valerenic acid in hairy root culture of Valeriana officinalis by elicitation. Cent. Eur. J. Biol. 9: 853-863
Torssell, K., Wahlberg, K., 1967. Isolation, structure and synthesis of alkaloids from Valeriana officinalis L. Acta Chem. Scand. 21:53-62.
Veith, J., Schneider, G., Lemmer, B., Willems, M., 1986. The effect of degradation products of valepotriates on the motor activity of light-dark synchronized mice. Planta Med. 3:179-183.
von der Hude, W., Scheutwinkel-Reich, M., Braun, R., 1986. Bacterial mutagenicity of the tranquilizing constituents of Valerianaceae roots. Mutat. Res. 169: 23-27
Vorbach, E., Gortelmayer, R., Bruning, J., 1996. Therapie von Insomnien: Wirksamkeit und Vertraglichkeit eines Baldrian-Praparates. Psychopharmakotherap. 3:109-115.
Wang, J., Zhao, J., Liu, H., Zhou, L., Liu, Z., Wang, J., Han, J., Yu, Z. and Yang, F., 2010. Chemical analysis and biological activity of the essential oils of two valerianaceous species from China: Nardostachys chinensis and Valeriana officinalis. Molecules 15: 6411 6422.
Weiss, R.F., 1988. Herbal medicine. 1st ed. (translated from the 6th German Ed.). AB Arcanum e Beaconsfield, Beaconsfield Publisher, Gothemburg, pp. 281-285
Zebarjadi, A.R., Najafi, S., Ghasempour, H.R., Motamedi, J., 2012. Establishment of a practical tissue culture for producing hairy roots of Valeriana officinalis L.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ via Agrobacterium rhizogenes. J. Med. Plants Res. 5: 4984-4992
Zhang, B.H., Meng, H.P., Wang, T., et al., 1982. Effects of Valeriana officinalis L. extract on cardiovascular system. Yao Hsueh Hsueh Pao 17: 382-384.
Hippocratic Journal of Unani Medicine 71 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 72 Some Depletive ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract Plants of he Kumaon Himalaya of Uttarakhand is rich in floristic Kumaon composition. It is a land of diverse cultures and ethnic groups. Traditionally the folk people utilize the vegetation of their ambient environment in the form of different Himalaya Used Tproducts as food, fuel wood, fibre, fodder, timber, medicine, etc. During in Unani ethnobotanical explorations of different forest divisions in this region, the authors collected a large number of information on local plants which are used as folk Medicine for drugs for treatment of a wide range of health related problems. This study has Treating Non- also yielded information on some medicinal plants that are becoming scarce in the area due to overexploitation, unsustainable harvesting practices and loss of Communicable habitat. A review of literature revealed that most of these plant species find use in Diseases and Unani medicine for treating various non- communicable diseases (NCDs). Hence, their an attempt has been made to highlight such species in this report. For each plant species are given the correct botanical and vernacular names, habit and habitat, Conservation the part used, medicinal use(s) and other observations. The threat of extinction of Strategies# these useful and commercially viable medicinal plants has been discussed and also the strategies for their conservation have been suggested.
*Zaheer Anwar Ali, Sarfraz Ahmad Keywords: Ethnobotanical explorations, Endangered medicinal plants, and Conservation, NCDs, Kumaon. Shariq Ali Khan
Survey of Medicinal Plants Unit, Introduction Regional Research Institute of Unani Medicine (CCRUM), Kumaon region of Uttarakhand is situated between 28o 43’ 55"- 30o 20’ 12" N Post Box 70, Aligarh - 202001 (U.P.) latitude and 78o 44’ 30"- 80o 18’ 45" E longitude in Western Himalayas. There is wide range of natural habitats which provide varied plant life including medicinal and aromatic plants. It is also the land of diverse culture and ethnic groups. This Himalayan region has always been reputed as a steady supplier of a good number ○○○○○○○○○○○○○○○○○○○○○○○○○○○ of potent medicinal herbs and also one of the leading regions in the use of herbal drugs. A number of reports on the use of native floras in traditional medicine of many cultures of this region have been published (Agnihotri et al., 2003; Arya and Prakash, 1999; Arya et al., 1999; Aswal, 1992; Bhatt and Gaur, 1992; Bhatt et al., 1987-88; Datt and Lal, 1993; Garbyal et al., 2005; Gupta, 1960; Joshi, 1993; Joshi et al., 1993; Kalakoti and Pangtey, 1988; Pandey and Pande, 1990; Pandey et al., 1989,1995; Pangtey et al., 1989; Pant and Pandey, 1998; Rawat and Pangtey, 1987; Shah and Gupta, 1976; Shah and Jain, 1988; Shah and Joshi, 1971; Singh and Ali, 1997, 1998; Singh, et al., 1980, 1987; Singh and Maheshwari,
#This paper is based on the data presented by the first author in National Seminar on “The role of Unani medicine in non-communicable diseases”, organized by Central Council for Research in Unani Medicine, New Delhi at New Delhi on 14-15 January 2015. * Author for correspondence
Hippocratic Journal of Unani Medicine 73 January - March 2015, Vol. 10 No. 1, Pages 73-84 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ 1990, 1993, 1994). Hence this area was selected for an extensive ethnobotanical study. The main objective of this field study, besides collecting folk medicinal claims prevalent among the indigenous communities, was to prepare an inventory of existing Unani medicinal plants and to record the current status as well as distribution of highly exploited medicinal taxa. Ethnomedicinal uses of plants collected during the present survey have been published by us (Ali et al., 2008, 2010a; 2010b, 2013a, 2013b, 2013c, 2014a, 2014b; Ali and Ahmad, 2008, 2010). In this communication an enumeration of some depletive medicinal plants of Kumaon is presented.
Fieldwork was carried out during the period 1999-2008. The study area includes different forest divisions of Bageshwar, Champawat, Nainital, Pithoragarh, and Udham Singh Nagar districts. In the course of this investigation, it was found that wild plants are still predominantly in use by the natives for their health purposes and all the basic raw drugs are collected from the forest without replenishing the growing stocks. Moreover, the natural habitats are being disturbed due to a variety of factors such as diversion of land for expansion of agriculture, dwellings and other developmental activities; recurring forest fire; soil erosion; invasion of some alien weed species; etc. Consequent to this, populations of some commonly used drug yielding plants have become reduced in this area. A literature survey was carried out. The Unani Pharmacopoeia of India, Part-I (2007-2009), National Formulary of Unani Medicine, Part-I (Anonymous, 1981) and other monographs published by the CCRUM (Standardization of Single Drugs of Unani Medicine, Part-I (1987), Part-II (1992) and Part-III (1997) were mainly consulted for general information as well as therapeutic uses and it was found that most of these plant species are mentioned for treating various non-communicable diseases. The present communication highlights such species along with pertinent information. Earlier authors have reported many threatened medicinal and aromatic plants that were extracted for commercial purposes from this region and suggested their ○○○○○○○○○○○○○○○○○○○○○○○○○○○ conservation (Pangtey and Samant, 1988; Shah, 1983; Singh, 1993; Shah and Kapoor, 1978; Sinha, 1975). This contribution is an addition to the above reports. The information presented herein is mainly based on our field observations and enquiries made with knowledgeable village elders belonging to various indigenous communities and officials of the forest department. The study might be useful for planning strategies on conservation, and cultivation of medicinal flora particularly threatened taxa of the region. In the following enumeration the plants are listed in alphabetic order by their botanical names with respective family, vernacular names, habit and habitat, medicinal uses and locality. This is followed by a remark on availability and threat categories. All voucher specimens were prepared and preserved in the herbarium of Survey of Medicinal Plants Unit, Regional Research Institute of Unani Medicine, Aligarh (U.P.), India.
Hippocratic Journal of Unani Medicine 74 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Enumeration
Acorus calamus L. (Araceae)
Vernacular names : Bach, Boj, Ghiroch, Ghurvach (L); Waj-e-Turki (U)
Habit & habitat : A semiaquatic, perennial herb with aromatic rhizome. Growing in wet places like edges of pond and streams near villages.
Medicinal uses : Rhizome is anthelmintic and also given for hoarse-ness of voice, dog bite and stomachache.
Locality : Champawat, Nainital, Pithoragarh, Udham Singh Nagar
Remark : Not observed in the forest but now, cultivated by the forest department.
Berberis aristata DC. (Berberidaceae)
Vernacular names : Kingor (L), Darhald (U)
Habit & habitat : An evergreen shrub. Found in association with other species of Berberis along roadsides.
Medicinal uses : Root is commonly used for controlling diabetes. Root extract is instilled in eye for redness.
Locality : Nainital, Pithoragarh
Remark : Overexploited in past for trade. It is an endangered taxon.
Bergenia ciliata (Haw.) Sternb. (Saxifragaceae) ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Vernacular names : Silphori, Patharphori, Pashanbhed (L), Pakhanbhed (U)
Habit & habitat : A perennial herb with thick rootstock, found in forest, especially on moist rocky slopes.
Medicinal uses : The root of this plant has from very early times been in much repute for its medicinal properties, particularly for kidney stones. It is also used for diabetes, anuria and furunculosis.
Locality : Bageshwar, Champawat, Nainital, Pithroragarh
Remark : Presently common, but the population of this taxon is on decline due to continuous over collection by the unauthorized collectors.
Hippocratic Journal of Unani Medicine 75 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Cinnamomum tamala Nees (Lauraceae)
Vernacular names : Kakaria (L), Sazaj Hindi (U)
Habit & habitat : An evergreen small tree, found in shady places in forest.
Medicinal uses : Leaves are used for cough and cold.
Locality : Nainital, Champawat, Pithoragarh
Remark : Leaves and stem bark are heavily extracted which have much value for condiments. Low-Risk Near Threatened. Now, it is cultivated.
Curculigo orchioides Gaertn. (Hypoxidaceae)
Vernacular names : Kali musli (L), Musli Siyah (U)
Habit & habitat : Herb with tuberous root stock. This species is characteristics in Sal trees in the Tarai-Bhabar belts.
Medicinal uses : Root is commonly used for sexual weakness and leucorrhoea.
Locality : Udham Singh Nagar
Remark : The tuberous root is heavily exploited and also greedily eaten by wild animals. It is a vulnerable taxon.
Dactylorhiza hatagirea (D. Don) Soo (Orchidaceae)
Vernacular names : Hathajari (L), Salab Panja (U)
Habit & habitat : A terrestrial orchid with tuberous roots which are digitate or finger shape. Rarely found in damp and shady places
○○○○○○○○○○○○○○○○○○○○○○○○○○○ in Oak forest.
Medicinal uses : Tubers are commonly used as general tonic and also believed as an aphrodisiac agent.
Locality : Bageshwar, Pithoragarh
Remark : It is a critically endangered taxon. Export already banned (Jain and Sastry, 1980). It is cultivated in Nandadevi Biosphere Reserve by the tribal (Maikhuri et al., 2002).
Drimia indica (Roxb.) Jessop (Liliaceae)
Vernacular names : Jangli Piyaz, Koli Kanda(L), Isqeel (U)
Habit & habitat : A scape-bearing herb rarely found in forests of Tarai.
Hippocratic Journal of Unani Medicine 76 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Medicinal uses : The bulb is commonly used for burning micturition.
Locality : Udham Singh Nagar
Remark : A vulnerable taxon.
Eulophia herbacea Lindl. (Orchidaceae)
Vernacular names : Salab Misri (L, U)
Habit & habitat : Herb with tuberous roots, rarely found in forest.
Medicinal uses : Root is commonly used for leucorrhoea and as a general tonic.
Locality : Nainital
Remark : An endangered taxon. Export already banned (Jain and Sastry, 1980).
Gloriosa superba L. (Liliaceae)
Vernacular names : Kalihari (L), Muleem (U)
Habit & habitat : A climbing herb with tuberous rootstock; climbing by means of leaves. It is readily recognized by its beautiful flowers. Rarely found in outskirts of villages in Sub-Himalayan forest tracts.
Medicinal uses : Tuber is used for joint pain.
Locality : Udham Singh Nagar
Remark : Due to removal of tubers, the whole plant is destroyed as such wild populations have been reduced very much. It is ○○○○○○○○○○○○○○○○○○○○○○○○○○○ an endangered taxon.
Hedychium spicatum Buch.-Ham. ex J.E. Sm. (Zingiberaceae)
Vernacular names : Kapoor kachri, Jangli Haldi (L), Kapura Kachri (U)
Habit & habitat : A robust herb with horizontal root, found in moist and shady places in forest.
Medicinal uses : Root is regarded by the natives as a remedy for inflammation and also used for urticaria and kidney stones.
Locality : Champawat, Nainital, Pithoragarh, Udaham Singh Nagar
Remark : It has become vulnerable.
Hippocratic Journal of Unani Medicine 77 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Picrorhiza kurrooa Royle ex Benth. (Scrophulariaceae)
Vernacular names : Kutki (L, U)
Habit & habitat : A perennial herb found in the forest between altitudes of 3300-4300m.
Medicinal uses : Root is given for leucorrhoea.
Locality : Pithoragarh
Remark : An endangered taxon.
Rauwolfia serpentina (L.) Benth. ex Kurz (Apocynaceae)
Vernacular names : Sarpgandha, Swaitbarua (L), Asrol (U)
Habit & habitat : A perennial under shrub. It is seen growing wild in shady situations in Sal forest of Sub-Himalayan forest tracts.
Medicinal uses : The root is believed to be an antidote to snake poison and also considered effective for fever and abdominal pain.
Locality : Udham Singh Nagar
Remark : Ramnagar forest is the natural habitat of this plant where it is still occurs in good quantity. This species has been depleted in other forest areas due to excessive exploitation for trade in past. It has poor regeneration. Now, it is under cultivation.
Swertia angustifolia Buch.-Ham. ex D. Don (Gentianaceae)
Vernacular names : Chiraita(L), Chiraeta Shirin (U) ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Habit & habitat : An erect herb, found in the forest up to 1800m.
Medicinal uses : Leaves are used for fever in pneumonia.
Locality : Nainital, Pithoragarh
Remark : It is highly threatened due to trade and destruction of natural habitat. Now, it is under cultivation.
Taxus baccata L. sbsp. Wallichiana (Zucc.) Pilger (Taxaceae)
Vernacular names : Thuner, Laventa (L), Talispatr (U)
Habit & habitat : An evergreen tree. Found in the evergreen forest at higher altitudes associated with other species of gymnosperms.
Hippocratic Journal of Unani Medicine 78 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Medicinal uses : Leaves and stem bark are commonly used to prevent cold.
Locality : Nainital, Pithoragarh
Remark : The plant yields taxol, a remedy for cancer. It is threatened to illegal trade and destruction of habitat. Now, it is cultivated by the forest department.
Valeriana hardwickii Wall. (Valerianaceae)
Vernacular names : Samoy (L), Taggar (U)
Habit & habitat : Herb with white flowers. Found in forest, but also grows along roadsides near villages.
Medicinal uses : Root is used for headache. It is also used as an insect repellent by the natives.
Locality : Bageshwar, Nainital, Pithoragarh
Remark : Often exploited commercially, in the past. It is vulnerable taxon.
L = Local name; U = Unani name
Discussion
This communication has brought to light 15 medicinal taxa which are dwindling in the forests of the present study area due to various causes. Among other factors, continued exploitation, non-sustainable harvesting practices and destruction of natural habitats are largely responsible for their depletion. It is predicted that some more plants of the area will be decreased in number soon if destruction of forests and their over-exploitation continue to occur. ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Medicinal plants wealth of a region is one of the vital resources having important bearing on human health, regions’ economy and environment. Therefore, preservation of this heritage along with original habitats must be protected on priority basis. Such observations are of special significance for the area where there is a threat to the natural habitats and vegetation owing to increasing human interference. In the situation prevailing here some measures for conserving diversity of medicinal plants are listed below:
1. Large scale cultivation and domestication of wild medicinal species used in pharmaceutical industries and local medicine should be encouraged by the local inhabitants who may reduce the pressure on existing wild population of the plants.
Hippocratic Journal of Unani Medicine 79 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ 2. Public awareness programmes on conservation of wild medicinal plants should be intensified.
3. Promoting the rationale and sustainable utilization of medicinal plants.
4. Standard methods of cultivation, i.e., agro-technology and preservation of high demand endangered medicinal plants of the area should be developed.
5. Sensitive habitats of the threatened flora should be protected on priority basis.
6. Illegal extraction of medicinal plants from the wild should be curbed.
7. Social forestry operation of fuel, fodder and fibres species should be encouraged.
8. In order to protect and propagate the threatened species, botanical gardens should be established at different agro-climatic zones.
9. Germplasm of rare species should be collected and maintained under protected conditions in the field, plantation, botanical gardens, clonal repositories, gene bank or in plant tissue culture repositories.
10. Endangered medicinal plants which have poor regeneration their breeding system should be studied. Through various tissue culture and micro propagation techniques plants can be regenerated.
11. Local medicine men should be involved in the conservation efforts since they use plant remedies in their homes and are generally respected by the villagers.
12. Harvesting or collection has very much importance. It should be done through trained and experienced plant collectors who could identify the ○○○○○○○○○○○○○○○○○○○○○○○○○○○ medicinal plants properly and aware of the methodology of preservation of plants and their parts used in medicine.
13. Training should be provided regularly to the persons engaged in the collection of crude drugs. They should be trained for proper and scientific methods of collection, right time of medicinally important plant parts without damaging whole plant. For the collection of different plant parts following points should be taken into consideration:
a. Seeds-should be collected after attaining full maturity; b. Leaves-after flowering stage; c. Stems-after leaf fall or fruiting stage; d. Flowers-during full flowering stage; e. Fruits- after full ripening; f. Bark-after raining season; g. Root or root bark- should be collected from the damaged tree or after attaining full growth and the plant should be replenished in nature.
Hippocratic Journal of Unani Medicine 80 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Acknowledgements
We are highly grateful to the Director General, Central Council for Research in Unani Medicine, New Delhi, for providing necessary facilities for carrying out this investigation. We would also record our gratitude to all the informants who cooperated in the collection of information presented herein.
References
Agnihotri, A.K., Sikarwar, R.L.S., Khatoon, S., Rawat, A.K.S. and Mehrotra, S., 2003. Some common medicinal plants used by the local people of Haldwani forest division of Uttaranchal. 2nd World Cong. on”Biotechnological Development of Herbal Medicine” (NBRI), Lucknow, U.P., India, p. 110.
Ali, Z.A. and Ahmad, S., 2010. A contribution to the ethnomedicinal flora of the Pithoragarh forests of Kumaon region, Uttarakhand. Hippocratic J. Unani Med.5(2):109-116.
Ali, Z.A., Ahmad, S. and Khan, I.U., 2008. A contribution to the ethnopharmacology Nainital forests of Kumaon region, Uttaranchal (India). Hippocratic J. Unani Med.3(1):35-45.
Ali, Z.A., Ahmad, S. Alam, M. and Khan, L.A., 2010a. Ethnopharmacological study of the Champawat forest of Kumaon region, Uttarakhand. Hippocratic J. Unani Med. 5(3): 39-49.
Ali, Z.A., Ahmad, S. and Khan, L.A., 2010b. Observations on Ethnomedicinal flora of Bageshwar forests in Kumaon Himalaya, Uttarakhand. Hippocratic J. Unani Med. 5(4): 155-165.
Ali, Z.A., Ahmad, S. Wasiuddin and Khan, L.A., 2013a. Ethnopharmacological survey of west tarai forest division, Ramnagar, Nainital in Kumaon region of ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Uttarakhand. Hippocratic J. Unani Med.8(1): 79-88.
Ali, Z.A., Wasiuddin and Zaidi, S.T.H., 2013b. Ethnopharmacological survey of central tarai forest division Haldwani, Nainital, Uttarakhand. Hippocratic J. Unani Med. 8(3): 141-148.
Ali, Z.A., Ahmad, S., Wasiuddin and Khan, L.A., 2014a. A contribution to the ethnomedicinal flora of Haldwani forest division, Nainital (Uttarakhand). Hippocratic J. Unani Med. 9(1): 143-149.
Ali, Z.A., Ahmad, S., Wasiuddin and Khan, L.A., 2014b. An ethnopharmacological study of Ramnagar forest division of Nainital district, Uttarakhand. Hippocratic J. Unani Med. 9(2): 153-160.
Hippocratic Journal of Unani Medicine 81 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Anonymous, 2008. Unani Medicinal Plants of Tarai Forests in Kumaon Region of Uttarakhand. Central Council for Research in Unani Medicine, New Delhi.
Anonymous, 1981. National Formulary of Unani Medicine. Part-I (First Edition). Ministry of Health & Family Welfare, New Delhi.
Anonymous, 2007-2009. The Unani Pharmacopoeia of India. Part-I, Vol. I-VI. Ministry of Health & Family Welfare, New Delhi.
Anonymous, 1987, 1992, 1997. Standardisation of Single Drugs of Unani Medicine, Part I-III. CCRUM, New Delhi.
Arya, K.R. and Prakash, V., 1999. Ethnomedicinal study of a remote tribal area of Almora district: A Survey report. Part-I. J. Econ. Tax. Bot. 23 (2): 247-252.
Arya, K.R., Pandey, P.C. and Prakash V., 1999. Ethnobotanical study on tribal areas of Almora district-II. Ethnobotany 11(1&2): 100-104.
Aswal, B.S., 1992. Less-known medicinal uses of three plants from Kumaon Himalaya (India). Indian J. For. 15(1): 76-77.
Bhatt, K.C. and Gaur, R.D., 1992. A contribution to the ethnobotany of Rajis in Pithoragarh district. Acta Bot. Indica 20: 76-83.
Bhatt, K.C., Bisht, M.K. and Gaur, R.D., 1987-88. Folk medicines of Byans Valley in Pithoragarh district of U.P. JOSHARD 11-12:117-123.
Datt, B. and Lal, B., 1993. Less-known medicinal uses of some plants from Pithoragarh district of Kumaon Himalayas U.P. Aryavaidyan 6 (4): 242-246.
Garbyal, S.S. and Aggarwal, K.K. and Babu C.R., 2005. Traditionally used medicinal plants in Dharchula Himalayas of Pithoragarh district, Uttaranchal. Indian J. Traditional Knowledge 4 (27): 199-207.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Gupta, R., 1960. Some useful and medicinal plants of Nainital in Kumaon Himalaya. J. Bombay Nat. Hist. Soc. 59: 309-329.
Jain, S.K. and Sastry, A.R.K., 1980. Threatened Plants of India “A state-of-the-art Report. BSI, Howrah.
Joshi, G.C., 1993. Notes on the vegetation types of inner Himalayan ranges of Kumaon Himalaya with special reference to herbal resources. J. Econ. Tax. Bot. 17:367-371.
Joshi, G.C., Tewari, K.C., Tewari, R.N., Pandey, N.K. and Pandey, G., 1993. Resource survey of the pharmaceutically important plants of Uttar Pradesh Himalaya. In: U. Dhar (ed.) Himalayan Biodiversity– Conservation Strategies. Gyanodaya Prakashan Nainital. pp. 279-291.
Hippocratic Journal of Unani Medicine 82 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Kalakoti, B.S. and Pangtey, Y.P.S., 1988. Ethnomedicine of Bhotia tribe of Kumaon Himalaya, U. P. Bull. Med. Ethnobot. Res. 9 (1-2): 11-20.
Maikhuri, R.K., Nautiyal, S., Rao, K.S. and Saxena, K.G., 2002. Medicinal plants cultivation in Nanda Devi Biosphere Reserve Buffer Zone Villages. In: V.K. Singh, J.N. Govil. G. Singh (eds.) Recent Progress in Medicinal Plants, Vol. 1. Ethnomedicine and Pharmacognosy. SCI TECH, Publishing LLC, U.S.A. pp.345-362.
Pandey, B., and Pande, P.C., 1990. Ethnobotanical studies on gymnospermic plants of Kumaon Himalaya. J. Econ. Tax. Bot. 23 (2): 253-256.
Pandey, G., Joshi, G.C., Pandey, N.K. and Tewari, K.C., 1995. Ethnobotanical studies on the medicinal flora of Tarikhet block, Kumaon Himalaya, Dt. Almora, U.P. part-IV. Aryavaidyan 8 (3): 154-164.
Pangtey, Y.P.S. and Samant, S.S., 1988. Observations on the threatened, rare and endangered flowering plants and fern in the flora of Kumaon Himalaya. Adv. For. Res. India 2:65-74.
Pangtey, Y.P.S., Samant, S.S. and Rawat, G.S., 1989. Ethnobotanical notes on the Bhotia tribe of Kumaon. Indian For. 12(3):191-196.
Pant, S.C. and Pandey, G.C., 1998. Ethnobotanical studies on medicinal flora of Tharu tribal pockets in Kumaon region in Uttar Pradesh. Bull. Med. Ethnobot. Res. 16: 1-10.
Rawat, G.S. and Pangtey, Y.P.S., 1988. A contribution to the ethnobotany of Alpine region of Kumaon. J. Econ. Tax. Bot. 11:139-148.
Shah, N.C. and Jain, S.K., 1988. Ethnomedicobotany of the Kumaon Himalaya, India. Social Pharmacol. 2: 359-380.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Shah, N.C. and Joshi, M.C., 1971. An ethnobotanical study of Kumaon region of India. Econ. Bot. 25: 414-422.
Shah, N.C. and Kapoor, L.D., 1978. Depletive medicinal plants of Kumaon Himalayas. J. Res. Indian Med. Yoga and Homoeo. 13:38-43.
Shah, N.C., 1983. Endangered medicinal and aromatic taxa of U.P. Himalaya. In: S.K. Jain and R.R. Rao (eds.) An assessment of threatened plants of India. B.S.I. Howrah, pp. 40-49.
Shah, S.C. and Gupta, L.K., 1976. Useful medicinal plants of Ranikhet-I. Indian Drugs 14: 47-52.
Singh, H. and Maheshwari, J.K., 1993. Phytotherapy for diphtheria by the Bhoxas of Nainital district, Uttar Pradesh, India. Ethnobotany 5 (1 & 2): 63-65.
Hippocratic Journal of Unani Medicine 83 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Singh, K.K. and Maheshwari, J.K., 1990. Plant wealth in the life and economy of the Tharus of Nainital district, U. P. Indian Forester 116: 636-642.
Singh, K.K. and Maheshwari, J.K., 1994. Traditional Phytotherapy of some medicinal plants used by the Tharus of the Naintial district, Uttar Pradesh, India. Int. J. Pharmacog. 32: 51-58.
Singh, K.K., Palvi, S.K., Singh, H.B., 1980. Survey of some medicinal plants of Dharchula Block in Pithoragarh district of U.P. Bull. Med. Ethnobot. Res.1(1): 1-7.
Singh, K.K., Saha S. and Maheshwari J.K., 1987. Observation on the ethnobotany of Boxa tribe of Bajpur block of Nainital district, Uttar Pradesh. Him. Res. Dev. 6 (I-II): 25-29.
Singh, V.K. and Ali, Z.A., 1997. Folk medicinal plants of the Garhwal and Kumaon forests of Uttar Pradesh, India. Hamdard Medicus 40: 35-47.
Singh, V.K. and Ali, Z.A., 1998. Herbal drugs of Himalaya: Medicinal plants of Garhwal and Kumaon regions of India. Aspect of Plants Science, Vol. 15. Today & Tomorrow‘s Printers and Publishers, Karolbagh, New Delhi.
Singh, V.K., 1993. Exploitation and threat to some medicinal plants and folk drugs of Garhwal and Kumaon regions of Uttar Pradesh, India. In: U. Dhar (ed.) Himalayan Biodiversity– Conservation Strategies. Gyanodaya Prakashan Nainital. pp. 407-412.
Sinha, R.L., 1975. Protection and planned exploitation of Indian medicinal plants in the hills of Uttar Pradesh. In: Proc. of Nat. Symp. on “Recent Adv. in the Dev. Prod. and Utilization of Med. and Arom. Plants”. CIMPO, Lucknow. ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 84 Quality ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract Evaluation of erbal products have become a major part in curing the Jawarish-e-Ood different kinds of human ailments. In India, the Unani system of medicine consists of large number of herbal products to cure the different type of diseases. Jawarish- Kibreet He-Ood Kibreet is a Unani herbal formulation prepared in combination of ingredients like Ood, Dana-e-Heel Khurd, Dana Heel Kalan, Sazaj Hindi, Post-e-Turanj, 1 *Rampratap Meena, Tabasheer, Nana Khushk, Darchini, Gul-e-Surkh, Zarambad, Qaranful, Jauzbuwa, 2D. Ramasamy, Bisbasa, Nabat Safaid and Arq-e-Kibreet. It is prescribed in the treatment of Zof- 2S. Mageswari, 2P. Meera Devi Sri, e-Meda (weakness of the stomach) and Zof-e-Ishteha (anorexia) disorders. 3Shamsul Arfin Evaluation of Unani formulations has become a fundamental requirement of the and research organizations. Based on the available sources, an attempt has been 3Aminuddin made to evaluate the drug Jawarish-e-Ood Kibreet. To evaluate the drug the parameters like macroscopic, microscopic, determination of moisture content, 1Drug Standardisation Research Institute, PLIM Campus, ash values, bulk density, pH values, extractive values, TLC and analysis of quality Kamla Nehru Nagar, control parameters viz. heavy metals, microbial content, aflatoxins and pesticide Ghaziabad-201002 residues were performed. The evaluated data shall help to lay down
2Regional Research Institute of pharmacopoeial standards for the drug Jawarish-e-Ood Kibreet and also in Unani Medicine, producing quality and efficacious products having batch to batch consistency. 1 West Mada Church Street, Chennai-600013 Keywords: Jawarish-e-Ood Kibreet, Powder microscopy, Physico-chemical, TLC 3Central Council for Research and WHO parameters in Unani Medicine, 61-65 Institutional Area, Janakpuri, New Delhi-110058 Introduction
Jawarish-e-Ood Kibreet (Anonymous, 2006) is one of the ancient commonly used Unani formulations. The poly herbal formulation is prepared using 15 ingredients (Table 1). The drug is prescribed for the treatment of Zof-e-Meda (Weakness of the stomach) and Zof-e-Ishteha (Anorexia) disorders. Evaluation of Unani ○○○○○○○○○○○○○○○○○○○○○○○○○○○ medicines with the perspective of safety, efficacy and quality will not only preserve the traditional heritage but also rationalize their uses of Unani medicines in the health care.
Due to lack of standards and quality control methods there are batch to batch variations in the similar formulations. The main requirement of a standardisation is to establish the presence of each ingredient in the formulations (Bandaranayake, 2006; Myers and Cheras, 2004). The present study is an attempt to evaluate the drug by applying modern parameters such as microscopical, physico-chemical, thin layer chromatography and WHO parameters viz., microbial load, aflatoxin, heavy metals and pesticide residue.
*Author for correspondence
Hippocratic Journal of Unani Medicine 85 January - March 2015, Vol. 10 No. 1, Pages 85-94 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 1: List of the raw drugs of Jawarish-e-Ood Kibreet Formulation
S. Unani name Botanical/ Part used Quantity No. English name taken for SOP 1. Ood Aquilaria agallocha Roxb. Heart wood 20 g. 2. Dana-e-Heel Elettaria cardamomum Seed 6 g. Khurd (L) Maton. 3. Dana Heel Amomum subulatum Seed 6 g. Kalan Roxb. 4. Sazaj Hindi Cinnamomum tamala Leaves 6 g. (Buch. Ham.) Nees & Eberm. 5. Post-e-Turanj Citrus medica Linn. Pericarp 6 g. 6. Tabasheer Bambusa bambos Druce. Bamboo Manna 6 g. 7. Nana Mentha viridis Linn. Aerial part 6 g. 8. Darchini Cinnamomum zeylanicum Inner stem bark 6 g. Blume. 9. Gul-e-Surkh Rosa damascena Mill. Flower 10 g. 10. Zarambad Curcuma zeodaria Li Rhizome 4 g. 11. Qaranful Syzygium aromaticum —4 g. (L.) Merr. L M Perry. 12. Jauzbuwa Myristica fragrans Houtt. Endosperm 4 g. 13. Bisbasa Myristica fragrans Houtt. Aril 4 g. 14. Nabat Safaid Sugar Crystals 350 g. ○○○○○○○○○○○○○○○○○○○○○○○○○○○ 15. Arq-e-Kibreet Sulphur extract Extract 600 ml.
Material and Methods
Collection of the raw drugs
Genuine raw drugs namely Ood, Dana Heel Khurd, Dana Heel Kalan, Sazaj Hindi, Post-e-Turanj, Tabasheer, Nana, Darchini, Gul-e-Surkh, Zarambad, Qaranful, Jauzbuwa, Bisbasa, Nabat Safaid and Arq-e-Kibreet of the formulation were procured from raw drugs dealers of Chennai. The raw drugs were identified using pharmacognostical methods and evaluated their pharmacopoeial standards.
Hippocratic Journal of Unani Medicine 86 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Preparation of the drug
The ploy herbal semisolid drug was prepared in different batches at Laboratory scale as per the ingredients composition and guidelines of NFUM, Part–IV (Table I).
Powder microscopy
Microscopical examination allows more detail of a drug and can be used to identify the organised drugs by their known histological characters viz., cell walls, cell contents, starch grains, calcium oxalate crystals, trichomes, fibres and vessels (Kokate et al., 2000).
The drug sample (5g) was weighed and mixed with 50ml of water in a beaker with gentle warming, till the sample completely dispersed in water. The mixture was centrifuged and decanted the supernatant. The sediment was washed several times with distilled water, centrifuged again and decanted the supernatant. A few mg of the sediment was taken and mounted in glycerine. Then few mg was taken in watch glass and added few drops of phloroglucinol and concentrated hydrochloric acid, mounted in glycerine. The microscopic salient features of the drug were observed in different mounts (Wallis, 1997 & Johansen, 1940).
Physico-chemical analysis
The physico-chemical methods viz., moisture content, ash values, solubility in different solvents, pH values, bulk density and sugar content etc., are useful tools in standardisation of a herbal product for maintaining batch to batch consistency. The drug samples were subjected for the standardisation of physico-chemical parameters and analysed as per the standards method (Anonymous, 1987). ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Thin layer chromatography
Thin Layer Chromatography is a physical method of separation in which the components to be separated are distributed between two phases; one of these is a stationary phase bed and other is a mobile phase which percolates through this bed. It offers the best method for recording the finger prints which can be reproduced anywhere at the same laboratory condition of a particular product.
The samples of the drug (2g) were soaked in chloroform and alcohol separately for 18 hours, refluxed for ten minutes on water bath and filtered. The filtrates were concentrated on water bath and made up to 5ml in a standard flask separately and carried out the TLC studies (Wagner et al., 1984).
Hippocratic Journal of Unani Medicine 87 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Quality control parameters
The usage of herbal products along with higher safety margins, WHO has taken necessary step to ensure quality control parameters with the modern techniques and application of suitable standards. The parameters such as microbial load and heavy metal were carried out as per the WHO guidelines (Anonymous, 1998). Aflatoxin and pesticide residues were carried out by standard methods (Anonymous, 2000).
Results and Discussion
Jawarish-e-Ood Kibreet is a dark brown semi-solid product with sweetish bitter in taste.
Microscopical observation
The salient features of the raw drugs used in the preparation of Jawarish-e-Ood Kibreet were evaluated using powder microscopic studies and the photographs are shown in Fig. 1. Pitted vessels with simple perforation plate upto 200μ, xylem parenchyma lignified with pitted walls, xylem ray parenchyma cells along with fibres, septate fibres (Ood); perisperm cells isolated or in groups with bulbous projections filled with starch grains and tiny prismatic crystal of calcium oxalate, elongated thin walled parenchyma cells from aril tissue, thick walled sclerenchyma cells in surface view (Dana Heel Khurd / Dana Heel Kalan); epidermal cells in surface view with sunken stomata (paracytic stomata) subsidiary cells not clear (Sazaj Hindi); epidermal cells in surface view with circular stomata and schizolysigenous oil glands (Post-e-Turanj); numerous, angular, sharp edged siliceous sandy particles (Tabasheer); epidermal cells (bigger cells) in surface view with wavy margin, diacytic stomata, prominent capitate glandular trichomes
○○○○○○○○○○○○○○○○○○○○○○○○○○○ upto 80μ in length with single basal cell and single head cell, labiaceous glandular trichomes with single basal cell and a head of 8 cells upto 80μ in diameter (Nana Khushk); fibres thick walled lignified with striated walls and narrow lumen of length upto 1000μ and breadth not over 30μ, stone cells with horse shoe shaped thickenings upto 100μ (Darchini); long simple unicellular covering trichome, pollen grains round to oval upto 35μ with three distinct germ pores (Gul-e-Surkh); numerous starch grains of various shapes and size upto 50μ (Zarambad); pollen grains tetrahedral spherical biconvex measuring upto 20ì in diameter, spindle shaped fibres, parenchyma cells with schizolysigenous oil glands, fragments of anther wall in surface view (Qaranfal); endosperm cells in surface view with numerous starch grains and crystalloid proteins, each crystalloid proteins upto 40μ, perisperm cells in surface view filled with reddish brown content (Jouzbuwa); thick walled elongated parenchyma cells in surface view upto 50μ wide (Bisbasa).
Hippocratic Journal of Unani Medicine 88 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 1: Powder microscopy of Jawarish-e-Ood Kibreet
Hippocratic Journal of Unani Medicine 89 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 1: Continued...
Chemical analysis
The physico-chemical data, moisture content was obtained in the drug 18.58%. The alcohol soluble extractive (58.50%) might be due to the extraction of polar chemicals constituents and the water soluble extractives (63.49%) indicate the presence of inorganic constituents. The obtained data are shown in Table - 2.
Thin Layer Chromatography analysis
The chloroform and alcohol extract of all the three batch samples showed identical
spots at UV – 254nm and 366nm ranges and the Rf values of both the extracts
Hippocratic Journal of Unani Medicine 90 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 2: Physico-chemical parameters
Parameters Analyzed Batch Number (n=3) I II III Extractives 58.52% 58.24% 58.76% Alcohol soluble matter 63.16% 63.48% 63.84% Water soluble matter Ash 1.82% 1.79% 1.86% Total ash 0.95% 0.89% 0.98% Acid insoluble ash pH values 5.59 5.74 5.41 1% Aqueous solution 4.34 4.41 4.42 10% Aqueous solution Sugar estimation 35.83% 35.61% 35.84% Reducing sugar 7.55% 7.74% 7.53% Non-reducing sugar Moisture 18.81% 18.26% 18.68% Bulk Density 1.3997 1.4087 1.4009
Table 3: Rf Values of chloroform extract
Solvent system Rf Values Toluene: Ethyl acetate (9 : 1) UV 254nm UV 366nm V. S. Reagent
0.93 Pink 0.90 Light blue 0.94 Blue
0.71 Light pink 0.72 Brown 0.82 Red
0.64 Pink 0.55 Blue 0.77 Grey ○○○○○○○○○○○○○○○○○○○○○○○○○○○
0.56 Light pink 0.51 Red 0.67 Grey
0.52 Pink 0.42 Fluorescent blue 0.59 Brown
0.46 Pink 0.34 Red 0.50 Brown
0.32 Yellowish green 0.14 Blue 0.42 Grey
0.14 Light pink 0.32 Violet
0.22 Light grey
B1 B2 B3 0.19 Violet Fig. 2 V. S. Reagent
are shown in Table 3 and 4. The plates were developed using vanillin-sulphuric acid and heated at 105ºC till appeared coloured spots shown in Fig. 2 and 3.
Hippocratic Journal of Unani Medicine 91 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 4: Rf Values of alcohol extract
Solvent system Rf Values Toluene: Ethyl acetate (6 : 4) UV 254nm UV 366nm V. S. Reagent
0.93 Yellowish green 0.93 Red 0.90 Brown 0.81 Pink 0.81 Blue 0.81 Brown
0.73 Light pink 0.78 Violet 0.69 Violet 0.60 Pink 0.56 Yellow 0.59 Green 0.57 Yellowish green 0.52 Blue 0.53 Light grey
0.47 Light pink 0.47 Brown 0.39 Blue 0.39 Light pink 0.40 Blue 0.26 Grey 0.27 Yellow 0.32 Fluorescent blue 0.14 Grey
0.15 Yellowish green 0.28 Red
B1 B2 B3 0.17 Red Fig. 2 V. S. Reagent
Quality control parameters
The evaluated quality control parameters such as microbial load and heavy metals were found within the permissible limit in the drug shown in Table 5 and 6. The
other parameters like aflatoxins B1, B2, G1 and G2 and pesticide residues - organo chlorine group, organo phosphorus group, acephate, chlordane, dimethoate, endosulphan, endosulfan, endosulfon, ethion, endosufon sulphate, fenthion, heptachlor, lindane, methoxychlor, phorate sulfoxide and phorate sulfone were not detected from the drug samples shown in Table 7 and 8.
Table 5: Analysis of Microbial load S.No. Parameter Analyzed Results WHO Limits 5
○○○○○○○○○○○○○○○○○○○○○○○○○○○ 1 Total Bacterial Count 2,000 CFU / gm 10 CFU / gm 2 Total Fungal Count < 10 CFU/ gm 103 CFU / gm 3 Enterobacteriaceae Absent / gm 103 CFU / gm 4 Salmonella Absent / gm Nil 5 Staphylococcus aureus Absent / gm Nil
Table 6: Estimation of Heavy Metals S.No. Parameter Analyzed Results WHO & FDA Limits 1 Arsenic Not detected 3 ppm 2 Cadmium Not detected 0.3 ppm 3 Lead 0.0023 10 ppm 4 Mercury Not detected 1.0 ppm
Hippocratic Journal of Unani Medicine 92 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 7: Estimation of Aflatoxins
S.No. Aflatoxins Results WHO Limits
1B1 ND 0. 5ppb
2B2 ND 0.1ppb
3G1 ND 0. 5ppb
4G2 ND 0.1ppb
ND = Not Detected
Table 8: Analysis of Pesticide Residues
S.No. Pesticide Residues Results Limits 1 Organo Chlorine group ND (DL 0.005mg/Kg) 2 Organo Phosphorus group ND (DL 0.005mg/Kg) 3 Acephate ND (DL 0.005mg/Kg) 4 Chlordane ND (DL 0.005mg/Kg) 5 Dimethoate ND (DL 0.005mg/Kg) 6 Endosulphan ND (DL 0.005mg/Kg) 7 Endosulfan ND (DL 0.005mg/Kg) 8 Endosulfon ND (DL 0.005mg/Kg) 9 Ethion ND (DL 0.005mg/Kg) 10 Endosufon sulphate ND (DL 0.005mg/Kg) 11 Fenthion ND (DL 0.005mg/Kg) 12 Heptachlor ND (DL 0.005mg/Kg)
○○○○○○○○○○○○○○○○○○○○○○○○○○○ 13 Lindane ND (DL 0.005mg/Kg) 14 Methoxychlor ND (DL 0.005mg/Kg) 15 Phorate sulfoxide ND (DL 0.005mg/Kg) 16 Phorate sulfone ND (DL 0.005mg/Kg)
ND – Not detected
Conclusion
The evaluated data such as powder microscopy, physico-chemical, TLC and quality control shows that the genuine raw drugs were added in the formulation and there is no variation in the batch to batch consistency of the drug.
Hippocratic Journal of Unani Medicine 93 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Acknowledgement
The authors are thankful to the Director General, CCRUM, New Delhi, for his valuable guidance, encouragement and providing necessary research facilities to carry out the studies.
Reference
Anonymous, 1987. Physico-chemical standards of Unani Formulations, Part – II. CCRUM, Min. of Health & Family Welfare, New Delhi, pp. 300 - 317.
Anonymous, 1998. Quality Control Methods for Medicinal Plant Materials. World Health Organization, Geneva, pp. 25 - 28.
Anonymous, 2000. Association of Official Analytical Chemists (AOAC), 17th Edition.
Anonymous, 2006. National Formulary of Unani Medicine, Part – IV. Min. of Health & Family Welfare, New Delhi, pp. 57 – 58.
Johansen, D.A., 1940. Plant Microtechnique. Mc. Graw Hill Book Company Inc., New York and London, pp. 181-186.
Kokate, C.K., Purohit, A.P. and Gokhale, S.B., 2000. Pharmacognosy, Nirali Prakashans, Pune, pp. 98 – 99.
Wagner, H., Bladt, S. and E.M., Zgainski, 1984. Plant Drug Analysis, A Thin Layer Chromatography Atlas (2nd Edition). Springer-Verlag, Germany.
Wallis, R.E., 1997. Text Book of Pharmacognosy, 5th Edition. CBS Publishers & Distributors, Delhi, pp. 494 – 496.
Bandaranayake, W.M., 2006. Quality control, screening, toxicity and regulation of herbal drugs. Modern Phytomedicines, pp. 25 - 57.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Myers, S.P., Cheras, P.A., 2004. The other side of the coin: safety of complementary and alternative medicine. Medical J. Australia (181): 222 - 225.
Hippocratic Journal of Unani Medicine 94 Standardization ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract of Unani Drug he peel of fruit or fruit rind of Punica granatum L. is used as Post-e-Anar herbal drug. In Unani medicine it has been used to cure many ailments and known as Post-e-Anar. It is anti-oxidant, anti-cancerous, anti inflammatory and anti- (Fruit rind of Tallergic. But its standardization has not been done so far, so keeping in mind the Punica medicinal properties of Post-e-Anar, its diagnostic characters has been identified by following the modern scientific quality control procedures. The study provides granatum L.) organoleptic, physico-chemical and thin layer chromatographic profile which will be helpful in laying down the standards and pharmacopoeial parameters. The *Nazish Siddiqui, qualitative screening for phytochemicals was carried out to facilitate desirable M. Masihuzzaman Ansari, therapeutic efficacy. Besides this, novel Infra-red (IR) spectral study of rind extract Alvia Khan, Mohd. Bilal Tafseer, has also been conducted by recording the IR spectrum of the extract, which can Saba Viquar be used as fingerprints for identity of drug Post-e-Anar. and Abdul Haleem Keywords: Standardization , Post-e-Anar, Physico-chemical, IR Spectrum, Fruit rind, Phytochemicals. Department of Ilmul Advia, A.K. Tibbiya College, Aligarh Muslim University, Aligarh 202002 Introduction The plant Punica granatum L. is a large deciduous shrub or a small tree (Anonymous, 2009). It belongs to the family Punicaceae. It is commonly known as Anar and considered to be native of Iran, Afghanistan and Baluchistan. It is also found growing wild in the Himalayas between 900 to 1800 m and also cultivated throughout India. Flowers occur during April to July, while fruiting takes place during July to September (Anonymous, 2007). Various parts of plants are used as drug. Fruit juice is excellent for curing diarrhoea, dysentary and other ailments such as colitis, anaemia, jaundice, high blood pressure, piles and arthritis (Anonymous, 2009; Dipak et al., 2012), decreases cholesterol absorption ( Julie ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Jurenka, 2004). The leaf, fresh seeds and root bark of Punica granatum have been studied and standardized. (Anonymous, 2009; Anonymous, 2007; Anonymous, 1997). The external leathery cover of fruit of Punica granatum L. is known as rind and in Unani medicine it is known as Post-e-Anar. The rind or peel of fruit contains allergic acid as one of the main constituents which interrupts free radicals induced damage and acts as anti-oxidant (Yunfeng et al., 2006; Amir et al., 2011 ). It also acts as anti-cancerous inhibiting the growth of tumours and is useful in cancer of colon, breast, prostate, skin and pancreas (Julie Jurenka, 2004; Amir et al., 2011; Negi et al., 2011). Besides this rind extract has been found to possess anti-allergic and anti-inflammatory activities also (Panichayupakaranant et al., 2010). The world is heavily polluted with bacteria,
*Author for correspondence
Hippocratic Journal of Unani Medicine 95 January - March 2015, Vol. 10 No. 1, Pages 95-103 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ viruses and fungi infections and these are the major cause of diseases in human beings. Further, due to indiscriminate use of synthetic anti-microbial drugs, most infective microorganisms have developed resistance to many antibiotics. Thus, there is an urgent need to discover novel herbal anti-microbials, as they have less or no side effects. The fruit rind extract of pomegranate also shows significant antibacterial and antifungal activities (Panichayupakaranant et al., 2010; Negi et al., 2003; Adollahzaden et al., 2011; Dahham et al., 2010), Apart from this, Post- e-Anar extract was found to exhibit antimalarial activity (Agli et al., 2010), It is also useful for prevention of cardio-vascular diseases, diabetes, alzheimer,s disease, arthritis, obesity, dental problem (Mohammad & Kashani, 2012; Negi et al., 2003) and possesses antidiarrhoeal activity (Akter et al., 2013) . Thus, keeping in mind the medicinal importance of drug, various physico-chemical and phytochemical studies on the fruit rind of Punica granatum (Post-e-Anar) were carried out with a view to standardize it and lay down standards for its purity, quality control & quality assurance, which has not been taken earlier.
Methodology
Collection of Plant Material
Red pomegranate fruits were collected . Longitudinal cuts were given to the peel of the fruit and peels were gently removed with the help of hand and knife. The seeds were removed and peel was cut into pieces of 1 inch size. It was then placed in a sunny spot during day light hours every day till the peel gets hard and loses all the moisture. Dried peels were powdered to get 60-mesh size using mixer grinder and placed in air tight container as ‘Post-e-Anar’ for study.
(i) Organoleptic Parameters: The colour, taste, odour, solubility were noted which provide first hand information. ○○○○○○○○○○○○○○○○○○○○○○○○○○○ (ii) Physico-chemical Parameters: Various physico-chemical studies like total ash, acid insoluble ash, water soluble ash, successive extractive values using soxhlet extraction method, water and alcohol soluble matter, pH, bulk density, moisture content were carried out as per guidelines of WHO and others (Anonymous, 1998; Anonymous, 1963),
(iii) Qualitative Analysis of Post-e-Anar: was conducted to identify the organic chemical constituents present in the drug (Overtone, 1963).
(iv) IR Spectroscopic Study: For this alcoholic extract of the drug was obtained by refluxing powdered drug (5.0 g) with absolute alcohol (50 ml) for 5 hrs and removing the solvent under reduced pressure. The IR spectrum of alcoholic extract was determined in KBr pellets with Perkin Elmer 1600 FTIR spectrometer.
Hippocratic Journal of Unani Medicine 96 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ (v) Thin layer chromatographic analysis: For this ethyl acetate and ethanolic 20,21 extract was conducted in percolated silica gel 60F254 TLC plates. The plates were visualized in day light and UV short wavelength respectively (Harborne, 1973; Stahl, 1969).
Observations and Results
Organoleptic characters: The powder of the fruit rind of pomegranate was brownish yellow with characteristic odour and taste, showed good solubility in water (Table 1).
Physico-chemical constants: Different physico-chemical constants were determined three times and their average values are depicted (Table 2).
Table 1 : Organoleptic characters of fruit rind of Punica granatum L.
S.No. Parameters Observations 1 Colour Brownish yellow 2 Odour Characteristic 3 Taste Characteristic 4 Solubility Good in water
Table 2 : Physico-chemical Constants of fruit rind of Punica granatum L.
S.No. Parameters Analytical values (%)* 1 Ash Value Total Ash (w/w) 4.21 Acid Insoluble Ash (w/w) 2.21 Water Soluble Ash (w/w) 2.9 2 Solubility Alcoholic Soluble Matter (w/w) 7.0
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Water Soluble Matter (w/w) 8.8 3 Successive Extractives Petroleum ether 0.53 Diethyl ether 5.0 Chloroform 0.49 Ethyl acetate 1.73 Absolute alcohol 32.62 Distilled water 25.88 4 pH Values 1% Aqueous solution 3.73 10% Aqueous solution 3.40 5 Moisture content (v/w) 12 6 Bulk density 0.49 *Values are average of three experiments
Hippocratic Journal of Unani Medicine 97 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Qualitative analysis of Post-e-Anar: The phytochemicals present in the drug were qualitatively analysed by different chemical tests and results are given (Table 3).
IR spectral study of the drug: Novel IR spectral study of the alcoholic extract of the drug was done by running the alcoholic extract in the IR range (4000-667cm-1) of the electro-magnetic spectra and major characteristic peaks were noted (Fig. 1, Table 4).
Thin layer chromatographic profile: Thin layer chromatographic analysis of the ethanolic and ethyl acetate extract of Post-e-Anar was carried out using Benzene : methanol : acetic acid (45 : 8 : 4) and ethyl acetate : acetic acid (20:4) respectively as solvent system. Rf values were calculated after the development of chromatogram. The Rf values in the given solvent are used to characterize the drug,s identity and purity. The results obtained are given in table-5.
Table 3: Qualitative Analysis of Phytochemicals of fruit rind of Punica granatum L.
S.No. Chemical Constituents Test / Reagents Inference
1. Alkaloids Dragendorff,s reagent +ve Wagner,s reagent +ve Mayer,s reagent +ve
2. Carbohydrates Molish Test +ve Fehling Test +ve Benedicts Test +ve
3. Flavonoids Mg ribbon and dil Hcl +ve
4. Glycosides NaOH Test +ve
○○○○○○○○○○○○○○○○○○○○○○○○○○○ 5. Tannins / Phenols Ferric chloride Test +ve
6. Protein Xanthoproteic Test +ve Biuret Test +ve
7. Steroids / Terpenes Salkowski Reaction +ve
8. Amino acids Ninhydrin Solution -ve
Table 4: IR Spectral Details of alcoholic extract of fruit rind of Punica granatum L.
Region of electromagnetic spectra Characteristic peaks
IR, γ (cm-1) 3412, 3329, 1706, 1615, 1512, 1448, 1345, 1222, 1190, 1048
Hippocratic Journal of Unani Medicine 98 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Figure 1
Hippocratic Journal of Unani Medicine 99 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 5 : TLC Profile of alcoholic extract of fruit rind of Punica granatum L.
Extract Solvent Visualizing No. of Rf Value System Agent Spots Ethyl Ethyl Acetate : Day Light 3 0.41, 0.65, 0.76 Acetate Acetic Acid (20:4) Ethanol Benzene : UV Short wave 3 0.47, 0.59, 0.88 Methanol : Acetic Acid (45:8:4)
Discussion
The interest in evaluating therapeutic effects of plants have increased as 80% of
world’s population depends on complementary and alternative medicine for their health care needs (Magee, 2005; Duraipandiyan, 2006). The quality of herbal drugs is sum of all the factors which contribute directly or indirectly to its safety, effectiveness and acceptability of the product. Standardization of drugs means confirmation of its identity, determination of its quality and purity (Ekka, 2008). Initially organoleptic studies were used to authenticate the drugs but some drugs require other physical and chemical studies too. So standardization of Post-e- Anar was carried out as it has an important role in folk medicine. The present standardization study has brought out many diagnostic characters of herbal drug Post-e-Anar (fruit rind of Punica granatum). Organoleptic characters provide preliminary information regarding colour, taste, odour and solubility which is necessary for authentification. Physico-chemical parameters were determined to ascertain the identity and purity. Ash values were found as total ash 4.21%, acid ○○○○○○○○○○○○○○○○○○○○○○○○○○○ insoluble ash 2.21% and water soluble ash 2.9%, if the drug is adulterated with siliceous or earthy matter ash values may change. Moisture content has been determined by toluene distillation method as 12%. If its value increases that may provide an evidence that the drug is not dried and stored properly. The pH value of 1% and 10% aq. solution was 3.73 and 3.40 respectively, which helps in knowing the drug receptor site interactions and also stability and therapeutic activity of the drug. The amount of extract that a drug yields to a particular solvent is an approximate measure of the amount of certain constituents or group of related constituents that the drug contains (Jenkins et al., 1967). Thus extractive values of Post-e-Anar in six organic solvents were calculated as an index of its purity, and these are shown in Table 2. Phytochemical screening of drug was done to know the chemical constituents present in Post-e-Anar (Table 3).
Hippocratic Journal of Unani Medicine 100 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Another important technique taken here to confirm the identity was Infra red (IR) spectroscopy. The IR spectrum of alcoholic extract of fruit rind of Punica granatum (as Post-e-Anar) was recorded in fig-1, table-4. Initially the IR spectroscopic method was restricted only for structural elucidation of isolated compounds from the herbal matrix but now it is also useful in phytochemical studies as a “fingerprinting” device for comparing a natural sample with synthetic sample. In IR spectrum, region 1430-910 cm-1 is known as finger print region, which can be compared with the finger prints of human beings, which differs from person to person. In the same manner this region would differ from drug to drug. In addition to this IR spectrum also provides information about functional groups present in the chemical constituents of the drug. To check the purity of drug IR spectrum may be compared with the authentic sample. If the characteristic bands are similar and identical, the test drug would be genuine. Apart from this thin layer chromatographic (TLC) study was also carried out. Now a days other instrumental chromatographic methods are also used like GC & HPLC but TLC provide first characteristic fingerprints of herbs and various pharmacopoeias still use TLC (Jain and Sharma, 2005). TLC was made here to construct the fingerprint of Post-e-Anar, as it is simple, versatile, sensitive and economic. Results are summarized in table-5. This study assumes great significance as it will provide a key of diagnostic characters which will serve as an important tool in laying down the standards for quality assurance.
Acknowledgement
The authors are grateful to the Chairman, Department of Ilmul Advia, AMU, Aligarh, for providing necessary research facilities. Facilities provided by SAP (DRS-1) by way of generous research support are duly acknowledged.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ References
Adollahzaden, S., Mashouf, R.Y., Mortazavi, H., Moghaddam, M.H., Roozbahani, N. and Vahedi, M. , 2011. Antibacterial and antifungal activity of Punica granatum peel extract against oral pathogens, J Dent (Tehran), 8(1): 1-6. Agli, M.D., Galli, G.V., Bulgari, M., Basilico, N., Romeo, S., Bhattacharya, D., Taramelli, D. and Bosisio, E., 2010. Ellagitannins of fruit rind of Pomegranate (Punica granatum L) antagonize in vitro the host inflammatory response mechanisms involved in the onset of malaria, Malaria Journal, 9: 208. Akter, S., Sarker, A. and Hossain, M.S., 2013. Antidiarrhoeal activity of rind of Punica granatum, International Current Pharmaceutical J, 2(5): 101-104. Amir, A., Motaal, Shaker, S., 2011. Anticancer and antioxidant activities of standardization of whole fruit, pulp and peel extract of Egyption pomegrate, The Open Conference Proceedings Journal 211(41): 41-45.
Hippocratic Journal of Unani Medicine 101 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Anonymous , 2009. Unani Pharmacopoeia of India, Part I, Vol. VI. Department of Ayush, Govt. of India, New Delhi, pp. 9-10.
Anonymous, 2007. Unani pharmacopoeia of India, Part I Vol II D/O Ayush govt of India ministry of health and family welfare New Delhi, pp. 5-6.
Anonymous, 2008. Quality control manual for Ayurveda, Siddha and Unani medicine, Govt of India, Dept of AYUSH, New Delhi, pp. 21-29.
Anonymous, 1997. Standardization of single drugs of Unani Medicine, Part III. CCRUM, New Delhi, pp. 250-254.
Anonymous, 1998. Quality control methods for medicinal plant materials. World Health Organization, Geneva, pp. 25-28.
Dahham, S.S., Ali, M.N., Tabassum, H. and Khan, M., 2010. Studies on antibacterial and antifungal activity of pomegranate (Punica granatum L), American- Eurasian J. Agric & Environ. Sci. 9(3): 273-281.
Dipak, G., Patel, A., Chakraborty, M. and Kanath, J.V., 2012. Physiochemical & pharmacological profile of Punica granatum L.: An overview, Int. Res. J. Pharmacy 3(2): 65-68.
Duraipandiyan, V., Ayyanar, M. and Ignacimuthu, S., 2006. Antimicrobial activity of some ethanomedicinal plants used by paliyar tribe from Tamilnadu India, BMC complement Altern Med 6: 35.
Ekka, N.R., Namdeo, K.P. & Samal, P.K., 2008. Standardization strategies for Herbal drugs- An overview, Research J. Pharm. and Tech (4): 310-312.
Growther, L., Sukirtha, K., Savitha, N. and Niren, Andrew, S., 2012. Antibacterial activity of Punica granatum peel extracts against shiga toxin proceeding E.coli, Int Journal of Life Science, Biotechnology and Pharma Research 1(4): 164- 172. ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Jain, M.K. and Sharma, S.C., 2005. Modern Organic Chemistry. Vishal Publishing Co, India, 1038.
Jenkins, G.L., Knevel, A.M. and Digangi, F.E., Quantitative pharmaceutical chemistry. Mcgraw Hill Book Company, 6th Ed, London, pp. 239-280.
Julie Jurenka, M.T., 2004. Therapeutic application of pomegrate (Punica granatum L): A review, Alternative Medicine Review 13(2).
Magee, K.A., 2005. Herbal Therapy: A review of potential health risks and medicinal interactions, Orthod Craniofac Res. 8(2): 60-74.
Mohammad, S.M. and Kashani, H.H., 2012. Review: Chemical composition of the plant Punica granatum L and its effect on heart and cancer, Journal of Med Plant Res, 6(40): 5306-5310.
Hippocratic Journal of Unani Medicine 102 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Negi, P.S., Jayaprakash, G.K. & Jena, B.S., 2003. Antioxidant and antimutagenic activity of pomegrate peel extracts. Food Chemistry 80 (3): 393-397.
Negi, P.S., Jayaprakash, G.K. & Jena, B.S., 2003. Antioxidant and antibacterial activity of pomegrate peel extracts. J of Food Sci. 68(4): 1473-1477.
Overtone, K.H., 1963. Isolation, purification & preliminary observation in elucidation of structures by physical & chemical methods. Bentley Interscience Pub, New York, pp. 34.
Panichayupakaranant, P., Tewtrakul, S. and Yuenyongsawad, 2010. Antibacterial, anti-inflammatory and antiallergic activity of standardized pomegranate rind extract. Food Chemistry, 123(2): 400-403.
Stahl, 1969. Thin layer chromatography: A Laboratory Handbook. Springer Verlag Student edition, Springer Verlag, Berlin, pp. 52-86,127-128,900.
Wagner, A., Bladt, S., Rickl, V., 1996. Plant Drug analysis, A thin layer chromatography Atlas, Springer Berlin, p. 2.
Yunfeng, Li., Changiang, G., Yang, J., Wei, J. and Xu, J., 2006. Evaluation of antioxidant properties of pomegranate peel exract in comparison with pomegrate pulp extract. Food Chemistry 36(2): 254-260. ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 103 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 104 A Study on ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract Diversity of series of medicinal plants collection trips to various forest Unani Medicinal regions of South Western Ghats of Tamil Nadu, have brought to light 53 plants species widely used in the study area to treat certain non-communicable diseases Plants Used for Aas per Unani text. All species collected have been analyzed in respect of their Non- diversity, life form and taxonomic category. The study reveals that many such species are either rare or endangered and needs immediate attention for their Communicable conservation and protection, through germiplasm collection initiatives before these Diseases in are lost for ever. All species have been listed providing information on their botanical name, family, Unani name, therapeutic uses as per Unani text. Further scientific Southern studies are suggested with a view to discover new drugs of plant origin to combat Western Ghats many such non-communicable diseases, hitherto incurable, in modern medicine. of Tamil Nadu Keywords: Medicinal Plants, Western Ghats, Non-Communicable disease, Unani
1*R. Murugeswaran, System of Medicine 1K. Venkatesan, 1 Aijaz Ahmed Introduction and 2Aminuddin The greatest public health challenge of 21st century is chronic non-communicable diseases (Anderson and Chu, 2007). Approximately 36 million deaths were 1Regional Research Institute of Unani Medicine, attributable to NCDs in the year 2008 due to diabetes and cardiovascular diseases 1, West Mada Church Road, alone (Alwan et al., 2011). India is endowed with rich wealth of medicinal plants Royapuram, Chennai-600013 which are widely used by all section of peoples either directly or as folk remedies
2Central Council for Research in different indigenous systems of medicine or indirectly in the pharmaceutical in Unani Medicine, preparations of modern medicines (Alagesaboopathy, 2011). Medicinal plants are 61-65, Institutional Area, distributed across diverse habitats and land scape elements. Around 70% of India’s Janakpuri, New Delhi - 110058 medicinal plants are found in to tropical areas mostly in the four types spread across the Western Ghats and Eastern Ghats, Vindiyas, Chotta Nagpur Plateau, ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Aravalis & Himalayas. (Sankar Murthy and Kiran, 2012).
India is experiencing great pressure on its resources due to growing demand of the herbal products in the domestic and global markets. During the past two decades the human activities on commercialization of plant based drugs and demand from the pharmaceutical industry for domestic needs and the export of herbal drugs have led scarcity of medicinal plants in forests and plains. The natural forests should be conserved seriously to protect many of the economically important medicinal plants. The utility and need of botanical exploration in the country is to identify and search the economically important medicinal plants which has to be propagated and conserved for future generation (Balakrishnan et al.,
*Author for correspondence
Hippocratic Journal of Unani Medicine 105 January - March 2015, Vol. 10 No. 1, Pages 105-116 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ 2009). In the present study, a large number of medicinal plants were collected and identified from the southern region in recent years.
The Survey of Medicinal Plants Unit of the Regional Research Institute of Unani Medicine, Chennai has been extensively involved in medicinal plants survey and collection trips in different parts of Tamil Nadu for over three decades. The present report provides information on some 53 important medicinal species as per Unani text and widely used for treating non-communicable and other diseases in the study area. Study also analysed the diversity of Unani medicinal plants flora in respect of their diversity and the predominant families for the study area.
The Study Area
Southern Western Ghats of Tamil Nadu are one of the important medicinal plants biodiversity hotspots in the country and spread over in the districts of Coimbatore, Nilgiris, Theni, Tirunelveli and Kannyakumari. The vegetation of this area comprise evergreen, deciduous, scrub jungles and shola forests. Many of the forest areas are rich in tribal populations. The Western Ghats mountain range runs parallel to the West coast of Peninsular India for about 1600km. It has very rich floristic diversity; about 4000 indigenous angiosperm species which includes 1500 endemic plant species.
Methodology
A series of medicinal plants surveys were conducted in different seasons at Southern Western Ghats forest region of Tamil Nadu. 53 species of wild medicinal plants species which are used in the Unani system of medicine are reported in the present paper with their therapeutic uses. The medicinal species have been identified through modern floras (Gamble, 1928; Kirtikar & Basu, 1933; Nair &
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Henry, 1983; Mathew, 1983) and confirmed at Botanical survey of India, Coimbatore. The plants are arranged alphabetically according to their botanical names with collection number, followed by family, Unani name, and their uses in the Unani system of medicine (Table-I).
Results and Discussion
In the present study 53 Unani medicinal plants used for non-communicable disease as per classical text have been collected (Fig. 4-9) from Western Ghats region of Tamil Nadu (Table-1) and analyzed for their diversity status. Of these, 32 species are common, 13 falls in sporadic distribution which is very much restricted to particular area, 8 vulnerable, 5 rare and 1 species each is in endangered category, respectively (Fig.1). In the life form analysis 21 species of herbs, 18 species of
Hippocratic Journal of Unani Medicine 106 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ trees, 9 species of climbers, twiners and creepers and 5 species of shrubs were collected and identified (Fig:2).
According to the systematic classification, the taxonomic hierarchy has also been analyzed in which 53 species, belonging to 51 genus and 32 family are recorded for the study area (Fig: 3). The study also revealed that largest families of the study area are Apocynaceae, Caesalpiniaceae and Euphorbiaceae with 4 species each, followed by Amaranthaceae, Solanaceae with each 3 species, similarly Asclepiadeceae, Combretaceae, Fabaceae, Liliaceae, Loganiaceae, Malvaceae and Rubiaceae possess 2 species each and other families possess 1 species each respectively.
Fig. 1: Analysis of diversity of Unani medicinal plants with respect to no. of species in the study area ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 2: Analysis of Unani medicinal plants life forms with respect to no. of species in the study
Hippocratic Journal of Unani Medicine 107 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 3: Analysis of taxonomical ranks of Unani medicinal plants with respect to number of species in the study
The medicinal plants are used in Unani system of medicine for various ailments like anthelmintic, arthritis, astringent, aphrodisiac, bleeding hemorrhoids, diarrhoea, dysentery, gastric ulcer, headache, inflammation and stomach disorders are among them. Some of the important plant species used are Amaltas (Cassia fistula L.), Amla (Phyllanthus embilica L.), Bijasar (Pterocarpus marsupium L.), Bhuiamla (Phyllanthus amarus L.), Chalmugrah (Hidnocarpus laurifolia (Dennst.) Sleumer.,Chungchi (Abrus precatorius L.), Dudhi (Euphorabia hirta L.), Gilo (Tinospora cordifolia (Willd.) Miers), Gul-e-abbas (Mirabilis jalapa L.), Gul-e-dahwa (Anogeissus latifolius (Roxb.ex DC.) Bedd.), Halela (Terminalia chebula L.), Inderjo shirin (Wrightia tinctorea R.Br.), Kamila (Mallotus philippensis (Lam.) Mull.-Arg.), Kasoos (Cuscuta reflexa Roxb.).Katai (Solanum virginianum L.), Lajwanti (Mimosa ○○○○○○○○○○○○○○○○○○○○○○○○○○○ pudica L.), Mainphal (Catunaregam spinosa (Thumb.) Tirveng., Marorphali (Helicteres isora L.) Muleem (Gloriosa superba L.), Neem (Azadirachta indica (L.) A. Juss), Panwar (Cassia tora L.), Patharphodi (Aerva lanata (L.) Jes, Qil qil (Cardiospermum halicacabum L.)Sadabhagar (Vinca rosea L.),Sambhalu (Vitex negundo L.), Santhal safaid (Santalum album L.), Sheetraj Hindi (Plumbago zeylanica L.), Talmakhana (Hygrophila auriculata (Schum.) Heine, Tarwar (Cassisa auriculata L.), Zarawand (Aristolochia indica L.) etc., were exemplified and have been discussed for their diversity status.
It has been observed that the plant species like Asgand (Withania somnifera Dunal), Chalmugrah (Hidnocarpus laurifolia (Dennst.) Sleumer.) Darchini (Cinnamomum verum Presl.), Ghungchi (Abrus precatorius L.), Kashim (Alstonia scholaris R.Br.), Gul-e-Dhawa (Anogeissus latifolius (Roxb. ex DC.) Wall. ex Guill. & Perr.), Chironji
Hippocratic Journal of Unani Medicine 108 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 1: List of Unani Medicinal Plants Used for Non-Communicable Diseases in Southern Western Ghats of Tamil Nadu, India
Sl. Botanical Name Family Unani Uses in the Unani Sta- No. & Collection No. Name System of Medicine tus
1 Abrus precatorius L. Fabaceae Ghungchi Seeds are used as anti- V SMPU, CH- 8611 inflammatory, and stimulant. Used in nerve disorders and ulcer. 2 Abutilon indicum (L.) Malvaceae Kanghi Leaves are used in bleeding C Sweet piles, constipation and used SMPU, CH-8511 as aphrodisiac and analgesic.
3 Achyranthes aspera L. Amaran- Chirchita Whole plant used in dropsy. C SMPU, CH 8491 thaceae 4 Aerva lanata (L.) Amaran- Pathar- Whole plant used as C A. Juss. Ex Schult. thaceae phodi diuretic and demulcent. SMPU, CH-8455 5 Alstonia scholaris Apocyna- Kashim Root used as carminative, S R.Br. SMPU, ceae digestive, stomachic, used CH-8590 to expel intestinal worms, dropsy and hemiplegia.
6 Amaranthus spinosus Amaran- Kateeli Root used as appetizer, C L. - SMPU, CH-8572 thaceae chauli carminative and anti- inflammatory. 7 Anogeissus latifolius Combreta- Gul-e- Flowers are used in . S (Roxb. ex DC.) ceae Dhawa constipation, diarrhea, Wall. ex Guill. & Perr. leucorrhoea, and intestinal SMPU, CH-8474 worms 8 Argemone mexicana Papavara- Satyanasi Plant used as diuretic. Juice C L. SMPU, CH-8561 ceae of shoot used as purgative, ○○○○○○○○○○○○○○○○○○○○○○○○○○○ and used as blood purifier. Seeds used in constipation, also used as antiseptic. Oil of seeds used as purgative and root used as tonic. 9 Aristolochia indica L. Aristolo- Zarawand Seeds and roots are used in R SMPU, CH-8689 chiaceae general debility, amenorrhea and wounds, Used as antidote for poison.
10 Asparagus recemosus Liliaceae Satawar Root powder used as R Willd. - SMPU, diuretic, aphrodisiac, CH-8682 antidiarrheal, nervine tonic and used in rheumatic complaints.
Hippocratic Journal of Unani Medicine 109 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 1: (Continued)
Sl. Botanical Name Family Unani Uses in the Unani Sta- No. & Collection No. Name System of Medicine tus
11 Buchanania lanzan Anacardia- Chironji Kernel used as aphrodisiac S Spreng. - SMPU, ceae and in sexual debility and CH-8542 body weakness.
12 Calotropis gigantea Asclepia- Madar Root bark used as C (L.) R.Br. ex Ait. deceae analgesic, digestive, anti- SMPU, CH-8592 dysenteric, expectorant, arthritis. Used in asthma, general debility, Flowers used as analgesic, stomachic and expectorant. Latex used in piles.
13 Cardiospermum Sapinda- Qil Qil Seeds used as aphrodisiac. C halicacabum L. ceae SMPU, CH-8444
14 Cassia auriculata L. Caesal- Tarwar Leaves, flowers and roots C SMPU, CH-8565 piniaceae are used in diabeties, diarrhea and cardiac tonic.
15 Cassia fistula L. - Caesal- Amaltas Fruit pulp, rind of the fruit C SMPU, CH-8454 piniaceae and leaves are used in constipation, asthma, rheumatism.
16 Cassia tora L. Caesal- Panwar Seeds used as blood C SMPU, CH-8589 piniaceae purifier. Used in asthma.
17 Catharanthus roseus Apocyna- Sadabahar Leaves and roots are used C (L.) G. Don. SMPU, ceae in haemorrhage and CH-8496 constipation.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ 18 Catunaregam spinosa Rubiaceae Mainphal Fruits used as purgative, C (Thumb.) Tiruveng. carminative and inflamma- SMPU, CH-8494 tion. Used in paralysis.
19 Cinnamomum verum Lauraceae Darchini Bark and oil are used as V Presl. SMPU, liver tonic, carminative, CH-8802 expectorant, used in piles and inflammations.
20 Citrus limon (L.) Rutaceae Neembu Fruit used in brain tonic, C Burm.f. SMPU, constipation, cardiac tonic, CH-8527 exhilarant, carminative and digestive.
21 Curcuma longa L. Zingibera- Zard Chob Rhizome used as anti- C/C SMPU, CH-8609 ceae Haldi inflammatory, expectorant, used in ulcers, asthma.
Hippocratic Journal of Unani Medicine 110 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 1: (Continued)
Sl. Botanical Name Family Unani Uses in the Unani Sta- No. & Collection No. Name System of Medicine tus
22 Cuscuta reflexa Roxb. Convol- Kasoos Seeds used as purgative, C SMPU, CH-8451 vulaceae carminative, diuretic, anti- inflammatory, blood purifier. Used in brain diseases, epilepsy, paralysis, arthritis. 23 Datura metel L. Solanaceae Jaoz Masil, Leaves used as analgesic, C SMPU, CH-8570 Dhatura and anesthetic, used in joint pain. 24 Euphorbia hirta L. Euphorbia- Dudhi Whole plant used as C SMPU, CH 8524 ceae demulcent, constipation, expectorant, and aphrodisiac. 25 Gloriosa superba L. Liliaceae Muleem Root used as astringent, S SMPU, CH-8622 expectorant. Used in bleeding piles. 26 Gymnema sylvestre Asclepia- Gurmarbuti Leaves used in polyuria, R R. Br. SMPU, daceae heart diseases and CH-8529 diabetes. 27 Helicteres isora L. Sterculia- Marorphali Root and bark are used in C SMPU, CH-8493 ceae stomach ache. 28 Hibiscus rosa- Malvaceae Gudhal Flowers, leaves, seeds and C sinensis L. SMPU, roots are used as exhilarant, CH- 8442 cardio tonic, demulcent and aphrodisiac. Used in dysuria, palpitation, weakness of spleen. 29 Hydnocarpus laurifolia Flacourtia- Chalmu- Seeds, oil used in wounds. V ○○○○○○○○○○○○○○○○○○○○○○○○○○○ (Dennst.) Sleumer ceae grah SMPU, CH-8791 30 Hygrophylla auriculata Acantha- Talma- Seeds, roots are used as V (Schum.) Heine ceae khana sedative, diuretic, arthritis, SMPU, CH- 8568 gonorrhea, renal calculus, and aphrodisiac. 31 Leonotis nepetiifolia Lamiaceae Dipmal Flowering heads are used S (L.) R.Br. - SMPU, as laxative, emmenagogue CH-8668 32 Limonia acidissima L. Rutaceae Kaith Fruit and leaves are used V SMPU, CH-8564 as cardio tonic, liver tonic, refrigerant, astringent, strengthening the gum and diuretic.
Hippocratic Journal of Unani Medicine 111 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 1: (Continued)
Sl. Botanical Name Family Unani Uses in the Unani Sta- No. & Collection No. Name System of Medicine tus
33 Mallotus phillipensis Euphorbia- Kamila Tree, fruit capsule, red- C (Lam.) Muell.-Arg. ceae glandular, common; fruits SMPU, CH- 8441 are used as antiseptic and in otorrhoea.
34 Mimosa pudica L. Mimosa- Lajwanti Leaves used as analgesic, C SMPU, CH-8456 ceae and in intestinal worms.
35 Mimosops elangi L. Sapotaceae Mulsari Root used as aphrodisiac, R SMPU, CH-8651 diuretic and astringent.
36 Mirabilis jalapa L. Nyctangi- Gul-e- Leaves used as anti- C SMPU, CH-8458 naceae Abbas inflammation, in dropsy and jaundice. Flowers used in piles. Root used in joint pain, and sexual debility. Seeds used in leucorrhoea.
37 Momodica charantia Cucurbita- Karaila Fruit used as nervine tonic, C/C L. SMPU, CH-8552 ceae aphrodisiac. Used in joint pain, gout, dropsy, intestinal worms Leaves used as anti-bilious.
38 Nerium oleander L. Apocyna- Kaner Root used as aphrodisiac C SMPU, CH-8539 ceae and blood purifier. Used in sexual debility. Leaves used as blood purifier. Used in paralysis.
39 Phyllanthus emblica L. Euphorbia- Amla Fruits used in weakness of C/C SMPU, CH-8873 ceae brain, amnesia, head ache, ○○○○○○○○○○○○○○○○○○○○○○○○○○○ gastric ulcer, and diarrhoea.
40 Piper nigrum L. Piperaceae Filfil Siyah Fruits used as expectorant, C/C SMPU, CH-8452 stomach tonic, aphrodisiac, appetizer, carminative. Root used as digestive, carminative. Used in weakness of stomach.
41 Plumbago zeylanica L. Plumbagi- Sheetraj Roots and leaves are used S SMPU, CH-8457 naceae Hindi in arthritis, and facial paralysis.
42 Pterocarpus Fabaceae Bijasar Gum used as anthelmintic V marsupium. Roxb. - and opthalmia. SMPU, CH-8755
Hippocratic Journal of Unani Medicine 112 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 1: (Continued)
Sl. Botanical Name Family Unani Uses in the Unani Sta- No. & Collection No. Name System of Medicine tus
43 Ricinus communis L. Euphorbia- Arand, Seeds used as anti- C SMPU, CH-8563 ceae Bedanjeer inflammatory, purgative. Leaves used as purgative. Oil of seed used as purgative and anti- inflammatory. Used in colic, constipation, worm infestations and joint pain.
44 Rubia cordifolia L. Rubiaceae Majeed Roots used in retention of C SMPU, CH-8447 urine, bleeding hemorrhoids and emmenagogue.
45 Santalum album L. Santala- Sandal Wood used as exhilarant, E SMPU, CH-8652 ceae Safaid antiseptic, expectorant, cardiac tonic and astringent. Oil of the wood used in gonorrhea.
46 Solanum virginianum Solana- Katai khurd Fruits and roots are used C L. SMPU, CH-8790 ceae as laxative, inflammation and anthelmintic.
47 Strychnos nux-vomica Logania- Kuchla Fruits are used as tonic, V L. SMPU, CH-8566 ceae aphrodisiac, diuretic and emmenagogue.
48 Strychnos potatorum Logania- Nirmali Seeds used as astringent, V L. SMPU, CH-8532 ceae aphrodisiac and diuretic.
49 Tamarindus indica L. Caesal- Tamar Fruits used as cardiac tonic C/C SMPU, CH-8445 piniaceae Hindi, Imli and sedative. Used in ○○○○○○○○○○○○○○○○○○○○○○○○○○○ stomatitis. Kernal of seeds used in sexual disorders. Stem bark used in anorexia.
50 Terminalia chebula Combreta- Halela Fruit used in constipation, S (Gaertn.) Retz. ceae expectorant, brain tonic and SMPU, CH-8453 appetizer. Used in diarrhea, asthma and headache.
51 Tinospora cordifolia Menisper- Gilo Stem used as blood purifier, C (Willd.) Miers ex maceae anti-inflammatory, diuretic Hook.f. & Thoms. - and constipation. Used in SMPU, CH-8602 joint pain.
52 Wrightia tinctoria L. Apocyna- Inderjo Bark and seeds used as C SMPU, CH-8465 ceae shirin aphrodisiac.
Hippocratic Journal of Unani Medicine 113 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 1: (Continued)
Sl. Botanical Name Family Unani Uses in the Unani Sta- No. & Collection No. Name System of Medicine tus
53 Withania somnifera Solanaceae Asgand Root used as aphrodisiac, R Dunal SMPU, diuretic, rheumatism, ulcer; CH-8427 leaves used for painful swelling sore eye; seed used for diuretic and coagulating milk.
C-Common, C/C- Common & Cultivated, S-Sporadic, V-Vulnerable, R- Rare, E- Endangered
Unani Medicinal Plants used in non-communicable diseases in the study area
Fig. 4: Bijasar (Pterocarpus marsupium Fig. 5: Sheetraj Hindi (Plumbago Roxb.) zeylanica L.) ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 6: Kataikhurd (Solanum Fig. 7: Halela (Terminalia chebula virginianum L. (Gaertn.) Retz.)
Fig. 8: Ghunghchi (Abrus precatorius L.) Fig. 9: Kuchla (Strychnonux-vomica L.)
Hippocratic Journal of Unani Medicine 114 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ (Buchanania lanzan Spreng.), Muleem (Gloriosa superba L.), Gurmarbuti (Gymnema sylvestre R. Br.), Dipmal (Leonotis nepetiifolia (L.) R.Br), Kaith (Limonia acidissima L.), Mulsari (Mimusops elangi L.), Talmakhana (Hygrophylla auriculata (Schum.) Heine), Satawar (Asparagus racemosus Willd.) Sandal Safaid (Santalum album L.) Kuchla (Strychnos nux-vomica L),. Kuchla (Strychnos potatorum L.) and Zarawand (Aristolochia indica L.), are very much restricted in their distribution and such plant species are recommended for cultivation and propagation.
The medicinal plants which are available in the natural sources need to be conserved and propagated seriously because many species are under threat in different bio-geographic regions of the country due to various external factors and many of the valuable medicinal plants species are under threat to become rare, endangered and some are on the verge of extinction. To avoid the depletion of such valuable medicinal plants, the only alternative way is to develop many herbal gardens in the suitable areas for large scale cultivation of important medicinal plants through modern agronomical techniques.
Acknowledgement
The authors thank the Director General, CCRUM, New Delhi, for providing necessary facilities, encouragement and financial sanctions to conduct this study. We also thank the District Forest Officers, for permission to conduct the study and concerned Forest Range Officers and staff of the study area for providing necessary field assistance during the study.
References
Alagesaboopathy, C., 2011. Ethno-medicinal plants used as medicine by the Kurumba tribals in Pennagaram Region, Dharmapuri district of Tamil Nadu,
○○○○○○○○○○○○○○○○○○○○○○○○○○○ India. Asian Journal of Exp. Biology Science 2 (1):140-142.
Alwan, A. Maclen D.R., Riley, L.M., d’Espaignet, E., Mathers, C.D., Stevens, G.A. et al., 2010. Monitoring and surveillance of chronic non-communicable diseases: progress and capacity in high-burden countries. Lancet 376(9755): 1861-8
Anderson, G.F., Chu, E., 2007. Expending priorities: Confronting Chronic Diseases in countries with low income. N. Engl. J. Med. 356(3): 309-11.
Balakrishnan, V., Prema, P., Ravindran, K.C., Philip Robinson, 2009. Ethno- botanical Studies among Villagers from Dharapuram Taluk, Tamil Nadu, India. Global Journal of Pharmacology 3 (1): 08-14.
Gamble, J.S. & C.E.C., Fisher, 1928. Flora of the Presidency of Madras. Adlard & Son Ltd.,
Hippocratic Journal of Unani Medicine 115 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Kirtikar, K.R. & Basu, B.D., 1933. Indian Medicinal Plants, Vol.1-4. Allahabad, India.
Mathew, K.M., 1983. Flora of Tamil Nadu Carnatic, Vol.1-3. Botanical survey of India, Howrah.
Nair, N.C., Henry, 1983. Flora of Tamil Nadu, Vol. 1-3. Botanical Survey of India, Coimbatore.
Sankar Murthy, K. and Kiran, B.R., 2012. Medicinal plants used as anti-diabetic drug in pharmaceutical industry and their conservation; an overview. International Research Journal of Pharmacy 3(10): 65-71. ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 116 Pharmacog- ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract nostical lthaea officinalis Linn. is equated as botanical source of Standardization Unani drug ‘Gul Khatmi’. This plant species is attributed for various medicinal properties and its every morphological part yields mucilage, â asparagin, betaine, of Flowers of Astarch and sugar. The present study is carried out on the pharmacognostical Althaea standardization of Althaea officinalis Linn. (Flower) including Thin- Layer Chromatographic profile which leads to develop quality standards for the drug. officinalis Linn. (Gul Khatami) Keywords: Althaea officinalis Linn., Pharmacognosy, Drug standardization.
1*Nitin Rai, Introduction 2Rampratap Meena, 3S. Mageswari, Althaea officinalis Linn. (Family - Malvaceae) in trade is known as ‘Gul Khatmi’ or 4Shamsul Arfin ‘Marsh Mallow’. The drug referred in common parlance as ‘Khatmi’ (root and seed) and in Ayurveda, Baikh khatmi (root) and Tukhme Khatmi (seed) in Unani and Althea 5Rajeev Kr. Sharma officinalis (root) in Homoeopathy. The generic name of drug plant species (Althea-
1Pharmacopoeial Laboratory for Greek-to heal) also reflects its medicinal utility. It is anti-inflammatory, alterative, Indian Medicine, antacid, anti-tussive, aphrodisiac, antidiarrhoeal, demulcent, diuretic, emollient, Ministry of Ayush, Govt. of India, expectorant, laxative, mucilaginous, nutritive, rejuvenative and soothing. The drug Kamla Nehru Nagar, Ghaziabad-201002 is traditionally used in the treatment for the irritation of mucous membranes, as a gargle for mouth and throat ulcers, gastric ulcers, relief to bronchial asthma, in 2 Drug Standardization Research intestinal disorder like colitis, diverticulitis. It also stimulates the immune system Institute (CCRUM), PLIM Campus, and production of white blood cells. Ghaziabad-201002 Methodology 3Regional Research Institute of Unani Medicine, Botanically identified genuine sample of Unani drug ‘Gule-e-Khatmi’ was subjected 1 West Mada Church Street, Royapuram,Chennai-600013 to macroscopic and microscopic study, powder analysis and Thin-layer-
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Chromatography. Hand sections were stained and mounted in canada balsam 4Central Council for Research for anatomical studies. Lignifications on smoothed cross-surface were studied in Unani Medicine, 61-65 Institutional Area, with phloroglucinol-HCl. For studying powder, Jackson and Snowdon (1992) was Janakpuri, New Delhi-110058 followed. To determine physico-chemical constants, Indian Pharmacopoeia (Anonymous, 1955 & 1966) was followed. Standard prescribed procedures for 5Pharmacopoeia Commission for Indian Medicine and Homeopathy, histochemical studies (Johanson, 1940; Youngken, 1951; Cromwell, 1955; Trease PLIM Campus, and Evans, 1978), Chromatography (Shellard, 1968; Stahl, 1969; Smith and Ghaziabad-201002 Feinberg, 1972) were adopted. The informatics has been compiled by reviewing the available literature.
Informatics
Drug Specification: The drug consists of dried flowers Althaea officinalis Linn.
*Author for correspondence
Hippocratic Journal of Unani Medicine 117 January - March 2015, Vol. 10 No. 1, Pages 117-127 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Systematics
Family: Malvaceae.
Genus: Althaea
Synonyms: Althaea sublobata Stokes, Althaea taurinensis DC, Althaea vulgaris Alef., Malva officinalis (l.) Schimp. & Spenn. ex Schimp. & Spenn.
A perennial herb that dies in the winter. Stem erect, 60-90 cm; leaves thick, three to five lobed with serrated edges and deep veins and covered with a fine down giving them a silvery shin and softness; stipules linear-subulate; Flowers peduncled, in axillary clusters, 2.25-5 diam, five separate petals, rosy to off white with purple lilac stamens; anther sub-globose; ovary many celled; ovules one in each cell, carpels numerous (Kirtikar and Basu, 1975) (Figure 1. A & B). ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig 1. Althaea officinalis Linn. (A. Plant in natural habitat and; B. Inflorescence)
Hippocratic Journal of Unani Medicine 118 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Flowering and Fruiting: July- September, Fruiting: August-October.
Distribution: The plant species is distributed in temperate regions of the world; it grows in salt marshes, damp meadows, by the sides of ditches, sea and on the banks of tidal rivers. In India, it occurs in Punjab and Kashmir and often it is cultivated
Observations & Results
I. Macroscopic Characteristics
A. Drug: Dried flowers are cream in colour, ebractetae, pedicellate, hairy, complete, regular, actinomorphic, bisexual, hypogynous, mucilaginous and cyclic flowers. Epicalyx 6 to 9 fused, green and hairy; Calyx 5 gamosepalous, companulate 5- fid, hairy and green; Corolla 5 petals, obovate with sinuate apex, mucilaginous and twisted; Androecium monoadelphous, epipetalous, staminal tube completely encircled the gynoecium, the staminal tube gives many free filaments that bear anthers at their tips, anthers monothecous, extrose; pollen grains large, multiporate and spinous; Gynoecium polycarpellary, syncarpous, superior, multilocular, placentation axile, style fused free above, style as many as carpels, stigma linear (Fig. 2. A & B).
B. Powdered Drug: The powdered drug is light or pale cream.
II. Micro-Morphological Characteristics
A. Drug:
(i) Pedicel: T.S. of pedicel shows circular in outline; epidermis consisting of single layer of parenchyma cells covered with thick cuticle; numerous trichomes present
○○○○○○○○○○○○○○○○○○○○○○○○○○○ on the surface of the epidermis, simple unicellular covering trichomes, stellate trichomes and glandular trichomes present; collenchyma consisting of 8 to 10 layers of cells followed by parenchyma cells; vascular cylinder consisting of 20 to 25 vascular strands; each vascular strand consisting of 2 to 6 rows of vessels
Fig. 2 A. Flower Fig. 2 B. L. S. of Flower
Hippocratic Journal of Unani Medicine 119 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ with phloem on the outer side; pith consisting of large parenchyma cells in the centre; numerous druses of calcium oxalate crystals present in almost all regions of the pedicel (Fig. 3. A & B).
(ii) Epicalyx: T.S. of epicalyx shows epidermis consisting of single layer of parenchyma cells covered with thick cuticle; simple unicellular covering trichomes, stellate trichomes and glandular trichomes present; mesophyll region consisting of only parenchyma cells filled with chloroplast; vascular strand consisting of 2 to
Fig. 3 A. T. S. of Pedicel (Diagrammatic) ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 3 B. T. S. of Pedicel (Cellular)
Hippocratic Journal of Unani Medicine 120 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ 4 rows of vessels with phloem on the lower side; numerous druses of calcium oxalate crystals present in almost all regions (Fig. 4. A & B).
(iii) Calyx: T.S. of calyx shows epidermis consisting of single layer of parenchyma cells covered with thick cuticle; simple unicellular covering trichomes, glandular trichomes and numerous stellate trichomes present only on the lower side; mesophyll region consisting of only parenchyma cells filled with chloroplast; vascular strand consisting of 2 to 5 rows of vessels with phloem on the lower side; numerous druses of calcium oxalate crystals present in almost all regions (Fig. 5. A & B).
Fig. 4 A. T. S. of Epicalyx (Diagrammatic) ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 4 B. T. S. of Epicalyx (Cellular)
Fig. 5 A. T. S. of Calyx (Diagrammatic)
Hippocratic Journal of Unani Medicine 121 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 5 B. T. S. of Calyx (Cellular)
(iv) Corolla (lower side): T.S. of corolla shows epidermis consisting of single layer of parenchyma cells covered with thick cuticle; vascular cylinder consisting of 10 to 15 vascular strands; each vascular strand consisting of 2 to 4 rows of vessels with phloem on the lower side; rest of the mesophyll region consisting of parenchyma cells with air spaces; numerous druses of calcium oxalate crystals present; unicellular covering trichomes present only at corners of the corolla (Fig. 6. A & B).
(v) Corolla: T.S. of corolla shows epidermis consisting of single layer of parenchyma
○○○○○○○○○○○○○○○○○○○○○○○○○○○ cells covered with thick cuticle; vascular strand consisting of 2 to 4 rows of vessels with phloem on the lower side; very few parenchyma cells present with large air spaces in the mesophyll region (Fig. 6. C & D).
Fig. 6 A. T. S. of Corolla towards lower end (Diagrammatic)
Hippocratic Journal of Unani Medicine 122 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 6 B. T. S. of Corolla towards lower end (Cellular)
Fig. 6 C. T. S. of Corolla (Diagrammatic) ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 6 D. T. S. of Corolla (Cellular)
Hippocratic Journal of Unani Medicine 123 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ B. Powdered Drug: Powdered drug consist of parenchyma cells, spiral vessels upto 20ì, simple unicellular covering trichomes, stellate trichomes anther wall in surface view, pollen grains large, multiporate and spinous upto 150μ and druses of calcium oxalate crystals upto 25μ (Fig. 7). ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Fig. 7. Cellular components of powdered drug
Hippocratic Journal of Unani Medicine 124 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ III. Physico-Chemical Constants
The analytical values in respect of physico-chemical constant of drug were established and results are reported in Table 1.
IV. Thin-Layer Chromatography
A. 2 g of drug sample extracted with 20ml of chloroform and separated alcohol and refluxed on a water bath for 30 min. Filtered and concentrated to 5ml to carry out the Thin- Layer chromatography. Chloroform extract applied on TLC plate. The plate was developed by using Toluene: Ethyl acetate (9: 1) as mobile phase. After development the plates were allow drying in air and examined under UV
(254nm) to record spots and Rf values. The plates were dip in vanilline sulphuric acid reagent followed by heating at 1100 for 5 min and observed under visible light and its shows different spots. Inferences are tabulated below (Table 2).
Table 1: Analytical Values of Physico-chemical Constants Sl.No. Physico-Chemical Constants Analytical values 1. Moisture content, % w/w 14.91% 4. Total Ash, % w/w 12.25% 5. Acid insoluble ash, % w/w 5.65% 6. Alcohol soluble extractive % w/w 4.59% 7. Water soluble extractive % w/w 34.81%
Table 2: TLC fingerprinting data
Drug Mobile Derivatizing Visualiza- No. Rf Values Phase/ Reagents tions of of bands Solvent Spots
○○○○○○○○○○○○○○○○○○○○○○○○○○○ System Althaea Toluene : 1% vanillin- Under UV 5 0.65 (Pink), 0.55, officinalis Ethyl sulphuric 254nm 0.34 (Light pink), Linn. acetate acid 0.21 (Green) and (9 : 1) reagent 0.10 (Pink) Under UV 5 0.65 (Red), 0.56 366 nm (Fluorescent green), 0.51, 0.20(Pale blue) and 0.10 (Fluorescent green) After 9 0.94(Grey), 0.87(Violet), derivatization 0.75(Grey), 0.63 (Blue), 0.54 (Yellow), 0.44 (Violet), 0.21(Grey), 0.13 (Green) and 0.10 (Violet).
Hippocratic Journal of Unani Medicine 125 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ B. The alcohol extract applied on TLC plate and developed by using Toluene: Ethyl acetate (1: 1) as mobile phase. After development, the plate was allowed to dry in air and examined under UV (254nm).
Table 3: TLC fingerprinting data
Drug Mobile Derivatizing Visualiza- No. Rf Values of Phase/ Reagents tions of bands Solvent Spots System
Althaea Toluene : 1% vanillin- Under UV 5 0.86 (Light pink), officinalis Ethyl sulphuric 254nm 0.62 (Yellowish green), Linn acetate acid reagent 0.58, 0.42 (Pink) and (1 : 1) 0.13 (Light pink)
Under UV 5 0.90 (Blue), 0.62 366 nm (Yellow), 0.56 (Violet), 0.51 and 0.14 (Blue).
After 8 0.90 (Grey), 0.84, derivatization 0.74 (Violet), 0.60 (Grey), 0.56 (Yellowish brown), 0.46 (Grey), 0.31(Violet) and 0.20 (Blue).
Discussion
The present pharmacognostical studies on Althaea officinalis Linn. highlight macro and microscopical characters which can be employed for authentication of drug.Physico-chemical data are useful in the assessment of purity and strength ○○○○○○○○○○○○○○○○○○○○○○○○○○○ of drug. Thin-layer chromatographic profile is important parameters to identify and assessing the presence of active and other phyto constituents in the drug. The standards developed are easy, reliable and cost effective tool for proper identification and detection of adulteration in the drug material claimed to be resourced from Althaea officinalis Linn.
References
Anonymous, 1955. Pharmacopoeia of India. Manager of Publications, Govt. of India, New Delhi.
Anonymous, 1966. Pharmacopoeia of India. Manager of Publications, Govt. of India, New Delhi.
Hippocratic Journal of Unani Medicine 126 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Cromwell, B.T., 1955. The Alkaloids. In: Modern Methods of Plant Analysis (Eds. Paeck, K. and M.V. Tracy) Vol. 4. Springer – Verlag, Heidelberg.
Jackson, B.P. and Snowdon, D.W., 1992. Powdered Vegetable Drugs. Churchill Ltd., London.
Johansen, D.A., 1940. Plant Microtechnique. Mc Graw Hill Book Co., New York.
Kirtikar, K.R. and Basu, B.D., 1975. Indian Medicinal Plants, Vol. 1, 3rd ed. L.M. Basu, Allahabad.
Shellared, E.J., 1968. Quantitative paper and thin-layer chromatography. Academic Press, London.
Smith, I. and Feinberg, J.G., 1972. Paper chromatography, Thin layer chromatography and Electrophoresis. Longmans, London.
Stahl, E., 1969. Thin-layer Chromatography. A Laboratory Hand book. (Translated by M.R.F. Ashworth). Allen and Unwin, London.
Trease, G.E. and Evans, W.C., 1978. Pharmacognosy, 11th edn. Bailliere Tindel, London.
Youngken, H.W., 1951. Pharmaceutical Botany, 7th ed., The Blackistan Company, Toronto. ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 127 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 128 Development of ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Abstract Standards of hysicochemical and Phytochemical standardization is Sapistan considered a pre-requisite for the assessment of biological activity or determination of biological standards of the plant material. It provides the analytical characteristics (Cordia Pwhich may prove to be useful in fixing the physicochemical standard for the Unani dichotoma herbal drugs. Forst.f.) Sapistan (Cordia dichotoma) belongs to the family Ehretiaceae; its fruits and leaves are used in respiratory diseases such as asthma, cough, diphtheria, pneumonia,
*Abdul Haleem, sore-throat, coryza. An effort has been made to carry out the physicochemical Abdul Latif, and phytochemical studies of plant. Physicochemical parameters as Extractive Abdur Rauf, Values: Petroleum Ether (1.9%), Di-ethyl ether (0.85%), Chloroform (0.59%), Nazish Siddiqui Acetone (0.12%), Alcoholic (6.13%), Aqueous (10.44%); Solubility: Water (9.44 and %) & Alcohol (1.16 %); Moisture contents (3.45 %), Total Ash values (7.188%), pH Sumbul Rehman of 1% (6.76) & 10% solution (6.16) and loss on drying (5.3%). Phytochemical Department of Ilmul Advia, analysis: These revealed the presence of almost all the phyto-constituents in the A.K. Tibbiya College, test drug sample i.e. alkaloid, flavonoids, glycoside, carbohydrate, tannin, protein, Aligarh Muslim University, Aligarh-202002 amino acids, starch and resins.
Keywords: Sapistan, Physicochemical, Phytochemical, Standardization, Cordia dichotoma Forst.f.
Introduction
Sapistan (Cordia dichotoma Forst.f.) belongs to the family Ehretiaceae (Anonymous, 2006). Use of the drug ‘Sapistan’ in Unani system of medicine dates back about a couple of thousands of years. It is mentioned by Theophrastus and was not disregarded through the Arabic, Persian and Urdu authors in their books.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ It was particularly mentioned by Razi (926 A.D.) Ibne Sina (1037 A.D.) Al- Harwi (10th C.A.D). Sebestan or Sapistan is an abbreviation of sag pistan, derived from sag, adog and pistan dugs. The fruit resembles bitches dugs in shape. Sapistan is used for cure of the respiratory diseases (Khory and Katrak, 1984).
Sapistan is a fruit of tree. The tree is of two types, one type of tree have a large fruits known as ‘Rai Gond’. And the other tree having small fruit called ‘Kath Gond’. This tree consists of many branches which are pale red in colour. Flowers are found in clusters form. Fruit get ripened in the month of May to July (Ghani, 2011).
Medicinally, the dried fruit is valued on account of its mucilaginous nature and demulcent properties; it is much used in coughs and chest affections, also in
*Author for correspondence
Hippocratic Journal of Unani Medicine 129 January - March 2015, Vol. 10 No. 1, Pages 129-136 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ irritation of the urinary passages; in larger quantities it is given in bilious affections as a laxative. Mahometan writers describe two kinds of Sapistan; the greater, the pulp of which is separable from the stone, and the lesser, the pulp of which is adherent.
Botany- brief description:
A middle-sized deciduous tree and usually with a crooked trunk. Bark grey, rough with shallow longitudinal furrows. Leaves thinly coriaceous, variable in shape and size, orbicular, ovate-elliptic. Flowers white, fragrant. Drupes ovoid, apiculate, pink or orange to reddish when ripe, filled with a viscid pulp; stone usually 1. Common in deciduous forests, flowers – March and April; fruits – May to July.
Therapeutic action and uses :
Fruits-astringent, anthelmintic, diuretic, demulcent, expectorant. Used in affection of urinary passages, diseases of the lung and spleen. Bark – used in dyspepsia and fever. Pulp – used in ringworm. Leaves – used in ulcer, headache (Ambasta, 1986; Chopra et al., 1956; Kirtikar and Basu, 1996; Nadkarni, 1989).
No work appears to have been reported regarding standardization of this drug so far. Keeping in mind the medicinal importance of this plant in Indian Systems of Medicine, a physico-chemical and phytochemical study of this drug was carried out in order to fix-up its pharmacopoeial standards.
Material and Method
Collection of plant material: The drug samples of Sapistan (fruits) were collected from Bara Duwari market of Aligarh city and were identified. Voucher specimens were preserved in the herbarium of Medicinal Plants Lab in the Department of ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Ilmul Advia, F/O Unani Medicine, Aligarh Muslim University, Aligarh (Voucher No. SC-0141/14).
Chemical parameters: First, the organoleptic characters were studied. The dried powder of the fruits of Sapistan was used for chemical analysis. Various physico- chemical studies like total ash, acid insoluble ash, water soluble ash, alcohol and water soluble matter, moisture content, successive extractive values using soxhlet extraction method, bulk density and pH studies were carried out as per guidelines of WHO (Anonymous, 1998, 2008). Qualitative analysis of the drug was conducted to identify the organic chemical constituents present in the drug (Overtone, 1963; Harborne, 1973).
The thin layer chromatographic analysis was conducted following Stahl (1969)
and Harborne (1973) on precoated silica gel 60F254 TLC plates. The plates were
Hippocratic Journal of Unani Medicine 130 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ visualized in day light, in short UV and Long UV. They were also derivatised using iodine vapors.
Observations
(a) Organoleptic characters: The powder of the fruits of Sapistan was brown with fruity smell any characteristic odour (Table 1).
(b) Physico-chemical constants: The analytical values of different physico- chemical constants were determined (Table 2).
Table 1: Organoleptic Characters of Cordia dichotoma Forst.f.
S.No. Organoleptic parameters Observations 1. Colour Brwon 2. Smell Fruity 3. Taste Disagreeable
Table 2: Physicochemical study of Powder of Sapistan
S.No. Parameters Percentage (w/w)* 1 Ash value Total ash 7.188 Acid insoluble ash 1.22 Water soluble ash 5.92 2 Soluble Part Ethanol soluble 1.66 Aqueous soluble 9.44 3 Successive Extractive Values Pet. Ether 1.72 ○○○○○○○○○○○○○○○○○○○○○○○○○○○ Di-ethyl ether 0.85 Chloroform 0.59 Acetone 0.12 Alcohol 6.13 Aqueous 10.44 4 Moisture content 3.45 5 Loss on Drying 5.3 6 pH values 1% water solution 6.76 10% water solution 6.16 7 Bulk density 0.67
*Note: Values are average of three experiments.
Hippocratic Journal of Unani Medicine 131 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ (c) Qualitative analysis of organic chemical constituents of drug: The phytochemicals present in the drug were identified on the basis of different chemical tests given for various plant constituents (Table 3).
(d) FTAR Analysis: Fluorescence analysis of the successive extract was studied under day light as well as Ultra Violet (short and long wave length) light, results have been summarized in Table-4. FTAR Analysis was also done of the powdered drug after reacting them with various chemical reagents (Table 4).
Table 3: Preliminary Screening of major Phyto-chemicals
S.No. Chemical Constituent Tests/Reagent Inference 1 Alkaloids Dragendorff’s reagent + Wagner’s reagent + Mayer’s reagent + 2 Carbohydrate Molisch’s Test + Fehling’s Test + Benedict Test + 3 Flavonoids Mg ribbon and Dil.Hcl + 4 Glycosides NaOH Test + 5 Tannins/Phenols Ferric Chloride Test + Liebermann’s Test + Lead Acetate Test + 6 Proteins Xanthoproteic Test + Biuret Test + 7 Starch Iodine Test +
8 Saponins Frothing with NaHCO3 + 9 Steroid/Terpenes Salkowski Reaction + 10 Amino Acids Ninhydrin Solution – ○○○○○○○○○○○○○○○○○○○○○○○○○○○ 11 Resin Acetic Anhydride test +
Indications: ‘–’ Absence and ‘+’ presence of constituent.
Table 4: FTAR Analysis of Sapistan
S.No. Extract Day Light UV Long UV Short 1. Pet. Ether Brown Black Greenish 2. Di-ethyl ether Greenish Bluish Greenish 3. Chloroform Light Green Black Light Green 4. Acetone Brown Bluish Greenish 5. Alcohol Brown Black Dark Brown 6. Aqueous Brown Black Black
Hippocratic Journal of Unani Medicine 132 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ (e) Thin layer chromatographic profile: Thin layer chromatographic analysis of successive extracts was carried out using different solvent systems and
visualizing agents and Rf values were calculated to standardize the drug for its identity and purity (Table 5).
Table 5: Fluorescence Analysis of Sapistan with different chemical reagents
S. Powdered drug + Day light UV short UV long No. Chemical Reagent
1. Powdered drug + Conc. HNO3 Golden Light Green Green 2. Powdered drug + Conc. Hcl Brown Light Green Dark Green 3. Powdered drug + Conc. Dark Brown Dark Green Black
H2SO4 4. Powdered drug + 2% Iodine Brown Green Black solution 5. Powdered drug + Glacial Light Brown Light Green Dark Green
Acetic acid + HNO3 6. Powdered drug + Glacial Pale Light Green Black acetic acid 7. Powdered drug + NaOH(10%) Brown Light Green Dark Green
8. Powdered drug + Dil. HNO3 Brown Green Green
9. Powdered drug + Dil. H2SO4 Brown Green Dark Green 10. Powdered drug +Dil. Hcl Light Brown Light Green Black 11. Powdered drug + Dark Brown Bright Green Green Dragendorff’s 12. Powdered drug + Wagner’s Brown Green Dark Green Reagent
○○○○○○○○○○○○○○○○○○○○○○○○○○○ 13 Powdered drug + Benedict’s Blackish Light Green Black reagent 14 Powdered drug + Fehling Brown Light Green Green reagent 15 Powdered drug + KOH (10%) Brown Green Black Methanolic
16 Powdered drug + CuSO4 (5%) Brown Light Green Black 17 Powdered drug + Ninhydrin Brown Green Black (2%) in Acetone 18 Powdered drug + Picric Acid Yellow Bright Green Black 19 Powdered drug + Lead Brown Light Green Black Acetate (5%)
Hippocratic Journal of Unani Medicine 133 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Table 6: Thin Layer Chromatography
Treatment Mobile phase: No of Rf value and colour of spots spots Petroleum Ether Extract Day Light Petroleum ether : 1 0.42 (Brown) UV Short Di-ethyl ether (4:1) 1 0.42 (Bluish) Iodine Vapour 1 0.42 (Yellowish) Aqueous Extract Day Light Butanol: Acetic 1 0.75 (Brown) UV Short acid: Water (5:1:4) 3 0.64 (Bluish), 0.78 (White), 0.85 (White) UV Long 4 0.21 (white), 0.42 (white), 0.78 (white), 0.85 (white) Iodine Vapour 1 0.75 (Pale Yellow) Alcoholic Extract Day Light Chloroform: 1 0.62 (Pale yellow) UV Short Methanol (9:1) 2 0.62 (Purple) , 0.75 (Purple) UV Long 2 0.62 (white), 0.75 (White) Iodine Vapour 1 0.62 (Pale Yellow)
Results and Discussion
Physico-chemical standardization is of prime importance in quality control of Unani drugs. As the efficacy of many drugs mainly depends upon its physical and chemical properties, therefore, the determination of physico-chemical characters for the authenticity of a drug is necessary before studying any medicinal property.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Phyto-chemical constituents present in the drug vary, not only from plant to plant but also among different samples of same species, depending upon various atmospheric factors, storage and drying conditions.
The present study is an attempt to fix-up pharmacopoeial standards of drug ‘Sapistan’ to ascertain its quality, identity, purity and strength. Powder of the drug has been used for study to bring out several standards like ash, solubility in alcohol and water, successive, extractive values and quality screening of physicochemicals, total alkaloids, total flavonoids, phenol, nitrogen, fatty matter, sterol/ terpenes, protein and carbohydrates.
Based on the various physico-chemical and phytochemical parameters, the drug ‘Sapistan’ has been standardized to ensure its use in manufacturing quality and genuine herbal preparations.
Hippocratic Journal of Unani Medicine 134 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○
Sapistan Fruits (Cordia dichotoma Forst.f.)
TLC of Alcoholic Extract of Sapistan TLC of Aqueous Extract of Sapistan ○○○○○○○○○○○○○○○○○○○○○○○○○○○
References
Afaq, S.H., Tajuddin, Siddiqui, M.M.H., 1994. Standardization of Herbal Drugs. Publication Division, AMU (Aligarh), pp. 33-34, 41-42, 100, 143-146.
Ambasta, S.P., 1986. The Useful Plants of India. PID, CSIR, New Delhi.
Anonymous, 1997. Standardization of Single Drugs of Unani Medicine, Part III. CCRUM New Delhi, pp. 124-128.
Anonymous, 1998. Quality control methods for medicinal plant materials. World Health Organization, Geneva, pp. 25 -28.
Hippocratic Journal of Unani Medicine 135 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ Anonymous, 2006. National Formulary of Unani Medicine, Part I. CCRUM, Dept. of AYUSH, New Delhi, p. 272.
Anonymous, 2008. Quality control manual for Ayurveda, Siddha and Unani medicine. Govt. of India, Dept. of AYUSH, New Delhi, pp. 21 – 29.
Anonymous, 2008. Unani Medicinal Plants of Tarai Forests in Kumaon Region of Uttarakhand. Central Council for Research in Unani Medicine, p. 60.
Baitar I., 1999. Al-jamiul-mufridat-ul-advia-wa-ul-aghzia. Urdu Translation. Part.III. CCRUM, New Delhi, pp. 23-24
Chopra, R.N., Nayar, S.L. and Chopra, I.C., 1956. Glossary of Indian Medicinal Plants, CSIR, New Delhi.
Dymock, 1891. Pharmacographica Indica, Part II. Published by Institute of Health and Tibbi Research, Hamdard National Foundation, Pakistan, pp. 518-519.
Ghani Najmul, 2011. Khazainul Advia. Idara Kitabul Shifa, New Delhi, pp. 787- 788.
Harborne, J. B., 1973. Phytochemical methods. Chapman and Hall. London, p. 70.
Ibn-e-Sina, 2007. Al-Qanoon-fi’l-tib, Vol. II. Urdu translation by Ghulam Hussain Kantoori. Idara Kitabul Shifa, New Delhi, p. 167.
Jenkins, G.L., Knevel, A.M. and Digangi, F.E., 1967. Quantitative Pharmaceutical Chemistry. 6th edition. The Blackiston Division. McGraw Hill Book Company, U.S.A., pp. 225, 235, 379, 425, 463, 492.
Khory, R.N. and Katrak, N.N., 1984. Materia Medica of India and their Therapeutics. Neeraj Publishing House, Delhi, p. 421.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ Kiritikar, K.R. and Basu, B.D., 1996. Indian Medicinal Plants, Vol. III. International Book Distributers (Dehradun), pp. 1674-1679.
Nadkarni, K.M., 1989. The Indian Materia Medica, Vol. II. Bombay Prakashan Pvt. Ltd., Bombay, pp. 379-380.
Overtone, K.H., 1963. Isolation, Purification and preliminary observation in elucidation of structures by physical and chemical methods. Bentley Interscience Pub., New York, p. 34.
Stahl, 1969. Thin Layer Chromatography: A laboratory handbook, Springer Verlag student edition. Springer Verlag, Berlin, pp. 52 – 86, 127 – 128, 900.
Hippocratic Journal of Unani Medicine 136 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ HIPPOCRATIC JOURNAL OF UNANI MEDICINE
Instructions to contributors
1. The paper(s) should be submitted in duplicate. Submission of a paper will be taken to imply that it is unpublished and is not being considered for publication elsewhere.
2. Papers should be written in English language and typed with double spacing on one side of A-4 size paper leaving top and left hand margin at least 1" (One inch) wide. Length of the paper should normally not exceed 12 pages.
3. Papers should be headed by a title, the initial(s) and surname(s) of author(s) followed by address.
4. Each paper should bear abstract, 2 to 5 keywords, introduction, methodology, observations, results and discussion followed by acknowledgements and references.
5. In all studies of plants or animals proper identification should be made as to the materials used.
6. While submitting the paper(s) for publication, Author(s) should decode the drugs specially in case of clinical studies.
7. Bibliographical references should be listed in alphabetical order of the author at the end of the paper. Authors should be cited in the text only by their surname(s) but their initial(s) should be shown in the bibliography.
○○○○○○○○○○○○○○○○○○○○○○○○○○○ 8. References to periodicals should include the name(s) and initial(s) of author(s), year of publication, title of the book, periodical, title of the article, volume number (Arabic numerals), issue number where appropriate, first and last page number. Reference to books should include name(s) and initial(s) of the author(s), year of publication, exact title, name(s) of publisher, place of publication, page number.
9. Reference should be cited in the text in parentheses by the name(s) of author(s) followed by the year of publication, e.g. “(Jain,1991)” except when the author’s name is part of the sentence, e.g. “Jain (1991) has reported that.” If there are more than two authors it is in order to put “ et al.” after the first name, e.g., Khan et al., 1981.
Hippocratic Journal of Unani Medicine 137 ○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○○ 10. Each table should be typed on a separate sheet of paper. Tables should be numbered consequently in Arabic numerals e.g. “Table 1, Table 2” etc., and attached to the end of the text. Tables should be provided with headings and kept as simple as possible and should be referred to in the text as “table 1” etc.
11. Figures (including photographic prints, line drawings on strong white or transparent paper, and maps) should be numbered consequently in Arabic numerals, e.g. “Fig. 1 etc.” and attached to the text behind the tables. Graphs and diagrams should be large enough to permit reduction to a required size, legends for figures should be listed consequently on a separate sheet of paper. Photographs should be on glossy printing paper.
12. The editors reserve the right to refuse any manuscript submitted, whether on invitation or otherwise, and to make suggestions and modifications before publication.
13. Paper accepted by the editorial board will become the property of the CCRUM. No article or any part thereof may be reproduced in whatever form, without the written permission of the Editor-in-Chief.
14. The editors and publisher are not responsible for the scientific contents and statements of the authors of accepted papers. ○○○○○○○○○○○○○○○○○○○○○○○○○○○
Hippocratic Journal of Unani Medicine 138