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Swertia chirata L. Scientific Name: Swertia chirata L. Synonyms: Gentiana chirayita Roxb. ex Fleming. Ophelia chirata, Swertia chrayita L. Family: Gentianaceae Genus: Swertia Species: chirata Common Name: Chiretta, Clearing Nut Tree, Bitter Stick, Chirette Indian, Dowa I Pechish, Indian Gentian Part used: Flower and above ground parts and Seeds Plant Description: Swertia chirata is an erect annual herb indigenous to the mountainous districts of Northern India. The drug consist chiefly of the stem, which is dark purplish, brown colour, flower is panicled small green. The name indicates that the plant originally came from Kirata Desha and is bitter. It has long been used in Ayurveda as a bitter tonic, febrifuge and anthelmintic. S. chirata was also recognized in the British and united state Pharmacopoeia. The stems are robust (0.6-1.5 m), branching leaves are opposite, broadly lanceolate, acute, lower leaf often much larger, sometimes petioled. Calyx and corolla are four-lobed. Corolla is green-yellow and tinged with purple. Chemical composition: Alkaloids (Gentianine, Gentiocrucine, Enicoflavine, Triterpene: Swertanone, Swertane, Kairatane and chiratane. Xanthone Derivative (Mangostin, isomangostin Mangostin triacetate and no. of other xanthones like 1, 5, 8-trihydroxy-3 methoxy-xanthoneetc).Bitter Glycoside (Amarogentin and Amaroswerin. Others: 1,3,7,8 tetrahydroxy-xanthone, 1,8-dihydroxy-3,7-Dimethoxy xanthone, 1-hydroxy-3,5,8-trimethoxyxanthone, β Amyrin, chiratanin, chiratogenin, Decussatin, Gentianine, Gentiocrucin, Gentiopicrin, lupeol, isobellidifolin, ophelic acid swechirin, swertianin, swertinin). Actions of herb: The plant is bitter, cooling, anthelmintic, antipyretic, anti-periodic, laxative, galactagogue, cures thirst, biliousness, leucoderma, inflammations, burning sensations, pain in the body, urinary discharges, ulcers, leucorrhoea, piles, bad taste in the mouth, good for vomiting in pregnancy. It is tonic, stomachic and febrifuge.Alternative, Antileishmaniak, Anticholinergic, Anticonvulsant, Antiedemic, Antiinflammatory, Antimalarial, Antipyretic, Antitubercular, Astringent, Bitter, Cardio stimulant, Cholagogue, Choleretic, CNS depressant, Emollient, Hepato-protective, Hypnotic, Hypoglycemic/antidiabetic Secretagogue, Stomachic, Tonic, Undersedative, Vermifuge. Medicinal Uses: Chirata is prescribed in a variety of forms and combination in chronic fevers and anaemia. It is also used as a special remedy for bronchial asthma liver disorder. It is credited with tonic, febrifugal, laxative, stomachic, anthelmintic, and anti-diarrheal properties in Indian medical systems. It is prescribed in dyspepsia, in the debility of convalescence and generally in cases in which corroborant measures are indicated. Alterative, Asthma, Atrophy, Bronchitis, Cachexia, Cough, Depurative, Fever, Gonorrhea, Gravel, Hypnotic, Laxative, Liqueur, Phthisis, Puerperium, Stomachic, Tonic, Tuberculosis, Vermifuge. It is used for the treatment of various skin problems. It is used to cures thirst, biliousness, leucoderma, inflammations, burning sensations, pain in the body, urinary discharges, ulcers, leucorrhoea, piles, bad taste in the mouth, good for vomiting in pregnancy.

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Structures of chemical constituents of S. chirata

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Table: Biological activity of S. chirata

Joshi P, Dhawan V (2005). Swertia chrayita – an overview. Current Sci., 89 (4): 635-640.

Isolation and Identification of secondary metabolites of S. chirata S. chirata well-known traditional uses have led to extensive chemical analysis resulting in isolation and identification of active principles. The isolated compounds and their chemical nature are listed in table below.

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Table: Secondary metabolites of S. chirata

Joshi P, Dhawan V (2005). Swertia chrayita – an overview. Current Sci., 89 (4): 635-640.

Dosage: Prescribed in decoction or infusion in the quantity of a small tea-cupful twice daily. Infusion: 10-15 ml b.i.d.; Root powder: 3-5gm b.i.d.; Mahasudarshan Churna: 2gm b.i.d.; Sudarshan Ghanavati: 2gm b.i.d. Powder 1-3 grams in divided dose in a day, water decoction / infusion – 50-100 ml in divided dose per day. Side Effects: vomiting Contraindications: Pregnancy, lactation, diabetes, intestinal (duodenal) ulcers. Organoleptic Study of S. chirata:

Table: Organoleptic Characters of powder of Swertia chirata L. S. No. Parameter Appearance 1. Colour Green 2. Smell Bitter 3. Taste Extremely bitter

Latif A, Rehman S (2014). Standardization of a herbal medicine – Swertia chirayita L. Pharmacophore., 5 (1), 98-108.

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Physico-chemical Analysis of S. chirata: Different physic-chemical factors were determined three times and their average values were depicted as shown in table below.

Table: Physicochemical Analysis of Swertia chirata L. S. No.

Physicochemical Parameter Results Mean±S.E.M. (S.D.)

1. Moisture Content Loss of Weight on Drying 8.69±0.01(0.02) Toulene Distillation Method 8.37±0.01(0.02)

2. Ash Value (%) Total Ash 2.40±0.00 (0.01) Acid Insoluble Ash 0.49±0.00 (0.01) Water Soluble Ash 3.12±0.00 (0.01) Sulphated Ash 0.86±0.01 (0.04)

3. pH Values (%) pH at 1% 49±0.00(0.01) pH at 10% 03±0.01(0.02)

4. Bulk Density (gm/ml) 0.67±0.01(0.02) 5. Melting Range 80-900C 6. Solubility (%)

Alcohol Soluble extractive 3.86±0.40(0.70) Water Soluble extractive 5.89±0.32(0.56)

7. Crude Fibre Content 4.80±0.07(0.04) 8. Total Alkaloid Estimation 11.53±0.15(0.08) 9. Extractive values in different organic

solvent

Petroleum ether (60-800) 0.92±0.02 (0.04) Diethyl Ether 4.12±0.02 (0.04) Chloroform 0.63±0.02 (0.05) Alcohol 4.35±0.11 (0.20) Aqueous 19.14±0.11 (0.20)

Latif A, Rehman S. 2014. Standardization of a herbal medicine, Swertia chirayita L. Pharmacophore., 5 (1), 98-108. Microscopic Examination of S. chirata: Powdered drug dark brown vessels with bordered pits, reticulate spiral thickening, fragment of epidermal cells of stem with parenchyma dorsiventral leaf, upper epidermis with straight anticlincal walls lower epidermis with wavy anticlincal walls stomata anomoytic and anisocytic, pollen 25 micron in diameter spherical three pores. Chemical Identification tests of S. chirata extract: The chemical identification tests showed positive results for the presence of triterpenes, tannin, alkaloids, carbohydrate and sterols. Florescence analysis: Florescence analysis under UV light is sometime very characteristic for a drug. As many drugs and the constituents present in the drug emit specific colour when they are exposed to ultraviolet radiations because the radiant energy excites the solution which emits that particular colour known as fluorescence. Hence the fluorescence analysis of the successive extracts and the powdered drug of Chirata treated with different chemical reagent were done and different change in the colour so appeared was observed and noted. The details are presented in following table.

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Table: Fluorescence Analysis of the successive extracts of Swertia chirayita L. Extracts Day Light UV Long UV Short

Petroleum ether Dark Green Light Green Black

Diethyl ether Green Light Green Dark Green

Chloroform Dark Green Dark Green Black

Alcohol Dark Green Green Brown

Aqueous Brown Dark Green Dark Brown Latif A, Rehman S. 2014. Standardization of a herbal medicine, Swertia chirayita L. Pharmacophore., 5 (1), 98-108. Thin-layer Chromatography of S. chirata extract: Thin layer chromatography of S. chirata extract was performed in two different solvent systems. Table 1 showed the result of TLC of S. chirata in ethylacetate-methanol-water solvent system in the ratio of 100:16.5:13.5 (254: 0.15, 0.36, 0.49, 0.73; 366: 0.38, 0.51, 0.7 & 0.85) and chloroform-methanol-water in the ratio of 80:20:2 (254: 0.73, 0.83, 0.89, 0.95; 366: 0.49, 0.55, 0.12) solvent systems. The TLC plates were observed under UV light at 254 and 366 nm. The distances of spots are measured after spraying with Vanillin-Sulphuric acid reagent.

Figure: TLC Chirata – Petroleum Ether extract TLC Chirata – Chloroform extract

TLC Chirata – Ethanolic extract

Latif A, Rehman S.2014. Standardization of a herbal medicine –Swertia chirayita L. Pharmacophore., 5 (1), 98-108.

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Fourier Transform Infra-red Spectroscopy: IR spectral study revealed prominent peaks at 3448.93, 2941.47,1609.39, 483.46 cm-1.

Sultana MJ, Ahmed FRS (2013) Phytochemical Investigations of the Medicinal Plant Swertia chirata Ham.Biochem

Anal Biochem 2 (4): 1-7. HPLC of S. chirata extract

Analgesic activity of S. chirata: In acetic acid induced writhing test aspirin 300 mg/kg was used as a reference compound. The crude extract of S. chirata showed significant dose related inhibition of number of writhes. In vehicle treated (control) animals, mean number of writhes induced by intra-peritoneal injection of acetic acid were 128, which was reduced to 43, 55 and 63 in animals treated with 500, 300 and 200 mg/kg oral dose of the test substance respectively. The results of the writhing test were highly significant and comparable with aspirin which produced 46.4 writhes. The % of inhibition of writhes with the three doses of crude extract were 66.4% (500mg/kg)57.03% (300mg/kg) and 50.8% (200mg/kg) whereas with aspirin it was 63.8% (300mg/kg). The results of hot plate activity were presented in table 2b. The readings were observed for 3 hours. It showed dose dependent increase in tail flick time. At 500mg/kg it showed 0.42 second flicking time at 3 hour whereas aspirin 300mg/kg produced delayed in pain response at 2 hour (0.43).

Table: Acetic acid induced writhing test Treatment Dose(mg/kg) Mean No. of writhes % of inhibition

Control 0.5ml saline 128±3.48 - Crude Extract 200 63±3.36 50.8* Crude Extract 300 55±2.16 57.03 Crude Extract 500 43±1.96 66.40

Aspirin 300 46.4±1.27 63.8* Each value is the mean ± S.E.M. of five determinations.* P < 0.05, Dunnet test as compared to control

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Table: Hot plate activity of crude extract of Swertia chirata variation flicking time (s) Group 0 1 2 3 Control 0.19±0.014 >0.22 >0.22 >0.22

500 0.19±0.04 0.35±0.05 0.41±0.06 0.45±0.01* 300 0.20 ±0.03 0.31±0.02 0.36±0.04 0.33±0.04 200 0.19±0.06 0.32±0.04 0.30±0.04 0.28±0.07 100 0.19±0.04 0.26±0.03 0.22±0.02 0.19±0.05

50 0.20±0.03 0.23±0.03 0.23±0.03 0.18±0.02 Aspirin 0.18±0.02 0.39±0.03* 0.44±0.04* 0.33±0.08

Each value is the mean ± S.E.M. of five determinations.* P < 0.05, Dunnet test as compared to control

Gross behavior and Neuro-pharmacological Studies on S. chirata extract The gross behaviour activity was observed for the the standard drug i.e. Diazepam 2mg/kg and Imipramine 15mg/kg as well as for test substance i.e. crude extract of S. chirata at the three different doses i.e. 200mg kg, 300mg/kg and 500mg/kg for at least 24 hours.

Parameters Dose mg/kg Diazepam Imiparamine 500 300 200 100 50 25 10 1 2mg/kg 10mg/kg

Nystagmus +3 +2 +1 +2 +2 1 +1 +1 +1 - Vocalization - - - - - - - - - - Piloerection +1 +1 - - - - - - - + Micturition + + + + + + +s +s +s + Irritability + + + + + + + + +s +

Disorientation/Staggeringgate + - - - - - - - - + Aggressiveness +1 + +s - - - - - +s

Motor activity(Decrease) + + + + + + + + + - Pain response + + + + + + + + + +

Ataxia - - - - - - - - - + Tail erection + + - - - - - - - +

Tremor +1 + - - - - - - - +2 The mean number of squares crossed by the mice was 131, for control group, 140 for 500mg/kg, 149 for 300mg/kg and 128 for 200mg/kg which showed that S. chirata is stimulant. The control, Diazepam (2mg/kg) and Imipramine (15mg/kg) treated groups showed 131, 21 and 153 squares traveled respectively. The results of head dip, cage cross and rearing activities were presented in table 3b. There was increased in the head dip and cage cross and rearing activity. In head dip test they showed 37, 33 and 24 times response for 500, 300 and 200 mg/kg respectively (control group: 31, Diazepam treated group: 05, Imipramine treated: 38). The results of cage cross activity were 29, 38, 30, 24, 09, 33 respectively for control, 500, 300, 200, 2(Diazepam) and 15mg/kg (Imipramine) treated groups. Similarly results of rearing activity also showed stimulatory action. The readings of rearing activity were 31, 39, 33, 27, 07, 41 respectively for control, 500, 300, 200, 2 (Diazepam) and 15mg/kg (Imipramine) treated groups. The time taken to travel iron rod was significantly increased at the dose of 500, 300 and 200 mg/kg which was 08 and 12 and 18 seconds respectively while it was 180 seconds for Diazepam and 06 seconds for Imipramine. The mean mobility time in water tub of animals with S. chirata was 3.34, 3.36, 3.37, 3.3, 1.3 and 4.2 minutes respectively for control, 500, 300, 200, 2 (Diazepam) and 15 (Imipramine) mg/kg treated groups.

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Table: Assesment of neuro-pharmacological activities Swertia chirata Treatment Dose mg/kg

orally Open field

Head dip

Cage cross Rearing test(min)

Mobility time(sec) *1

Traction test

Control 0.5ml 159.8±4.6 34±3.75 35.67±1.54 39.17±0.93 3.45±1.12 22 ±0.05 Crude Extract 500 40±3.85 15±1.56 15±0.56 12.17±1.51 2.41±1.45 28± 0.07 Crude Extract 300 51±2.34 20±2.24 17± 1.58 18± 0.51 2.56±1.03 50±0.02 Crude Extract 200 65±3.34 18± 1.14 20±2.71 12±1.52 2.34±1.01 33±1.13

Diazepam 2 31±1.12 05±0.09 09±1.12 14±1.12 2.1±1.3 180±3.12 Imipramine 15 166±4.2 45±1.42 39±2.3 55±2.5 4.4±2.1 06±0.09

*1: Above 1mg (5 to 300 mg/kg mice fall, f: fall, * significant, Anti-Parkinsonian Activity The two isolated compounds from S. chirata were administered in 100mg/kg i.p dose to both the male and female mice. The following hypokinesia and locomotor activity were observed.

Table: CNS profile of compounds

Hypokinesia = reserpine reversal test p < 0.001; Locomotor activity = L-Dopa potentiation test p < 0.001

Table: Anti-Parkinsonian activity of the compounds

Srivastava S, Mishra N, Misra U (2010). Neurological studies of novel compounds from Swertia chrayaita. J. Chem.

Pharm. Res., 2 (1): 125-134. Anti-Parkinsonian activity was evaluated by observing reduction in tremor induced by oxy-tremorine (0.5 mg/kg i.p.), Reserpine 5mg/kg i.p) induced rigidity and hyperkinesia in mice. References Anon. 1982. In The Wealth of India: Raw Materials, Publication and In-formation Directorate, CSIR, New Delhi.

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