Institute for Clinical Chemistry, University Hospital Zurich, Zurich, Switzerland.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Feb 1;883-884:189-97. doi: 10.1016/j.jchromb.2011.08.022. Epub 2011 Aug 24.
The two main matrices for screening are urine or serum and heparinized plasma. Whereas urine has the advantage of usually higher concentrations and longer detection windows, serum or heparinized plasma represent the current systemic drug exposure of a patient.
An online extraction LC-MS(n) method using a MS(2) and MS(3) spectral library for the identification of substances has been developed and validated to screen serum and heparinized plasma. Extraction was performed by online turbulent flow chromatography under alkaline conditions. Chromatographic separation was achieved using a phenyl/hexyl column with acidic eluents. For detection, a linear ion trap, equipped with an APCI interface, was used and the different compounds were identified using a MS(2) and MS(3) spectral library containing 453 compounds. From 47 patients, urine and heparinized plasma samples were analyzed and the results compared.
The validation of the method gave satisfactory results. Only 3% of the compounds showed a matrix effect>10% in serum. For all other substances and heparinized plasma, the quantitative matrix effect was <10%. 78% of the compounds where a therapeutic range was described in the literature had a limit of identification below the therapeutic range in heparinized plasma and 77% in serum, respectively. In urine and heparinized plasma samples, a total of 168 substances (identified as 86 different compounds) could be identified. In 20 out of 47 cases (43%), the results were identical. On a substance level, the agreement between urine and heparinized plasma was in average 71% with a range of 0-100%.
The presented method allows a fast identification of 453 substances in serum and heparinized plasma. If plasma or serum is used for toxicological screening, the current systemic exposure of a patient can be monitored.
尿液或血清和肝素化血浆是筛选的两种主要基质。虽然尿液具有浓度通常更高和检测窗口更长的优点，但血清或肝素化血浆代表患者当前的全身药物暴露情况。
开发并验证了一种在线提取 LC-MS(n) 方法，该方法使用 MS(2) 和 MS(3) 谱库来鉴定物质，以筛选血清和肝素化血浆。提取是在碱性条件下通过在线湍流流色谱进行的。采用酸性洗脱液的苯基/己基柱进行色谱分离。用于检测，使用配备 APCI 接口的线性离子阱，并使用包含 453 种化合物的 MS(2) 和 MS(3) 谱库来鉴定不同的化合物。从 47 名患者中分析了尿液和肝素化血浆样本，并比较了结果。
该方法的验证结果令人满意。仅 3%的化合物在血清中表现出基质效应>10%。对于所有其他物质和肝素化血浆，定量基质效应<10%。在文献中描述了治疗范围的 78%的化合物在肝素化血浆中的鉴定限低于治疗范围，分别为 77%。在尿液和肝素化血浆样本中，共鉴定出 168 种物质（鉴定为 86 种不同的化合物）。在 47 例中的 20 例（43%）中，结果相同。在物质水平上，尿液和肝素化血浆之间的一致性平均为 71%，范围为 0-100%。
所提出的方法允许快速鉴定血清和肝素化血浆中的 453 种物质。如果使用血浆或血清进行毒理学筛选，可以监测患者当前的全身暴露情况。
