双波长HPLC法同时测定穿心莲浸膏中穿心莲内酯和脱水穿心莲内酯的含量

Simultaneous determination of andrographolide and dehydroandrographolide in extractum andrographis by dual-wavelength HPLC

目的建立双波长高效液相色谱法同时检测穿心莲浸膏中穿心莲内酯和脱水穿心莲内酯的含量测定方法。方法采用Intersil C18色谱柱（250 mm×4.6 mm,5μm）,以甲醇—水（60∶40）为流动相；检测波长穿心莲内酯为225 nm,脱水穿心莲内酯为254 nm；柱温为35℃；流速为1.0 mL·min-1。结果穿心莲内酯进样量在0.1043～1.0430μg范围内与峰面积呈良好的线性关系（r=1.000 0,n=5）,脱水穿心莲内酯进样量在0.099 9～0.999 0μg范围内与峰面积呈良好的线性关系（r=1.000 0,n=5）；穿心莲内酯的平均加样回收率为98.53%（RSD=1.27%,n=6）,脱水穿心莲内酯的平均加样回收率为101.51%（RSD=1.10%,n=6）。结论该方法灵敏度高,重现性好,能准确测定穿心莲浸膏中穿心莲内酯和脱水穿心莲内酯的含量。

Objective To establish a dual-wavelength HPLC method for the determination of andrographolide and dehydroandrographolide in extractum andrographis.Methods The analysis was performed on Intersil C18column（250 mm × 4.6 mm,5 μm）,using methanol-water（60∶40） as mobile phase,the UV detector wavelength set at 225 nm for andrographolide and 254 nm for dehydroandrographolide.Column temperature was maintained at 35 ℃ and the flow rate was 1.0 mL·min-1.Results There was good linearity within the range of 0.104 3 ～ 1.043 0 μg（r = 1.000 0,n = 5） for andrographolide and 0.099 9 ～ 0.999 0 μg（r = 1.000 0,n = 5） for dehydroandrographolide.The average recoveries of andrographolide and dehydroandrographolide were 98.53%（RSD = 1.27%,n = 6） and 101.51%（RSD = 1.10%,n = 6）.Conclusions This method is highly sensitive,rapid and accurate for determination of andrographolide and dehydroandrographolide in extractum andrographis.