HPLC法测定血浆中烟酸浓度的研究

STUDY ON DETERMINATION OF NIACIN IN RAT PLASMA BY HPLC

目的:建立反相高效液相色谱测定大鼠血浆中烟酸浓度的方法。方法:血浆样品用甲醇提取,过滤后进样20 μl。色谱柱为Shim-pack VP-ODS柱(4.6 mm×250 mm,5 μm),流动相为甲醇-异丙醇-0.005 mol辛烷磺酸钠水溶液(7:2:91,v/v),流速1.0 ml/min,紫外检测波长261 nm,柱箱温度45℃。内标物为咖啡因。结果:本法最低检测浓度为0.02 μg/ml,线性范围为0.22-42.6 μg/ml,r=0.9996。日内精密度<4％,日间精密度<7％,烟酸的绝对回收率(％)为96.93±4.9。结论:本法快速、准确、灵敏,可用于烟酸的体内代谢动力学研究。

Objective: To develop a method for determination of niacin in rat plasma using RP-HPLC. Methods: The plasma was extracted with methanol. Shim-pack VP-ODS column (250 mm×4.6 mm, 5μm) was used as stationary phase and methanol-isopropyl alcohol-sodium octanlysulfonate water solution (7:2:91, v/v) as mobile phase, with flow rate 1.0 ml/min, UV wavelength at 261nm, column temperature at 45℃, and caffeine as internal standard. Results:The minimal detectable level was 0.02 μg/ml. The calibration curve was linear from 0.22 μg/ml to 42.6 μg/ml (r=0.9999) . Inter-day RSD was <4% and intra-day <7% . The recovery rate was (96.93±4.9)%. Conclusion: This method is economic, simple, rapid, sensitive and accurate for determination of niacin in plasma.