高效液相色谱荧光检测法测定人血浆中盐酸曲马多浓度

HPLC-fluorescence method for determination of tramadol hydrochloride in human plasma

目的：建立测定人血浆中盐酸曲马多的高效液相色谱-荧光检测法。方法：血浆样品经液-液萃取，以氟康唑为内标，在Diamonsil C18柱（迪马公司，200mm×4．6mm，5μm），以磷酸二氢纳缓冲液（0．04mol·L^-2，pH3．5）-乙腈（77：23）为流动相，荧光检测，激发波长为275nm，发射波长为302nm。结果：曲马多的线性范围为5～1000μg·L^-1，最低检测限为1μg·L^-1。低、中、高3种浓度的日内日间RSD均小于7．7％，提取回收率分别为（85．2±4．3）％，（80．6±3．5）％和（83．6±3．6）％。结论：该法重复性好、准确、简便、快速，适用于人血浆中盐酸曲马多浓度的测定及临床药动学研究。

OBJECTIVE To develop an HPLC fluorescence rriethod for determination tramadol hydrochloride in human plasma. METHODS The method involved a single liquid-liquid extraction procedure and aspecific determination with fluorescence detector, in witch fluconazol was used as internal standard. HPLC instrument was used with the column C18 （Diamonsil, 250 mm×4.6mm,5μm）, the mobile phase was composed of phosphate buffer solution（0. 04 mol·L^-1 , pH 3. 5）-acetonitrile（77： 23）. The fluorimetric excitation and emission wavelength were set at 275 nm and 302 nm, respectively. RESULTS Linear range was 5- 1 000 μg·L^-1. The lower limit of quantitation was 1.0 μg·L^-1. The within-day and between-day precision of variation was less than 7. 7%. The average extraction recoveries for tramadol were （85.2 ±4. 3） %, （80. 6 ±3. 5） % and （83. 6 ±3. 6）%, respectively. CONCLUSION The method is reproducible, sensitive, convenient and rapid. It is applicable to the determination of the concentration of tramadol in human plasma.