HPLC法测定新型酪氨酸酶抑制剂UP302乳膏的含量及皮肤表面残留量

目的: 建立测定UP302乳膏中UP302含量的HPLC方法,并应用于研究UP302乳膏在大鼠皮肤局部给药后的皮肤残留百分率。 方法: 采用Phenomenex Luna 5u C₁₈(150 mm×4.6 mm,5 μm)色谱柱,柱温35 ℃;流动相为甲醇-水,以1.0 mL·min^-1梯度洗脱;检测波长280 nm;检测时间10 min。UP302对照品用甲醇溶解稀释,乳膏和空白乳膏基质中UP302用甲醇提取。 结果: UP302在5~100 μg·mL^-1的浓度范围内线性关系良好(r=0.9999)。本方法日内日间准确度在98.5%~100.6%,日内日间精密度小于2.4%。 结论: 本方法专属性强,灵敏度高,重现性好,耐用性好,操作简便,结果准确,可用于UP302乳膏的含量测定,以及乳膏皮肤表面残留量的测定和制剂透皮吸收研究。

Objective: To establish an HPLC method for the determination of UP302 in cream,and quantify residual amount of UP302 after cream applied to rat skin. Method: The chromatographic separation was achieved within 10 min on Phenomenex Luna 5u C₁₈(150 mm×4.6 mm,5 μm) column with gradient mobile phase containing a mixture of methanol and water at a flow rate of 1.0 mL·min^-1.The detection wavelength was 280 nm and column temperature was 35 ℃.The methodology involved extraction of the analyte from UP302 cream by methanol. Results: The calibration curves were linear in a range of 5-100 μg·mL^-1 with correlation coefficient more than 0.999.The intra-and inter-day accuracy was between 98.5% and 100.6%,and precision was less than 2.4%. Conclusion: The method is simple,sensitive,precise,accurate and suitable for the estimation of UP302 in cream.The method is applied to quantify the residual amount of UP302 cream on rat skin.,and can be used for study on skin absorption of drug preparation.

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