High-throughput determination of theophylline and caffeine in human serum by conventional liquid chromatography-mass spectrometry
High-throughput determination of theophylline and caffeine in human serum by conventional liquid chromatography-mass spectrometry
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DOI：
10.1007/s11419-008-0058-6
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发表时间：
2009-02-01
影响因子：
2.2
通讯作者：
Seno, Hiroshi
中科院分区：
文献类型：
--
作者：
Arinobu, Tetsuya;Hattori, Hideki;Seno, Hiroshi
建立了反吹柱切换自动高效液相色谱/质谱法（HPLC-MS）超快速测定茶碱和咖啡因的方法。进样400 μ l用超纯水稀释的血清样品，并转移至用作预柱的Oasis HLB柱中进行提取。在切换阀门后，在反冲洗模式下洗脱预柱中捕获的分析物，并用Chromolith Performance RP-18 e柱（C（18）-键合的整体式二氧化硅）分离;然后通过大气压化学电离（APCI）-MS检测柱流出物中的化合物。本方法成功地提供了在2分钟内对茶碱和咖啡因的高通量测定。令人满意的线性，可获得用于分析两种化合物的治疗和毒性水平的再现性和灵敏度。由于该方法操作简单、通量高，在法医毒理学和急救医学领域具有广阔的应用前景。
Automated high-performance liquid chromatography/mass spectrometry (HPLC-MS) with backflush column-switching was established for ultra-fast determination of theophylline and caffeine. A 400-mu l portion of serum sample diluted with ultrapure water was injected and transferred to an Oasis HLB cartridge used as a precolumn for extraction. After switching the valves, the analytes trapped in the precolumn were eluted in the backflush mode and separated with a Chromolith Performance RP-18e column (C(18)-bonded monolithic silica); the compounds in column effluents were then detected by atmospheric pressure chemical ionization (APCI)-MS. The present method successfully provided high-throughput determination of theophylline and caffeine within 2 min. Satisfactory linearity, reproducibility, and sensitivity could be obtained for analysis of therapeutic and toxic levels of both compounds. Because of the very simple procedure and high throughput using the conventional HPLC system, the present method seems to have high potential in the fields of forensic toxicology and emergency medicine.