Abinash kumar sahu, et al., 2020/ Phytochemical Evalution and In-vitro Anthelmintic
RESEARCH ARTICLE
International Research Journal of Pharmaceutical and Biosciences
Pri -ISSN: 2394 - 5826 http://www.irjpbs.com e-ISSN: 2394 - 5834
Phytochemical Evalution and In-vitro Anthelmintic Activity of Oldenlandia corymbosa
Abinash Kumar Sahu*, Amiya Ranjan Padhan, Nishigandha Sa, Anjan Kumar Padhan The Pharmaceutical College, Tingipali, Barpali, Bargarh, B.P.U.T Odisha.
Article info Abstract
ABSTRACT: The plant Oldenlandia corymbosa belongs to family Article history: “Rubiaceae”. The plant is used as anthelmintic, antirheumatic, depurative, Received 15 JULY 2020 Accepted 26 JULY 2020 diaphoretic, digestive, diuretic, febrifuge, pectoral , stomachic, treat jaundice and other liver conditions and to treat viral infections, cancer, acne, boils, appendicitis, hepatitis, eye problems and bleeding etc Aim; The aim of the study is to investigate Phytochemical screening of the
Petroleum ether and Aqueous extract of Oldenlandia corymbosa and the presence of different secondary metabolites responsible for the therapeutic values of the drug and also to find out the anthelmintic activity study by in-vitro test species Pheretima posthuma responded towards the plant extracts by showing the sign of paralysis and death finally Results; The % of yield of petroleum ether extract and Aqueous extract were found to
10.92% w/w and 5.45%w/w. In fluorescence analysis the powder drug was treated with different reagent which showed different colours in U.V light. *Corresponding author: Conclusion: The % yields of both extracts were found to 10.92% w/w and [email protected] 5.45%w/w. The preliminary phytochemical screening showed the presence of alkaloids, carbohydrate, saponins, Proteins & amino acid, Phenolic & Copyright 2020 irjpbs flavonoids compounds and shows the absence of Phytosterol. In fluorescence analysis the powder drug was treated with different reagent which showed different colours in U.V light. The data revealed that the aqueous extract has a better wormicidal effect than Petroleum Ether extract.
KEY WORD: Oldenlandia corymbosa, medicinal use, phytochemical
screening, Phytochemical, Albendazole, Anthelmintic Activity, Pheretima posthuma.
International Research Journal of Pharmaceutical and Biosciences (IRJPBS) 5 (4) 1 Abinash kumar sahu, et al., 2020/ Phytochemical Evalution and In-vitro Anthelmintic INTRODUCTION Plants have been used in medicines since time immemorial. India has a rich heritage of using medicinal plants in traditional medicines, as in the Ayurveda, Siddha and Unani systems besides folklore practices. The earliest inscription of the medicinal uses of plants is found in the “Rigveda”, which is one of the oldest repositories of human knowledge. Fairly comprehensive information on the curative properties of some herbs has been found recorded in “Charak Samhita” and “Sushruta Samhita”. The plant kingdom is a virtual goldmine of biologically active compounds and it is estimated that only 10-15% of existing species of higher plants have been surveyed. Many plants have been successfully used in the treatment of various diseases. The ancient record is evidencing their use by Indian, Chinese, Egyptian, Greek, Roman and Syrian dates back to about 5000 years. In India, around 20,000 medicinal plant species have been recorded recently, but more than 500 traditional communities use about 800 plant species for curing different diseases [1]. Currently, 80% of the world population depends on plant-derived medicine for the first line of primary health care for human alleviation.
TAXONOMY AND ETHNO MEDICINE OF OLDENLANDIA CORYMBOSA
Scientific Classification [2] Rank Scientific Name Kingdom :Plantae Subkingdom :Tracheobionta Superdivision :Spermatophyta Phylum :Angiosperms Division :Magnoliophyta Class :Magnoliopsida Subclass :Asteridae Order :Rubiales Family :Rubiaceae Subfamily :Rubioideae Genus :Oldenlandia L. Species :Oldenlandia corymbosa L. Synonyms :Hedyotis corymbosa, Hedyotis biflora, Gerontogea corymbosa, Gerontogea biflora, Gerontogea herbacea. In different parts of India Oldenlandia corymbosa is known by different names[3] Sanskrit : Parpata, Parpataka English : Diamond flower Hindi : Daman pappar, Pitpapra Malayalam : Parpatakapullu, Parpatakam Gujarati : Parpat, khet-papra Marathi : Papti, Phapti, khet-papda, paripat International Research Journal of Pharmaceutical and Biosciences (IRJPBS) 5 (4) 2
Abinash kumar sahu, et al., 2020/ Phytochemical Evalution and In-vitro Anthelmintic Tamil : Parpatagam, kattucayaver, pappanpuntu Telugu : Verrinella- vemu Kannada : Parpatahullu, Kallasabatrasige Bengali : Khet-papra
GEOGRAPHICAL DISTRIBUTION [4]
Oldenlandia corymbosa is native to India, but also found throughout Malaysia, Sri Lanka, tropical East Asia to Java, Africa and Phillipines. It is a weed habitually growing in sandy lands and waste places. It is dispersed all over India, Baluchistan, Ceylon, tropical and subtropical Asia and Africa.
PLANT PROFILE: [5]
Oldenlandia corymbosa is known as diamond flower which is an annual, terrestrial, dichotomous, slender herb with ascending or erect stems up to 50 cm long ridged, glabrous or scabridulous or pubescent on the ribs which are 4-angled. Linear-oblong or narrowly elliptic, almost stalkless leaves are 1-3 to 2 cm long and by 0.8 -3 mm wide, rough on margins. The lower leaves are often broader than upper ones, linear, acute, glabrous, usually with recurved margins. Midrib of the leaf is prominently visible. Flowers are borne in 2-8 flowered cymes in leaf axils. Flowers are white in pairs or in threes or faintly pinkish- purplish, on slender stalks 4-8 mm long and usually on solitary axillary peduncles longer than the calyx. Flower tube is about 2 mm long, with 4 petals. Stamens are inserted just above the base of the tube. Capsule is about 2 x 2 mm, flattened at apex, slightly laterally compressed. Fruits are loculicidal capsules, globose and the seeds are minute, pale brown, angular, testa teticulate. These capsules are about 2 x 2 mm, exalate, flattened at apex, slightly laterally compressed Diamond Flower is found in found all over the tropical world. It is also found in the Himalayas.
Figure.1– Oldenlandia corymbosa herb Habit: This Oldenlandia corymbosa herb is sparsely to very densely branched near the base and mainly found in grassland, crop field, mountain scrub, shallow soil on rocks, lawn, garden, waste places, roadsides, cultivated and disturbed ground (Fig. 1), at elevations from sea level to 2000 meters [6].
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Macroscopical characters [7, 8, 9, 10] Leaves: leaves are subsessile, linear-lanceolate, acute, recurved and hairy margins, stipulate, stipules membranous, truncate with a few bristles. Leaves are opposite, linear lanceolate, 1-3 cm long and 2-4 cm wide. They are sessile, acute at the apex, entire at the margins, smooth, pale green beneath and dark green above. leaf is entire, secondary veins are sometimes or not at all visible; stipules are fused to petiole bases. Stem: Stem of the plant is 4-angled to flat glabrous and angles are thick to wing. Stem is woody at the base, more or less angled, slender and glabrous. Inflorescence: Axillary solitary. Is a corymbose-umbel with 2-4 flowers on a peduncle up t0 2 cm long, occurring in leaf axils all along the stem. Flowers: The flowers are White, small peduncles, pedicels 2-3 on the top, bract single below the pedicels. Calyx teeth 4, triangular, corolla lobes 4, acute, valvate. Stamens 4, in the throat of the corolla. Ovary 2-celled, ovules many, stigmas 2, linear. white, 2cm in diameter on pedicels up to 1 cm long. Calyx: Calyx-tube ellipsoid, 0.7–1 mm. long; lobes triangular, 0.5–1.8 mm. long, setulose on the margins. Corolla: Corolla white or tinged blue, pink or purple or with 2 pink stripes on each lobe; tube 0.6–1 mm. long; lobes ovate to oblong, 0.5–1.2 mm. long. Capsule: Capsule ovoid, 1.2–2.2 mm. tall, (1–)1.8–2.8 mm. wide, the beak scarcely raised. globose, these capsules are with two carpels. Flowering and Fruiting Time: August - October Seed: Seeds are, smooth, numerous minutes, round and brownish pale brown, ellipsoid or very obtusely depressed conic, ± 0.3 mm. long, reticulate. Fruit: Fruit capsular, sub-globose, ovoid, 1.2-2×1.2-2.2 mm size, dehiscent through flat to broadly rounded apex, beak when present to 0.5 mm, peduncles and pedicels usually extending promptly and prominently as the fruit develops.
CHEMICAL CONSTITUENTS:
The chemical constituents reported in different parts of Oldenlandia corymbosa are mentioned beneath. 1) Different phytochemical studies show the presence of proteins, carbohydrates, phenols, tannins, flavanoids, saponins, steroids, terpenoids, coumarin and glycosides. Some of the isolated compounds from whole plants are Geniposide, iridoid glycosides, 6 alpha – hydroxygeniposide, scandoside methyl ester (6 beta - hydroxygeniposide), 10-o-benzoylscandoside methyl ester, asperulosidic acid, asperuloside, deacteylasperuloside, 10-o-p-hydroxy benzoylscandoside methyl ester, rutin and (+)- lyoniresinol-3-alpha -o-beta glucopyranoside [11,12,13].
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2) The plant also contains ursolic acid, oleanolic acid and γ-sitosterol. The air-dried plant contains 0.12% of alkaloids – bifloron and biflorin, these two alkaloids are interconvertible. It also contains 13.55% of inorganic ash, which is mainly responsible for its cooling effect [14]. 3) An aqueous extract of the plant yielded a polysaccharide, composed of rhamnose, arabinose, zylose, mannose, galactose and glucose. 4) The methanol extracts of Oldenlandia corymbosa showed the presence of flavonols such as Quercetin, 3‟-Methoxy quercetin and 3‟, 4‟-Dimethoxy quercetin. Phenolic acids like vanillic, syringic acid, melilotic acid, p-hydroxy benzoic, p-coumaric, ferulic and caffeic acids are also present. Anthocyanidins like cyanidin and pelargonidin are present. Irridoids and alkaloids are also present [15]. The freshly prepared crude extract was qualitatively tested for the identification of chemical constituents, such as, alkaloids, flavonoids, steroids, glycosides, saponins, terpenoids, gums and tannins. [16]
TRADITIONAL USES [17, 18, 19, 20] 1. The leaves are pounded, soaked in warm water and the liquid drunk to treat stomach disorders 2. They are used externally as a poultice to treat sores and sore eyes 3. The roots are reported to have vermifuge properties. They are often used as a tincture. 4. The roots yield a green dye known as 'gerancine' after treatment with a mordant. 5. The entire plant is used in decoction as an anthelmintic, antirheumatic, depurative, diaphoretic, digestive, diuretic, febrifuge, pectoral and stomachic 6. It is also used to treat jaundice and other liver conditions. 7. In Chinese medicine it is used to treat viral infections, cancer, acne, boils, appendicitis, hepatitis, eye problems and bleeding 8. The entire plant is used in helminthiasis, leprosy, skin diseases, cough, bronchitis, necrosis and nerve depression 9. In Africa it is used to facilitate child birthatic, depurative, diaphoretic, digestive, diuretic, febrifuge, pectoral and stomachic.
The plant is known to clear heat and toxins, activate blood circulation, promote diuresis and relieve strangury. It is also known to act on lymphosarcoma and carcinoma of the liver and larynx. It is also active against appendicitis, hepatitis, pneumonia, cholecystesis, urinary infection, cellulites and snake bite. Chinese folk medicine describes the plant to treat skin sores, ulcers, sore throat, bronchitis, gynecologic infections and pelvic inflammatory diseases [21, 22, 23, 24].
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Abinash kumar sahu, et al., 2020/ Phytochemical Evalution and In-vitro Anthelmintic A scrutiny of literature revealed some notable pharmacological activities of the plant such as hepatoprotective, cytotoxic anti-oxidant, oxytocic and anti-malarial activity [25]. The plant is widely used in treating various diseases associated with kidney.
MATERIALS AND METHODS
The following drugs and chemicals were used for the different experimental study. The Mayer's, Hager’s, Barfoed’s, Benedict’s and millon’s reagent were purchased from S.D. Fine Chemical, Mumbai. The solvents petroleum ether, Chloroform was purchased from Hi Media Laboratories Pvt. Ltd., Mumbai. Methanol and Petroleum ether was purchased from Qualigens chemicals. Mumbai. And all others chemicals, solvents and reagents were of analytical grade and procured from authorized dealer.
Plants collection, Identification and processing:
The stems of Oldenlandia corymbose were collected from adjoining area of Barpali (Dist-Bargarh, Orissa) in the month of October 2019. The plant was identified by and authenticated By Prof. (Dr.) Santosh Kumar Dash, Retired Professor and H.O.D., P.G Dept. of Biosciences, C.P.S, Mohuda, Berhampur, Ganjam, Odisha. on dated 02-10-2019. The plant was washed properly with water to remove the mud or dust, and then it was dried in sun light for 1 hour and kept in shade dried and powdered by the help of mechanical process. The coarse powder has stored in air tight container for further studies.
PREPARATION OF EXTRACTS
Preparation of the extracts of Oldenlandia corymbosa powdered of entire plant by using following solvents:
(a) Pet ether Extract (b) Aqueous Extract
(a) Pet ether Extract
The shade dried course powder of the entire plant (200 gm) was packed well in soxhlet apparatus and was subjected for continuous hot extraction with Pet ether extract. The extract was filtered while hot and the resultant extract was distilled in vacuum under reduced pressure in order to remove the solvent completely. Dried and kept in a desiccator till experimentation. Obtained extract (dark wine red) was weighed and percentage yield was calculated in terms of air-dried powdered crude material.
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Abinash kumar sahu, et al., 2020/ Phytochemical Evalution and In-vitro Anthelmintic (b) Aqueous extract The shade dried course powder of the entire plant (200 gm) was packed well in soxhlet apparatus and was subjected to continuous hot extraction with distilled water until the completion of extraction. The extract was filtered while hot and the resultant extract was distilled in vacuum under reduced pressure in order to remove the distilled water completely. It was finally dried and kept in a desiccator till experimentation. Obtained extract (dark reddish brown) was weighed and percentage yield was calculated in terms of air-dried powdered crude material. The yield and % yield of both Petroleum ether and aqueous extracts of powdered of entire plant Oldenlandia corymbosa were reported below.
Table-1 Extraction values of Hydro-alcoholic and aqueous extracts of Oldenlandia corymbosa
Sl. No. Extracts % Yield (w/w) Colour of extract
1. Petroleum ether 10.92% Dark wine red
2. Aqueous 5.45% Dark reddish brown
PRELIMINARY PHYTOCHEMICAL SCREENING: The plant may be considered as a biosynthetic laboratory, not only for the chemical compounds such as Carbohydrates, Protein and Lipids that are utilized as food by human being but also for a multitude of compounds like Glycosides, Alkaloids, Volatile oils, Tannins etc., that exerts a physiological effect. The compounds that are responsible for therapeutic effect are usually the secondary metabolites. A systemic study of a crude drug embraces through consideration of both primary and secondary metabolites derived as a result of plant metabolism. The plant material may be subjected to preliminary phytochemical screening for the detection of various plant constituents. The different qualitative chemical tests can be performed for establishing profile of given extract for its chemical composition. The following tests may be performed on extracts to detect various phytoconstituents present in them [25]. Methods:
Qualitative phytochemical studies of extract will be studied for the presence of different secondary metabolites responsible for the therapeutic values as per the standard procedures mentioned in Pharmacopoeia [26,27]. The solvent free extract of about 50 mg was stirred with few ml of dilute hydrochloric acid filtered. The filtrate was tested carefully for different constituents.
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Table-2Qualitative Phytochemical analysis of Petroleum ether and aqueous extracts of Oldenlandia corymbosa. Phytochemical test Petroleum ether Aqueous Extract Extract Alkaloid test Mayer’s test Present Present Wagner’s test Present Absent Hager’s test Present Present Dragendorff’s test Present Present Carbohydrates Molish’s test Present Present Fehling’s test Present Absent Barfoid’s test Absent Absent Benidict’s test Present Absent Borntrager’s test Present Present Saponins Foam test Absent Present Proteins & amino acid Millon’s test Present Present
Biuret’s test Present Absent Ninhydrin test Present Present Phenolic compounds & flavonoids Ferric chloride test Absent Absent Lead acetate test Present Absent Alkaline test Present Positive Phytosterol : Libermann-Burchard’s Absent Absent test
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Table-3 Powder analysis with chemical reagent
Reagents Colour of powder
Powder as such Light Brown Powder + conc. HCI Light yellow
Powder + conc. HNO3 Yellowish brown
Powder + conc. H2SO4 Dark brown Powder + glacial acetic acid Brown
Powder + dil. HCI Light brown Powder + NaOH sol. Light brown
Powder + FeCl3 Yellowish green
Powder + picric acid Yellow Powder + ammonia Light brown
Powder +Iodine Brown
Table-4Fluorescence analysis of powder drug Reagent Colour Colour Colour observed observed (U.V observed (U.V (naked eye) short wave long wave length) length) Powder as such Light Brown Light Brown Dark brown
Powder + 1N NaOH Yellowish Light green Dark brown in methanol brown
Powder + NaOH in water Light brown Light brown Dark brown
Powder + 50% HCl yellowish Light green Black brown
Powder +50% H2SO4 Light brown Light green Black
Powder +50% HNO3 Light brown Green Dark brown Powder + petroleum ether Light brown Light brown Dark brown
Powder + chloroform brown Green Dark brown Powder + picric acid yellow Green Black
Powder + 5% FeCl3 Yellow green Green Black Powder + 5% iodine solution Dark green Dark green Black Powder + methanol Light brown Dark brown Dark brown
Powder + (HNO3 + NH3) Light brown Yellow green Dark brown
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DETERMINATION OF ANTHELMINTIC ACTIVITY [ 28] :
The anthelmintic study was done by using one in-vitro species adult earthworms Pheretima posthuma. Earthworms were collected near the swampy water in our locality. The average size of the round worm was 5-7 cm; average size of the earthworm was 8-9 cm. These earthworms were identified and services of veterinary practioner were utilized to confirm the identity of worms. The suspensions of various extracts were prepared in 2% gum acacia solution to obtain 1, 2.5 and 5% concentrations. Solutions of similar concentrations of the standard drug albendazole were also prepared in distilled water.
Two ml of each concentration of various extracts of Oldenlandia corymbosa and standard drug albendazole were diluted to 10 ml separately with normal saline and poured in petridishes. 2ml of 2% gum acacia solution was diluted to 10ml with normal saline to serve as control. Six earthworms of nearly equal size were placed in each Petridis at room temperature.
Table 5: Anthelmintic effect of oldenlandia corymbose extracts. Group Concentration Time in minutes (Mean ± SEM) of Paralysis time Death time (Min) Extract (%) (Min) Albendazole 10 mg/ml 14min,18 sec ±17 18min,18 sec ±42 (standard) 30 mg/ml 10min,21 sec ±12 14 min,12 sec ±10 50 mg/ml 8 min,12 sec ±14 12min,40 sec ±11 Petroleum 15 mg/ml 27min,16 sec ±17 32min,15 sec ±48 ether extract 30 mg/ml 23min,26 sec ±12 30 min,26 sec ±12 50 mg/ml 21 min,48 sec ±14 24 min,48 sec ±14 15 mg/ml 12min,19 sec ±17 20min,15 sec ±48 Aqueous extract 30 mg/ml 12min,26 sec ±12 16 min,26 sec ±12 50 mg/ml 10 min,48 sec ±14 13min,14 sec ±10 Control - - - Results are expressed as mean ± SEM from six observations, Control worms were alive up to 24 hrs. of observation, N/A= No Activity shown within 24 hours.
Time was recorded at the time of releasing the earthworms to each concentration. The time taken (minutes) for the complete paralysis and death were recorded. The mean paralysis time for each sample was recorded. The anthelmintic activity was evaluated on adult Indian earthworm Pheritima posthuma due to its anatomical and physiological
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Abinash kumar sahu, et al., 2020/ Phytochemical Evalution and In-vitro Anthelmintic resemblance with the intestinal round worm parasites of human beings. Paralysis was said to occur when the worms did not revive even in normal saline. Death was concluded when the worms lost their motility followed by fading away of their body colour.
RESULTS AND DISCUSSIONS
The percentage yield of Petroleum ether and aqueous extract were found to 10.92%w/w and 5.45%w/w (Table-1). The preliminary phytochemical screening on the leaf extract was carried out by subjecting the different extracts to qualitative test for the identification of various plant constituents. It showed the presence of alkaloids, glycosides, saponins, anthraquinone, terpenoids, phlobatanin like compounds (Table-2) but do not shows the presence of carbohydrate in Petroleum ether, phenolic and flavonoid. The Powder analysis and fluorescence was observed in ultra-short, ultra-long and visible. The results were shown in (Table-3 & 4).
The results (Table-5) depict the time taken for paralysis and death of earthworms after the treatment with the test extracts at the selected concentrations. The data revealed that the aqueous extract has a better wormicidal effect than Petroleum ether extract with compared with the standard drug Albendazole. Further study is required to find out the novel phytoconstituents responsible for anthelmintic action against various helminthes.
ACKNOWLEDGEMENTS:
The authors sincerely thanks to the Principal and Management of The Pharmaceutical College, Barpali, Bargarh for providing all the facilities to carry out the study.
REFERENCES:
1.Kamboj VP. Herbal medicine. J Cur Sc 2000;78(1):35-9. 2.Khastgir H N, Sengupta S K and Sengupta P. Note on the constituents of the Indian medicinal plant Oldenlandia corymbosa linn. Journal of the American Pharmaceutical Association.1960;49(8):562– 563. 3. Sivarajan VV, Indira B (1994) Ayurvedic Drugs and their Plant Sources. Oxford and IBH Publishing Co. Pvt. Ltd, Delhi. 4.Anonymous, Wealth of India- Volume III, CSIR, New Delhi. 5.Kirtikar KR, Basu BD (1994) Indian Medicinal Plants. Bishensingh Mahendrapalsingh, Dehradun. 6.Ruffo CK, Birnie A, Tengnas B. Edible Wild Plants of Tanzania, Regional Land Management. Nairobi, Flora of China. 2002; 19:149-155, 160-161. 7. Flora of China, Vol. 19; P 149, 155, 160, 161. 8. Wagner WL, Herbst DR, Sohmer SH (1999) Manual of the Flowering Plants of Hawaii. University of Hawaii and Bishop Museum Press.
International Research Journal of Pharmaceutical and Biosciences (IRJPBS) 5 (4) 11
Abinash kumar sahu, et al., 2020/ Phytochemical Evalution and In-vitro Anthelmintic
9. Wagner, Warren LH, Derral R, Sohmer SH. Manual of the Flowering Plants of Hawaii. Revised edition. Bernice P. Bishop Museum special Publication. University of Hawai’i Press. Bishop Museum Press, Honolulu, 1999. 10. Chopra RN, Nayar SL, Chopra IC. Glossary of Indian Medicinal Plants, Council of Scientific and Industrial Reasearch, New Delhi, 1986. 11. Otuska H, Yoshoimura K, Yamasaki K, Cantoria MC. Isolation of Acyl Iridoid Glycosides from a Philippine Medicinal Plants Oldenlandia Corymbosa (L). Chemical and Pharmaceutical bulletin. 1991; 39:2049-2052. 12. Nesamony S, Medicinal Plants. State institute of languages, Nalanda, Thiruvananthapuram, 2001; 342-345. 13. Khushbu Pandey, Pramod K. Sharma, Rupesh Dudhe, Anticancer Activity of Parthenium hysterophorus Linn and Oldenlandia corymbosa Lam by SRB Method.2012;1(6):12, 1-3. 14. Ghosh P, Das P, Mukherjee R, Banik S, Karmakar S, Chatterjee S. Extraction and Quantification of Pigments from Indian Traditional Medicinal Plants: A Comparative Study between Tree, Shrub, and Herb, International Journal of Pharmaceutical Sciences and Research. 2018, 9(7):3052-3059, 0975-8232. 15. Mammen D, Daniel M, Sane RT, 2011, Identification of pharmacognostic and phytochemical biomarkers to distinguish between Hedyotis corymbosa (L.) Lam. and its adulterant, Glinus oppositifolius (L.), Research Journal of Pharmaceutical, Biological and Chemical Sciences, 2, 649. 16. Noiarsa P, Ruchirawat S, Otsuka H, and Kanchanapoom T. Chemical constituents from Oldenlandia corymbosa L. of Thai origin. J Nat Med. 2008;62(2):249-50. 17. Ruffo CK, Birnie A, Tengnas B. Edible Wild Plants of Tanzania, Regional Land Management. Nairobi, Flora of China. 2002; 19:149-155, 160-161. 18. Nguyen VD. Medicinal Plants of Vietnam, Cambodia and Laos, Mekong Printing, Vietnam, 1993. 19. Journal of the Arnold Arboretum, Harvard University, 1987, 68. 20.Kirtikar KR, Basu BD (1994) Indian Medicinal Plants. vol. 2. Bishen Singh Mahendrapal Singh, Dehradun. 21. Chang HM, But PPH (1986) Pharmacology and Applications of Chinese Materia Medica. World Scientific, Singapore. 22. Bensky D, Gamble A (1990) Chinese Herbal Medicine: Materia Medica. Eastland Press, Seattle, WA. 23. Chang Minyi (1992) Anticancer Medicinal Herbs. Human Science and Technology Publishing House, Changsha. 24. Ou Ming (1990) An Illustrated Guide to Antineoplastic Chinese Herbal Medicine. The Commercial Press, Hong Kong.
International Research Journal of Pharmaceutical and Biosciences (IRJPBS) 5 (4) 12
Abinash kumar sahu, et al., 2020/ Phytochemical Evalution and In-vitro Anthelmintic
25.Himangshu Sekhar Maji, Sushomasri Maji, Kausik Bhar, Jaishree Chandra Manik Baral, Pranabesh Chakraborty and Sujata Ghosh Dastidar. Anthelmintic, Free-Radical Scavenging Property and Potent Antibacterial Activity of Flavonoid Fraction Isolated from the Whole Plant of Oldenlandia corymbosa L. International Journal of Biomedical and Pharmaceutical Sciences. 2011; 5 (1):43-48. 26.Harborne JB, 1984, Phytochemical Method, A Guide to Modern Technique of Plant Analysis, 2nd edition, Chapman and Hall, London, Pg, 4. 27.Khandelwal K.R. Practical Pharmacognosy. Techniques and Experiments, Nirali Prakashan 2nd edition, Pg-149. 28.Ajaiyeoba EO,Onocha PA,Olarenwaju OT.In-vitro Anthelimintic properties of Buchholziacoriaceae and Gynandropsisgynanndra extract.Pharm Biol.2001;39:217-20
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