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目的:建立HPLC梯度洗脱法同时测定蜂胶牙痛酊中甲硝唑和丁香酚的含量的新方法。方法:HPLC法,采用Agilent Eclipse XDB-C18(5μm,4.6 mm×150mm)色谱柱,流动相组成:A相为甲醇,B相为水,进行梯度洗脱;流速1.0mL.min(1,柱温:28℃,二极管阵列检测器(DAD),检测波长293nm,以外标法峰面积定量。结果:甲硝唑、丁香酚与阴性空白样品溶液分离良好,甲硝唑在16.26～162.6μg内呈良好的线性关系(r=0.9995),丁香酚在80.16～801.6μg内呈良好的线性关系(r=1.0000),样品溶液在24h内稳定,平均加样回收率甲硝唑为101.0%,RSD=1.06%(n=9),丁香酚为101.0%,RSD=0.84%(n=9)。结论:该方法简便、快速,测定结果准确可靠,重现性好,可用于蜂胶牙痛酊的质量控制。
OBJECTIVE: To establish a new method for simultaneous determination of metronidazole and eugenol in Propolis Tortulent Tincture by HPLC gradient elution. METHODS: The HPLC method was performed on an Agilent Eclipse XDB-C18 column (5 μm, 4.6 mm × 150 mm). The mobile phase consisted of methanol with phase A and water with gradient elution. The flow rate was 1.0 mL · min Temperature: 28 ℃, diode array detector (DAD), detection wavelength of 293nm, external standard peak area quantification.Results: Metronidazole, eugenol and negative blank sample solution was well separated, metronidazole in 16.26 ~ 162.6μg was (R = 0.9995). There was a good linear relationship between eugenol and 80.16 ~ 801.6μg (r = 1.0000). The sample solution was stable within 24 hours. The average recoveries of metronidazole were 101.0% with RSD = 1.06% (n = 9), eugenol was 101.0%, RSD was 0.84% (n = 9) .Conclusion: The method is simple and rapid, the method is accurate and reproducible and can be used for the quality control of prophylactic tincture of propolis .