An optimal LC-MS/MS method for determination of azithromycin in white blood cells: application to pediatric samples
An optimal LC-MS/MS method for determination of azithromycin in white blood cells: application to pediatric samples
复制标题
DOI：
10.4155/bio.14.81
复制
发表时间：
2014-01-01
期刊：
影响因子：
1.8
通讯作者：
Pruvost, Alain
中科院分区：
文献类型：
--
作者：
Legrand, Tiphaine;Elie, Valery;Pruvost, Alain
背景：研究表明阿奇霉素 (AZM) 的特殊抗菌和抗炎特性可能与其在白细胞 (WBC) 中的广泛积累有关。然而，测定白细胞中 AZM 的可用方法需要大量血容量，不适合儿科环境。因此，开发了一种 LC-MS/MS 方法来测定 WBC 中的 AZM。结果：通过裂解红细胞从500μl全血中分离出WBC。然后，用含有 AZM-d(3)-C-13 作为内标的甲醇/缓冲液进行 WBC 裂解。反相LC后，在多反应监测条件下以正电喷雾模式进行检测。每个 WBC 样品的线性范围为 0.5 至 200 ng。运行内准确度和整体准确度和精确度分别为 95.3% 至 101.1% 和 1.6% 至 4.7%。所有验证参数均满足国际要求。结论：该方法可以轻松地在小样本上进行，并提供可靠的数据，包括儿童和新生儿。
Background: Studies suggest that particular antimicrobial and anti-inflammatory properties of azithromycin (AZM) can be related to its extensive accumulation in white blood cells (WBCs). However, available methods for determination of AZM in WBCs require large blood volumes unsuited to a pediatric context. Therefore, an LC-MS/MS method was developed for determination of AZM in WBCs. Results: WBCs were isolated from 500 mu l of whole blood by lysing red blood cells. Then, lysis of WBCs was performed with methanol/buffer containing AZM-d(3)-C-13 as internal standard. After reversed phase LC, detection was performed under multiple reaction monitoring conditions in positive electrospray mode. Linearity ranged from 0.5 to 200 ng per WBC sample. Within-run and overall accuracy and precision ranged from 95.3 to 101.1% and from 1.6 to 4.7%, respectively. All validation parameters fulfilled international requirements. Conclusions: This method can be easily performed on small samples and provides reliable data, including in children and neonates.