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Development of bioanalytical parameters for the standardization of Zingiber officinale

2013-06-15 17:47:44DineshPatelDhanabal
Journal of Acute Disease 2013年2期

Dinesh K. Patel, S. P. Dhanabal

1Department of Pharmaceutics, Indian Institute of Technology, Banaras Hindu University, Varanasi-221005, India

2J.S.S. College of Pharmacy, Ooty-643 001, India

Development of bioanalytical parameters for the standardization of Zingiber officinale

Dinesh K. Patel1,2, S. P. Dhanabal2*

1Department of Pharmaceutics, Indian Institute of Technology, Banaras Hindu University, Varanasi-221005, India

2J.S.S. College of Pharmacy, Ooty-643 001, India

HPTLC

Phytochemical analysis

Standardization

Zinziber officinale

Objective: To develop noval bioanalytical parameters for standardization of Zingiber officinale (Z. officinale) extract. Methods: Phytochemical analysis, solubility test, heavy metal analysis, antimicrobial study and fingerprint analysis through HPTLC method were carried out in the present investigation. Moreover quantitative analysis of rutin and quercetin in Z. officinale extract through HPTLC techniques were also performed in the present investigation. Results: Phytochemical analysis showed the presence of alkaloid, carbohydrate, tannin, steroid, triterpenoid, glycoside, triterpenoid, saponin, flavonoid, amino acid and protein. The total phenol and flavonoid content were found to be 0.59% and 4.50%. For quantitative analysis through HPTLC techniques, optimization of solvent system was done and the content of quercetin and rutin in the extract was found to be 3.22% (w/w) and 0.65% (w/w) respectively. Conclusion: These above mentioned parameters can be used as an important tool for the standardization of Z. officinale extract.

1. Introduction

Zingiber officinale (Z. officinale) Roscoe (family, Zingiberaceae), commonly known as ginger, is consumed worldwide as spice and flavoring agent. It is commercially cultivated in India, China, South East Asia, West Indies, Mexico, Africa, Fiji and Australia[1]. Biochemically, the main active components of Z. officinale is gingerol and shogaol, while zingiberene is obtained from ginger oil[2].

In Ayurveda, Z. officinale is considered as valuable medicine due to rubefacient, antiasthmatic and stimulant to the gastrointestinal tract. It possesses antioxidant, hypoglycaemic, hypolipidaemic effects[1]. Z. officinale has a long history of use in ailments such as nausea, respiratory disorders, cardiovascular and rheumatic disorders. Z. officinale also has immunomodulatory properties andis reported to inhibit various inflammatory mediators such as prostaglandins and proinflammatory cytokines[3]. Ginger extract has antibacterial, anticonvulsion, analgesic, antiulcer, antitumour, antifungal, antiallergen, antiplatlet, antioxidant, antirhinoviral, antihepatotoxicity and antiarthritic activity[2,4].

2. Material and methods

2.1. Development of bioanalytical parameters

Crude plant extract of Z. officinale was procured from Garlico Herbal Concentrate (M.P.), India. Phytochemical screening was conducted on the Z. officinale extract to conferm the presence of different phytoconstituents[5]. The presence of phytoconstituents in the extract was also analysed through thin layer chromatography (TLC) analysis[6].

Parameters such as solubility in water, pH, moisture content, heavy metal analysis and microbiological analysis were also performed in the present investigation as perstandard official methods[7,8]. Total phenol and flavonoid content were also determined according to the standard methods[9,10]. The quantification of rutin and quercetin in Z. officinale were determined through high performance thin layer chromatography (HPTLC) techniques (Table 1).

Table 1.Bioanalytical parameters of analysis.

3. Results

Phytochemical analysis showed the presence of alkaloid, carbohydrate, tannin, steroid, triterpenoid, glycoside, triterpenoid, saponin, flavonoid, amino acid and protein. TLC analysis showed two prominent spots with Rf0.20 and 0.90 value in n-hexane: diethyl amine (40:60) solvent system. However in the fingerprint analysis through HPTLC showed seven prominent spots in ethyl acetate: formic acid: glacial acetic acid: H2O (100:11:11:26) solvent system (Table 2). Moisture content and solubily in alcohal was found to be 6.94% and 32.10%. The total phenol and flavonoid content were found to be 0.59% and 4.50%. Further heavy metal level (lead, arsenic, mercury, cadmium) were found to be under the limit. Microbiological assay showed that E. coli and Salmonella was found to be absent whereas total bacterial count and yeast & moulds contents were found to be below the limit. For quantitative analysis through HPTLC techniques, optimization of solvent system was done and ethyl acetate: formic acid: glacial acetic acid: water (100:11:11:26) was found to be most suitable solvent system. The content of quercetin and rutin in Z. officinale was found to be 3.22% (w/w) and 0.65% (w/w) respectively.

Table 2.HPTLC fingerprint analysis of Z. officinale extract.

4. Discussion

Physicochemical and phytochemical analysis was generally performed to ensure the identity, purity and quality of the drug. These parameters are also used to check the genuine nature of the crude drug, thus it plays an important role in preventing the possible steps of adulteration. Phytoconstituents obtained from natural sources have been gaining importance in the day by day due to the health promoting activity. So it is necessory to check the quality safety and efficacy of herbal drugs before its consumption. In the last few decades, an HPTLC technique has gained much popularity for standardization of the herbal drugs and formulations due to analysis of several samples simultaneously with small quantity of marker compound and mobile phase with very less time[11].

TLC and HPTLC techniques can be used to solve many qualitative and quantitative analytical problems in a wide range of fields, including medicine, pharmaceuticals, chemistry, biochemistry, food analysis, toxicology and environmental analysis[12,13]. Quality evaluation and standardization of the herbal preparation is a fundamental requirement of industry and other organization dealing with ayurvedic and herbal products. According to WHO guidelines, an herbal product needs to be standardized with respect to safety before releasing it into the market[14]. Heavy metals are important environmental pollutants and many of them are toxic even at very low concentrations. Heavy metal accumulations in edible plants can directly affect plant growth and an excess dietary intake of contaminated plants could also be dangerous for the health of humans and animals[15].

These above mentioned bioanalytical parameters can be utilizes for the simultaneous analysis of different phytoconstituents present in the Z. officinale plant material. In future, this information may be useful as a standard to identify the adultrants and other related species.

Conflict of interest

The authors declare they have no conflict of interests.

Acknowledgements

This work was financially supported by the TIFAC CORE in Herbal Drugs by DST in J.S.S. College of Pharmacy Ooty, T.N., India.

[1] Bhandari U, Kanojia R, Pillai KK. Effect of ethanolic extract of Zingiber officinale on dyslipidaemia in diabetic rats. J Ethnopharmacol 2005; 97(2): 227-230.

[2] Ahmad N, Sulaiman S, Mukti NA, Murad NA, Hamid NAA, Yusof YAM. Effects of ginger extract (Zingiber officinale Roscoe) on antioxidant status of hepatocarcinoma induced rats. Malays J Biochem Mol Biol 2006; 14: 7-12.

[3] Daswani PG, Brijesh S, Tetali P, Antia NH, Birdi TJ. Antidiarrhoeal activity of Zingiber officinale (Rosc.). Curr Sci 2010; 98(2): 222-229.

[4] Darbar S, Bose A, Chattaraj TK, Pal TK. Protective role of Zingiber officinale roscoe on aceclofenac induced oxidative stress in rat liver. Int J Pharm Tech Res 2010; 2(1): 495-501.

[5] Khandelwal KR. Practical pharmacognosy techniques and experiments. 17th Ed. Pune: Nirali Prakashan; 2007.

[6] Wagner H, Bladt S. Plant drug analysis, A thin layer chromatography atlas. Germany: Springer-Verlog Berlin Heidelberg; 1996.

[7] Government of India, Ministry of Health and Family Welfare. Indian Pharmacopeoia. Delhi: Controller of publications publishers; 1996, p. 54.

[8] World Health Organization. Traditional medicine[Online]. Available from: http.//www.who.int/mediacentre/factsheets/ fs134/en/. [Retrieved April 12, 2009].

[9] Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic 1965; 16(3): 144-158.

[10] Chang CC, Yang MH, Wen HM, Chern JC. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 2002; 10(3): 178-182.

[11] Kshirsagar VB, Deokate UA, Bharkad VB, Khadabadi SS. HPTLC method development and validation for the simultaneous estimation of diosgenin and levodopa in marketed formulation. Asian J Res Chem 2008; 1(1): 36-39.

[12] Choudhary N, Sekhon BS. An overview of advances in the standardization of herbal drugs. J Pharm Educ Res 2011; 2: 55-70.

[13] Patel DK, Patel K, Dhanabal SP. Phytochemical standardization of Aloe vera extract by HPTLC techniques. J Acute Dis 2012; 1(1): 47-50.

[14] Jain A, Lodhi S, Singhai AK. Simultaneous estimation of quercetin and rutin in Tephrosia purpurea Pers by high performance thinlayer chromatography. Asian J Traditional Med 2009; 4: 104-109.

[15] Rai S, Sharma DK, Arora SS, Sharma M, Chopra AK. Concentration of the heavy metals in Aloe vera L. (Aloe barbadensis Miller) Leaves collected from different geographical locations of India. Ann Biol Res 2011; 2(6): 575-579.

JAD obtained the CODEN (JADOCO) of CA

CODEN (JADOCO) has been provided for Journal of Acute Disease (ISSN 2221-6189) by International CODEN Service Centre.

CODEN is a six character, alphanumeric bibliographic code that provides concise, unique and unambiguous identification of the titles of periodicals and non-serial publications from all subject areas. It is particularly common in the scientific community as a citation system for periodicals cited in technical-as well in chemistry-related publications and as a search tool in many bibliographic catalogues.

1 March 2013

*Corresponding author: Dr. S. P. Dhanabal, Professor & Head, Department of Phytochemistry & Phytomedicine, JSS college of Pharmacy, Ooty- 643 001, India.

Tel: 09443095697

E-mail: dhanskanaksahil@rediffmail.com

ARTICLE INFO

Article history:

Received in revised form 24 March 2013

Accepted 20 May 2013

Available online 20 June 2013

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