丹磺酰氯柱前衍生-超高效液相色谱-串联质谱法测定人体尿样中的环己胺

建立丹磺酰氯柱前衍生-超高效液相色谱-串联质谱法测定人体尿样中环己胺的方法。冷冻样品经解冻、离心后,用丹磺酰氯衍生,固相萃取小柱净化。目标化合物采用 Waters ACQUITY CSHTM C18色谱柱(50 mm×2.1 mm,1.7μm)分离,以甲醇和0.002 mol/L乙酸铵溶液为流动相梯度洗脱,采用电喷雾离子源电离、正离子多反应监测模式质谱检测。环己胺在2.5~200μg/L浓度范围内有较好的线性关系,相关系数大于0.999,回收率为98.7%~102.3%,精密度为3.1%~5.2%,检出限和定量限分别为1.0和3.0μg/L。结果表明,本方法操作简单、准确可靠,可适用于人体尿液中环己胺的定量分析。应用本方法测定200份学生尿液样品,环己胺检出率为34.5%。

Ultra Liquid Chromatography-Tandem Mass Spectrometry with Dansyl Chloride Pre-column Derivatization

A method was developed for the quantification of cyclohexylamine in human urine samples by ultra performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry ( UPLC-ESI-MS/MS ) with pre-column derivatization. After the refrigerated centrifugation of urine samples, cyclohexylamine in supernatant was derived with dansyl chloride and then its derivant was purified with solid phase extraction (SPE). The target compounds were separated on a Waters ACQUITY CSHTM C18 column (50 mm× 2. 1 mm, 1. 7 μm) with the gradient elution of methanol and water ( containing 0. 002 mol/L ammonium acetate ) and were determined in electrospray positive ionization ( ESI+) and multiple reaction monitoring ( MRM) modes. The linear range was 2. 5-200 μg/L for cyclohexylamine with the correlation coefficient more than 0 . 999 . The recovery ranged from 98 . 7% to 102 . 3%, with the relative standard deviations (RSDs) of 3. 1%-5. 2%. The limit of detection (LOD) and the limit of quantitation (LOQ) were 1 μg/L and 3 μg/L. The results indicated that the method was accurate and reliable, and suitable for the detection of cyclohexylamine in urine samples. The urine samples of 200 primary school students were analyzed by this method and the positive rate of cyclohexylamine was 34 . 5%.