Pharmaceutico-analytical Study of Patolakaturohinyadi Ghanavati - A Poly Herbal Formulation

Introduction

Patolakaturohinyadi Kashaya is a very famous classical Ayurvedic formulation explained in shodhaniya adhyaya of Astanga Hridaya Sutrasthana, 15th chapter. The ingredients of Patolakatrohinyadi Kashaya are as follows: Patola, Katurohini, Chandana, Madhusrava, Guduchi, Paata. Phalashruti is as follows: kaphapitta nihanti, kushta, jwara, visha, chardi, arochaka, kamalahara.¹ It is a potent antitoxic medicine used for RGUHS Journal of AYUSH Sciences liver detoxification. Even though this kashaya is potent, the palatability of the kashaya is very difficult because of its tiktarasa pradanata. Probably for this reason, Acharyas came up with a secondary preparation from panchavida kashaya kalpanas. In the market, kashaya tablets are available to overcome this palatability issues. The primary preparation Kashaya (decoction) is used as a base in the preparation of medicated oils, ghee, arista, rasakriya, etc.² With the concept of rasakriya, classical formulation of Patolakaturohinyadi Kashaya was modified into a ghanavati form for better efficacy and palatability.

This kashaya is further boiled to form the Rasakriya/ Ghana. ³ Ghana or Rasakriyas are solidified decoctions or swarasa of a drug or drugs. In other words, they are water soluble extracts in solid form. All water-soluble principles of a drug can be extracted and preserved for more days compared to decoction, powder, and other preliminary preparations. The dose of a drug could be minimized in this form. It becomes easy to consume bitter drugs in this form.⁴ The ghanavaties as they are in concentrated form of Kashaya, the dose of the drug may be reduced by preparing the ghanavaties.

In the present study, effort was made to prepare Patolakaturohinyadi Ghanavati as per classical reference. The organoleptic, physicochemical and phytochemical, high-performance thin layer chromatography (HPTLC) and microbial analysis was done by considering the parameters mentioned for standardization of vati/gutika/ modaka as the preparation is in vati form.

Materials and Methods

Collection of raw drugs

All the required raw drugs except Chandana were collected from KLE pharmacy and authentication of raw drugs was done in Central research facility, Shri BM K Ayurveda Mahavidyalaya, Belgaum. Chandana was collected from the Srigandha kote, Shivamogga and authentication was done in Central research facility, Shri BM K Ayurveda Mahavidyalaya, Belgaum. The list of ingredients are mentioned in Table 1.

All the individual drugs were taken in equal parts, kept for soaking overnight by adding 1/4th quantity of water (i.e 432 L) from the total quantity of the water. On the next day, after adding the remaining amount of water (i.e 1296 L), it was kept over low flame till the volume of water reduced to 1/4th. During the entire process of preparation of Kashaya, continuous stirring was maintained. To maintain uniform low temperature, the biogas masifier was used.

After complete preparation of the Kashaya, container was taken out of fire and allowed for complete cooling (swanga sheeta). The content was filtered to another vessel through a fine clean cloth. The Kashaya thus obtained was Patolakaturohinyadi Kashaya. This Kashaya was again subjected for reheating under continuous low flame. After about 1/4th of water evaporated, the consistency of the liquid started to slowly thicken. The heating process was continued till a semisolid mass was obtained and care was taken to avoid overheating and sticking of the mass to the container. Then the obtained cooled semisolid paste was spread very thin on the plastic sheet in a tray with the help of a spatula, was subjected for drying naturally under sun. It took almost more than a week for complete drying. After drying, it became very hard and was made into small pieces by breaking. The powder of the Ghana of Patolakaturohinyadi Kashaya was obtained which was about 4 kg. This Ghana powder was then punched into tablet form which measured about 500 mg each. Thus the prepared tablets were stored in an air tight jar.

Physiochemical and phytochemical parameters

The finished drug was analyzed by using qualitative and quantitative parameters at the Central research facility, Shri B M Kankanawadi Ayurveda Mahavidyalaya, Belgaum. Physical tests like organoleptic characteristics, physicochemical, phytochemical analysis was carried out.

HPTLC

One gram of sample of Patolakaturohinyadi Ghanavati was dissolved in 10.0 mL of ethanol, kept overnight and filtered. Six µL of each of the above extract was applied on a pre-coated silica gel F254 on aluminum plates to a band width of 7 mm using Linomat 5 TLC applicator. Patolakaturohinyadi Ghanavati sample plate was developed in toluene:ethyl acetate:formic acid (5.0:3.5:0.5). The developed plates were visualized in short UV, long UV and then derivatized with vanillin sulphuric acid (VSA) reagent, subsequently scanned under UV 254 nm, 366 nm and 620 nm (after derivatization). Rf, colour of the spots, densitometric scan and 3-D chromatograms were recorded.

Microbial load analysis

Preparation of Casein Soya Bean Digest Agar Medium (CSDAM):

Casein peptone (15g), soya peptone (5g), sodium chloride (5g) were taken and dissolved in 990 mL distilled water, pH was adjusted to 7.3±0.2 and the volume was made up to 1000 mL. Finally, 15 g of agar was added to the media and autoclaved at 121˚C for 20 minutes. HPTLC and microbial load analysis was done at Sri Dharmasthala Manjunatheshwara Centre for Research in Ayurveda and Allied Sciences, Kuthpady, Udupi.

Results

Organoleptic evaluation

Various parameters such as colour, odour, taste, etc. of Patolakaturohinyadi Ghanavti were observed and recorded. The results are mentioned in Table 3.

Physicochemical analysis

Physicochemical analysis was carried out with the following parameters i.e., loss on drying, total ash, acid insoluble ash, water soluble extract, alcohol soluble extract, average weight, hardness, friability, disintegration, pH. The results are mentioned in the following table (Table 4).

Phytochemical analysis

Preliminary phytochemical analysis was done and the results are mentioned in Table 5.

HPTLC Study

The given sample of Patolakaturhinyadi ghanavati was standardized analytically by HPTLC as per testing protocol mentioned in Ayurvedic Pharmacopoeia of India. Rf values, densitometric scan results are presented in Table 6 and Figures 3-5.

Microbial load

The given sample of Patolakaturohinyadhi Ghanavati was tested against microorganisms. The results are mentioned in Table 7.

Discussion

The preparation of ghanavati is a very tedious procedure. Most of the manufacturing companies are not preparing the ghanavati as the yield is very less. For producing one kg of ghana, minimum 10 L of Kashaya needs to be subjected for boiling, thus resulting in just 10 percent yield. Large amounts of raw drugs are needed for this to be produced in a large scale. 

One more difficulty is the procurement of Shweta Chandana in Karnataka which is expensive The Government of Karnataka sells Chanadana for public at three places in Karnataka Shivamogga, Dharwad and Mysore. For this, strict rules and regulations are followed and prior approval from the Deputy Conservator of Forests (DCF) of that division is necessary. An individual is allowed to buy a maximum of 3 kg of Chandana. Considering all this, procurement in large quantities is a challenge.

Madhusrava or murva is one of the controversial drugs. Many species are available in the name of murva. In the present study, murva with botanical name Marsdenia tenacissima was selected for the preparation

The vati were found to have an average weight of 479 mg ± 10% range of weight variation.⁵ Bioavailability of the medicine depends on the hardness and disintegration of vati. Patolakaturohinyadi Ghanavati was found to have a hardness of 6.0 kg/cm and required 25 min for disintegration which was found to be within normal limits. Moisture content (loss on drying) was found to be 6.268% which is also within normal limits. If the moisture content is high, it could be more susceptible for microbial contamination.

Ash value is the criteria for considering the purity of crude drug. Patolakaturohinyadi Ghanavati contained 46.222% of total ash and 1.925% of acid insoluble ash. The 77.972% w/w of water soluble extractive and 28.557% w/w of alcohol soluble extractive were present which indicated good solubility of drugs in water. The HPTLC of Patolakaturohinyadi Ghanavati revealed maximum four spots at short UV and long UV.

The microbial load analysis showed a result of 38 CFU/ mL by direct method which was in the acceptable range for microbial contamination.⁶ Therefore, further serial dilution method was not followed.

In the phytochemical analysis, the steroids, tannins, saponin glycosides were found to be present in the water extract, and while the presence of flavonoids, tannins, cardiac glycosoids, anthraquinone, and saponin glycosides was observed in the alcohol extract. Medicinal property of the preparation could be attributed to the presence of the above mentioned phytochemicals.

Conclusion

From the present analytical study, it can be concluded that the formulation of Patolakaturohinyadi Ghanavati contains all good characteristics of an ideal vati, and also the formulation is of good quality and purity. API standards are not mentioned for this formulation. Hence the obtained results of the present study may serve as reference standards in the preparation of drug formulation and may also help in further clinical research.

Conflict of Interest

None