Separation and determination of β-casomorphins by using glass microfluidic chip electrophoresis together with laser-induced fluorescence detection

✍ Scribed by ZongWen Wang; WenJian Wang; Wei Wang; LiangJun Xu; GuoNan Chen; FengFu Fu

• Publisher: John Wiley and Sons
• Year: 2010
• Tongue: English
• Weight: 328 KB
• Volume: 34
• Category: Article
• ISSN: 1615-9306
• DOI: 10.1002/jssc.201000634

✦ Synopsis

Abstract

A simple, reliable and reproducible method for the separation and determination of five β‐casomorphins (β‐CMs, namely TPGN, PGPI, TPGI, TPGP and TPPG) based on glass microfluidic chip electrophoresis and laser‐induced fluorescence detection is first described in here. The microfluidic chip electrophoresis and laser‐induced fluorescence detection system consisted of a home‐made glass “double‐T” microchip and a simple LIF detector with excitation and emission wavelengths of 473 and 525 nm, respectively. Fluorescein isothiocyanate (FITC) was used as the precolumn derivatization reagent to label fluorophore on five β‐CMs, and the optimum conditions of FITC‐derivatization reaction and MCE separation were investigated in detail. Under optimum conditions, five β‐CMs were completely separated and detected within 30 min with a detection limit of 18.7–75.1 nmol/L and an RSD (n=5) of 3.0–5.9%, respectively. The proposed method has been successfully used to detect β‐CMs in real cheese sample with a recovery of 89–109%, suggesting that our method is sensitive and reliable. These features, as well as its low cost, operation convenience, stability and reusability, make it a promising alternative to β‐CMs detection methods.