Standardization and Comparative Evaluation of Shivakshar Pachan Churna: A Traditional Ayurvedic Formulation

Varun Chaddha¹*, Avinash Singh Kushwah², Vaibhav Srivastava³

¹Shri Ramnath Singh Institute of Pharmaceutical Science and Technology, Sitholi, Gwalior

²Ramnath Singh Mahavidyalya , Gormi, Bhind

³NIPS Sitholi, Gwalior

*Corresponding Author E-mail: nurav86@gmail.com

ABSTRACT:

The aim of this study is to improve the acceptability of Shivakshar Pachan Churna, across the world by developing certain quality control parameters and fingerprinting method according to WHO guidelines for traditional formulations. Two batches of Shivakshar Pachan Churna (SPC) were purchased from the market for standardization and to check uniformity in batches. All batches were physiochemically evaluated for Extractive value, Phyto-constituents, Moisture Content and Micromeritic parameters. In present communication Chromatographic TLC (Thin Layer Chromatography) was developed for the standardization of Shivakshar Pachan Churna an Ayurvedic formulation. Spectroscopic studies were carried out to develop the spectrum of the formulation and validated by Overlain and Linearity study (Beer`s law). We have developed a simple scheme for the standardization and authentication of Shivakshar Pachan Churna (SPC) an Ayurvedic formulation. The present study signifies the use of TLC, Spectroscopic Fingerprint profiles for deciding the identity, purity and strength of the Shivakshar Pachan Churna (SPC) an Ayurvedic formulation.

KEYWORDS: Shivakshar Pachan Churna (SPC) ; Shivakshar Pachan Churna Dabur Pharmaceuticals (SPCD), Shivakshar Pachan Churna Sharmayu Pharmaceuticals (SPCS), U.V. Spectroscopy; Thin Layer Chromatography (TLC)

INTRODUCTION:

Ayurveda is a holistic healing science which comprises of two words, Ayu and Veda. Ayu means life and Veda means knowledge or Science. So the literal meaning of the word Ayurveda is the science of life. The quality of herbal medicine that is the profile of the constituents in the final product has implication in efficacy and safety. Due to the complex nature and inherent variability of the chemical constituents of the plant based drugs, it is difficult to establish quality control parameters and modern analytical techniques are expected to help in circumventing this problem. Shivakshar Pacha Churna (SPC) an Ayurvedic formulation , consist of Hingvashtaka churna, Haritaki (Terminalia Chebula), and Sarji-kshara ingredients in churna (powder) form .It is one of the most widely used Ayurvedic product. Shivakshar Pachan Churna is the fortified form of Hingvastaka churna.

Here Shiva stands for Haritaki (Terminalia chebula) it is one of the best known digestive rejuvenator; kashar is for Sarjikshar it loosens and clears out the feces from the bowel; Pachan is for Hingwashtak Churna and also tells about its rejuvenating effect on all aspects of the digestive system digestion, assimilation and excretion of wastes and toxins. Shivakshar Pachan Churna is useful in managing all the digestive disturbances irrespective of the dosha involvement. Shivakshar Pachan Churna ignites the digestive fire and ensures timely bowel clearance. Shivakshar pachan churna digests Aama deposited in body channels, it improves the functioning of liver. Therefore an attempt has been made to standardize Shivakshar Pachan Churna (SPC) an Ayurvedic formulation based on their TLC, and U.V. Spectroscopic Fingerprint profile. The marketed formulations were subjected to TLC and U.V. Spectroscopic

MATERIALS AND METHODS:

All the chemicals used in the experiment were of analytical grade. The U.V. spectra were recorded on the Shimadzu-UVPharmaspac-1700spectrophotometer. The TLC was performed on a silica Gel G plate. Two products of Shivakshar Pachan Churna (SPC) from two different manufacturers one from Dabur Pharmaceuticals (SPCD) and other from Sharmayu Pharmaceuticals (SPCS) were purchased from the market of District Jhansi (U.P).

Table1. Formulation composition of Shivakshar Pachan Churna:

1. Haritaki (Terminalia chebula) 33.33%

2. Hingvashtaka churna: 33.33%

Ginger (Zingiber officinale) Rz, 1 part

Ajamoda (Apium leptophyllum) Fr. 1 part

Sveta jiraka (Cuminum cyminum) Fr. 1 part

Caraway (Carum carvi) Fr. 1 part

Black pepper (Piper nigrum) Fr. 1 part

Pippali (Piper longum) Fr. 1 part

Hing (Ferula foetida) Exd. 1 part

Saindhava lava (Black Salt) 1 part

3. Sarji-kshara 33.33%

1. Physico-Chemical Evaluation

A. Extractive values: Extractive values of SPCD and SPCS (5gm) were obtained in methanol, chloroform, solvent ether of different batches were obtained in methanol, chloroform, ether, n-hexane and distilled water as per the method given in Indian Pharmacopoeia and WHO guidelines. Results shown in table-1.^(1,3)

B. Organoleptic properties:

All two batches of SPC were observed for color, odor, taste and appearance.⁽²⁾

C. Moisture Content:

For determining the moisture content 2gm of SPCD and SPCS were taken in a China Dish weighed. Dried in the oven at 105°C for 5 hrs. Cool in a dessicator and weight was measured till the constant weight. Results shown in table-1.⁽²⁾

D. Total Ash value:

For determining the ash value 2 gm of ground air-dried material, accurately weighed. Spread the material in an even

layer and ignite it by gradually increasing the heat at 450oc until it is white, indicating the absence of carbon. Cool in a dessicator and weigh. ⁽²⁾

Acid insoluble ash:

For determining the acid insoluble ash the ash was boiled with 25 ml. of 2M hydrochloric acid for 5 minutes, collected the insoluble matter in an ash less filter paper, washed with hot water, and ignite cool in a dessicator and weight was measured. Calculated the percentage of acid insoluble ash with reference to the air-dried drug. ⁽²⁾

WATER SOLUBLE ASH:

For determining the water soluble ash the total ash was boiled with 25 ml. of 2M hydrochloric acid for 5 minutes, collected the insoluble matter in an ash less filter paper, washed with hot water, and ignite for 15 min. at a temperature not exceeding 450˚C. Substrated the weight of the insoluble matter from the weight of the ash; the difference of weight represented the water-soluble ash. Calculated the percentage of water-soluble ash with reference to the air-dried drug.⁽²⁾

Table 2. Physiochemical evaluation of SPCD and SPCS

Analytical Parameters Mean of SPCD Mean of SPCS

Extractive Values

Water soluble 51.191 ± 0.101 57.472 ± 0.283

Methanol soluble 46.345 ± 0.167 28.657 ± 0.189

Chloroform soluble 14.404 ± 0.01 12.799 ±0.056

n-hexane soluble 8.089 ± 0.1626 16.107 ±0.096

Ether soluble 4.892 ± 0.3095 4.997 ± 0.0775

Moisture Content 10.066 ± 0.1061 5.3794±0.0814

E. Phytochemical Evaluation:

The methanol, chloroform, water, n-hexane, and ether extract of SPCD and SPCS respectively were subjected to phytochemical test as per the method given in standard reference book. Results shown in table-3.⁽⁴⁾

Table 4. Phyto-constituents present in SPCD

*S.No*.	*Phytoconstituents*	*SPCD*					*SPCS*
*S.No*.	*Phytoconstituents*	*Wt.*	*Et.*	*CHCL₃*	*MeOH*	*n-hexane*	*Wt.*	*ET.*	*CHCL₃*	*MeOH*	*n- hexane*
1.	Alkaloids	+	-	-	-	-	+	-	-	+	-
2.	Glycosides	+	-	+	+	-	+	+	+	+	+
3.	Steroids	-	-	+	-	+	-	+	+	+	+
4.	Protein	+	+	-	+	-	+	+	+	+	+
5.	Carbohydrates	+	+	-	+	-	+	-	-	+	-
6.	Saponins	+	-	-	+	+	+	+	-	-	-
7.	Tannins	+	+	+	+	-	+	-	-	+	-
8.	Flavanoids	+	+	-	-	+	+	+	-	-	+

2. Micromeretic Parameters:

The physical characteristics of the formulations were determined in terms of bulk density, compressibility index and angle of repose in accordance the method given in a Pharmacopeia of India ⁽⁵⁾. Results shown in table- 3.

Table 3. Micromeretic Parameters of SPCD and SPCS

S.No Parameters Mean of SPCD Mean of SPCS

1. Bulk Density 0.7107±0.0077 0.5598±0.0035

2. Tapped Density 1.12167±0.1476 0.8070±0.0123

3. Compressibility 36.2335±2.6986 30.590±1.1309

index

4. Angle of repose 26.5185±0.3356 38.2672± 1.215

3. Spectrophotometric Studies:

SPCD and SPCS were extracted separately in distilled water, chloroform, methanol, ether and n-hexane. The extracts were dried under reduced pressure and the extract of all the two batches were studied spectrophotometrically for spectrum study. The Overlain spectrum and linearity studies were performed with only distilled water and methanol soluble extract of the formulations. The U.V. spectra were recorded in a Shimadzu-U.V. Pharmaspac-1700 spectrophotometer. This study was performed to find out the reproducible peaks in U.V. range 200-400nm (Overlain study) and content uniformity (linearity study) in finished products.⁽⁶⁾

Spectrum study:

All the extract from each batch of SPC were dissolved in corresponding solvent. Their U.V- spectrum were recorded at the range of 200-400nm.In each case the baseline were cleared against the solvent in which the particular solution of extract is prepared. The scanned spectrums were recorded and the peak of the maximum absorbance was noted. ⁽⁶⁾Results have been shown in table-5.

Overlain spectrum study:

The overlain UV- spectrums were recorded only aqueous and alcohol soluble extract of SPCD and SPCS were recorded.⁽⁶⁾

Linearity Study:

The linearity studies were carried out with aqueous and alcoholic solutions only. SPC (1.25 gm) from each batch were dissolved separately in 25ml, of alcohol and distilled water for 1 hour at 100rpm in cold conditions at 25°±1°C, then filter the mixture. The prepared solution of the formulation were diluted in the concentration range of 0.2-1 ml (of the formulation) for aqueous at 256.20nm and 0.2-1ml (of the formulation) for alcoholic at 277.20 nm respectively. The linear correlation between these concentrations (X- axes) and absorbance (Y- axes) were graphically presented and the slope (b), intercept (a) and correlation coefficient (r) were calculated out for linear equation by regression analysis by Microsoft Excel.⁽⁶⁾

Fig. 5 Linearity of Aq. Extract of SPCD

Fig. 6 Linearity of Aq. Extract of SPCS

Fig. 5 Linearity of Aq. Extract of SPCD

Fig. 6 Linearity of Aq. Extract of SPCS

Table 7. Regression data for linearity Studies

S.No Extract Regression

SPCD SPCS

1. Wtr. Extract y=2.564x+0.006 y=2.242x+0.345

Regression=0.985 Regression= 0.995

2. Alc. Extract y=2.181x+0.250 y=0.955x+0.001

Regression=0.989 Regression= 0.997

4. Chromatographic studies⁽³⁾:

Chromatographic studied was performed for the TLC fingerprinting of SPC of different companies.

Test solution: 0.6gm of powdered churna was taken in an iodine flask. Than 12.5ml of distilled water is added and 6hrs continuous shaked and kept for 24hours. Water extract was filtered and kept for drying.

Solvent system: n-butanol: acetic acid: water (4:1:5)

Procedure: Applied one spot of test solution of different companies on precoated Silica Gel G plate of uniform thickness of the plates were developed in the solvent system.

Visualization: The plates were examined under iodine vapour chamber.

Evaluation: Different Rf value of different companies have been seen. SPCD gives Rf 0.41 while SPCS gives Rf 0.35.

RESULTS AND DISCUSSION:

The SPCD were sour and salty in taste, yellowish brown in color, and characteristic odor. While SPCS were sour and salty in taste, brownish in color and characteristic odor. All results are presented in mean ±SEM. The loss on drying at 105°C is 10.066 ± 0.1061 for SPCD and SPCS were found to be 5.3794±0.0814 respectively. The parameters like extractive values of SPCD in water, methanol, chloroform, hexane and ether were found to be respectively indicated the presence of almost polar and semipolar components in the formulation, where the plant material is used. The results of preliminary phytochemical analysis also show the presence of alkaloids, glycosides, protein, carbohydrate, saponin, tannins, flavanoids in water extract of SPCD, in ether extract of SPCD showed the presence of protein, carbohydrate, tannins and flavanoids, chloroform extract of SPCD shows the presence of glycosides, steroids and tannins, in methanol extract of SPCD showed the presence of glycosides, protein, carbohydrate, saponins and tannins and the n- hexane extract of SPCD showed the presence of steroids, saponins and flavanoids while the presence of alkaloids,glycosides,protein,carbohydrate,saponin,tannins,flavanoids in water extract of SPCS, in ether extract of SPCS showed the presence of glycosides, steroids, protein, saponin, and flavanoids, chloroform extract of SPCS shows the presence of glycosides, steroids and protein, in methanol extract of SPCS showed the presence of alkaloids , glycosides, steroids, protein, carbohydrate and tannins and the n- hexane extract of SPCS showed the presence of glycosides, steroids, protein, and flavanoids . Evaluation of Micromeretic parameters like bulk density, tapped density, Compressibility index and angle of repose were done to check the flow properties of SPCD and SPCS. The result shows that the two samples have good flow properties. The TLC was performed for fingerprinting of SPCD and SPCS. The results of TLC shows that the Rf value of SPCD is 0.41 and 0.35 for SPCS. For fingerprinting of the SPCD and SPCS an attempt has been made with the help of spectroscopic studies because of its simplicity. The U.V-Spectrums of solutions form of SPCD and SPCS indicates prominent and identical peaks for aqueous and alcoholic extracts respectively. The peaks are sharper at lower concentration while diffuse as the concentration is increase. An overlain spectrum reflects the studied extract to have some component in same ratio.

The absorption maximum at designated wavelength can be utilized to evaluate consistency of the product and constituents. It may be utilized for product evaluation for adulteration where addition of excess of inorganic substance and substituted constituents, etc.

The prepared sample dilution for aqueous (0.20-1ml) and alcoholic (0.20-1ml) solution of SPCD and SPCS shows linearity in the absorbance at nm and nm respectively. The absorption maxima at designated wavelengths can be utilized to evaluate consistency of the product and constituents. Linearity studies make feasible quantitative estimation of components in SPC contributing for generating the maxima at designated wavelength. The regression data for linearity studies of SPCD water extract shows 0.985 and for SPCD alcohol extract shows 0.989 while of SPCS water extract shows 0.995 and for SPCS alcohol extract shows 0.997 respectively. Static analysis shows significant difference in data obtained among all batches.

CONCLUSION:

Ayurvedic medicine Shivakshar Pachan churna has been standardized by intervention with modern scientific quality control measures and for the development of finger printing methods. The results obtained of SPCD and SPCS found to be comparable with each other like in extractive value, micromeretic properties, Rf values, Phytochemical tests and U.V analysis. These finding may be applicable for quality control and finger printing of different companies of same Ayurvedic products. So far since no attempt has been made to standardization and comparable studies of an Ayurvedic formulation of Shivakshar Pachan churna of different companies products. This is the small step towards the development of quality control methods and finger printing of different companies same products. This will also helps to produce uniform standard product and will be helpful in detecting adulteration or substandard product in traditional Ayurvedic formulation.

The TLC analysis also showed as a comparable finger printing of two companies of a SPCD and SPCS. Hence these parameters and developed methods for their determination may be considered as a tool for quality control, and identification methods of traditional formulations. This will also assist the regulatory authorities, scientific organizations and manufacturers in identification and comparing their product with other company same product.

REFFERENCES:

1. The Ayurvedic Formulary of India, 1, 2, the controller of publication, Delhi.

2. Parihar Singh Sandeep "Standardization of Ashokarista Formulation by TLC method”, International Journal of Pharm Tech Research Vol.2, No.2, pp 1427-1430.

3. Singh Himmat "Standardization of Arjunarishta formulation by TLC method” International Journal of Pharmaceutical Sciences Review and Research, Vol 2, Issue 1, May- June 2010 pp25-28.

4. Shukla, Karunakar "Development of Quality Control Parameters of Bhaskar Lavan Churna: A Traditional Ayurvedic Formulation” Taiwan Pharmaceutical Journal, 2007, 59, 47-56.

5. Indian Pharmacopoeia, Vol.3, Government of India, Ministry of Health and Family welfare. Ghaziabad: Indian Pharmacopoeia Commission, 2007.

5. Anonymous, Quality Control methods for medicinal plant material (World Health Organization), 2002.

6. Khandewal K.R, Practical Pharmacognosy, (Nirali Prakashan Pune), 2000,149.

Received on 20.12.2013 Modified on 15.01.2014

Res. J. Pharmacognosy & Phytochem. 6(2): April-June 2014; Page 70-74