QUALITY STANDARDS OF’ _. INDIAN MEDICINAL PLANTS Volume 230 | Terminalia arjuna (Roxb.) Wight & Arn. (Kakubhah, Parttha) Drug consists of dried stem bark of Terminalia arjuna (Roxb.) Wight & Arn. (Plate 30.1 A & B), Iant is a large evergreen deciduous tree common in plains and foothills Fam. Combretaceae!, T of Himalayas and-also planted for sha €. Pieces of stem bark Plate 30.1 Terminalia arjuna (Roxb.) Wight & Arn.244 Quality Standards of Indian Medicinal Plants-Vol 2 Other Names Assam.- Arjun Beng. - Aghan Guj. - Sadado Hind. - Arjuna Kan, - Maddi, Bibimatti Mar. Sanmadat Ori. Arjuna Pun). - Anon ‘Tam. - Vellamatta, Marudam Tel. - Yeramaddi Description Macroscopic Flat or slightly curved, varying in size, up to 15 em or more in length, 10.¢m in width, 0.5 to 1.5 cm in thickness. Outer surface smooth, pale greenish yellow; inner surface finely longitudinally striated and pinkish in colour; fracture laminated (Plate 30.1 C). Taste astringent Microscopic Cork consists ot a tew layers of tangentially running and radially elongated cells: phellogen, 2 to 4 celled thick, phetloderm narrow, consisting of 4 to 6 rows of tangentially elongated and radially arranged cells. Phloem, very broad, traversed by uniseriale medullary rays running straight and parallel, ‘occasionally becoming slightly curved near the rosette crystal; groups of phloem fibres, lignified, thin- walled, tangentially arranged, associated with idioblasts containing clusters and rosettes of calcium ‘oxalate. Some parenchymatous cells of cortex and secondary phloem contain reddish brown pigment and some cells contain starch grains (Fig. 30.1). Powder Reddish brown. Fibrous powder exhibits parenchymatous cells of the cortex and phloem containing clusters and rosettes of calcium oxalate erystals, starch grains, tannins and dark reddish brown pigment: fragments of thin-walled phloem fibres associated with idioblasts containing rosettes and clusters of calcium oxalate and longitudinal radially cut medullary rays (Fig. 30.2).Terminalia arjuna (Roxb.) Wight &Ara. 245 Figure 30.1 Microscopy of Terminalia arjuna stem bark. A. Diagrammatic TS of stem bark (x18); B. TS of siem bark showing cork and cortex (x130); C. TS of stem bark showing inner phloem region (x130); cep, cell containing pigment; ck, cork; cr, cluster of calcium oxalate crystals; ct, cortex; mr, medullary rays; pd, phelledcrm; pg, phellogen; phf, phloem fibres; ro, rosette of calcium oxalate; sg, starch grains.246 Quality Standards of Indian Medicinal Plants-Vol 2 ph ey f 30 Sot Figure 30.2 Microscopy of Terminatia arjuna stem bark powder, a, cork fragment; b, parenchyma tusettes of calcium oxalate, starch grains and tani arch graing; d, rosettes and clusters of calcium oxalate; e, crystal fibre; f, phloem fibres; g, radially cut fragment of phloem region; cep, cell containing pigment; clr, cluster of calcium oxalate crystals; phf, phloem fibre; ro, rosette of calcium oxalate; sg, starch grains, Chemical constituents Major Hydrolysable and condensed tannins. ellagic acid, arjunin, arjanic acid. arjunetin, arjunolitin, friedelin, terminoic acid, arjungenin, arjunglucoside I, IP°'5*".Terminalia arjuna (Roxb.) Wight &Arn. 247 Others f-Sitosterol and a flavone arjunotone* Termineic wid Arjunic acid HG Cs ne i ; - AS ee caf Hy T° Cin re an oc A 0 e 0 L ne “a; mle | on Aciunetin Ou 4 Bilagh: acid OH Asjunolone on TH ‘Arjunotitin OH248 Quality Standards of indian Medicinal Planis Vol 2 TLC Identity Test Test solution 1 Extract | g of powdered drug with methanol (3 x 15 ml) under reflux on a water bath. Filter the methanolic extract and evaporate. Take chloroform soluble portion of methanolic extractive and make up the volume to 10 1 Test solution Ut Extract 0.5 g of powdered drug with methanol (3 x 15 ml) under reflux on a water bath. Filter the methanolic extract and concenirate and make up the volume to 25 mi with methanol Standard solution Dissolve 5 mg of ellagie acid# in methanol and make up the volume with methanol to 50 ml in a ‘volumetric flask. Solvent systems 1. Toluene : Ethy! acetate : Kormic acid > Methanol (6 : 3. 0.1 . 1.0). Il. Toluene : Ethyl formate : Formic acid (5 : 5 : 2). Procedure Apply 10 411 of test solution I on precoated silica gel 60 Fa, plate (B. Merck) of uniform thickness of 0.2 mm. Develop the plate in solvent system | (0 a distance of 8 em. Apply 10 il each of test solution UI and standard solution on precoated silica gel 60 Foss plate (E. Merck) of uniform thickness of 0.2 mm. Develop the plate in solvent system I! to a distance or cm. Visualization Spray the plate developed in solvent system I with anisaldehyde-suiphuric acid reagent and heat at 110° for 10 min. Note the R, and colour of the bands resolved (Table 30.1, Plate 30.29). Dorivatize the plate developed in salvent system TI with 5 per cent methanolic ferric chloride solution and note the R, of the greyish blue bands resolved (Table 30.1, Plate 30.28) Evaluation ‘A band (R, 0.43) corresponding to ellagic acid is visible in both standard and test solution tracks, © Billagie acid from Natural Remedios Pvt, Ltd., Dangalore, India.Terminalia arjuna (Roxb.) Wight &Arn, 249, ‘Table 30.1 TLC Details of Test Solution of Terminalia arjuna Stem Bark. Derivatized Test R, value Colour of R, value the band 0.079 Grey 0.17 0.19 ish biue 0.26 023 Dark blue 0.34 00 Blue 043 (Bllagic acid) oat Dark blue 092 0.45 Grey 055 0.65 Grey on Greyish blue 080 Dark pink Assay / Analytical Methods ‘TLC densitometric estimation of ellagic acid. TLC plates Plate 20.2 TLC profiles of test solution of Terminalia arjuna stem bark A. Test solution 1 (derivatized with anisaldehyde culphuric reagent); B, Test solution II (derivatized with ferric chloride solution) 11: Test solution Ll; 2: Ellagic acid standard. Precoated plates of silica gel 60 Fis: (BE. Merck) of uniform thickness of 0.2 mm. Solvent system Toluene : Ethyl formate : Formic acid (9 : 9 : 2). Scanning Under UY light at 381 am, Test solution Extract | g drug powder with 2N hydrochloric acid (2 x 25 ml) by vefluaing each Gane for 2 b. Filter and extract the filtrate with diethyl ether Evaporate the combined ether extract and dissolve the residue in methanol. Make up the volume to 50 ml with methanol.250 Quality Standards of Indian Medicinal Plants-Vol 2 Standard solution Dissolve 10 mg of ellagic acid in 100 ml of methanol in « volumetric flask. From this stock solution prepare standard solutions of 15 to 35 pg/ml by transferring aliquots (7.5 to 17.5 ml) of stock solution to 50 ml volumetric flasks and adjusting the volume with methanol Calibration curve Apply 10 ut! of the standard solutions (ranging in concentration from 150 to 350 ng per spot) on a precoated TLC plate. Develop the plate in the solvent system in twin trough chamber to a distance of 8. cm. Scan densitometrically at 381 nm. Record the peak area and prepare the calibration curve by plotting peak area vs concentration apy Estimation of ellagic acid in the drug Apply 10 pil of the test solution in triplicate on a precoated TLC plate, Develop the plate in the solvent syotci anal icvuid thie peak aiva of the chromatogram as described above for the calibration curve, Calculate the amount of ellagic acid present in the sample from the calibration curve of ellagic acid (Fig, 30.3). ‘The percentage of tolai ellagic acid content ranges from 0.13 to 0.33 in the samples analyzed. 10 100 so B 381 ow ua Figure 30.3 TLC denstlometric sean at 381 nm. A. Test solution of stem bark of Terminatia arjunu: B. Ellagic acid standard. Quantitative Standards Foreign matter: Not more than 2.0 per cent. Ash: Not more than 27.0 per cent. Acid-insoluble ash: Not more than 2.0 per cent. Ethanol-soluble extractive: Not less than 16.0 per cent. Water- soluble extractive: Not less than 17.0 per centTerminalia arjuna (Roxb.) Wight &Arn. 251 Adulterants / Substitutes An the market, slem bark of 13 different species are being sold as Arjuna. ‘These are: Terminalia bialata Steud., T. bellirica (Gaertn.) Roxb., T. alata Heyne ex Roth (syn. T: tomentosa (DC.) Wight & Amn), T. manii King, T. myriocarpa Heurck & Muell.-Arg., 7. chebula (Gaertn.) Retz... T. catappa Linn., 7. calamansanai (Blanco) Rolfe (syn. T. pyrifolia (Presl) Kurz), 7. travancorensis (DC) Wight & Am., 7: pallida Brandis, T. citrina Roxb. ex Flem. and 7. paniculata Roth. Among the above, the bark of 7: alata is the most common adulierant, the main distinguishing characters of which include. cough, cracked or fissured browa coloured external surface and longitudinally striated inner surface**, Pharmacology “The stem bark of 7: arjuna is a potent antihyperchotesterolacmic agent”. 1 reduced the total cholesterol, LDL, VLDL, free fatty acids, phospholipids and triglycerides and elevated HDL in hypercholesterolaemic rabbits", Seventy per cent ethanolic extract of the stem bark showed hypotensive and bradycardiac effects in experimental animais"", Aqucous and 9S per cent ethanolic extracts. exhibited myocardial depressant property". Polar fraction of ethanolic extract relaxed norepinephrine contracted rat aorta'*. Stem bark increased endogenous superoxide dismutase and reduced glutathione, catalase and thiobarbituric acid reactive substances in the heart. It also protected the heart from oxidative stress during ischaemic-reperfusion injury in vitro! Clinically stem bark extract demonstrated antianginal activity'”"* and ametiorative effect in hypertension and heart failure”, Ethanolic extract showed antifertility activity in albino rats and antifungal activity against dermatophytes”. Methanolic extract and ellagic acid isolated from the stem bark showed antimutagenic activity, Major Therapeutic Claims Antihypertiptdaemic'™'""*, antihypertensive”, n heart diseases! Safety Aspects ‘The drug used traditionally in prescribed doses may be considered safe. LDgo of 70 per cent ethanolic extract : 2.5 g/kg ip in mice”, Dosage 3 to 6 g of stem bark powder'. RRWERENCES 1. The Ayurvedic Pharmacopoeia of India, Part-t, vol. IL. 1" edn. New Delhi: Government of India, Ministry of Health & Family Welfae, Department of Indian Systems of Medicine and Homeopathy; 1999, p. 17-18. 2. Row LR, Murty PS, Subba Rao GSR, Sastry CSP, Rao KI. Chem ation of Terminalio specter: Part XU Isotation and structure determination of agjunic acid, a new wilydeoxyliterpene carboxylic acid from Terminalia arjuna bark. indian J Chem 1970; 8: 116-721 3. Row LR, Murty PS, Subba Rao GSR, Sastry CSP, Rao KV. Chemical examination of Terminalia species: Past XIU ~ Isolation and structure determination of arjunetin from Terminatia arjuna, Indian 4 Chem 910: 8: 772-775 4. Sharma PN, Shoeb A. Kapil RS, Popli SP. Arjanolone - A new flavone from stem hark af Terminalia arjuna. Indian F-Chem 1982; 2B: 263-264, 5. Abmed MU, Mullah KB, NérinT, Ulla J-K, Terminoic acid, a new tikydroxytriterpene carboxylic acid from back of Terminalia arjuna, Indian J Chem 1983; 22B: 738-740,252. 25. 26. Quality Standards of Indian Medicinal Planis-Vol 2 Anjaneyutu B, Babu Rao ¥, Ganguly AK, Govindachaci TR, Joshi BS, Kamat VN, Manmade AH, Mohamed PA, Rahimtula AD, Saksena AK, Varde DS, Viswanathan N, Chemical investigation of some Indian plants, Indian J Chem 1965; 3: 237-238, Tripathi VK, Pandey VB. Udupa KN, Rucker G. Arjuoolitia, a titerpene glycoside from Terminalia arjuna, Phytochemistry 1992; 31: 349-451 Pharmacognosy of Indigenous Drage, val. 1. Compiling & ed, Raghunath for Research in Ayurveda & Siddha; 1999: p. 84-86. K, Mitra R Now Dethi: Ce Cow Sarin YK. Mllustvated Manuat of Herbal Drugs Used i" ayurveda New Delhi Councih of Sesemtific and fiutustiiel Research and Indian Councit of Medical Research; 1996: p. 118-419. |. Tiwari AK, Gode JD, Dubey GP. Btkect of Terminalia axjuno on lipas profiles of rabbits ted bypercholesterotaemuc diet. Int J Crude Drug Res 1990: 28: 43-47 |. Pathak SR, Upadhyaya L, Singh RH, Dubey GP, Udupa KN. Effect of Terminatia arjuna W. & A. on autocotdal and lipid profiles of rabbits. (ndian Drugs 1990; 27: 221-229. Shaita HP, Udupa SL, Udupa AL, Naie NS. Bllect of Terminalia arjune on experimental hypes ia in abuts Int J Pharmacog \997; 35: 126-129 Singh N. Kapur KK, Singh SP, Shanker K, Sinha JN, Kohli RP. Mechanism of cardiovascutar action of Terminatia tarjuna, Planta Med 1982; 45: 102-104 Srivastava RD, Dwivedi S, Siccnivasue KM, Chandrsshelthar OM Cardiowscentar aAtaote of Tarminatia species of plaots, Indian Drugs 1992; 29: 4d ids. |. Tipati YB, Zerminalia arjuna eatract modulates dhe conuaction oF vat avila juduced Uy KCI and nusepinepbrine Phytother Res 1993; 7: 320-322. Gauthaman K. Maulik M, Kumari ®, Manchanda SC, Dinds AK, Mautik SK. Effect of chronic treatment with bart. ‘of Terminalia arjuna * a study on the isolated ischaemic-reperlused rat heart. J Ethnopharmacol 2001: 73: 197-201 Dwivedi $, Chansouria JPN, Somani PN, Udupa KN. Effect of Terminatio arjuna on ischaemic heart disease Alternative ‘Med 1989; 3: 115-122. Bharani A, Gangutt A. Mathur 1.K, Jamea Y, Raman PG. Efticacy of Terminatio orien ‘double-blind placebo-controlte Heart J 2002; 54: 170-175 Verma SK, Bordia A. Effect of Terminatia avjana bark (Arjun chhal) in patients of congestive heart failure and hypertension. J Ree Educ Indian Med 1988; 7: 31-36 Bharani A, Ganguly A, Bhargava KD. Salutary effect of Terminalia arjun failure, fms J Cardiot 1995; 49: 191-199. Prakash AO, Mathur R. Screening of Indian plants for antifertilty activity. Indian J Exp Biol 1976; 14: 623-626. Dutta BK, Rahman 1, Das TK. Terminatia (Myrobalan) and its effect on dermatophytes. Geobios 1999; 20: 43-49. Jain AK, Shima: K. Nakamura Y, Tomita 1. Kada , Preliminary study on the desmutagenic and aniinutagen ‘of some natural products. Curr Sei 1987; 56: 1266-1269. Kaur 8, Grover IS, Kamar 8. Antimutagenic potential of Biol 1997; 35: 478-482. Kaur K, Arora S, Kumar S, Nagpal A, Modutatory effeet of phenolic fractions of Terminatia arjuna on te in Ames assay. J Env Pathol Toxicol Oncol 2002; 21: 45-56. Pasquini R, Seassetlati-Sforzolini G, Villarini M, Moretti M, Marcarelli M, Fatigoni C, Kaur S, Kumar $, Grover IS. In vitro protective effects of Tern J Eav Pathol Toxicol Oncol 2002; 24: 33-44 in chronic stable angina’ 9 | crossover sludy comparing Tevminalia axjuna with isosorbide mononitrate. fadian patients with severe refractory heart lagic acid isolated from Terminalia arjuma. Indian 1 Esp