Zhao Boxin, Liu Sijia, Liu Yuan, Li Guofeng, Zhang Qing
Department of Pharmacy/Rational Medication Evaluation and Drug Delivery Technology Laboratory, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2020 Mar 30;40(3):342-345. doi: 10.12122/j.issn.1673-4254.2020.03.08.
To investigate the effect of heat inactivation (56℃for 30 min) of SARS-CoV-2 on the results of therapeuticdrug monitoring (TDM) of voriconazole by liquid chromatography tandem mass spectrometry (LC-MS/MS).
We collected clinical blood samples from voriconazole-treated patients in heparinized tubes and sterilized the surface of the tubes with 75% ethanol. The whole blood samples were centrifuged to separate the plasma with or without prior heat inactivation, or only the separated plasma was heat inactivated. Heat inactivation of the samples was carried out at 56 ℃ for 30 min followed by protein precipitation with acetonitrile or ethanol. The plasma standard and quality control samples were inactivated in an identical manner and tested with LC-MS/MS along with the treated samples.
The optimized method showed a high imprecision (with mean intra- and inter-day imprecisions of 3.59% and 2.81%, respectively) and a high accuracy (mean 97.37%) for detecting voriconazole in the inactivated samples at different concentration levels. Sample preparation with acetonitrile or ethanol resulted in a high mean recovery (100.56% or 95.90%) with minimal mean matrix effect (102.85% or 93.62%). The measured voriconazole concentrations in inactivated whole blood, inactivated plasma and the samples without inactivation all showed good linear correlations with correlation coefficients all greater than 0.99.
Heat inactivation at 56 ℃ for 30 min combined with ethanol sample preparation only has limited effects to affect LC-MS-based voriconazole concentration measurement in whole blood samples collected in heparinized tubes, and can be used for therapeutic drug monitoring of voriconazole during the ongoing COVID-19 pandemic.
研究严重急性呼吸综合征冠状病毒2（SARS-CoV-2）热灭活（56℃，30分钟）对伏立康唑液相色谱串联质谱法（LC-MS/MS）治疗药物监测（TDM）结果的影响。
我们从接受伏立康唑治疗的患者中收集肝素抗凝管中的临床血样，并用75%乙醇对试管表面进行消毒。全血样本离心分离血浆，部分样本先进行热灭活，部分不进行热灭活，或仅对分离出的血浆进行热灭活。样本在56℃下热灭活30分钟，然后用乙腈或乙醇进行蛋白沉淀。血浆标准品和质量控制样品以相同方式灭活，并与处理后的样品一起用LC-MS/MS进行检测。
优化后的方法在检测不同浓度水平的灭活样品中的伏立康唑时，显示出高精密度（日内和日间平均精密度分别为3.59%和2.81%）和高精度（平均97.37%）。用乙腈或乙醇进行样品制备，平均回收率高（分别为100.56%或95.90%），平均基质效应最小（分别为102.85%或93.62%）。灭活全血、灭活血浆和未灭活样品中测得的伏立康唑浓度均显示出良好的线性相关性，相关系数均大于0.99。
56℃热灭活30分钟并仅结合乙醇样品制备对基于LC-MS的肝素抗凝管中采集的全血样品中伏立康唑浓度测量的影响有限，可用于正在进行 COVID-19 大流行期间伏立康唑的治疗药物监测。
