HPLC/MS of Antibiotics in Biological Samples

生物样品中抗生素的 HPLC/MS

• 批准号: 09670454
• 负责人: SATO Keizo
• 金额: $ 2.3万
• 依托单位: Showa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670454/
• 关键词: Liquid Chromatography; Mass Spectrometry; Fast Atom Bombardment; Antibiotics; Cephem Antibiotics; Aminoglycoside Antibiotics; Ion-pair HPLC; Solid-Phase Extraction; 固相抽出; イオンベアー逆相HPLC; 薬物ショック死

We analyzed two representative antibiotics, cephems and aminoglycosides, by our recently established system of capillary high-performance liquid chromatography (HPLC)/fast atom bombardment (FAB)-mass spectrometry (MS). Cephems could be analyzed by conventional reversed-phase HPLC, but aminoglycosides required ion-pair reversed-phase HPLC.As a matrix for FAB-MS, diethanolamine (DEA) along with glycerin (GLY) were used. The authentic compounds of 24 cephems and 8 aminoglycosides were analyzed by our capillary HPLC/FAB-MS system. Whend a suitable matrix was used for each antibiotic, quasi-molecular peak together with adequate fragment peaks were obtained from all 32 antibiotics, though DEA provided more information on molecular mass of a compound than did GLY for some cephems. The range of the detection limits of the mass spectral measurements was rather wide (10 ng - 2.5 mug on column), but sufficiently sensitive to detect therapeutic levels of antibiotics in human serum for most compounds under investigation.The extraction of the antibiotics from human serum was studied using solid-phase extraction cartridges. In general, Bond Elut C_<18> cartridges were effective for the isolation of cephems and aminoglycosides under investigation. With this isolation procedure and the capillary HPLC/FAB-MS in the positive mode, cefazolin and ribostamycin, representative cephem antibiotic and aminoglycoside, respectively, could be identified in sera from cadavers who suddenly died after injection of them. The capillary HPLCIFAB-MS was semi-quantitative for all compounds under investigation. However, we believe that accurate quantifation can be made by SIM of FAB-MS with a suitable isotopic internal standard. We also devised a simple method for quantitation of some other drugs in human serum by UV monitoring of the capillary HPLC.These analytical methods devised in this study would seem to be useful in clinical pharmacology as well as in forensic toxicology.

采用新建立的毛细管高效液相色谱(HPLC)/快速原子轰击(FAB)-质谱（MS）联用技术对头孢菌素和氨基糖苷类抗生素进行了分析。传统的反相高效液相色谱法可以分析头孢类，而氨基糖苷类则需要离子对反相高效液相色谱法。采用二乙醇胺（DEA）和甘油（GLY）作为FAB-MS的基质。采用毛细管高效液相色谱- FAB-MS联用系统对24种头孢醚和8种氨基糖苷类化合物进行了分析。当每种抗生素使用合适的基质时，所有32种抗生素都获得了准分子峰和足够的片段峰，尽管DEA比GLY提供了更多关于化合物分子质量的信息。质谱测量的检出限范围相当宽（柱上10纳克- 2.5马克），但对大多数被调查化合物的人类血清中抗生素的治疗水平检测足够敏感。采用固相萃取筒对人血清中抗生素的提取进行了研究。总的来说，Bond Elut C_<18>试剂盒对所研究的头孢类和氨基糖苷类的分离是有效的。采用该分离方法和阳性模式下的毛细管高效液相色谱/FAB-MS，可在注射后突然死亡的尸体血清中分别鉴定出头孢唑林和核糖素这两种具有代表性的头孢唑林抗生素和氨基糖苷类药物。毛细管hplc - ms对所研究的所有化合物均为半定量。然而，我们相信在合适的同位素内标下，FAB-MS的SIM可以进行准确的定量。我们还设计了一种简便的毛细管高效液相色谱紫外监测法测定人血清中其它药物的含量。本研究设计的这些分析方法在临床药理学和法医毒理学中似乎是有用的。

项目成果

Xiao-Pen Lee: "Simple extraction of alcohols in human body fluids by headspace solid-phase microextraction(SPME)" 6th Indo Pacific Congress on Legal Medicine and Forensic Sciences. 6. 820-823 (1999)

李晓鹏：“通过顶空固相微萃取（SPME）简单提取人体体液中的醇”，第六届印度太平洋法律医学和法医学大会。

X.-P.Lee, T.Kumazawa T.Kurosawa, K.Akiya, Y.Akiya, S.Furuta and K.Sato: "Simple extraction of methanol in human whole blood by headspace solid-phase microextraction (SPME)." Jpn.J.Forensic Toxicol.16. 64-68 (1998)

X.-P.Lee、T.Kumazawa T.Kurosawa、K.Akiya、Y.Akiya、S.Furuta 和 K.Sato：“通过顶空固相微萃取 (SPME) 简单提取人全血中的甲醇。”

Tomoko Taguchi: "High-performance liquid chromatographic determination of benzodiazepines in serum with 0.3mm I.D.columns" Proceedings of the 6th Indo Pacific Congress on Legal Medicine and Forensic Sciences. (in press).

Tomoko Taguchi：“用 0.3mm I.D. 柱高效液相色谱测定血清中苯二氮卓类药物”第六届印度太平洋法律医学和法医学大会论文集。

Toshiyuki Sano: "Quantitation of midazolam in serum by column-switching capillary HPLC/FAB-MS using a deuterium-labeled internal standard" Current Topics in Forensic Science. 2. 235-236 (1997)

Toshiyuki Sano：“使用氘标记内标，通过柱切换毛细管 HPLC/FAB-MS 定量血清中的咪达唑仑”法医学当前主题。

Xiao-Pen Lee: "Detection of cresol isomers and phenol in human whole blood by headspace solid-phase micro extraction(SPME)and capillary gas chromatography" Jpn.J.Forensic Toxicol. 15. 21-28 (1997)

李晓鹏：“通过顶空固相微萃取（SPME）和毛细管气相色谱法检测人全血中的甲酚异构体和苯酚”Jpn.J.Forensic Toxicol。