Department of Laboratory Medicine, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China.
Shanghai Trace Source Biotechnology Co., Ltd, Shanghai, China.
J Clin Lab Anal. 2021 Mar;35(3):e23658. doi: 10.1002/jcla.23658. Epub 2020 Nov 20.
To study the prevalence of the exposure of pregnant women to antimicrobials, a sensitive and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to determine nine antimicrobials, namely sulfadimidine, sulfapyridine, sulfadiazine, sulfathiazole, ofloxacin, ciprofloxacin, norfloxacin, tetracycline, and lincomycin, in human serum.
The sample preparation procedure included protein precipitation followed by a cleanup step with solid phase extraction (SPE). Separation was carried out using a CORTECS T3 column (100 × 2.1 mm, 2.7 µm) by gradient elution with a runtime of 8.0 min. Detection was performed on a triple quadruple tandem mass spectrometer with scheduled multiple reaction monitoring (sMRM) in positive ion scan mode.
The calibration curves were linear over the concentration range of 0.5-50 ng/ml, and the limit of quantitation was between 0.01 and 0.2 ng/ml. For each level of quality control samples, the inter- and intra-assay precision values were less than 12.0%, and the accuracy ranged from 86.1% to 109.0%. No significant matrix effect or carryover was observed. The antimicrobials of interest were stable under all investigated conditions. The validated method was applied to analyze clinical samples from pregnant women in China, and 10 out of 500 samples showed the presence of antimicrobial residues. Moreover, compared with the time-resolved fluoro-immunoassay (TRFIA) method, the developed method showed greater sensitivity and specificity.
This study provides a simple and rapid LC-MS/MS method for the simultaneous measurement of nine antimicrobials in serum samples, which could be a useful tool in clinical utilization.
为了研究孕妇接触抗生素的情况，开发并验证了一种灵敏、可靠的液相色谱-串联质谱（LC-MS/MS）方法，用于测定人血清中的九种抗生素，即磺胺嘧啶、磺胺吡啶、磺胺甲恶唑、磺胺噻唑、氧氟沙星、环丙沙星、诺氟沙星、四环素和林可霉素。
样品制备过程包括蛋白沉淀，然后用固相萃取（SPE）进行净化。采用 CORTECS T3 柱（100×2.1mm，2.7μm）进行梯度洗脱，运行时间为 8.0min。采用三重四极串联质谱仪，在正离子扫描模式下，采用定时多重反应监测（sMRM）进行检测。
校准曲线在 0.5-50ng/ml 浓度范围内呈线性，定量下限在 0.01-0.2ng/ml 之间。对于每个质控样品水平，批内和批间精密度值均小于 12.0%，准确度范围为 86.1%-109.0%。未观察到明显的基质效应或交叉污染。在所有考察条件下，目标抗生素均稳定。该方法已应用于分析中国孕妇的临床样本，在 500 个样本中有 10 个显示存在抗生素残留。此外，与时间分辨荧光免疫分析法（TRFIA）相比，该方法具有更高的灵敏度和特异性。
本研究提供了一种简单、快速的 LC-MS/MS 方法，用于同时测定血清样品中的九种抗生素，可作为临床应用的有用工具。
