Standardization of Proprietary Ayurvedic Laxative Tablet

Formulation – Vasulax

 

Madhavi Patel¹, Shivangi Chauhan¹, Vishal Patel^2*, Hardik Soni², Vikram Trivedi²

¹Parul Institute of Pharmacy, Faculty of Pharmacy, Parul University, Vadodara, Gujarat, India.

²Vasu Research Centre (A Div. of Vasu Healthcare Pvt. Ltd.), Vadodara, Gujarat, India.

 

ABSTRACT:

Traditional systems of medicine gain upsurge with the estimated 80% preference for maintenance of healthcare in the entire world. As a rich source for the treatment of diseases, Ayurvedic medicine’s consumption have increased day by day which has led to an increase in various forms of adulteration in raw materials. In a view of increasing demand, it is very crucial to maintain batch-to-batch consistency and efficacy avoiding adulteration and safety issues. Thus, standardization of Ayurvedic medicine in all aspects including phytochemical profiling becomes crucial for the manufacturer. So, the present study portrays standardization of an Ayurvedic proprietary laxative tablet comprising Yashtimadhu, Swarnpatri and Aragvadha as major components. General parameters of tablet were evaluated as per Indian Pharmacopoeia method for quality control purpose. For quantification of phytoconstituents, Sennoside-B was selected as a phytomarker of Swarnpatri & Aragvadha and Monoammonium glycyrrhizinate was selected as a phytomarker of Yashtimadhu. Both the markers were effectively quantified using HPTLC method. The Sennoside B and Monoammonium glycyrrhizinate contents in formulation were found to be 5.10 % and 0.634 % w/w, respectively. These standardization tools will help in maintaining the quality and consistency of this vital Ayurvedic formulation.

 

 

1. INTRODUCTION:

India is blessed with very rich biodiversity and Ayurveda correctly utilizes these resources for health management from past. Ayurveda, being the oldest medicine system in India is the preference of many people who are suffering from chronic diseases, got tired from taking allopathy medicine and are facing with side effects of synthetic medicines. This legacy and trust of people can be maintained by providing quality medicine which must prove themselves as consistent in therapeutics. World Health Organization (WHO) also stresses for quality maintenance of medicinal plants using modern methods¹ which needs to be followed for standardization of Ayurvedic formulations for reproducible therapeutic effect so that it gains the importance².

 

Establishing quality control parameters of the Ayurvedic and other poly herbal formulations is quite challenging especially for analysis of the chemical markers^3,4. A commercial proprietary Ayurvedic laxative tablet formulation, Vasulax was selected for development of its standardization parameters with special emphasis on quantification of phytomarkers using HPTLC. Major active plant material ingredients incorporated in Vasulax are Yashtimadhu, Swarnpatri, and Aragvadha.

 

Yashtimadhu commonly known as Liquorice (Glycyrrhiza glabra L.), is the preferred herb from ancient times of Ayurveda as expectorant, laxative, demulcent, antiulcer, anti-tussive and sweetner^5,6. Monoammonium glycyrrhizinate is an ammonium salt of glycyrrhizin, one of the major triterpenoids saponin present in the plant showing laxative action and also one of the natural sweetners⁷. The active constituent in Swarnpatri called as senna (Cassia angustifolia M. Vahl) and Aragvadha (Cassia fistula L.) is anthraquinone glycoside, sennoside. Both are widely used in treatment of constipation from years^8-14.

The study was aimed to standardize a proprietary laxative Ayurvedic formulation - Vasulax for its general formulation parameters as per Indian Pharmacopoeia and its major markers quantification by HPTLC. HPTLC is simple chromatographic fingerprint to evaluate qualitative and quantitative profile of markers present in drugs. Here, the separation and resolution are better, reliable and reproducible¹⁵. Densitometric scanning provides facility for quantitative measurement of phytoconstituents which is applied precisely to Vasulax tablet to determine the amount of Sennoside B and Monoammonium glycyrrhizinate present in it.

 

2. MATERIALS AND METHODS:

2.1. Physico-chemical analysis:

The tablet was evaluated for size (diameter & thickness), uniformity of weight, hardness, friability and disintegration test as per the Indian Pharmacopoeia guideline^16-18.

 

Size:

Twenty tablets were selected at random from formulation and diameter & thickness were measured using Vernier calipers (Mitutoyo, Japan). Percent deviation was calculated and documented.

 

Uniformity of weight:

Twenty tablets were selected at random from formulation and weighed individually and altogether in analytical balance (Shimadzu, Japan). Percent deviation was found and documented.

 

Hardness:

The hardness was determined for six randomly selected tablets by measuring the pressure required to break the tablet with its diametrical axis using a Monsanto type tablet hardness tester (Campbell, India). The result was expressed as an average of six readings in terms of kg/cm².

 

Friability:

10 tablets were selected at random from the product. Initial weight was determined. Tablets were put in friability test apparatus (Veego, India) and friabilator was run for four minutes for 100 revolutions. Final weight was determined after dusting and % loss was calculated and documented.

 

Disintegration time:

Disintegration time studies were carried out on disintegration test apparatus (Veego, India). The test was carried out at 37 ± 2⁰ C in distilled water. The time for disintegration was noted in the product for 6 tablets.

 

 

 

2.2. Assay by HPTLC^19-23

Chemicals:

The Phytomarkers Monoammonium glycyrrhizinate and Sennoside-B were purchased from Natural Remedies Pvt. Ltd. (Bangalore, India). Methanol, chloroform and HPLC-grade water and other chemicals (Merck, Germany) used were of HPLC-grade only.

 

Materials:

Proprietary Ayurvedic formulation-Vasulax, Yashtimadhu dry extract, Swarnpatri dry extract and Aragvadha dry extract were provided as a gift sample by Vasu Research Centre (A Div. of Vasu Health Care Pvt. Ltd.), Vadodara, Gujrat., India.

 

Preparation of standard solutions:

10 mg of Monoammonium glycyrrhizinate was accurately weighed, transferred to a 10 mL volumetric flask and volume was made up to 10 mL using methanol. 5 mg of Sennoside-B was accurately weighed, transferred to a 5 mL volumetric flask and volume was made up to 5 mL using methanol.

 

Preparation of sample solution:

500 mg of each dry extract were accurately weighed in 250 mL conical flask, and to it 25 mL of methanol was added. Then, it was refluxed for 30 min on water bath, cooled & filtered through Whatman filter paper no. 1 and then concentrated up to 10 mL. 2 gm of tablet was accurately weighed, powdered and transferred in 250 mL iodine flask, and to it 25 mL of Methanol was added. Then, it was refluxed for 30 min on water bath, cooled & filtered through Whatman filter paper no. 1 and then concentrated up to 10 mL.

 

HPTLC Chromatographic condition:

Chromatography was performed on 6 cm x 20 cm aluminium foil plates coated with 0.2 mm thickness silica gel 60 F₂₅₄ (E. Merck, Germany). The standard and samples were applied on the plates as 8 mm wide bands and the space between two spots was kept at 6 mm (10 mm from the bottom and 10 mm from the sides) under a flow of nitrogen gas providing delivery speed 150 μL/s from the syringe using Linomat 5 HPTLC applicator (Camag, Switzerland). 10 μL of the solution were spotted on the plate. These parameters were kept constant throughout the analysis.

 

Detection and Quantification:

After the sample application was completed, the plate was developed in a twin trough glass tank presaturated with a mobile phase of n-Butanol: Water: Glacial Acetic acid (7: 2: 1) for Yashtimadhu dry extract and n-Propanol: Ethyl acetate: Water: Glacial Acetic acid (4: 4: 2.9: 0.1) for Swarnpatri & Aragvadha dry extract. TLC runs were performed under laboratory conditions of 25 ± 2°C. After development, the plates were removed from the chamber, dried in air for 5 min and spots were visualized under UV light using a UV viewer cabinet (Camag, Switzerland). For the densitometric scanning, the plate was observed under UV light at 254 nm for Monoammonium glycyrrhizinate and fluorescent light at 366 nm for Sennoside-B.  Scanning was performed with an HPTLC Scanner-4 (Camag, Switzerland) operated by winCATS software1.4.1. The slit width was 6 mm x 0.45 mm. The evaluation was based on the peak area with linear regression. The identification of Monoammonium glycyrrhizinate and Sennosides-B were confirmed by superimposing the UV spectra of samples and standards within the same R_f window.

 

3. RESULTS AND DISCUSSION:

The data for size measurement of the tablet was used for the quality control of the commercialized product. The uniformity of weight, hardness, friability & disintegration test data of Vasulax were within limits as prescribed in IP (Table 1).

 

Table 1: Physico-chemical quality control data of proprietary Ayurvedic laxative formulation-Vasulax tablet.

Sr. No.	Parameters	Result
1	Diameter	11.34 ± 0.063 mm
2	Thickness	5.92 ± 0.019 mm
3	Uniformity of weight	575.16 ± 0.577 mg
4	Friability	0.012 ± 0.001 %
5	Disintegration test	35 ± 1.132 min

 

The desired resolution of Monoammonium glycyrrhizinate and Sennoside-B together with symmetrical, high resolution and reproducible peaks were obtained by using the mobile phase n-Butanol: Water: Glacial Acetic acid (7: 2: 1) for Monoammonium glycyrrhizinate and n - Propanol: Ethyl acetate: Water: Glacial Acetic acid (4: 4: 2.9: 0.1) for Sennoside-B. HPTLC fingerprinting profiles of Monoammonium glycyrrhizinate, Sennoside-B, Yashtimadhu dry extract, Swarnpatri dry extract, Aragvadha dry extract and proprietary Ayurvedic tablet formulation-Vasulax shown in Figure 1. The presence of Monoammonium glycyrrhizinate and Sennoside-B were visualized in dry extracts and sample of proprietary Ayurvedic formulation (Figure 1).

 

Figure 1: HPTLC fingerprinting profile of (A) Monoammonium glycyrrhizinate-MAG, Yashtimadhu dry extract-YD &Vasulax tablet-VT, (B) Sennoside-B-SB, Swarnpatri dry extract-SE &Vasulax tablet-VT and (C) Sennoside-B-SB, Aragvadh dry extract-AE &Vasulax tablet-VT.

 

Monoammonium glycyrrhizinate and Sennoside-B when quantitatively estimated were found to be 7.246 % w/w, 12.07 % and 9.33 % respectively in the dry extract of Yashtimadhu, Swarnpatri and Aragvadha dry extracts. The proprietary Ayurvedic formulation-Vasulax contains 0.634 % w/w of Monoammonium glycyrrhizinate and 5.10 % of Sennoside-B when analyzed by HPTLC. Results obtained for active content analysis of the tablet are shown in table 2.

 

Table 2: Sennoside-B and Monoammonium glycyrrhizinate content in proprietary Ayurvedic laxative formulation-Vasulax tablet & its ingredients by HPTLC

Name	% Monoammonium glycyrrhizinate	% Sennoside-B
Yashtimadhu dry extract	7.246 ± 0.82 %	---
Swarnpatri dry extract	---	12.07 ± 1.01 %
Aragvadha dry extract	---	9.33 ± 0.89 %
Vasulax Tablet	0.634 ± 0.18 %	5.10 ± 0.58 %

 

HPTLC chromatogram of Monoammonium glycyrrhizinate, Yashtimadhu dry extract & Vasulax tablet are shown in Figure 2 and HPTLC chromatogram of Sennoside-B, Swarnpatri dry extract, Aragvadha dry extract & Vasulax tablet were shown in Figure 3.

 

 

Figure 2: HPTLC chromatogram of (A) Monoammonium glycyrrhizinate (B) Yashtimadhu dry extract and (C) Vasulax Tablet.

 

 

Figure 3: HPTLC chromatogram of (A) Sennoside-B (B) Swarnpatri dry extract (C) Aragvadh dry extract and (D) Vasulax tablet.

 

4. CONCLUSION:

In conclusion, obtained physico-chemical data of the tablets will be useful in the evaluation of the formulation and in maintaining the manufacturing processes. Both the makers were effectively quantified using HPTLC method. The Sennoside-B and Monoammonium glycyrrhizinate contents in formulation were found to be 5.10 % and 0.634 % w/w, respectively which is equivalent to the theoretical value calculated from the result of raw materials. This standardization tool will help in maintaining the quality and batch-to-batch consistency of this important Ayurvedic formulation.

 

5. ACKNOWLEDGEMENTS:

Authors are thankful to Vasu Research Centre, a division of Vasu Healthcare Pvt. Ltd., Vadodara, Gujarat, for providing support to carry out this research work. Authors are also thankful to Parul University, Vadodara, Gujarat, for providing essential support throughout the work.

 

6. COMPETING INTERESTS:

The authors declare no competing interests.

 

7. ABBREVIATIONS:

AE: Aragvadh dry extract; HPLC: High Pressure Liquid Chromatography; HPTLC: High Performance Thin Layer Chromatography; MAG: Monoammonium glycyrrhizinate; SB: Sennoside-B; SE: Swarnpatri dry extract; TLC: Thin Layer Chromatography; VT: Vasulax tablet; WHO: World Health Organization; YD: Yashtimadhu dry extract.

 

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Accepted on 28.02.2022           © RJPT All right reserved

Research J. Pharm. and Tech 2022; 15(11):5206-5210.