Zhang Panpan, Zhang Fucheng, Wang Zhaohong, Jiang Ye, Lu Yongjiang
School of Pharmacology, Hebei Medical University, Shijiazhuang 050017, China.
Se Pu. 2013 Mar;31(3):211-7. doi: 10.3724/sp.j.1123.2012.10018.
A rapid, specific and sensitive ultra performance liquid chromatography-mass spectrometry (UPLC-MS/MS) method was developed for the simultaneous determination of four trace alkaloids, including aconitine (AC), hypaconitine (HA), mesaconitine (MA) and yunaconitine (YA) in human urine samples. UPLC-MS-MS system coupled with an electrospray ionization (ESI) source was performed in multiple reaction monitoring (MRM) mode. The sample preparation was performed with hollow fiber microextraction (HF-LPME) prior to the analysis. The enrichment factors of the four alkaloids were 102 - 301. The separation was applied on a Waters ACQUITY UPLC BEH C18 column (50 mm x 2.1 mm, 1.7 micro m) with a gradient elution of acetonitrile and 10 mmol/L NH4HCO3, as mobile phase. The retention times , were less than 3 min. This method significantly improved the detection sensitivity, and the limits of quantitation were from 0. 01 to 0.1 ng/L. The calibration curves were linear over the ranges of 0.01 - 10 ng/L for AC, MA and YA, 0.1 - 100 ng/L for HA in human urine samples, and the correlation coefficients were 0. 998 1, 0.998 4, 0.999 5 and 0.998 6, respectively. The method was proved to be rapid and sensitive for aconitum alkaloid analysis in urine samples.
建立了一种快速、特异且灵敏的超高效液相色谱-质谱联用（UPLC-MS/MS）方法，用于同时测定人尿液样本中的四种痕量生物碱，包括乌头碱（AC）、次乌头碱（HA）、中乌头碱（MA）和云乌头碱（YA）。UPLC-MS-MS系统与电喷雾电离（ESI）源联用，采用多反应监测（MRM）模式。分析前采用中空纤维微萃取（HF-LPME）进行样品制备。四种生物碱的富集因子为102 - 301。分离在Waters ACQUITY UPLC BEH C18柱（50 mm×2.1 mm，1.7μm）上进行，以乙腈和10 mmol/L NH4HCO3的梯度洗脱作为流动相。保留时间小于3分钟。该方法显著提高了检测灵敏度，定量限为0.01至0.1 ng/L。人尿液样本中AC、MA和YA的校准曲线在0.01 - 10 ng/L范围内呈线性，HA在0.1 - 100 ng/L范围内呈线性，相关系数分别为0.998 1、0.998 4、0.999 5和0.998 6。该方法被证明对尿液样本中乌头生物碱的分析快速且灵敏。
