测定肿瘤细胞内外5-氟尿嘧啶含量的方法研究

目的建立了高效液相色谱法测定A549组胞内外5-氟尿嘧啶含量的方法，为探索5-氟尿嘧啶的作用机理奠定基础。方法色谱柱采用Hypersil ODS柱（4．6mm×250mm，5μm）；流动相为甲醇·水=2：98（V／V）；流速：0．8ml／min；柱温：30℃；检测波长：265nm；进样量为5μl。结果细胞内液中5-氟尿嘧啶在0.1～50．0μg/ml范围内线性关系良好（r=0．9996），平均回收率为97．65％，RSD为0．40％，日内和日间精密度分别为1．30％，1．04％；细胞跨液中5-氟尿嘧啶在1．0～250．0μg/ml范围内线性关系良好（r=0.9999），平均回收率为102.34％，RSD为2．11％，日内和日间精密度分别为1．32％，1．06％。结论该色谱方法简便易行，准确度高，重现性好，可用于细胞内外5-氟尿嘧啶浓度的动态变化规律研究。

Method Research for Determination of Intracellular and Extracellular 5-Fluorouracil

Objectlve A HPLC method for the determination of 5-FU in intracellular and extraeenular is established so as to understand more about antitumor mechamism of 5-FU. Methods The Hypersil ODS C18 column （250 mm × 4.6 mm,5 μm）was applied, and the parameters are as following： the mobile phase was methanolwater（2： 98, V/V） ; the flow rate was 0. 8 ml/min;the column temperature was 30℃ and the detection wavelength was 265 nm; the injection volume was 5 μl. Results The linear range of intracellular 5-FU was 0. 1 - 50. 0 μg/ml （ r = 0. 999 8） ;the average recovery was 97.65% ,and RSD was 0. 40% ;the precision of intra-day and inter-day was 1.30% and 1.04% respectively. And the linear range of extracellular 5-FU was 1.0 - 250. 0 μg/ml （ r = 0. 999 9 ）, the average recovery was 102.34%, RSD was 2. 11% ,the precision of intra-day and inter-day was 1.32% and 1.06%. Conclusion The method is simple,accurate and of good reproduc- ibility which can be used for the dynamic study of intracellular and extracellular 5-FU.