The Selection and Evaluation of Matrix for the Determination of Fat-soluble Vitamin Calibrators by Liquid Chromatography Tandem Mass Spectrometry
Objective To select and evaluate a simulated substrate for the determination of standard curves of fat-soluble vitamins by liquid chromatography-tandem mass spectrometry.Methods The standard curve of fat-soluble vitamin was prepared by using pure methanol and 4％bovine serum albumin(BSA)as simulated matrix.The simulated matrix was evaluated by linear experiment,accuracy evaluation experiment and matrix effect mixed experiment.Two standard curves were drawn to quantify serum samples from 44 clinical cases and the results were compared according to CLSI EP9-A3 file.Results Using pure methanol as the substrate,the correlation coefficients of the corresponding standard curves of vitamin A(VA),25-hydroxyvitamin D3[25(OH)D3],25-hydroxyvitamin D2[25(OH)D2]and vitamin E(α-VE)were 0.999,0.997,and 0.999,respectively,while the recoveries were 90.0％-110.1％.The relative matrix effect ranges from-7.40％to 7.90％.Using 4％BSA as matrix,the correlation coefficients of VA,25(OH)D3,25(OH)D2 and α-VE corresponding standard curves were 0.999,0.997,0.997,and 0.999,respectively.The recoveries were 91.1％-111.2％and the relative matrix effects were-8.84％-7.60％.Two standard curves were applied to linearly fit the quantitative results of 44 clinical samples.The developed regression equations of VA,25(OH)D and α-VE were as follows:Y=0.9083X-3.7026,Y=1.0406X-3.0114,and Y=1.018X-72.786,respectively.The estimated deviations at medically determined levels were within acceptable limits.Conclusion Both pure methanol and 4％BSA can be used as simulated substrates for fat-soluble vitamin calibrators.The applicability of simulated substrates should be evaluated in laboratories to ensure the accuracy of clinical test results.
Fat-soluble vitaminSimulated matrixLiquid chromatography tandem mass spectrometry
万方数据
维普
