Materials and Methods

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Materials and Methods 4. MATERIALS AND METHODS
4.1 PLANT MATERIAL The plant material was bought from Botanical Source of India, Jodhpur and powdered in grinder, to get coarse powder for extraction. 4.2 EXTRACTION AND FRACTIONATION The plant material was extracted by soxhlet with 50% alcohol and concentrated. A fresh batch of the plant material was successively extracted with petroleum ether, ethyl acetate, n-butanol and water (in the increasing order of polarity) and was concentrated in same order in a rotary evaporator at reduced pressure. Extraction methodology: 50% extract –PDE (A1) screened for the activity (using base model) Inflammation inflammation model Diuretic – urine volume and electrolyte balance Drug powder Petroleum ether – Acute, sub-acute and Chronic

Petroleum ether Fraction – PDP (A)

Defatted powder (dry) Ethyl acetate

Ethyl soluble Fraction - PDEA (B)

Powered drug n- butanol

n-butanol Fraction - PDB (C)

Powered drug Water Water Fraction - PDW (D)

A, B, C, and D were screened for base models. Fraction with highest activity was screened for mechanism of action.

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Phytopharmacological action of Pergularia daemia with special reference to its actions and mechanism of action as diuretic and anti-inflammatory agent

Materials and Methods

4.3 STANDARDIZATION OF PLANT MATERIAL The quality control of herbal crude drug and bioconstituents is of paramount important for their acceptability in modern system of medicine. One of the major problems faced by user in industry is non availability of rigid quality control profile for herbal raw material and their formulation with advanced of analytical technique and sophisticated instrument technology; it is possible to suggest a practicable quality assurance profile for a crude drug or its bioactive constituents. 4.3.1 ASH VALUE 4.3.1.1 Determination of Total Ash: 2 to 3 gm accurately weighed leaves powder was placed in silica crucible, previously ignited and weighed. Incinerated by gradually increasing the heat-not exceeding 4500C until free from carbon, cool and crucible was weighed. Calculated the percentage of ash with reference to the air-dried drug. Ash% = Loss in Weight x 100 W 4.3.1.2 Determination of Acid-insoluble Ash: Boiled the ash for 5 to 10 minutes with 25 ml of dilute hydrochloric acid. Collected the insoluble matter in a dry crucible or an ashless filter paper, washed with hot water, ignited and weighed. Calculated the percentage of acidinsoluble ash with reference to the air-dried drug. Ash% = Loss in Weight x 100 W 4.3.1.3 Determination of Water Soluble Ash: Boiled the ash for five minutes with 25 ml of water, collected the insoluble matter in a dry crucible, washed with hot water and ignited to constant weight at a low temperature. Subtracted the weight of insoluble matter from the weight of ash; difference in the weight represents the water-soluble ash. Calculated the percentage or water-soluble ash with reference to airdried drug. Ash% = Loss in Weight x 100 W

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Phytopharmacological action of Pergularia daemia with special reference to its actions and mechanism of action as diuretic and anti-inflammatory agent

Materials and Methods

4.3.2

EXTRACTIVE VALUES: 4.3.2.1 Determination of Alcohol-soluble Extractive: Coarsely powdered 5 gm of the air-dried drug macerated with 100 ml of alcohol in a conical flask for 24 hours, shaked frequently for six hours and allowed to stand for 18 hours. Filtered rapidly taking precaution against loss of alcohol evaporated 25% of the filtrate to dryness in a tarred bottomed shallow dish dried at 1050C and weighed. Calculated the percentage of alcohol soluble extractive with reference to the air-dried drug. 4.3.2.2 Determination of Water-Soluble Extractive: Proceeded as directed for the determination of alcohol soluble extractive, used chloroform water, I.P., instead of alcohol. 4.3.2.3 Loss on Drying: About 1.5 g of powder / drug was accurately weighed in a petridish and...
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