Phytochemical and Physico-Chemical analysis of Siddha herbo-mineral Herbal Preparation Kalladaippu Chooranam

Arul priya S, Sudha V

Siddha physician, Agathiyar’s Siddha clinic, Chennai, Tamilnadu, India.

ABSTRACT:

Kalladaippu Chooranam is a herbo-mineral siddha formulation for the treatment of Urolithiasis. Aim and objective: To do the physico-chemical and phytochemical analysis for Kalladaippu Chooranam. Materials and methods: the drug was prepared as per mentioned in the classic Siddha literature. The physico-chemical analysis such as loss of drying, total ash, insoluble ash, water soluble ash, water soluble extractive and alcohol soluble extractive and phytochemical analysis such as detection of alkaloids, carbohydrates, glycosides, saponins, phytosterols, phenols, flavonoids, proteins & amino acids, diterpenes, gum & mucilage and fixed oils & fats were carried out. Results: The physico-chemical analysis revealed that the loss on drying was 8.3%, total ash was 73.53%, acid insoluble ash was 17.96%, water soluble ash was 50.55%, water soluble extraction was 72.15% and alcohol soluble extraction was 2.96%. The phytochemical analysis revealed that the presence of carbohydrates, glycosides, tannins, saponin, phenols, flavonoids and gum & mucilage.

Keywords: Physico-chemical, Phytochemical, Siddha system, Urolithiasis, Kalladaippu Chooranam

1. INTRODUCTION:

Siddha medicine is mainly practiced in Southern part India¹. It is the oldest system of medicines, and the same is not only followed as rituals and norms of daily life but also as a medical practice. The main motto and concept of Siddha is “Food must be taken as medicine or else medicine must be taken as food”². These kinds of medicine are easily and readily available. The standardization of a medicine is very essential and mandatory requirement, so that the quality of medicine can be achieved as per the standard of modern scientific society. Standardization is a vast area to cover, that can be achieved by evaluating the medicine with the required modern standardization rules and regulations³.

Medicinal plants and minerals are beneficial for urolithiasis but there is a requirement of standardization to find out its efficacy. Kalladaippu Chooranam is one of the herbo -mineral formulation mentioned in Siddha Literature for the treatment of urolithiasis. This drug was analysed physico-chemically and phytochemically.

2. AIM AND OBJECTIVE:

The aim of the study is to do physico-chemical analysis and preliminary phytochemistry for the drug Kalladaippu chooranam

3. MATERIALS AND METHODS:

3.1. Collection and identification of drug:

The herbals and minerals of Kalladaippu Chooranam were identified and authenticated (certificate no: 17051801 - 03) by Research officer, Siddha Central Research of India, Central Council for Research in Siddha, Chennai.

3.2. Ingrediants of Kalladaippu Chooranam:

Purified padigaram (alumen alum)

Purified venkaram (borax - sodium biborate)

Purified induppu (rock salt - sodium chloride)

Purified savukkaram (sodium carbonate) equal ratio

Ashes of Nayuruvi (achyranthes aspera)

Ashes of Panangathir (borassus flabellifermis)

Ashes of Vazhai mattai (musa paradisiaca)

3.3. Preparation of drug:

Equal amount of purified padigaram, venkaram, induppu and savukkaram were made into fine powder. Equal amount of ashes of nayuruvi, panangathir and vaazhai mattai made it also into fine powder. Then mixed both the powder contents and sieved by pure cloth (vasthirakayam), and stored it into an air tight container.

3.4. Administration:

Dosage: 1 gm/ BD/ p.c

Adjuvant: honey⁴

3.5. Methodology:

The physico-chemical and phytochemical analysis were carried out in The Tamilnadu Dr.M.G.R Medical University, Guindy, Chennai, Tamilnadu.

I. PHYSICOCHEMICAL ANALYSIS OF KALLADAIPPU CHOORANAM:

1. Loss on Drying:

An accurately weighed 2g of Kalladaippu Chooranam formulation was taken in a tarred glass bottle. The crude drug was heated at 105⁰ C for 6 hours in an oven till a constant weight. The percentage moisture content of the sample was calculated with reference to the shade dried material.

2. Determination of Total Ash:

Weighed accurately 2g of Kalladaippu Chooranam formulation was added in crucible at a temperature 600⁰ C in a muffle furnace till carbon free ash was obtained. It was calculated with reference to the air dried drug.

3. Determination of Acid Insoluble Ash:

Ash above obtained, was boiled for 5 min with 25ml of 1M Hydrochloric acid and filtered using an ash less filter paper. Insoluble matter retained on filter paper was washed with hot water and filter paper was burnt to a constant weight in a muffle furnace. The percentage of acid insoluble as was calculated with reference to the air dried drug.

4. Determination of Water Soluble Ash:

Total ash 1g was boiled for 5 mins with 25 ml water and insoluble matter collected on an ash less filter paper was washed with hot water and ignited for 15 min at a temperature not exceeding 450⁰ C in a muffle furnace. The amount of soluble ash is determined by drying the filtrate.

5. Determination of Water Soluble Extractive:

5 g of air dried drug, coarsely powdered Kalladaippu Chooranam was macerated with 100 ml of distilled water in a closed flask for twenty-four hours, shaking frequently. The solution was filtered and 25 ml of filtered was evaporated in a tarred flat bottom shallow dish, further dried at 100 ⁰ C and weighed. The percentage of water soluble extractive was calculated with reference to the air dried drugs.

6. Determination of Alcohol Soluble Extractive:

2.5 g of air dried drugs, coarsely powdered Kalladaippu Chooranam was macerated with 50 ml alcohol in closed flask for 24 hrs with frequent shaking, it was filtered rapidly taking precaution against loss of alcohol. 10 ml of filtrate was then evaporated in a tarred flat bottom shallow dish, dried at 100⁰ C and weighed. The percentage of alcohol soluble extractive was calculated with reference to air dried drug.

II. PRELIMINARY PHYTOCHEMICAL SCREENING:

1. Detection of Alkaloids:

Extracts were dissolved individually in dilute Hydrochloric acid and filtered.

a) Mayer’s Test : Filtrates were treated with Mayer’s reagent (Potassium Mercuric Iodide). Formation of a yellow colored precipitate indicates the presence of alkaloids.

b) Wagner’s Test: Filtrates were treated with Wagner’s reagent (Iodine in Potassium Iodide) formation of brown / reddish precipitate indicates the presence of alkaloids.

c) Dragendroff’s Test: Filtrates were treated with Dragendroff’s reagent (Solution Of Potassium Bismuth Iodide) formation of red precipitate indicates the presence of alkaloids.

d) Hager’s Test: Filtrates were treated with Hager’s reagent with Hager’s reagent (Saturated Picric Acid Solution) presence of alkaloids confirmed by the formation of yellow colored precipitate.

2. Detection of Carbohydrates:

Extracts were dissolved individually in 5 ml distilled water and filtered. The filtrates were used to test for the presence of carbohydrates.

a) Molishch’s Test:

To 2 ml of plant sample extract, two drops of alcoholic solution of α – naphthol are added. The mixture is shaken well and few drops of concentrated sulphuric acid is added slowly along the sides of test tube. A violet ring indicates the presence of carbohydrates.

b) Benedict’s Test:

Filtrates were treated with Benedict’s regent and heated gently. Orange red precipitate indicates the presence of reducing sugars.

3. Detection of Glycosides:

Extracts were hydrolyzed with diluted HCL and then subjected to test for glycosides.

a) Modified Borntrager’s Test: Extracts were treated with ferric chloride solution and immersed in boiling water for about 5 minutes. The mixture was cooled and extracted with equal volumes of benzene. The benzene layer was separated and treated with ammonia solution. Formation of rose-pink color in the ammonical layer indicates the presence of anthranol glycosides.

b) Cardiac Glycoside (Keller- Killiani Test):

Extract was shaken with distilled water (5ml). to this, glacial acetic acid (2ml) containing a few drops of ferric chloride was added, followed by H ₂SO₄ (1 mL) along the side of the test tube. The formation of brown ring at the interface gives positive indication for cardiac glycoside and a violet ring may appear below the brown ring.

4. Detection of Saponins:

a) Froth Test : Extracts were diluted with distilled water to 20 ml and this was shaken in a graduated cylinder for 15 minutes. Formation of 1 cm layer of foam indicates the presence of saponins.

b) Foam Test: 0.5 g of extract was shaken with 2 ml of water. If foam produced persists for ten minutes it indicates the presence of saponins.

5. Detection of Phytosterols:

a) Salkowski’s Test: extracts were treated with chloroform and filtered. The filtrates were treated with few drops of Conc. Sulphuric acid, shaken and allowed to stand. Appearance of golden yellow color indicates the presence of triterpenes.

6. Detection of Phenols Ferric Chloride Test:

Extracts were treated with 3-4 drops of ferric chloride solution. Formation of bluish black color indicates the presence of phenols.

7. Detection of Tannins Gelatin Test:

The extract is dissolved in 5 ml of distilled water and 2 ml of 1% solution of gelatin containing 10% NaCl is added to it. White precipitate indicates the presence of phenolic compounds.

8. Detection of Flavonoids:

a) Alkaline Reagent Test: Extracts were treated with few drops of sodium hydroxide solution. Formation of intense yellow color, which becomes colorless on addition of dilute acid, indicates the presence of flavonoid.

b) Lead Acetate Test: Extracts were treated with a few drops of lead acetate solution. Formation of a yellow color precipitate indicates the presence of flavonoid.

9. Detection of Proteins and Amino acids:

a) Xanthoproteic Test: The extracts were treated with a few drops of Conc. Nitric acid. Formation of yellow color indicates the presence of proteins.

b) Ninhydrin Test: The extract, 0. 25% w/v Ninhydrin Reagent was added and boiled for a few minutes. Formation of blue color indicates the presence of Amino Acid.

10. Detection of Diterpenes Copper Acetate Test:

Extracts were dissolved in water and treated with 3-4 drops of copper acetate solution. Formation of emerald green color indicates the presence of Diterpenes.

11. Gum and Mucilage:

To 1 ml of extract add 2.5 ml of absolute alcohol and stirring constantly. Then the precipitate as dried in air and examine for its swelling properties. Swelling was observed that will indicate presence of gum and mucilage.

12. Test for Fixed Oils and Fats:

a) Spot Test: a small quantity of extract is pressed between two filter papers. Oil stain on the paper indicates the presence of fixed oils.

13. Test for Quinones:

Extract was treated with sodium hydroxide blue or red precipitate indicates the presence of Quinones.

4. RESULTS AND DISCUSSION:

4.1. Physico - Chemical analysis:

The result of the physicochemical parameters given in Table 1.

Table 1. Physico-chemical parameters

Loss on drying indicates the low moisture content which reveals the stability and its shelf life. The high total ash value indicates the presence of inorganic compounds because the nature the drug was consisting inorganic compounds also. High water soluble ash and extractive denotes solubility of water⁵.

4.2. Preliminary Phytochemical analysis

The preliminary phytochemical studies of aqueous extract of Kalladaippu Chooranam were done using standard procedures. The results were presented in Table 2. The present study reveals that the bioactive compounds were present in all the extracts of Kalladaippu Chooranam.

Table 2: Preliminary Phytochemical analysis

+ve/-ve present or absent if component tested

Flavonoids can exert their anti-oxidant activity by scavenging the free radicals, by chelating metal ions or by inhibiting enzymatic systems responsible for free radical generation. Flavonoids are immunomodulator ⁵.

The antioxidant activity of phenolic compounds is attributed to the capacity of scavenging free radicals⁶. The phenols are anesthetic or pain reliever⁵.

Saponin has adaptogenic, adjuvant, analgesic, anticancer, antioxidant, anticholesterol, antispasmodic and diuretic activity⁷

Tannins can effective in protecting kidneys⁸

Gums and mucilage used in the field of novel drug delivery systems and pharmaceutical applications such as stabilizing agents, thickening agents, emulsifiers, binding agents etc⁹.

5. CONCLUSION:

The current study investigation is done to meet the standardization of WHO and other scientific committees. The present analysis revealed that physico-chemical analysis such as loss of drying, total ash, insoluble ash, water soluble ash, water soluble extractive and alcohol soluble extractive and phytochemical analysis such as detection of alkaloids, carbohydrates, glycosides, saponins, phytosterols, phenols, flavonoids, proteins & aminoacids, diterpenes, gum & mucilage and fixed oils & fats were carried out. Based on the above results, it can be assumed the drug Kalladaippu chooranam has validated the traditional claim. This research work is done for, further studies to be carried out in future with different analytical, qualitative chemical properties and also by in vivo methods.

FOOTNOTES:

Declaration of conflicting interests: The author declared no potential conflicts of interest with respect to the research and publication of this article.

Funding: The author received no financial support for the research and publication of this article

Acknowledgement: The author cordially thanks to The Tamilnadu Dr. M.G.R University for support doing Physico-chemical and Phytochemical analysis.

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