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Dushyant Patel Extractive Spectrophotometric Method

S.K.P.C.P.E.R., Kherva 72 M .Pharm. Thesis

separating funnels. Buffer solution (2 ml) and BCG solution (in excess) were added to

each. Shaken well and extracted with 10 ml of chloroform. Later the extracts were taken

into 10 ml volumetric flasks, treated with anhydrous sodium sulphate and made up the

volume with chloroform. The absorbances were measured periodically at an interval of

30, 60, 90, 120, 180, 240, 300 and 360 minutes at 416 nm. Same procedure was applied

for the BTB method using buffer solution (2.5 ml) and BTB solution (in excess), and

then absorbances were measured at 421 nm. Finally it was found that BCG-GBP

complex was stable at least for 6 hrs, whereas BTB-GBP complex was stable at least for

4 hrs.

5.2.5 METHOD VALIDATION:

a) Linearity:

Method I (BCG Method)

¾ From the working standard solution, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 ml were

transferred to a series of separating funnels and buffer solution (2 ml) was added to each

separating funnels, then BCG solution was added in excess and shaken well, and then 10

ml of chloroform was added to each and shaken well and kept for few minutes. Later the

extracts were taken into 10 ml volumetric flasks, treated with anhydrous sodium sulphate

and made up the volume with chloroform, and then absorbance of the solution was

measured at 416 nm (Figure 5.3) against reagent blank. Final concentrations of analyzed

solutions were 10 µg/ml to 120 µg/ml. The standard calibration plot was prepared to

calculate the amount of the analyte drug in unknown samples.

Figure 5.3 Representative absorption spectra of GBP-BCG showing λmax at 416 nm

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