On-Line Flow-Injection Chemiluminescence Determination of Bovine Serum Albumin Using Surfactant-Enhanced Dichlorofluorescein-Hypochlorite System

Abstract

A flow injection technique combined with a chemiluminescence method was established for the determination of bovine serum albumin (BSA). Strong chemiluminescence was observed when BSA-dichlorofluoresce (DCF) complex was oxidized by sodium hypochlorite (NaClO) in an alkaline medium and in the presence of cetyltrimethylammonium bromide (CTAB). The reaction conditions of the chemiluminescence were carefully optimized. Under the optimal conditions, the method had a linear range of 0.01–20.0 μg/mL, with a detection limit of 0.007 μg/mL for BSA (3σ). The relative standard deviation of 1.0 μg/mL BSA (n = 8) is 1.4%. The method was applied to determine BSA in milk samples and it worked well.

Keywords:

• Bovine serum albumin
• Cetyltrimethylammonium bromide
• Chemiluminescence detection
• Dichlorofluorescein
• Flow injection

Abbreviations:

• BSA: bovine serum albumin
• CL: chemiluminescence
• CTAB: cetyltrimethylammonium bromide
• DCF: dichlorofluorescein
• FIA: flow-injection analysis
• HCl: hydrochloric acid
• NaClO: sodium hypochlorite

This work was supported by the Open Fund of Ministry-of-Education Key Laboratory for the Synthesis and Application of Organic Functional Molecules Foundation, Hubei University (No. 2006-KL-012), and the Natural Science Foundation of Hubei Province (No. 2007ABA377).

Notes

*C_DCF = 8.0 × 10⁻⁶ mol/L, BSA = 0.8 µg/ml, C_CTAB = 6.0 × 10⁻⁴ mol/L, C_NaClO = 0.12 mol/L, pH = 9.0.

*C_DCF = 8.0 × 10⁻⁶ mol/L, C_CTAB = 6.0 × 10⁻⁴ mol/L, C_NaClO = 0.12 mol/L, pH = 9.0.