Physico-chemical investigation of a polyherbal formulation - Vidangatandulaadi choorna

Vidangatandulaadi Choorna is a polyherbal formulation consisting of seven ingredients. Trivrith (Operculina turpethum (Linn.) Silva Manso) is the chief ingredient responsible for the purgative action of the formulation. This yoga ismentioned inKalpasthana of Ashtangahridaya, intended for virechana (Purgation). It is useful in Kapha-vatha disorders. Even though many kinds of research have been done to identify the physicochemical constituents of individual drugs in the formulation, no studies were done to identify the physicochemical properties of the formulation. This analysis helps in understanding the mechanism for different pharmacological actions of the formulation. Hence, Physico-chemical study of Vidangatandulaadi Choorna along with high-performance thin-layer chromatography (HPTLC) ingerprinting is done to ix the standards. All the drugs included in the formulation is identi ied by the botanist and is prepared according to Standards mentioned for the preparation of Choornamentioned in Ayurvedic Pharmacopoeia of India. The formulation is least encountered, but it has shown its signi icant action in Dyslipidemia in folklore practices. As there are no Standards mentioned for this formulation, the result observed in the present study may be considered suitable. The data obtained from Physicochemical investigation, highperformance thin-layer chromatography pro ile and ICP-MS (Inductively Coupled Plasma Mass Spectrometry) could be used as the standards for the present formulation under study.

High-performance thin-layer chromatography, Inductively Coupled Plasma -Mass Spectrometry, Physico-chemical, Vidangatandulaadi Choorna A Vidangatandulaadi Choorna is a polyherbal formulation consisting of seven ingredients. Trivrith (Operculina turpethum (Linn.) Silva Manso) is the chief ingredient responsible for the purgative action of the formulation. This yoga is mentioned in Kalpasthana of Ashtangahridaya, intended for virechana (Purgation). It is useful in Kapha-vatha disorders. Even though many kinds of research have been done to identify the physicochemical constituents of individual drugs in the formulation, no studies were done to identify the physicochemical properties of the formulation. This analysis helps in understanding the mechanism for different pharmacological actions of the formulation. Hence, Physico-chemical study of Vidangatandulaadi Choorna along with high-performance thin-layer chromatography (HPTLC) ingerprinting is done to ix the standards. All the drugs included in the formulation is identi ied by the botanist and is prepared according to Standards mentioned for the preparation of Choorna mentioned in Ayurvedic Pharmacopoeia of India. The formulation is least encountered, but it has shown its signi icant action in Dyslipidemia in folklore practices. As there are no Standards mentioned for this formulation, the result observed in the present study may be considered suitable. The data obtained from Physicochemical investigation, highperformance thin-layer chromatography pro ile and ICP-MS (Inductively Coupled Plasma -Mass Spectrometry) could be used as the standards for the present formulation under study.

INTRODUCTION
Vidangatandulaadi Choorna is one of the clinically signi icant formulations used in the management of Dyslipidemia. It is mentioned in Ashtangahridaya, kalpasthana as a Nityavirechaka (mild laxative) (Ashtangahridaya of Vagbhata, 2002).
The said medicine is used as a traditional method in managing Dyslipidemia. No studies have been published related to Vidangatandulaadi Choorna exploring its Physico -Chemical properties and Pharmacological effects. So an attempt is made to understand the Physico -Chemical properties of the drug along with HPTLC pro ile to ix the standards of the drug, which may lay a future scope for further studies related to this drug. The result of the study can be used to explain the therapeutic bene its of the medicinal formulation.

MATERIALS AND METHODS
All the drugs were procured from Amrita Life, the manufacturing unit under Amrita School of Ayurveda, Vallikavu, Kollam. The authenticity of the drug was con irmed by botanist and experts in Department of Dravyaguna, Amrita School of Ayurveda, Vallikavu, Kollam. Choorna was prepared based on API. Ingredients of the Choorna is mentioned in Table 1. Physico-Chemical parameters like a loss on drying, Water-soluble extractive, Alcohol soluble extractive, pH of 10% solution of Total ash, Acid insoluble ash were done as per API standard guidelines (Department Of Ayush, 2007b). HPTLC ingerprinting was done. ICP -MS was used to estimate the heavy metal contents in the prepared drug.

Determination of pH
Procedure -Ten gram of total ash was dissolved in 100ml of demineralised water. The pH of this 10% solution was measured with a digital pH meter.

Determination of Water-soluble extractive
Procedure -Five gram powdered drug was placed in a round bottom lask and mixed with 100 ml chloroform water. It is kept for 24 hrs with occasional shaking. After that, it was iltered and the iltrate collected in a tarred clean beaker. The residue was weighed after evaporating to dryness.

Determination of Alcohol soluble extractive
Procedure -Five gram powdered drug was placed in a round bottom lask. It was mixed with 100ml alcohol. It was kept for 24 hours with occasional shaking. It was iltered and the iltrate collected in a tarred clean beaker. The iltrate was evaporated to complete dryness, and then the remnant was weighed.

Determination of Total ash
Procedure -Two-gram air-dried drug was weighed with accuracy. It was placed in a tared crucible. It was heated in an incinerator gently, and then the drug was incinerated to ash until it was free from any organic matter. The crucible was kept in a desiccator and cooled and weighed with the contents. Percentage of ash concerning the air-dried drug was calculated.

Determination of Acid insoluble ash
Procedure -Total ash was prepared with two gram dried drug. The whole total ash was dissolved in 25ml diluted HCl. It was boiled for 5minutes. The insoluble portion was iltered through an ashless ilter paper. It was washed with demineralised water. Dried ilter paper with the contents was selected. It was incinerated in a tarred silica crucible with the contents. The incinerated content with the crucible was weighed. Percentage of acid-insoluble ash was calculated.

ICP-MS (Inductively Coupled Plasma -Mass Spectrometry)
Procedure -About 200-500 mg of the sample was accurately weighed and transformed in a cleaned microwave digesting system(MDS)tube. An adequate amount of Con.HNO 3, Con. HCl and a few drops of H 2 O 2 were added to the MDS tube. The MDS tube was further kept in microwave digestive system for complete digestion of solid samples to liquid for one hour at a temperature of 180 0 C. The resultant liquid sample was carefully transferred to a 50ml standard lask and diluted to 50ml. The diluted sample was directly aspirated into the ICP-MS instrument, and the result was obtained (Wilschefski and Baxter, 2019).

Observation on Physicochemical parameters of Vidangatandulaadi Choorna
The analysis of Physicochemical parameters of Vidangatandulaadi Choorna revealed the following observations. It appears as a dark brown powder. The loss on drying is estimated to 3%w/w. The pH of 10% solution of content is 5.23. The water-soluble extractive is 28.23% w/w, Alcohol soluble extractive is 31.58% w/w, Ash value is 5.80% w/w and acid insoluble ash is below the detection limit.

Observation on HPTLC analysis of Vidangatandulaadi Choorna
Peak display (Densitogram) of Vidangatandulaadi Choorna sample at 254nm is shown in Figure 1. Peak display (Densitogram) of Vidangatandulaadi Choorna sample at 366nm is shown in Figure 2 Table 3.

Observations on ICP-MS (Inductively Coupled Plasma -Mass Spectrometer)
The presence of heavy metals observed by Inductively Coupled Plasma -Mass Spectrometer is given as follows. Lead accounts the maximum value reaching about 1.36mg/kg and Mercury about 0.65mg/kg. The presence of Arsenic is 0.18mg/kg and Cadmium is 0.12mg/kg.

Physico -Chemical study
Degradation time of the plant material indicates the quantity of moisture content in it. The powdered plant material degrades quickly due to the growth of microbes and fungus if the moisture content is high. The loss on drying was only 3% for the present sample, which ensure a reasonable period of shelf life for it. Presence of minerals and silica in the plant material was indicated by the Total ash value, which was obtained as 5.80%. The amount of acid-insoluble siliceous matter present in the plant was under the detectable limit. 28.23% w/w water-soluble extractive value indicated that Vidangatandulaadi Choorna

HPTLC Analysis
HPTLC ingerprinting pro ile of Vidangatandulaadi Choorna was developed in Toluene: Ethyl acetate: Formic acid: Methanol (7:5:1:0.5) solvent system. This was taken as the standard Densitogram of Vidangatandulaadi Choorna. Densitogram showed ten peaks at 254nm and eight peaks at 366nm. Each peak represents a chemical entity. Almost four spots are seen repeated in 254 and 366nm. There is a scope for further analysis for inding out the chemical compounds represented by the peaks. To ind out the chemical nature represented by the peaks, TLC-MS may be utilised, in the present solvent system, Toluene: Ethyl acetate: Formic acid: Methanol (7:5:1:0.5). At 366nm there are luorescent spots with green, blue, red and pink colours. Three distinct brown spots are there; it may indicate compound with unsaturation.

ICP-MS (Inductively Coupled Plasma -Mass Spectrometer)
The procedure estimates the number of heavy metals present in the given sample. As per WHO permissible limit of heavy metals like Mercury is 1ppm, lead is 10 ppm, Cadmium is 0.3 ppm, and arsenic is 3 ppm (Department Of Ayush, 2007c). Here in our study, we got all these values under the normal limit, which increases the authenticity of the sample.

CONCLUSION
Vidangatandulaadi Choorna was studied for understanding its physicochemical parameters and HPTLC pro ile. As there are no standards mentioned for this formulation, the physicochemical data and HPTLC pro ile evolved from the present study could be used as standardisation parameters of the formulation.