Biosensor-based determination of riboflavin in milk samples
Biosensor-based determination of riboflavin in milk samples
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DOI：
10.1021/ac034876a
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发表时间：
2004-01-01
影响因子：
7.4
通讯作者：
Wahlström, L
中科院分区：
文献类型：
--
作者：
Caelen, I;Kalman, A;Wahlström, L
利用表面等离子体共振原理和芯片检测技术，建立了乳制品中核黄素(RF)的定量分析方法。定量是通过测量过量的RF结合蛋白(RBP)间接完成的，RBP在与样品溶液中最初存在的RF分子络合后保持自由。用共价固定化的RF修饰芯片，以过量结合RBP。通过化学修饰，在天然RF的N-3位引入活性酯基，结合芯片表面的氨基。通过将一系列射频标准溶液与最佳浓度的RBP混合来制备校准溶液。然后，通过将响应与校准进行比较来测量奶基产品中的RF含量。所得结果与官方的高效液相色谱-荧光法的结果非常接近。当进样量为160MU/L时，定量下限为234MU/L，检出限为70MU/L。
An assay for quantification of riboflavin (Rf) in milk-based products has been developed using the principle of surface plasmon resonance with on-chip measurement. The quantification was done indirectly by measuring excess of Rf binding protein (RBP) that remains free after complexation with Rf molecules originally present in the sample solution. The chip was modified with covalently immobilized Rf in order to bind the RBP in excess. A chemical modification was performed to introduce a reactive ester group at the N-3 position of the natural Rf to bind amino groups present on the chip surface. Calibration solutions were prepared by mixing a range of Rf standard solutions with an optimized concentration of RBP. The Rf content in the milk-based products was then measured by comparison of the response against the calibration. Results obtained were very close to those from an official HPLC-fluorescence procedure. The limit of quantification was determined to 234 mug/L and the limit of detection to 70 mug/L by an injection volume of 160 muL.