Total Guggulusterones (By UV)

Method for the estimation of Total Guggulusterones content 

Reagent Required:

Methanol LR Grade

Glass wares, Apparatus & Instruments required:

Volumetric Flasks          100mL, 50mL, 250mL

Beakers                        100mL, 50mL,

Pipettes –                     5mL, 2mL

U-V Spectrophotometer

STANDARD PREPARATION:

Weigh accurately about 25mg of working standard into a 50mL volumetric flask. Add about 25mL of methanol. Ultrasonicate till completely dissolved and make up the volume with methanol (Solution A). Further dilute 5mL solution A   to 50mL with methanol (Solution B). Further dilute 5mL solution B to 50mL with methanol.

SAMPLE PREPARATION:

Weigh accurately about 100mg of sample into a 50mL volumetric flask. Add about 25mL of methanol. Ultrasonicate till completely dissolved and make up the volume with methanol, warming if necessary. Make up the volume with methanol. Further dilute above 5mL solution to 50mL with methanol. Ensure that the active concentration in the sample is approximately similar to that of standard concentration.  Shake well and filter through what man No.1 filter paper.

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PROCEDURE:

Determine absorbance of both standard and sample solution at wavelength of 237.2nm.

Percentage of Total Guggulusterones content is calculated as:

                                          Absorbance of sample X Concentration of std X   purity of std

% of Guggulusterones =----------------------------------------------------------------------------------------------

                                             Absorbance of standard X Concentration of sample 

For Raw Material: -

Weigh accurately about 5.0gm Raw herb of Guggul in 250 mL R.B.Flask. Add about 75mL of methanol and reflux for one hour. Cool and filter the solution. Rinse the residue in the flask with small quantity of methanol & filter through same filter paper and make up the volume with methanol. Further dilute above 5mL solution to 50mL with methanol. Further dilute above 5mL solution to 100mL with methanol. Ensure that the active concentration in the sample is approximately similar to that of standard concentration.  Shake well and filter through what man No.1 filter paper.