反相高效液相色谱法测定人体尿样中蝙蝠葛碱的方法学研究

目的建立测定蝙蝠葛碱在人尿液中浓度的RP-HPLC法。方法尿液样品经0．1mol·L^-1NaOH碱化后用乙醚提取，以原阿片碱为内标。色谱柱为Diamonsil C18（4．6mm×200mm，5μm）；流动相为乙腈-4％的冰醋酸溶液-三乙胺（70：300：1．2，v／v／v）；流速1．0mL·min^-1；检测波长为282nm；柱温35℃；进样量50μL。结果人尿中内源性杂质不干扰蝙蝠葛碱和内标原阿片碱的测定，蝙蝠葛碱的线性范围为40．00～4000ng·mL^-1，提取回收率为79．8％～84．6％，最低定量下限为40ng·mL^-1。结论本方法简便、快速、专属性强，可用于健康人体尿样中蝙蝠葛碱含量的测定。

Determination of dauricine in human urine by RP-HPLC

Objective To establish a reversed phase high pressure liquid chromatography （RP-HPLC） method for the determination of daurieine concentration in human urine. Methods Alkalified urine samples were extracted by diethyl ether solvent with internal standard of protopine. A Diamonsil C18 （4. 6 mm ×200 mm, 5 μm） column was used with mobile phase consisting of acetonitrile-4% acetic acid solution-triethylamine （70 ： 300 = 1.2, v/v/v） at 1.0 mL·min^-1. The column temperature was 35 ℃ and the detect wavelength was 282 nm. Results Endogenous urine impurities did not interfere the determination of dauricine and internal standard protopine. The linear range of dauricine was 40. 00-4 000 ng·mL^-1. The extraction recoveries at low, middle and high levels were 82. 9%, 79. 8% and 84. 6%, respectively. The lowest quantitative limit of dauricine was 40 ng·mL^-1. Conclusion The reliable, simple and sensitive RP-HPLC method can he used to determine dauricine in human urine.