Evaluation of surrogate matrices for serum 25-hydroxyvitamin D calibrators by LC-MS/MS

doi:10.3969/j.issn.1673-8640.2021.02.020

Abstract

Abstract:

Objective To explore the selection and evaluation of a suitable surrogate matrix for preparing 25-hydroxyvitamin D [25（OH）D] calibrators by liquid chromatography tandem-mass spectrometry（LC-MS/MS）. Methods 4% bovine serum albumin（BSA） buffer and 30% ethanol-water were used to prepare 25（OH）D calibrators. They were thoroughly assessed by comparing the peak areas of internal standards in different matrices,testing their matrix effects and accuracy validation. Results There was no significant difference in response between 4% BSA matrix and serum（P>0.05）,while there was statistical difference between 30% ethanol water and serum（P<0.05）. The validation of matrix effect showed that 4% BSA-based calibrator was acceptable,while 30% ethanol-water-based calibrator was not. The external quality assessment（EQA） samples quantified by 4% BSA-based calibrator were well consistent with the target values,but the EQA samples quantified by 30% ethanol-water-based calibrator did not meet accuracy requirements. The bias between the two samples was over 25%. Conclusion 4% BSA can be used as substitute calibrator matrix for the measurement of serum 25（OH）D by LC-MS/MS.

Key words: Liquid chromatography-tandem mass spectrometry, Serum 25-hydroxyvitamin D, Surrogate matrices for calibrators

CLC Number:

R446.1

LI Bingling, CHENG Yating, ZHAO Beibei, SHE Xuhui, DONG Heng, YU Mujun. Evaluation of surrogate matrices for serum 25-hydroxyvitamin D calibrators by LC-MS/MS[J]. Laboratory Medicine, 2021, 36(2): 213-218.

Figures/Tables 8

References 14

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基质类型	例数	d6-25（OH）D₂/ ×10⁵cps	d6-25（OH）D₃/ ×10⁵cps
4% BSA	7	6.750±0.359	15.000±0.317
30%乙醇水	7	3.700±0.265	8.004±0.609
血清	6	6.580±0.667	15.000±1.080

基质类型	例数	d6-25（OH）D₂/ ×10⁵cps	d6-25（OH）D₃/ ×10⁵cps
4% BSA	7	6.750±0.359	15.000±0.317
30%乙醇水	7	3.700±0.265	8.004±0.609
血清	6	6.580±0.667	15.000±1.080

血清（A）样本编号	A:B/C 混合比例	AB混合样本响应比值				AC混合样本响应比值
血清（A）样本编号	A:B/C 混合比例	可接受范围	平行1	平行2	平行3	可接受范围	平行1	平行2	平行3
A1	3:1	0.179~0.269	0.221	0.217	0.227	0.179~0.268	0.234	0.226	0.216
A2	3:2	0.314~0.471	0.364	0.394	0.368	0.313~0.469	0.409	0.394	0.404
A3	1:1	0.378~0.567	0.469	0.462	0.466	0.377~0.565	0.469	0.476	0.486
A4	2:3	0.396~0.594	0.510	0.482	0.484	0.395~0.592	0.485	0.506	0.514
A5	1:2	0.526~0.789	0.665	0.647	0.670	0.524~0.786	0.771	0.778	0.792^①
A6	2:5	0.533~0.799	0.643	0.669	0.657	0.531~0.796	0.683	0.691	0.665

血清（A）样本编号	A:B/C 混合比例	AB混合样本响应比值				AC混合样本响应比值
血清（A）样本编号	A:B/C 混合比例	可接受范围	平行1	平行2	平行3	可接受范围	平行1	平行2	平行3
A1	3:1	0.179~0.269	0.221	0.217	0.227	0.179~0.268	0.234	0.226	0.216
A2	3:2	0.314~0.471	0.364	0.394	0.368	0.313~0.469	0.409	0.394	0.404
A3	1:1	0.378~0.567	0.469	0.462	0.466	0.377~0.565	0.469	0.476	0.486
A4	2:3	0.396~0.594	0.510	0.482	0.484	0.395~0.592	0.485	0.506	0.514
A5	1:2	0.526~0.789	0.665	0.647	0.670	0.524~0.786	0.771	0.778	0.792^①
A6	2:5	0.533~0.799	0.643	0.669	0.657	0.531~0.796	0.683	0.691	0.665

血清（A）样本编号	A:B/C混合比例	AB混合样本响应比值				AC混合样本响应比值
血清（A）样本编号	A:B/C混合比例	可接受范围	平行1	平行2	平行3	可接受范围	平行1	平行2	平行3
A1	3:1	0.181~0.271	0.239	0.233	0.228	0.194~0.291	0.252	0.242	0.253
A2	3:2	0.292~0.439	0.363	0.362	0.362	0.314~0.471	0.405	0.390	0.408
A3	1:1	0.358~0.536	0.461	0.440	0.431	0.384~0.576	0.506	0.486	0.459
A4	2:3	0.452~0.677	0.614	0.566	0.558	0.484~0.725	0.624	0.610	0.634
A5	1:2	0.495~0.743	0.614	0.608	0.615	0.531~0.796	0.772	0.727	0.739
A6	2:5	0.568~0.852	0.691	0.747	0.758	0.606~0.909	0.783	0.774	0.750