TRADITIONAL KNOWLEDGE IJATM.2020;2(2):20-25. International Journal of Ayurveda and Traditional Medicine Physico-chemical Evaluation of an Ethnomedicinal Formulation Karmaranga (Averrhoa ​ ​ ​ Carambola linn.) Fruit Oil. ​ 1 1 2 P Shetty Shifa, ​ Faisal Mohammed, P​ rabhu Suchitra 1 ​ ​ ​ D​ ept. of PG Studies in Dravyaguna Vijnana, Sri Dharmasthala Manjunatheshwara College of Ayurveda and 2 Hospital, Kuthpady, Udupi. S​ ri Dharmasthala Manjunatheshwara Centre for Research in Ayurveda and Allied ​ Sciences, Udupi. ABSTRACT Introduction: Processing of crude drugs helps the transformation of the pharmacological action of a drug bestowing it with better efficacy and greater bioavailability. Lesser explored traditional practices have many such single drug therapies that can ​ be brought into the limelight. One such folklore practice is Taila Paka of Karmaranga (Averrhoa carambola Linn.) Phala ​ ​ ​ ​ which is used traditionally in treating joint disorders. Standardization of drugs is of paramount importance in order to deliver a promising treatment with respect to safety and efficacy, with this background the oil prepared using Karmaranga fruits is ​ taken for its analytical study. Methods: The present study exclusively deals with the physico-chemical analysis of Karmaranga Taila by standard oil ​ ​ ​ ​ parameters including High-performance thin-layer chromatography (HPTLC). ​ ​ Results: The sample was liquid at room temperature with a reddish-brown color and agreeable odor. As the fruits are rich in ​ vitamin C, HPTLC was done along with ascorbic acid running parallel to the test sample. There were 3 and 7 bands respectively observed in short UV and long UV. Simultaneously ascorbic acid showed a blue fluorescent band at Rf 0.48, after derivatization with vanillin sulphuric acid spraying reagent, there were two bands evident in A.carambola oil at Rf 0.48 ​ and 0.57 (all purple). Conclusions: This study can serve as a standard reference for the preliminary identification of Karmaranga Taila. ​ ​ ​ _________________________________________________________________________________ ​ Keywords: HPTLC; Karmaranga Taila; Physico-chemical analysis; Traditional formulation. ​ ​___________​ ______________________​ _____________________________​ __________________ ​ INTRODUCTION Tremendous amount of traditional knowledge hidden Averrhoa carambola Linn., belonging to the family Oxalidaceae,2 commonly called star fruit in Indian within tribals on herbal medicine is unknown to the outer ​ world. Karmaranga is a plant drug mentioned under markets. Edible fruits of this plant are commonly used as 3 ​ ​ 1 a home remedy for cough and cold. Phalavarga of Bhavaprakasha Nighantu. F​ ruits have ​ ​ ​ ​ Madhura Amla Rasa, Snigdha Ruksha Guru Guna, and Correspondence:​ ​Dr. Shifa Shetty P. Post-Graduate Amla Vipaka, Sheeta Virya according to Bhavaprakasha ​ ​ Scholar, Department of Dravyaguna, Sri Dharmasthala and Ushna Virya according to Rajanighantu. It is ​ ​ ​ ​ Manjunatheshwara College of Ayurveda and Hospital, attributed with Grahi, Hridya, Bala Pushtikara Karma. ​ Kuthpady, 574118 Udupi, Karnataka, India. Email: With all the above qualities it is one of the best [email protected]. Phone: +91-8073898748. Vata-Kaphahara Dravya. Botanically it is identified as ​ IJATM I VOL 2 I ISSUE 2 I JUNE 2020 ​ ​ ​ ​ ​ ​ Free Full Text Articles are Available at https://ijatm.org/ 20 ​ ​ Shifa Shetty P. et.al. Physico-chemical Evaluation of an Ethnomedicinal Formulation Karmaranga (Averrhoa Carambola linn.) Fruit Oil. ​ ​ ​ Siddhi Laxana were obtained, the heat source was turned A lesser-known folklore use of this fruit is in the form of ​ oil in various joint pains.4 Standardization is the process off, and later the oil was packed in airtight containers. ​ of implementing and developing technical standards This oil was further analyzed as per standard protocol at based on the consensus of different parties that include SDM center for Research in Ayurveda and Allied firms, users, interest groups, standard organization and Sciences, Udupi. governments.5 With a growing demand for ​ Analytical Study: evidence-based medicine, any formulation classical or Following physico-chemical parameters were tested to folklore has to undergo standardization techniques as per standardize Karmaranga Taila as per the ASU testing API before administration. Fruits of Karmaranga have ​ ​ ​ ​ protocols prescribed by API and laboratory procedures for been studied extensively for its nutritional benefits but, testing Siddha and Ayurvedic medicines by CCRAS.7 currently, no medicinal formulation is in practice. This ​ paper exclusively deals with the physico-chemical Refractive index, Specific gravity, determination of acid analysis of an Anuktayoga- Karmaranga Taila prepared value, determination of saponification value, ​ ​ ​ using a drug that is classically described by various determination of rancidity, determination of iodine value, Nighantukaras with reference to its standard oil determination of the unsaponifiable matter, peroxide ​ parameters in order to form a basis for further clinical value, and viscosity along with chromatographic analysis trials. The expected outcome of the study was to do were carried out. preliminary physico-chemical analysis of Karmaranga ​ Sample preparation for High-performance thin-layer Taila. chromatography (HPTLC) : A sample obtained in the procedure for the determination METHOD of the unsaponifiable matter is dissolved in 10 ml of As the preparation methods varied from one ethno group chloroform this was followed for the sample of to another, to maintain the uniformity general method of A.carambola, and chloroform soluble portion was used ​ preparation of Taila was followed as per Sharangadhara for HPTLC. 6 ​ ​ Samhitha. (​ Figure 1) ​ ​ 3.75 kgs of Karmaranga Phala Kalka was added to 15 HPTLC: ​ liters of Tila Taila, to this 60 liters of Phalarasa was 3, 6, 9µl of the above sample was applied on a precoated ​ ​ ​ added and heated over Mandagni. The color and odor of silica gel F on aluminum plates to a bandwidth of 8 mm ​ ​ ​254 Taila gradually started changing by the end of the first using Linomat 5 TLC applicator. The plate was developed day. Taila Paka was done for 3 days as the Dravadravya in Toluene – Ethyl acetate (9:1) and the developed plates ​ ​ ​ used was Swarasa. On the third day, oil was were visualized under UV 254 and 366 nm, and after ​ ​ continuously tested by subjecting the Kalka to fire until derivatization in vanillin-sulphuric acid spray reagent and ​ the hissing sound disappeared and also was rolled scanned under UV 254 and 366 nm. Rf, the colour of the ​ ​ between the fingers to form a wick. Once the Sneha spots, and the densitometric scan were recorded ​ 1a 1b 1c 1d Figure 1a. Fruits of A.carambola . Figure 1b. .Kalka .Figure 1c. Swarasa. Figure 1d. Subject to Taila Paka. ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ IJATM I VOL 2 I ISSUE 2 I JUNE 2020 21 Free Full T​ex​ t Articles a​ re​ Available at h​ tt​ ps://ijatm.org/ ​ Shifa Shetty P. et.al. Physico-chemical Evaluation of an Ethnomedicinal Formulation Karmaranga (Averrhoa Carambola linn.) Fruit Oil. ​ ​ < Table 1. Standardization parameter of A. carambola ​ ​ RESULT fruit oil. Results of standardization with respect to parameters like Parameter Results n = 3 %w/w refractive index, saponification value, iodine number, etc Averrhoa carambola were tabulated (Table 1). Results of HPTLC (Table 2) Linn. fruit oil showed that in the short UV of the test sample there were Refractive index 1.47032 3 bands identified at Rf of 0.25, 0.37, 0.48 (all green). In Specific gravity 0.9204 long UV, there were 7 bands identified at 0.27, 0.44, 0.48, Acid value 1.10±0.00 0.57, 0.67, 0.79, 0.91(all fluorescent blue). Saponification value 218.52±0.01 Simultaneously ascorbic acid showed a blue fluorescent Iodine value 188.94±2.46 band at Rf 0.48, after derivatization with vanillin sulphuric acid spraying reagent, there were two bands Unsaponifiable matter (%) 3.78 evident in A.carambola oil at R 0.48 and 0.57 (all purple) Peroxide value 0.19±0.01 ​ ​f (Figure 2-5). Viscosity 64.38 Rancidity Oil is not oxidized Table 2. Standardization parameter of A. carambola fruit oil. ​ ​ ​ Short UV Ascorbic acid Long UV Ascorbic acid After Ascorbic acid derivatization 0.25 (Green) - - - - - - - 0.27 (F. blue) - - - 0.37 (Green) - - - - - - - 0.44 (F. blue) - - - 0.48 (Green) - - - 0.48 (Purple) - - 0.50 (F. blue) - - - 0.57 (F. blue) - 0.57 (Purple) - - - 0.67 (F. blue) - - - - - 0.79 (F. blue) - - - - - 0.91 (F. blue) - - - 2a 2b 2c Figure 2. HPTLC Photodocumentation of a sample of Chloroform fraction of fruit oil of A.carambola Linn. ​ ​ ​ Figure 2a. Short UV . Figure 2b.Long UV . Figure 2c. After derivatization. ​ ​ ​ ​ ​ ​ ​ ​ Track 1: Fruit oil of A.carambola - 3µl Track 2: Fruit oil of A.carambola - 6µl, Track 3: Fruit oil of A.carambola - 9µl, Track 4:Ascorbic ​ ​ ​ ​ ​ ​ acid- 6µl, Solvent system- Toluene: Ethyl acetate (9.0: 1.0) 22 IJATM I VOL 2 I ISSUE 2 I JUNE 2020 ​ ​ ​ ​ ​ ​ Free Full Text Articles are Available at https://ijatm.org/ ​ Shifa Shetty P. et.al. Physico-chemical Evaluation of an Ethnomedicinal Formulation Karmaranga (Averrhoa Carambola linn.) Fruit Oil. ​ ​ ​ 3a 3b 3d 3c Figure 3. Densitometric scan of the sample of fruit of A.carambola Linn. at 254nm . Figure 3a and 3c sample of A. carambola. Figure ​ ​ ​ ​ ​ ​ ​ ​ 3a and 3c Ascorbic acid. ​ 4b 4a 4d 4c Figure 4. Densitometric scan of the sample of fruit of A.carambola Linn. at 366nm . Figure 4a and 4c sample of A. carambola. Figure ​