高效液相色谱法测定血浆中的吉西他滨及其血浆蛋白结合研究

Determination of gemcitabine concentration in plasma and its plasma protein binding rate by HPLC

采用高效液相色谱法建立了检测血浆中吉西他滨浓度的方法,并采用平衡透析法测定其血浆蛋白结合率。Aglient ZORBAX SB-C18色谱柱（4.6 mm×250 mm,5μm）,流动相为50 mmol/L磷酸氢二钾溶液（p H 3.0±0.1）-甲醇-乙腈（80∶10∶10）,检测波长为268 nm,柱温为30℃,流速为1.0 m L/min。吉西他滨在0.2~20.0μg/m L浓度范围内呈现良好的线性关系（r2=0.995 6）。吉西他滨不同浓度在血浆中的蛋白结合率分别为（13.27±2.48）%,（12.64±5.49）%和（14.97±3.97）%。该方法对吉西他滨进行分离,方法简便、灵敏、稳定,其血浆中的血浆蛋白结合率较低。

To establish a HPLC method for determination of gemcitabine in plasma and to study its plasma protein binding rate by equilibrium dialysis. The chromatographic condition included a Aglient ZORBAX SB-C18（ 4. 6 mm × 250 mm,5 μm） column and the mobile phase consisting of 50 mmol / L dipotassium hydrogen phosphate solution（ p H 3. 0 ± 0. 1）-methanol-acetonitrile（ 80 ∶ 10 ∶ 10）,the detection wavelength was at 268 nm and the flow rate was 1. 0 m L / min. The column was 30 ℃. The calibration curve of gemcitabine show good linearity（ r2= 0. 995 6） in the range of 0. 2 ~ 20. 0 μg / m L. The plasma protein binding rates of gemcitabine with different concentrations were（ 13. 27 ± 2. 48） %,（ 12. 64 ± 5. 49） %and（ 14. 97 ± 3. 97） %,respectively. The method of determining the concentration of gemcitabine by HPLC is simple,sensitive and stable. The plasma protein binding rate of gemcitabine was very low.