HPLC波长切换法同时测定经典名方槐花散中5种成分的含量

Simultaneous Determination of 5 Components in the Classical Formula Huaihua San by HPLC Wavelength Switching Method

目的:建立同时测定经典名方槐花散中芦丁、柚皮苷、新橙皮苷、槲皮素和胡薄荷酮等5种成分含量的方法。方法:采用高效液相色谱波长切换法。色谱柱为Cosmosil C18,流动相为乙腈-0.05%磷酸溶液(梯度洗脱),检测波长依次为257 nm(芦丁)、283 nm(柚皮苷、新橙皮苷)、254 nm(槲皮素)、252 nm(胡薄荷酮),流速为1.0 mL/min,柱温为30℃,进样量为10μL。结果:芦丁、柚皮苷、新橙皮苷、槲皮素、胡薄荷酮检测质量浓度的线性范围分别为21.7-2170、46-4600、22.3-2230、0.96-96、2.7-270μg/mL(r均大于0.999);精密度、稳定性(24 h)、重复性试验的RSD均小于2%(n=6);平均加样回收率分别为100.70%、99.31%、101.10%、100.03%、99.63%(RSD均小于2%,n=9)。3批槐花散样品中,上述5种成分的含量分别为20.055-22.615、25.557-27.806、11.428-13.250、0.350-0.478、2.372-4.011 mg/g。结论:所建含量测定方法操作简便、结果准确、重复性好,可用于槐花散中5种成分的同时测定。

OBJECTIVE:To develop a method for simultaneous determination of 5 components in classical formula Huaihua san,including rutin,naringin,neohesperidin,quercetin and pulegone.METHODS:HPLC wavelength switching method was adopted.The determination was performed on Cosmosil C18 column with mobile phase consisted of acetonitrile-0.05%phosphoric acid solution(gradient elution)at the flow rate of 1.0 mL/min.The detection wavelengths were set at 257 nm for rutin,283 nm for naringin and neohesperidin,254 nm for quercetin,252 nm for pulegone,respectively.The column temperature was set at 30℃,and sample size was 10μL.RESULTS:The linear range was 21.7-2170μg/mL for rutin,46-4600μg/mL for naringin,22.3-2230μg/mL for neohesperidin,0.96-96μg/mL for quercetin,2.7-270μg/mL for pulegone(all r>0.999),respectively.RSDs of precision,stability(24 h)and reproducibility tests were all lower than 2%(n=6).Average recoveries were 100.70%,99.31%,101.10%,100.03%and 99.63%(all RSD<2%,n=9).Among 3 batches of Huaihua san samples,the contents of above 5 components were 20.055-22.615,25.557-27.806,11.428-13.250,0.350-0.478,2.372-4.011 mg/g,respectively.CONCLUSIONS:Established method is simple,accurate and reproducible,and could be used for the simultaneous determination of 5 components in Huaihua san.