干血斑能否用于准确测量维生素D代谢物？

Can dried blood spots be used to accurately measure vitamin D metabolites?

作者信息

Binks Michael J, Bleakley Amy S, Rathnayake Geetha, Pizzutto Susan, Chang Anne B, McWhinney Brett, Ungerer Jacobus

机构信息

Child Health Division, Menzies School of Health Research, Charles Darwin University, Darwin, Northern Territory, Australia.

出版信息

Clin Chim Acta. 2021 Jul;518:70-77. doi: 10.1016/j.cca.2021.03.003. Epub 2021 Mar 10.

DOI:10.1016/j.cca.2021.03.003

PMID:33713691

Abstract

BACKGROUND

Where conventional blood sampling is challenging, dried blood spots (DBS) provide a practical sample alternative for measuring vitamin D levels. Our study aimed to develop and evaluate a clinical pathology service-based assay suitable for measuring vitamin D in batches of DBS samples collected remote to the testing site.

METHODS

A high throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with derivatisation was developed to measure 25-hydroxyvitamin D metabolites (25OHD3, 25OHD2 and 3-epi-25OHD3) in DBS samples. The assay was validated using paired DBS and plasma samples from 37 healthy adults.

RESULTS

The assay reproducibly (<11.5% coefficient of variation) quantified 25OHD3 (range 1-300 nmol/L), 25OHD2 (range 2-300 nmol/L) and 3-epi-25OHD3 (range 1-200 nmol/L) in DBS samples. The 25OHD3 metabolite was detected in all DBS samples, 3-epi-25OHD3 in six plasma (range 2.1-6.3 nmol/L) and paired DBS samples, and 25OHD2 was not detected. Concentrations of 25OHD3 were highly correlated between paired samples: capillary DBS and venous plasma (r = 0.92), venous DBS and venous plasma (r = 0.93), and capillary DBS and venous DBS (r = 0.97). Ordinary least squares regression was used to characterise (β = 0.81) and correct the systematic bias in DBS data (compared to paired plasma). Thereafter, Bland-Altman analysis demonstrated robust agreement between sample-methods.

CONCLUSION

This simple and rapid DBS-based LC-MS/MS assay accurately quantified serum vitamin D metabolites using a paired-sample 'bridging strategy' to correct for the inherent sample-method bias.

摘要

背景

在传统血液采样具有挑战性的情况下，干血斑（DBS）为测量维生素D水平提供了一种实用的样本替代方法。我们的研究旨在开发并评估一种基于临床病理服务的检测方法，该方法适用于测量在远离检测地点采集的一批批DBS样本中的维生素D。

方法

开发了一种带有衍生化的高通量液相色谱 - 串联质谱（LC-MS/MS）方法，用于测量DBS样本中的25 - 羟基维生素D代谢物（25OHD3、25OHD2和3 - 表 - 25OHD3）。使用来自37名健康成年人的配对DBS和血浆样本对该检测方法进行了验证。

结果

该检测方法可重复性地（变异系数<11.5%）对DBS样本中的25OHD3（范围1 - 300 nmol/L）、25OHD2（范围2 - 300 nmol/L）和3 - 表 - 25OHD3（范围1 - 200 nmol/L）进行定量。在所有DBS样本中均检测到25OHD3代谢物，在六个血浆样本（范围2.1 - 6.3 nmol/L）及其配对的DBS样本中检测到3 - 表 - 25OHD3，未检测到25OHD2。配对样本中25OHD3的浓度高度相关：毛细血管DBS与静脉血浆（r = 0.92）、静脉DBS与静脉血浆（r = 0.93）以及毛细血管DBS与静脉DBS（r = 0.97）。使用普通最小二乘法回归来表征（β = 0.81）并校正DBS数据中的系统偏差（与配对血浆相比）。此后，Bland - Altman分析表明样本 - 方法之间具有稳健的一致性。