采用 LC-HRMS 测定系统性红斑狼疮患者血液中的羟氯喹-全血、血浆和血清作为样本基质的评估。
Measurement of hydroxychloroquine in blood from SLE patients using LC-HRMS-evaluation of whole blood, plasma, and serum as sample matrices.
机构信息
Department of Medical Sciences, Clinical Chemistry, Uppsala University, Uppsala University Hospital, Entrance 61, 3rd floor, Dag Hammarskjölds Väg 18, SE-751 85, Uppsala, Sweden.
Department of Medical Sciences, Rheumatology, Uppsala University, Uppsala, Sweden.
出版信息
Arthritis Res Ther. 2020 Jun 1;22(1):125. doi: 10.1186/s13075-020-02211-1.
BACKGROUND
Hydroxychloroquine (HCQ) is the standard of care in the treatment of systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and other inflammatory rheumatic diseases and potentially for the treatment in COVID-19 patients. Determination of HCQ for therapeutic drug monitoring (TDM) can be performed in whole blood (WB), serum, and plasma. Direct comparisons of WB, serum, and plasma levels of HCQ in patients with SLE have not previously been reported. We describe a method for the determination of HCQ in human blood using liquid chromatography-high-resolution mass spectrometry (LC-HRMS) and compare the suitability of the three sample matrices.
METHODS
A method for the determination of HCQ in human blood using LC-HRMS was developed, validated, and applied for the determination of HCQ levels in WB, serum, and plasma from 26 SLE patients. The reproducibility of the method, in the three matrices, was evaluated using quality control samples and repeated preparations and measurements of patient samples. The performance of the developed method for HCQ measurement in serum was further evaluated by comparison with two previously reported extraction methods.
RESULTS
The performance of the presented method demonstrated high accuracy and precision. A large range of HCQ concentrations was observed for the SLE patients in all three matrices (WB, serum, and plasma). The mean levels in WB were approximately two-fold the levels in serum and plasma (813 ng/mL compared to 436 ng/mL and 362 ng/mL, respectively). Spiked quality controls showed high reproducibility for all matrices (coefficient of variation, CV, approx. 5%), whereas in patient samples, equally high-precision was only found using WB as the matrix (CV 3%). The CV for serum and plasma was 14% and 39%, respectively. Two alternative methods applied to serum samples did not demonstrate improved precision.
CONCLUSIONS
A LC-HRMS method for the measurement of HCQ in human blood was developed and validated. Whole blood was found to be the superior sample matrix in terms of sample reproducibility. Thus, whole blood samples should be used for HCQ analysis when patients are monitored for HCQ treatment effects. The assay is in clinical use to monitor levels of HCQ in patients.
背景
羟氯喹（HCQ）是治疗系统性红斑狼疮（SLE）、类风湿关节炎（RA）和其他炎症性风湿病的标准治疗药物，并且可能对 COVID-19 患者的治疗有效。治疗药物监测（TDM）中可以测定全血（WB）、血清和血浆中的 HCQ。以前没有报道过 SLE 患者 WB、血清和血浆中 HCQ 水平的直接比较。我们描述了一种使用液相色谱-高分辨率质谱（LC-HRMS）测定人血中 HCQ 的方法，并比较了三种样本基质的适用性。
方法
建立、验证了一种使用 LC-HRMS 测定人血中 HCQ 的方法，并应用于 26 例 SLE 患者 WB、血清和血浆中 HCQ 水平的测定。使用质控样品和患者样品的重复制备和测量，评估了该方法在三种基质中的重现性。通过与两种先前报道的提取方法的比较，进一步评估了该方法在血清中测定 HCQ 的性能。
结果
所提出的方法表现出很高的准确度和精密度。三种基质（WB、血清和血浆）中 SLE 患者的 HCQ 浓度范围很大（WB 中的平均水平约为血清和血浆中的两倍，分别为 813ng/ml、436ng/ml 和 362ng/ml）。在所有基质中，添加的质控样品都表现出很高的重现性（变异系数，CV，约为 5%），而在患者样品中，只有使用 WB 作为基质时才能达到同样的高精度（CV 为 3%）。血清和血浆的 CV 分别为 14%和 39%。应用于血清样本的两种替代方法并未显示出更好的精密度。
结论
建立并验证了一种用于测定人血中 HCQ 的 LC-HRMS 方法。全血在样本重现性方面是更好的样本基质。因此，在监测 HCQ 治疗效果时，应使用全血样本进行 HCQ 分析。该检测方法目前正在临床中用于监测患者 HCQ 水平。
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