GAIA Research Center, Bioanalytical Department, The Goulandris Natural History Museum, 13 Levidou Street, Kifissia 14562, Greece.
J Pharm Biomed Anal. 2011 Aug 25;56(1):78-85. doi: 10.1016/j.jpba.2011.04.019. Epub 2011 Apr 27.
A rapid, simple and accurate analytical method based on ultra performance liquid chromatography (UPLC) combined with electrospray ionization (ESI) tandem mass spectrometry (MS/MS) on a hybrid q TOF instrument has been developed and fully validated for the quantification of daptomycin (DPT) in human plasma. The samples were analyzed after simple pretreatment involving protein precipitation, while chromatographic separation of DPT and the internal standard (reserpine) was achieved on an Acquity BEH C18 column (100 mm × 2.1 mm, 1.7 μm) using gradient elution with 0.1% aqueous formic acid (FA) and acetonitrile with 0.1% FA (with DPT eluting at 2.60 min). The method presented good fit (r>0.999) over the quantification range of 0.01-10 μg mL⁻¹ with the lower limit of quantitation (LLOQ) being 0.01 μg mL⁻¹ of human plasma for DPT. The intra- and inter-day precision, measured as % relative standard deviation, was less than 11% for DPT. The validation results showed that the developed method demonstrated adequate selectivity, sensitivity, precision and accuracy and therefore was successfully applied to the analysis of clinical samples following intravenous (iv) administration of 5.4 mg kg⁻¹ DPT to patients suffering from post-traumatic osteomyelitis induced by methicillin-resistant Staphylococcus aureus (MRSA). The developed methodology is the first report of an accurate mass tandem MS method for the analysis of this potent antibiotic in human plasma and can be used to further study pharmacokinetic, bioequivalence and even metabolic aspects related to this drug.
建立了一种基于超高效液相色谱（UPLC）结合电喷雾串联质谱（ESI-MS/MS）在混合 qTOF 仪器上的快速、简单、准确的分析方法，并对其进行了充分验证，可用于定量测定人血浆中的达托霉素（DPT）。样品经过简单的预处理，包括蛋白质沉淀，然后在 Acquity BEH C18 柱（100mm×2.1mm，1.7μm）上进行色谱分离，采用 0.1%甲酸水溶液（FA）和含 0.1%FA 的乙腈进行梯度洗脱，DPT 在 2.60min 洗脱。该方法在 0.01-10μgmL⁻¹的定量范围内具有良好的线性（r>0.999），DPT 的定量下限（LLOQ）为 0.01μgmL⁻¹人血浆。DPT 的日内和日间精密度（以相对标准偏差表示）小于 11%。验证结果表明，该方法具有良好的选择性、灵敏度、精密度和准确度，因此成功应用于静脉注射（iv）5.4mgkg⁻¹ DPT 治疗耐甲氧西林金黄色葡萄球菌（MRSA）引起的创伤后骨髓炎患者后，分析人血浆中的临床样品。所建立的方法是第一个用于分析人血浆中这种强效抗生素的精确质量串联 MS 方法的报告，可用于进一步研究与该药物相关的药代动力学、生物等效性甚至代谢方面。
