Practical Pharmaceutical Techniques

1609086

WARDA SAHAL

5PY015

PRACTICAL PHARMACEUTICAL TECHNIQUES

BLOCK 1: PROTEIN ANALYSIS

Experiment 1: Protein assay using the “Bradford” method

Abstract:

Materials and Methods:

Refer to practical schedule.

Results and Calculations:

PART 1:

Table 1: Calculating the quantity/test (µg/test) using a standard protein solution of known concentration

The following equation was used to calculate the quantity of protein (µg/test):

[pic 1]

For example, the calculations for the quantity/test for tube number 3 are as follows:

Quantity/test = 50 µg/ml x 0.04 ml

Quantity/test = 2 µg/test

The results in table 1 were used to prepare diagram 1, ‘Standard Calibration Curve 1’ below:[pic 2]

The gradient of the line on the graph can be calculated using the following equation:[pic 3]

[pic 4]

m = 0.325 – 0.08 / 8 – 2

m = 0.041 Absorbance at 595nm per µg protein

PART 2:

Table 2: Absorbance values obtained from the diluted samples of the unknown protein

* This absorbance value is the most suitable as it is ½ way between the minimum and the maximum absorbance values from the ‘Standard Calibration Curve 1’.

PART 3:

Table 3: Calculating the quantity (in µg) of protein in each of cuvettes 1 – 8

* These values were calculated using the y = mx equation where y = A595, m is the gradient which was 0.0413 and x is the unknown. With cuvette 1, the calculations are as follows:

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