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目的建立豚鼠外耳淋巴液和血浆中甲泼尼龙含量的测定方法。方法应用HPLC法测定甲泼尼龙的含量。HPLC色谱条件为ZORBAX Eclipse XDB-C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相为7.56 mmol·L~(-1)硫酸铵-乙腈(68:32,V/VV),流速1.0 mL·min~(-1),柱温20℃,紫外检测波长243 nm。结果本研究建立的HPLC法,用于测定豚鼠外淋巴液和血浆中甲泼尼龙含量时的线性范围分别为0.05～5.0 mg·L~(-1)(r=0.999 6)和0.10～5.0 mg·L~(-1)(r=0.999 5)。给药后2 h鼓阶外淋巴液中甲泼尼龙的质量浓度为(0.14±0.02)mg·L~(-1),血浆质量浓度为(1.67±0.19)mg·L~(-1)。结论方法灵敏、准确,重现性好,可用于血浆和外淋巴液中甲泼尼龙的测定。
Objective To establish a method for the determination of methylprednisolone content in the outer ear lymph of guinea pigs and plasma. Methods HPLC method for the determination of methylprednisolone content. The chromatographic conditions of HPLC were ZORBAX Eclipse XDB-C 18 column (250 mm × 4.6 mm, 5 μm) with a mobile phase of 7.56 mmol·L -1 ammonium sulfate-acetonitrile (68:32, V / VV) The flow rate was 1.0 mL · min ~ (-1) and the column temperature was 20 ℃. The UV detection wavelength was 243 nm. Results The linear range of the method for the determination of methylprednisolone in peripheral blood and plasma of guinea pigs was 0.05-5.0 mg · L -1 (r = 0.999 6) and 0.10-5.0 mg · L -1 (r = 0.999 5). The concentration of methylprednisolone was (0.14 ± 0.02) mg · L -1 and the plasma concentration was (1.67 ± 0.19) mg · L -1 at 2 h after administration. Conclusion The method is sensitive, accurate and reproducible. It can be used for the determination of methylprednisolone in plasma and perilymph.