HPLC法测定酸橙枳壳和枸橘枳壳中黄酮苷的含量

目的：建立酸橙枳壳和枸橘枳壳中异柚皮苷、柚皮苷、橙皮苷、新橙皮苷和枳属苷同时测定的HPLC方法。方法：采用Dikma C₁₈（2）柱（150 mm×4.6 mm，5 μm），以乙腈（A）-0.1%磷酸水（B）为流动相，梯度洗脱（0~10 min，20% A；10~11 min，20% A→30% A；11~25 min，30% A），流速1.0 mL·min^-1，检测波长283 nm，柱温25℃。结果：异柚皮苷、柚皮苷、橙皮苷、新橙皮苷和枳属苷进样量分别在0.864 0~6.912 μg（r=0.999 5）、418.0~3 344 μg（r=0.999 1）、18.21~145.6 μg（r=0.999 6）、422.1~3 376 μg（r=0.999 7）、204.2~1 633 μg（r=0.999 1）的范围内呈良好的线性关系；平均回收率（n=6）分别为102.5%、99.60%、101.4%、100.1%和99.61%，其RSD分别为1.5%、2.1%、1.9%、0.34%和0.23%。17批酸橙枳壳中异柚皮苷含量均为0.01%，柚皮苷、橙皮苷、新橙皮苷和枳属苷的含量范围分别为5.73%~12.2%、0.10%~0.61%、2.18%~7.27%和1.03%~1.90%。9批枸橘枳壳中异柚皮苷含量均为0.02%；柚皮苷和枳属苷含量范围分别为1.90%~7.34%和1.11%~4.14%；部分枸橘枳壳中橙皮苷含量低于定量限，最高为0.40%；新橙皮苷含量均低于定量限。酸橙枳壳中柚皮苷和新橙皮苷含量与枸橘枳壳相比均较高，异柚皮苷、橙皮苷和枳属苷含量与枸橘枳壳相比均较低。结论：本法可用于酸橙枳壳和枸橘枳壳中异柚皮苷、柚皮苷、橙皮苷、新橙皮苷和枳属苷的含量测定。

Objective: To develop an HPLC method for simultaneous determination of narirutin,naringin,hesperidin,neohesperidin,and poncirin in Fructus Ponciri Trifoliatae and Fructus Aurantii.Methods: A Diamonsil C₁₈ column(150 mm×4.6 mm,5 μm) was adopted and the mobile phase consisted of acetonitrile(A) -phosphoric acid aqueous(B) with gradient elution(0-10 min,20%A;10-11 min,20%A→30%A;11-25 min,30%A) at the flow rate of 1.0 mL·min^-1.The detection wavelength was 283 nm and the column temperature was 25℃.Results: The contents of narirutin,naringin,hesperidin,neohesperidin,and poncirin had good linear relationship in the range of 0.864 0-6.912 μg(r=0.999 5),418.0-3 344 μg(r=0.999 1),18.21-145.6 μg(r=0.999 6),422.1-3 376 μg(r=0.999 7),and 204.2-1 633 μg(r=0.999 1),respectively.And the average recoveries(n=6) of the four components were 102.5% with RSD of 1.5%,99.60% with RSD of 2.1%,101.4% with RSD of 1.9%,100.1% with RSD of 0.34% and 99.61% with RSD of 0.23%,respectively.The content of narirutin,naringin,hesperidin,neohesperidin and poncirin were 0.01%,5.73%-12.2%,0.10%-0.61%,2.18%-7.27% and 1.03%-1.90% in 17 batches of Fructus Aurantii.The content of narirutin,naringin and poncirin were 0.02%,1.90%-7.34% and 1.11%-4.14% in Fructus Ponciri Trifoliatae.The contents of hesperidin in some Fructus Ponciri Trifoliatae samples were lower than the limit of quantification with the highest content of 0.40%.Contents of neohesperidin in all samples were lower than the limit of quantification.Contents of naringin and hesperidin in Fructus Aurantii were higher than those in Fructus Ponciri Trifoliatae while contents of narirutin,neohesperidin,and poncirin in Fructus Aurantii were lower than those in Fructus Ponciri Trifoliatae.Conclusion: The methold can be applied to the determination of narirutin,naringin,hesperidin,neohesperidin,and poncirin in Fructus Ponciri Trifoliatae and Fructus Aurantii.