Spectrophotometric method for routine quality control of Ayurvedic formulation Drakshadi gutika

 

V. Jain¹*, S.J. Daharwal¹, Tripti Jain², Swarnlata Saraf¹ and S. Saraf¹

¹University Institute of Pharmacy, Pt. Ravi Shankar Shukla University, Raipur (C.G.), 492 010

²Chhattisgarh Food and Drug Administration, Raipur (C.G.) 492 010

*Corresponding author: vj_rsofiop@rediffmail.com

ABSTRACT:

The Drakshadi gutika is effective for amlapitta, hrddahakanthadaha, trsna, murccha, agnimandaya and amavata is official in Ayurveduc formulary in India. Quantification of active principles through modern analytical tools is essential for establishing the authenticity, creditability, prescription and usage of Ayurvedic medicines/herbal formulations. The Ayurvedic formulation Drakshadi gutika has been prepared as per Ayurvedic formulary of India was estimated spectrophotometrically for its tannic acid content. Three-laboratory batch of Drakshadi gutika were estimated for their tannic acid contents against standard tannic acid solution on double beam UV-Visible Spectrophotometer at lmax 276 nm. The concentration of tannic acid present in raw material was found to be 8.057± 0.29w/w in Terminalia belerica, and 1.283%±0.64 w/w in vitis vinifera and in three identical laboratory batch of Drakshadi gutika DG-I, DG-II and DG-III, was found to be 3.132±0.396, 3.124±0.396, 3.119±0.682w/w respectively. The tannic acid content of all the three batches is found to be in close proximities with each other and recovery studies are indicative of reproducibility of method. Hence the present method is simple, sensitive, precise and accurate and can be adopted for routine quality control of Drakshadi gutika. 

 

 

INTRODUCTION:

The most of the Ayurvedic formulation are lacking in their defined quality control parameters and method of its evaluation. ¹ The World Health Organization (WHO) in its resolution WHA 31.33 (1978), WHA 40.33 (1987), WHA 42.43 (1989) has emphasized the need to ensure the quality of medicinal plant products by using modern controlled technique and applying suitable standards^2, . The present paper is an effort to develop the quality control parameter of  Drakshadi gutika by spectrophotometric determination using tannic acid as an internal standard.

 

Drakshadi gutika is effective for amlapitta, hrddahakanthadaha, trsna, murccha, agnimandaya and amavata is official in Ayurveduc formulary in India. Main ingredients include Terminalia belerica (Vibhitaka), vitis vinifera (draksha), and Jaggery (Khanda) from Soccharum officinarum ³.

 

In this connection an attempt has been made to develop the method of estimation of tannic acid which is chemically hydrolysable tannin corresponding to a complexity of pentadigalloyl glucose⁴C₇₆ H₅₂ O₄₆ an important content in Drakshadi gutika. Present study is based on UV spectrophotometric analysis, which is a simple, precise, and accurate method that can be considered as quality control method for routine analysis.

 

EXPERIMENTAL:

Plants: All the crude drugs were purchased from local market Raipur (C.G.), India and identified on the basis of morphological and microscopical characters and compared with standard Pharmacopoeial Monograph.^5-,8

 

Chemicals: All the chemicals and solvents were used of A.R. Grade.

 

Preparation of Drakshadi gutika : Drakshadi gutika, three batch name DG-I, DG-II, DG-III, were prepared in laboratory using method described in Ayurvedic Formulary¹⁰. The individual fruit of terminalia belerica, and vitis vinifera was also powdered. These three batches of  Drakshadi gutika and powdered terminalia belerica, and vitis vinifera were estimated for their tannic acid contents against standard tannic acid solution on UV-Visible Spectrophotometer (Shimadzu, UV-1700, Pharmaspec).

 

Preparation of extract of Drakshadi gutika:

The tannic acid extract for each batch of  Drakshadi gutika and separately powdered amla, and draksha were prepared as per the method described by Jain et.al^9,10.

 

Preparation of standard solution:

Tannic acid stock solution was prepared by dissolving 100 mg of accurately weighed tannic acid in 0.1N hydrochloric acid and made up the volume up to 100 ml. Further, 2 ml of this solution was diluted up to 100 ml with 0.1N hydrochloric acid to give 20 mg/ml tannic acid stock solution

 

Calibration Curve of tannic acid; A series of calibrated 10 ml volumetric flask were taken and appropriate aliquots of the working standard solution of tannic acid were withdrawn and diluted up to 10 ml with 0.1 N hydrochloric acid. The absorbance was measured at absorption maxima 276 nm, against blank prepared in similar manner without the tannic acid. The absorption maxima and Beer^’s law limit were recorded and data that prove the linearity and obey Beer^’s law limit were noted.

 

The linear correlation between the concentrations (X-axis) and absorbance (Y- axis) were graphically presented and the slope (b), intercept (a), and correlation coefficient (r²) were calculated out for linear equation (Y= bx + a) by regression analysis using the method of the least square, Table -1.

 

Determination of limit of quantitation and limit of detection

The limit of detection (LOD) is the lowest amount of analyte in a sample which can be detected but not necessarily quantitated as an exact value. The limit of quantitation (LOQ) is the lowest amount of analyte which can be quantitatively determined with suitable precision. The LOD and LOQ of the developed method were determined by injecting progressively low concentration of the standard solution and the lowest concentrations assayed.

 

Estimation of tannic acid:

The appropriate aliquots from tannic acid extract of each batch of  Drakshadi gutika & separately amla and draksha were withdrawn in 10 ml volumetric flask separately absorbance for each aliquots was noted at 276 nm. The corresponding concentration of tannic acid against respective absorbance value was determines using the tannic acid calibration curve. The statistical analysis for checking uniformity in batches is also performed (Table 2).

 

Precision and accuracy

The method was validated for precision and accuracy, by performing the recovery studies.

 

The recovery studies performed at three levels were done by adding known amount of tannic acid to extract of Drakshadi gutika of which the tannic acid content have been already estimated. The observations recorded and recovery was calculated (Table 3).

 

RESULTS AND DISCUSSION:

Tannic acid was found obey Beer Lambert’s law in concentration range 2-20mg/ml at l_max 276 nm. The correlation coefficient (r²) was calculated where the r² value 0.9995 indicates the good linearity between the concentration and absorbance. Under the developed UV conditions, the limit of quantitation was determined to be 0.362μg/mL with triplicate of the sample. Also, the limit of detection was found to be 0.129 μg/mL.

 

The estimation of tannic acid content of Drakshadi gutika (three identical laboratory batch) and powdered terminalia belerica, and vitis vinifera was carried out separately. The concentration of tannic acid present in raw material was found to be 8.057± 0.29w/w in terminalia belerica, and 1.283%±0.64 w/w in vitis vinifera and in three identical laboratory batch of Drakshadi gutika DG-I, DG-II and DG-III, was found to be 3.132±0.396, 3.124±0.396, 3.119±0.682w/w respectively.

 

Table 1: Validation Parameter of Tannic Acid

S. No.	Parameter	Value
1 2 3   4 5 6 7 8 9 10	Absorption Maxima Beer’s Law limit Regression equation (y= bx+a) Intercept (a) Slope (b) Correlation coefficients (r2) Precision (n=6, % RSD) Accuracy (%) LOQ LOD	276 nm 2-20mg/ml y= 0.0417x + 0.0128   0.0128 0.041682 r2 = 0.9995 0.251 99.30 0.362μg/mL 0.129μg/mL

 

 

Table 2:  Estimation of Tannic Acid Content in Drakshadi gutika

S.  no.	Name		Tannic acid content %w/w	Confidence level (95%)
1	Terminalia belerica		8.057± 0.29	±0.248
2	Vitis vinifera		1.283±0.64	±0.592
3	Drakshadi gutika	DG-I	3.132±0.396	±0.416
		DG-II	3.124±0.396	±0.612
		DG-III	3.119±0.682	±0.744

Mean ± SD of six determinations, DG-I:  Drakshadi gutika Batch I,  DG- II:  Drakshadi gutika Batch II, DG -III:  Drakshadi gutika Batch III 

 

 

Table 3: Compilation of Data of Recovery Study

Mean ± SD of six determinations, RSD =Relative Standard Deviation, SE = Standard Error

 

 

 

The recovery studies were carried out for the precision and accuracy parameter. The study was carried out at three levels. To the preanalysed formulation, the standard drugs of tannic acid were added at 50% 100% and 150% levels; dilutions were made, and analyzed by the method. The experiment was repeated six time at three level (Table 3) and result shows 99.48%±0.32, 99.12%±0.29 and 100.24±0.62 recovery of tannic acid at all level with mean value 99.61%±0.41 which prove reproducibility of the result. This shows significant Precision of methods with 95% confidence level. The %Relative Standard Deviation (% RSD) value was found to be0.261, 0.242 and 0.167 with mean 0.233 at all three level while the Standard Error was 0.159, 0.195 and 0.171 with mean 0.175 respectively. From the data’s it is obvious that the present method of Spectrophotometric determination of tannic acid is simple, precise, accurate and suitable fir routine analysis of tannic acid in Drakshadi gutika.  

 

As Drakshadi gutika is a good source of tannic acid, these findings can be taken as one of the parameter, along with other parameters, for Quality control of Drakshadi gutika.

 

ACKNOWLEDGEMENT:

The authors are grateful to DST under FIST and UGC New Delhi for providing financial assistance under MRP (39-169/2010(SR) and SAP Scheme.

 

REFERENCES:

1.     Jain V., Saraf S., and Saraf S. (2007) Spectrophotometric Determination of Piperine in Trikatu Churna: An Ayurvedic Formulation, Asian Journal of Chemistry, 19, (7), 5331-5335.

2.     World Health Organization, Quality Control Methods For Medicinal Plants Materials, Geneva, 1998, 1-15.

3.     The Ayurvedic Formulary of India, Part II, 1^st english  edition, Govt. of India, Ministry of Health and Family Planning, Dept. of Indian system of medicine and Homeopathic, Delhi, 2000,  177.

4.     Khan NS, Hadi SM, Structural features of tannic acid important for DNA degradation in the presence of Cu(II), Mutagenesis,1998 May;13(3):271-4.

5.     The Ayurvedic Pharmacopoeia of India, Part I, Volume I, First edition, Ministry Of Health And Family Welfare, Government Of India, Department of Health, New Delhi, 1986, 4,5,26,47,48.

6.     Indian Herbal Pharmacopoeia, Volume II, Regional Research Laboratory Jammu, Indian drug Manufacturing Association Mumbai, 1999, 50-57.

7.     Quality Standards of Indian Medicinal Plants, Volume I, Indian Council of Medicinal Research, New Delhi, 2003, 198-212.

8.     Mukherjee, P., Pharmacological Screening of Herbal Drug, Quality Control of Herbal Drug: An Approach to Evaluation of Botanicals; Eastern Publishers (Business Horizontal Ltd.), New Delhi, 2002, 539-541.

9.     Jain V., Saraf S., and Saraf S. (2007) Standardization of Triphala Churna: Spectrophotometric Approach, Asian Journal of Chemistry, 19, (2), 1406-1410

10.   Jain V., Saraf S., and Saraf S. (2007) Quantification Of Tannic Acid In Ayurvedic Formulation Bhuvneshavara Vati: Spectrophotometric Approach For Routine Quality Control , Natural Product: An Indian Journal, 3, (2), 107-110.

 

 

Received on 25.09.2011          Modified on 22.10.2011

Accepted on 12.01.2012         © RJPT All right reserved