色谱

色谱 ›› 2012, Vol. 30 ›› Issue (05): 445-451.DOI: 10.3724/SP.J.1123.2011.11036

• 研究论文 • 上一篇    下一篇

超高效液相色谱-串联质谱法同时测定全血中青霉素G及其主要代谢产物

陈聪1, 2, 3, 严慧1, 沈保华1, 卓先义1*   

  1. 1. 司法部司法鉴定科学技术研究所 上海市法医学重点实验室, 上海 200063; 2. 上海市疾病预防控制中心, 上海 200336; 3. 复旦大学上海医学院法医系, 上海 200032
  • 收稿日期:2011-11-21 修回日期:2012-02-05 出版日期:2012-05-28 发布日期:2012-06-07
  • 通讯作者: 卓先义,研究员,主要从事毒物化学研究. Tel: (021)52352955, E-mail: zhuoxianyi126@126.com.
  • 基金资助:

    上海市法医学重点实验室项目（11DZ2271500, 2009-2）.

Simultaneous determination of penicillin G and its major metabolites in blood using ultra performance liquid chromatography-tandem mass spectrometry

CHEN Cong1,2,3, YAN Hui1, SHEN Baohua1, ZHUO Xianyi1*   

  1. 1. Institute of Forensic Science of Ministry of Justice, Shanghai Key Laboratory of Forensic Medicine, Shanghai 200063, China; 2. Shanghai Municipal Center for Disease Control & Prevention, Shanghai 200336, China; 3. Forensic Medicine Department of Shanghai Medical College of Fudan University, Shanghai 200032, China
  • Received:2011-11-21 Revised:2012-02-05 Online:2012-05-28 Published:2012-06-07
  • Contact: ZHUO Xianyi

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298

被引次数

31

摘要： 建立了同时检测全血中青霉素G及其2种主要代谢产物青霉噻唑酸和脱羧青霉噻唑酸的超高效液相色谱-电喷雾串联质谱分析方法。血样经简单的蛋白沉淀提取后,目标化合物经BEH C18色谱柱(50 mm×2.1 mm, 1.7 μm),以含0.1%甲酸溶液-乙腈为流动相,梯度洗脱分离后,选用正离子电喷雾多反应监测(MRM)模式检测。方法检出限(信噪比(S/N)为3计)为0.1～2.0 ng/mL,定量限(S/N=10)为0.5～5.0 ng/mL。各被测物的线性相关系数均大于0.9974,准确度为92.3%～105.5%,日内精密度小于10%。考察了不同储存温度(18、4、-18、-80 ℃)下全血中青霉素G及其代谢物的稳定性,结果表明随着储存温度升高和时间的延长,青霉素G的质量浓度下降明显。应用建立的方法检测了给予20 mg/kg青霉素G后的大鼠血液样本,血液中青霉素G原形药物在给药后0.5 h即消除完全(低于检出限),而其代谢物的体内消除时限可延长至36 h。所建方法对司法鉴定的适用性得到了进一步扩大,同时对食品残留检测也具有一定的参考价值。

关键词: 超高效液相色谱-串联质谱, 代谢产物, 青霉素G, 全血

Abstract: A fast method for the quantitative determination of penicillin G (PEN G), penicilloic acid and penilloic acid in blood with ultra performance liquid chromatography-electrospray tandem mass spectrometry was developed. A simple deproteinization of the blood was used with a mixed solution of acetonitrile and water (4:1, v/v) as extraction solvent. The blood extract was directly injected onto an LC column. The chromatographic separation of the components was performed on a BEH C18 column (50 mm×2.1 mm, 1.7 μm) using acetonitrile and water containing 0.1% formic acid. The mass spectrometer was operated in positive electrospray ion mode. Finally, the analysis was carried out with multiple reaction monitoring (MRM) mode. The limits of detection (LODs) for these three compounds were in the range of 0.1 to 2.0 ng/mL and the limits of quantification (LOQs) in the range of 0.5 to 5.0 ng/mL. Within the linear range, the correlation coefficients (r) of PEN G and its metabolites were all more than 0.9974. Accuracies for these targeted compounds were ranged from 92.3% to 105.5%, and the within-day precisions were less than 10%. The stabilities of the components were evaluated in the temperature range from 18 to ~80 ℃, and the mass concentration of penicillin G was decreased significantly with the extensions of storage temperature and storage time. Biological samples of the rats medicated with PEN G were analyzed using the developed method. The results show that PEN G can just be detected at 0.5 h after administration. However, the detection time limitation of penicilloic acid can be extended to 36 h. The established method has been further expanded for the applicability of forensic identification, and has a reference value for the detection of penicillin G residue in food.

Key words: blood, metabolites, penicillin G, ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)

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