Selection of an analytical method for evaluating bovine serum albumin concentrations in pharmaceutical polymeric formulations

New trends in reversed-phase liquid chromatographic separations of therapeutic peptides and proteins: Theory and applications

“…All of the most common casein and protein genetic variants were separated in less than 40 min [53]. Umrethia et al compared different techniques for the evaluation of bovine serum albumin concentrations in pharmaceutical polymeric formulations and found that RPLC was the most suitable method [54]. A conventional HPLC method that preserved the heterodimer of human follicle-stimulating hormone (hFSH) preparations (pituitary, urinary, and two commercial recombinant hFSH preparations) was established for the qualitative and quantitative analysis of hFSH.…”

A polycarboxylic/amino functionalized hyaluronic acid derivative for the production of pH sensible hydrogels in the prevention of bacterial adhesion on biomedical surfaces

“…In particular 1 ml of BSA solution in DPBS pH 7.4 (0.3 mg/ml) was added into the wells of a 24-well plate previously covered with 100 ml of hydrogel. At fixed time points (0, 1, 4, 12 and 24 h), the entire volume was withdrawn and the concentration of BSA was read using an HPLC method (Umrethia et al, 2010). After 24 h, the amount of BSA adsorbed on the hydrogel was quantified.…”

Development of an Analytical Method for the Rapid Quantitation of Peptides Used in Microbicide Formulations

“…For example, the commercially available BCA/Bradford assay based on spectrophotometry lacks specificity and sensitivity. These assays cannot distinguish between different proteins/peptides, and the lower limit of quantification (LLOQ) for the BCA assay that can be achieved without interference is approximately 1.23 µg mL −1 [11]. Furthermore, the accuracy of these assays are significantly reduced when approaching the LLOQ and the results can be inconsistent due to interference by impurities or the presence of other substances, e.g., excess salts, surfactants, polymers, etc.…”

“…Furthermore, the accuracy of these assays are significantly reduced when approaching the LLOQ and the results can be inconsistent due to interference by impurities or the presence of other substances, e.g., excess salts, surfactants, polymers, etc. [11]. Western blot has good specificity in identifying the target protein/peptide.…”

“…ELISA is a reliable method with both specificity and accuracy, but the sample analysis is generally time-consuming requiring optimization of appropriate antibody/antigen incubation and can be expensive due to the need of antibodies with high specificity, thereby affecting its efficiency for analyzing large quantities of pharmaceutical samples. In contrast, reversed-phase high-performance liquid chromatography (RP-HPLC) has good precision (<2 %), acceptable selectivity and sensitivity (as low as 0.33 μg mL −1 ), relatively inexpensive (e.g., does not require the use of antibodies) and is less time-consuming than traditional protein analysis methods (e.g., gel electrophoresis) [11, 12]. …”