枳壳HPLC特征图谱及6个黄酮苷类成分含量测定

目的： 采用高效液相色谱法建立枳壳的特征图谱，并同时测定6个黄酮苷类成分的含量。方法： 以新北美圣草苷、芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷和枳属苷为指标性成分，采用Kinetex? 2.6 μ C₁₈ 100（100 mm×4.6 mm，2.6 μm）色谱柱，以乙腈为流动相B，0.1%甲酸为流动相A，梯度洗脱（0~10 min，15% B→15% B；10~15 min，15% B→20% B；15~20 min，20% B→25% B；20~30 min，25% B→40% B），流速1.0 mL·min^-1，检测波长为283 nm，柱温30℃。结果： 建立枳壳特征图谱，30 min内枳壳的主要色谱峰能够达到完全分离，同时标定6个黄酮苷类成分，对12批饮片中的6个成分进行含量测定。新北美圣草苷、芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷和枳属苷的质量浓度分别在1.77~70.8 μg·mL^-1（r=0.999 8，n=5）、11.3~452.0 μg·mL^-1（r=1.000 0，n=5）、34.0~1 360 μg·mL^-1（r=0.999 9，n=5）、13.6~544 μg·mL^-1（r=1.000，n=5）、27.6~1 104 μg·mL^-1（r=0.999 6，n=5）、14.9~596 μg·mL^-1（r=1.000，n=5）范围内线性关系良好，方法的平均回收率（n=6）分别为96.9%（RSD=1.8%）、104.6%（RSD=1.7%）、97.4%（RSD=1.11%）、107.4%（RSD=1.1%）、93.2%（RSD=1.1%）、107.8%（RSD=1.5%）；12批枳壳中上述6个成分含量分别为0.11%~0.45%、0.17%~0.59%、4.85%~9.09%、0.37%~0.74%、3.24%~6.17%、0.05%~0.56%。结论： 该方法操作简单，重复性好，并能同时测定6个黄酮苷类成分含量，可为枳壳的质量标准研究提供更加全面的依据。

Objective: To establish an HPLC characteristic chromatogram and quantitative method for six components in Aurantii Fructus.Method: With neoeriocitrin,narirutin,naringin,hesperidin,neohesperidin and poncirin as reference substances,HPLC analysis was performed on a Kinetex? 2.6 μ C₁₈ 100 A column(100 mm×4.6 mm,2.6 μm)with the gradient elution of acetonitrile(B)-0.1% formic acid solution(A)(0-10 min,15%B→15%B;10-15 min,15%B→20%B;15-20 min,20%B→25%B;20-30 min,25%B→40%B). The flow rate was 1.0 mL·min^-1,the detection wavelength was 283 nm and the column temperature was 30℃. Results: HPLC characteristic chromatogram of Aurantii Fructus was established and six characteristic peaks were marked.Six characteristic peaks were simultaneously determined by HPLC within 30 min.The linear ranges of neoeriocitrin,narirutin,naringin,hesperidin,neohesperidin and poncirin were 1.77-70.8 μg·mL^-1(r=0.999 8,n=5),11.3-452.0 μg·mL^-1(r=1.000,n=5),34.0-1 360 μg·mL^-1(r=0.999 9,n=5),13.6-544 μg·mL^-1(r=1.000,n=5),27.6-1 104 μg·mL^-1(r=0.999 6,n=5)and 14.9-596 μg·mL^-1(r=1.000,n=5),respectively.The average recovery rate(n=6)were 96.9%(RSD=1.8%),104.6%(RSD=1.7%),97.4%(RSD=1.1%),107.4%(RSD=1.1%),93.2%(RSD=1.1%)and 107.8%(RSD=1.5%),respectively.The contents of the six components in 12 batches of Aurantii Fructus were 0.11%-0.45%,0.17%-0.59%,4.85%-9.09%,0.37%-0.74%,3.24%-6.17% and 0.05%-0.56%,respectively.Conclusion: The proposed specific HPLC characteristic chromatogram and quantitation method of six components for Aurantii Fructus are simple and repeatable,which offer reference for quality control of the drug.

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