Optimization and validation of analytical conditions for bovine serum albumin using capillary electrophoresis.
A quick and reproducible capillary electrophoresis method was optimized and validated for the assay of bovine serum albumin (BSA). The effects of various parameters such as pH of buffer, concentration of buffer, capillary dimensions, use of coated capillaries, and additives such as surfactants and protein solubilizers were evaluated. The capillary coatings or additives did not give any advantage in reducing the surface adsorption of BSA on the capillary walls. The optimized conditions include use of borate buffer, pH 8.5 having a concentration of 150 mM in a 27 cm capillary with an aperture window of 100 x 200 microns for detection. The optimized method for the detection of BSA was validated. The interday and intraday coefficient of variation was not greater than 7.59% at BSA concentrations of 25-1000 micrograms/ml. The method developed was reproducible and accurate.
Pande
P G
PG
Department of Pharmaceutics, School of Pharmacy, University of Maryland, Baltimore 21201.
Nellore
R V
RV
Bhagat
H R
HR
D000818:Animals; D002021:Buffers; D002417:Cattle; D004586:Electrophoresis / Q000379:methods* / Q000706:statistics & numerical data; D005069:Evaluation Studies as Topic; D006863:Hydrogen-Ion Concentration; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000032:analysis*
D002021:Buffers; D012710:Serum Albumin, Bovine