Strychnos nux-vomica Seeds:
(Pharmacognostical standardization, extraction and anti diabetic activity)
34194754352925Index
Abstract
Introduction
Objective
Material and methods
Result
Discussion
Conclusion
References
Strychnos Nux-Vomica Seeds
ABSTRACT:
BACKGROUND:
Strychnos nux-vomica, commonly known as kuchla, contains strychnine and brucine as main constituents. In the seeds minor alkaloids are present i.e. protostrychnine, vomicine, oxy strychnine, pseudo strychnine, isostrychnine, chlorogenic acid and a glycoside. Traditionally seeds are used to treat diabetes, asthma, aphrodisiac and to improve appetite.

OBJECTIVE:
The present study was aimed to evaluate the various pharmacognostical characters and anti diabetic activity of S. nux-vomica seeds
MATERIALS AND METHODS:
WHO guideline performed the pharmacognostic characters. Extraction was carried out in petroleum ether, chloroform, alcohol, hydro alcohol, aqueous and phytochemical constituents were detected by different chemical tests present in the extracts. On the basis of extractive yield and phytoconstituents, hydro alcoholic, aqueous extracts were evaluated for anti diabetic activity. Gliclazide is used as standard in alloxan-induced diabetic rats.

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RESULTS:
Various analytical values of S. nux-vomica extract were established. Phytoconstituents present in S. nux-vomica extracts were detected.

CONCLUSION:
In experimental animals S. nux-vomica extracts show anti hyperglycemic activity.

INTRODUCTION:
Strychnos nux-vomica Linn (family: Loganiaceae) is a medicinally important toxic plant, commonly known as kuchla or poison nut, has diverse clinical and therapeutic applications. In different part of the world such as European Union, Guangxi, United States, Fujian, Guangdong, North Australia, Taiwan, Hainan and throughout Tropical Asia, this plant is commercially cultivated.

Presently, 60 formulations of Indian system utilized nux-vomica, of which 30 formulations are used in disorders of Vata Doha.

Kuchla contain 2.6-3 % total alkaloids out of which 1.25-1.5 % is strychnine, 1.7 % is brucine and rests are igasurine and vomicine. Alpha-colubrine, 3-methoxyicaine, novacine, pseudo strychnine, beta-colubrine, protostrychnine, n-oxy strychnine, isostrychnine, glycoside, lignin and chlorogenic acid are some other minor alkaloids. Strychnine was analyzed and identified in detoxified kuchla seeds using liquid chromatography- electro spray mass spectrometry. Time of flight mass spectrometry (TOFMS) and Matrix-assisted laser desorption/ionization (MALDI) analysis of crude and processed S. nux-vomica seeds allowed rapid screening of alkaloid components. Brucine and strychnine were separated and quantified in S. nux-vomica by non aqueous capillary electrophoresis.

Kuchla as a crude drug used in the treatment of anemia, lumbago, bronchitis, diabetes, skin disease, muscle weakness, asthma, constipation, appetite loss and malarial fever. It is also used for nerve conditions, depression, migraine, head-ach, symptoms of menopause and a blood vessel disorder called Reynaud’s disease. Man us nux-vomica for erectile dysfunction (ED). In manufacturing nux-vomica is used as rat poison that is, because it contains two deadly compounds; strychnine and brucine. In mice suppressive effect of S. nux-vomica observed on induction of ovalbumin-specific IgE antibody response. In alloxan-induced diabetic rats the anti diabetic activity was determined and gliclaide was used as a positive control. This study was aimed to explore anti diabetic potential of aqueous and hydro alcoholic extracts prepared from seeds and established pharmacognostical parameters of S. nux vomica.

MATERIALS AND METHODS:
PLANT MATERIAL:
From market of Jodhpur the seeds of nux-vomica were purchased. Dr. S. N. Mishra, senior scientist authenticated this plant material and aromatic plants project, K.N.K college of Horticulture, Madhya Pradesh, India and deposited at the department of Pharmacognosy, Jodhpur National University, and Jodhpur India.

PHARMACOGNOSTICAL PARAMETERS:
S. nux-vomica were estimated as per WHO guidelines for pharmacognostical parameters, namely, powder characteristics, ash values, extractive values, loss on drying (LOD) and florescence analysis.

EXTRACTION AND PHYTOCHEMICAL INVESTIGATION:
The plant material was ground in mixer, powder pass through sieve no: 10 and retained on sieve no:60 was used for extraction. The powdered drug was packed Soxhlet extractor and extracted using solvents like chloroform, ethanol, ethanol 50%, petroleum ether and water. Phytoconstituents present in extracts were detected by different chemical tests.To find the inorganic content in the sample determine different ash values.Seeds powder was done of nux-vomica by flourescence analysis.

Table: 1 Fluorescence analysis of S. nux-vomica seeds powder
DRUG+SOLVENT VISIBLE SHORT UV LONG UV
Drug No no green
Drug + ethanolic sodium hydroxide solution No no green
Drug + dil. Nitric acid No no no
Drug + petroleum ether no no green
Drug + potassium hydroxide solution Green green no
Drug + acetic acid No green green
Drug + ethanol No no green
Drug + dil. Sulfuric acid No no no
Drug + hydro alcoholic acid No no no
Drug + chloroform No no no
Table: 2 Different Ash values were determined to find inorganic content in the sample.

Total Ash 4.23+, -0.25% w/w
Acid-Insoluble Ash 3.06+, -0.05% w/w
Swelling Index 53.3+, -11.54% v/v
Water-Soluble Ash 0.93+, -0.05% w/w
Loss of Drying 7.83+, -0.28% w/w
Table: 3 Solvents of different polarity were used to find the extractive value for seeds and rhizome of S. nux-vomica.

SOLVENT PETROLEUM ETHER CHLOROFORM ETHANOL EYHANOL 50% WATER
Table: 4 Detection of phytoconstituents present in various extracts of S. nux-vomica.

PHYTO
CONSTITUENT CHEMICAL TEST OBSERVATION PETROLEUM ETHER EXTRACT CHLORO
FORM ETHER EXTRACT
Carbohydrate Molisch`s test + + Fehling test – – Steroid Salkowski test – – Resin HCl test – – FeCl3 test – – Alkaloid Mayer`s test – + Tannic acid test – – Dragendroof `s test – – Strychnine Sulphuric acid +Potassium dichromate – – Compound Lead acetate – – Tanin ; Phenolic 5% FeCl3 – – Saponin Glycosides Foam test – – Brucine Nitric acid – – Fats ; Oil Filter paper spot test + + Protein Biuret test – – Ninhydrin – – (-)=Absent, (+)=Present
RESULTS:
-660405194300The powder characteristics show the presence of lignified trichomes, aleurone grains, oil globules and endosperm with plasmodesma.

Powder characteristics (a) Aleurone grain; (b) vessel; (c) Fiber; (d) Starch grains; (e) Endosperm.

Various fluorescence analysis and analytical values are presented in table 1 and 2 respectively. Extractive values and extractive yields of S. nux-vomica seeds in solvents of different polarity are reported in table 3. Carbohydrate, protein, oil, steroid, alkaloids, resin, brucine and strychnine were detected in phytochemical screening in table 4.

High-performance liquid chromatography analysis with chemical controls and thin layer chromatography may be conducted in ongoing studies for through phytochemical investigation.

DISCUSSION:
Medicinal plants have been widely claimed useful and found effective in the treatment of diabetes mellitus in various traditional systems of medicine. Various species of Strychnos have been reported traditionally for the treatment of diabetes.  Considering these facts, the present study deals with the evaluation of diabetic activity of the aqueous and hydroalcoholic (50% ethanol) extract of seed powder of S. nux-vomica in alloxan-induced diabetic rats.

Earlier reported literature suggests that the glycosides, flavonoids, steroids, tannins, and alkaloids are responsible for anti diabetic activity. Preliminary phytochemical studies revealed the presence of steroids, alkaloids, and glycosides in hydroalcoholic and aqueous extracts of S. nux-vomica (strychnine, brucine, vomicine, lignin, and others).

The anti diabetic effect produced by the extract of S. nux-vomica may be due to the presence of these active ingredients. In our studies, significant reduction of blood glucose was observed on day 4 and 10 of treatment in groups III, IV, and V, which proved significant anti diabetic activity with the gliclazide, 50% ethanol extract of S. nux-vomica, and aqueous extract of S. nux-vomica when compared with group II. Both the extracts were found effective and significantly (P;0.05) superior in reducing the blood glucose when compared with standard (gliclazide) control drug on day 10.  S. nux-vomia possesses antioxidant property, which prevents lipid per oxidation that leads to protein modification and damage.

The serum insulin and cholesterol levels were not significantly modified upon treatment with the plant extract, suggesting that the hypoglycemic activity of S. potato rum is independent of insulin secretion.

CONCLUSIONS:
S. nux-vomica seeds were estimated for Pharmacognostic parameters, which are helpful in identification and standardization. The aqueous and hydroalcoholic extracts showed the presence of steroids, alkaloids, and glycosides in S. nux-vomica seeds. Aqueous and hydroalcoholic extracts were superior to positive diabetic control in controlling blood glucose level on day 10. In conclusion, the present study indicates that hydroalcoholic and aqueous S. nux-vomica seed extracts are effective in controlling diabetes.

REFERENCES:
Warrier PK, Nambiar VP, Ramankutty C. Vol. 5. Hyderabad: Orient Longman; 1996. Indian medicinal plants: A compendium of 500 species; p.202
Chitra V, Verma PV, Raju KA, Prakash KJ. Study of antidiabetic and free radical scavenging activity of the seed extract of strychnos nux-vomica. Int J Pharma Sci. 2010;2
Yin W. Wang T-S, Yin F-Z, Cai B-C. Analgesic and anti-inflammatory properties of brucine and brucine N-oxide extracted from seeds of nux-vomica. J Ethnopharmacol 2003;88;205-14 PubMed
Nadkarni RM, 3rd ed. Vol.2. Mumbai: Popular Prakashan; 1954. Dr.K.M. Nadkarni`s Indian material medica: p.1179
Geneva: World Health Organization; 1998. WHO Quality control methods for medicinal plant materials; pp.10-34
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Singh S, Gupta SK, Sabir G, Gupta MK, Seth PK, A database for anti-diabetic plants with clinical/experimental trials. Bio-information. 2009;4:263-8 Pmc Free article PubMed
Mukherjee PK. 1st ed. New Delhi: Business Horizons; 2002. Quality control of herbal drugs. An approach to evalution of botanicals; pp.114-24
Mathuram LN, Samanna H, Ramasamy VM, Natarajan R. Studies on the hypoglycemic effects of Strychnos potato rum and Acacia anarbica on alloxan diabetes in rabbits. Cheiron. 1981;10:1-5
Dhasarathan P, Theriappan P. Evalution of anti-diabetic activity of Strychnos Potato rum in alloxan induced diabetic rats. J Med Sci. 2011;2:670-4
Koti BC, Biradar SM, Karadi RV, Taranalli AD, BEnade VS. Effect of Bauhinia Variegate bark extract on blood glucose level in normal and alloxanised diabetic rats. J Nat Remedies 2009,9:27-34

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