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目的:建立多肽卵泡刺激素-4(FSH-4)的高效液相色谱(HPLC)法的检测方法,并考察了其方法学。方法:采用HPLC法,色谱柱为Phenomenex C_(18)柱(250 mm×4.60 mm,5μm);流动相为纯水(0.1%三氟乙酸)(A)-乙腈(0.1%三氟乙酸)(B),按梯度洗脱;柱温为25℃;检测波长为218 nm。结果:该法的线性范围介于10~400μg/mL;回归方程为A=7937.2ρ+132.08,r~2=0.999 6,血浆中含卵泡刺激素2.8μg/mL;日间和日内相对标准偏差(RSD)<5.0%;血浆样品检测提取回收率为85.0%~90.0%。结论:本法简便、准确、重现性好,适用于测定FSH-4的含量。
Objective: To establish a method for the determination of follicular stimulating hormone-4 (FSH-4) by high performance liquid chromatography (HPLC) and investigate its methodology. Methods: The mobile phase was Phenomenex C_ (18) column (250 mm × 4.60 mm, 5 μm) by HPLC. The mobile phase consisted of pure water (0.1% trifluoroacetic acid) B), gradient elution; column temperature was 25 ℃; detection wavelength was 218 nm. Results: The linear range of this method was 10-400μg / mL. The regression equation was A = 7937.2ρ + 132.08, r = 2 = 0.999 6, and the plasma contained follicle-stimulating hormone 2.8μg / mL. The relative standard deviations (RSD) <5.0%. The recovery rate of plasma samples was 85.0% -90.0%. Conclusion: This method is simple, accurate, reproducible, suitable for the determination of FSH-4 content.