Laboratory Medicine, University Hospital Basel, University of Basel, Petersgraben 4, 4031, Basel, Switzerland.
Anal Bioanal Chem. 2020 Jan;412(3):715-725. doi: 10.1007/s00216-019-02285-0. Epub 2020 Jan 3.
An analytical method using 2D high-performance liquid chromatography followed by tandem mass spectrometry for the quantification of the beta-lactam antibiotics amoxicillin, flucloxacillin, piperacillin, benzylpenicillin, the beta-lactamase inhibitors clavulanic acid, and tazobactam, as well as the macrolide antibiotic clindamycin, is presented. All analytes were measured in human plasma, while amoxicillin, clavulanic acid, flucloxacillin, and clindamycin were also analyzed in human tissue samples. Because of its high-protein binding, additionally, the free fraction of flucloxacillin was measured after ultrafiltration. As internal standards, deuterated forms of the beta-lactams were used. Sample preparation for all matrices was protein precipitation followed by online extraction on a TurboFlow MAX column, while sample separation was performed on an Accucore XL C18 column. Calibration curves were linear over 0.2-25 mg/kg for the tissue samples and 0.05-20 mg/l for the free fraction of flucloxacillin. In plasma, the calibration curves for amoxicillin and piperacillin were linear over 3.125-125 mg/l, for clavulanic acid and tazobactam over 1-40 mg/l, for benzylpenicillin 0.25-40 mg/l, and for flucloxacillin and clindamycin over 1.5-60 mg/l and 0.05-8 mg/l respectively. In plasma and plasma ultrafiltrate, inaccuracy and imprecision for any analyte were always less than 15%. In tissue, the accuracy and precision varied up to 16%, respectively, 20%, when various tissues were analyzed using a calibration in water. Graphical abstract.
一种使用二维高效液相色谱法和串联质谱法对β-内酰胺类抗生素阿莫西林、氟氯西林、哌拉西林、苄青霉素、β-内酰胺酶抑制剂克拉维酸和他唑巴坦以及大环内酯类抗生素克林霉素进行定量分析的方法。所有分析物均在人血浆中进行测量，而阿莫西林、克拉维酸、氟氯西林和克林霉素也在人组织样本中进行分析。由于其具有高蛋白结合性，因此还测量了超滤后氟氯西林的游离分数。作为内标，使用了β-内酰胺的氘代形式。所有基质的样品制备均为蛋白沉淀，然后在线提取到 TurboFlow MAX 柱上，而样品分离则在 Accucore XL C18 柱上进行。组织样本的校准曲线在 0.2-25mg/kg 范围内线性，游离氟氯西林的校准曲线在 0.05-20mg/L 范围内线性。在血浆中，阿莫西林和哌拉西林的校准曲线在 3.125-125mg/L 范围内线性，克拉维酸和他唑巴坦在 1-40mg/L 范围内线性，苄青霉素在 0.25-40mg/L 范围内线性，氟氯西林和克林霉素在 1.5-60mg/L 和 0.05-8mg/L 范围内线性。在血浆和血浆超滤物中，任何分析物的准确度和精密度始终小于 15%。在组织中，当使用水中的校准来分析各种组织时，准确度和精密度分别变化至 16%和 20%。
