替代对照品法测定连翘中连翘酯苷A和连翘苷的含量

目的: 建立HPLC替代对照品法同时测量连翘中连翘酯苷A和连翘苷的含量。 方法: 选取连翘苷作为连翘含量测定对照品连翘酯苷A的替代对照品,在不同条件下测定替代对照品相对对照品的校正因子,利用校正因子和替代对照品连翘苷进行连翘中连翘酯苷A的含量测定。色谱条件:采用Agilent ZORBAX SB-C₁₈₍₂₎(5 μm,4.6 mm×250 mm)色谱柱,以乙腈(A)-0.3%的醋酸水溶液(B)为流动相,梯度洗脱[0~5 min,A-B(15:85)→A-B(17:83);5~10 min,A-B(17:83)→A-B(20:80);10~15 min,A-B(20:80)→A-B(23:77);15~20 min,A-B(23:77)→A-B(25:75)],流速0.8 mL·min^-1,柱温25℃,检测波长230 nm。 结果: 连翘酯苷A和连翘苷进样浓度分别在0.08~0.62 mg·mL^-1(r=0.9995)和0.012~0.08 mg·mL^-1(r=0.9995)内与峰面积呈良好的线性关系,测得校正因子f=0.4761,用替代对照测定方法测得回收率(n=6)为99.20%。 结论: 本法使用替代对照品法测定了连翘中连翘酯苷A的含量,结果证明了替代对照品法用于连翘药材质量控制可行、实用。

Objective: To establish an HPLC method for determination of forsythoside A and forsythin in Forsythiae Fructus by substitute reference substance. Methods: Forsythin was selected as the substitute reference substance for the reference substance forsythoside A for determination of Forsythiae Fructus.The correction factor was measured of forsythoside A and forsythin under different conditions.The content of forsythoside A in Forsythiae Fructus was determined by the substitute reference substance and the correct factor.Chromatographic conditions:Agilent ZORBAX SB-C₁₈₍₂₎(5 μm,4.6 mm×250 mm)column was adopted,the mobile phase consisted of acetonitrile(A)-0.3% acetic acid solution(B)with gradient elution [0-5 min,A-B(15:85)→A-B(17: 83);5-10 min,A-B(17: 83)→A-B(20: 80);10-15 min,A-B(20: 80)→A-B(23: 77);15-20 min,A-B(23: 77)→A-B(25: 75)]was at the flow rate of 0.8 mL·min^-1,the column temperature was 25℃,and the detection wavelength was 230 nm. Results: The linear response ranges of forsythoside A and forsythin were 0.08-0.62 mg·mL^-1 (r=0.9995)and 0.012-0.08 mg·mL^-1 (r=0.9995),respectively.The correction factor(f)was 0.4761.The average recovery(n=6)was 99.20%. Conclusion: The substitute reference substance is adopted in HPLC determination of Forsythiae Fructus. The assay demonstrates that the method is feasible and useful for quality control of Forsythiae Fructus.