| Abstrakt: |
Aiming at the complicated operation, time-consuming, incomplete hydrolysis and unstable hydrolysis products of enzymatic hydrolysis, we have established a microwave hydrolysis-liquid chromatography-atomic fluorescence spectrometry method for the determination of selenoamino acids in selenoproteins. After microwave hydrolysis with HC1(6 mol/L),the sample was adjusted to pH value of 7.5, then the selenocystine (SeCys2), Se-methylselenocysteine (MeSeCys) and selenomethionine (SeMet) were separated by anion chromatography with pH = 6.0,40 mmol/L (NH4)2HPO4 solution as mobile phase, flow rate at 0.7 mL/min and isocratic elution for 12 min, and the corresponding concentration were detected with an atomic florescence spectrometer. The microwave hydrolysis conditions are optimized by star point design-response surface method. The optimal hydrolysis conditions are as follows: weighing 20 mg, hydrolysis temperature 150 °C, hydrolysis time 40 min. The SeCys2, MeSeCys and SeMet showed a linear relationship in the concentration range of 0-100 ng, the correlation coefficients were all greater than 0.999, the detection limit was 0.07, 0.03 and 0.14 mg/kg, respectively, and the RSD was 5.6%, 4.6%, 3.7% (n= 6), respectively. It has the advantages of short time-consuming, low cost, complete hydrolysis, simple operation and stable hydrolysate. It can scientifically evaluate the composition of selenoamino acids in selenoproteins, and provides a scientific method for the quality control and traceability of selenoprotein products. The method is worthy of promotion and application. [ABSTRACT FROM AUTHOR] |
