HPLC波长切换联合梯度洗脱法同时测定荆防败毒丸中10个成分的含

目的：HPLC 波长切换联合梯度洗脱法同时测定荆防败毒丸中的5-O-咖啡酰莽草酸、落新妇苷、花旗松素、黄杞苷、白藜芦醇、洋川芎内酯H、洋川芎内酯I、洋川芎内酯A、瑟丹酸内酯和藁本内酯含量。方法：采用Shiseido C₁₈ 色谱柱（4.6 mm×250 mm，5 μm），以甲醇-乙腈（3：1）（A）与0.1% 冰醋酸水溶液（B）为流动相，梯度洗脱，流速1.2 mL·min^-1，柱温30 ℃，5-O-咖啡酰莽草酸、落新妇苷、花旗松素、黄杞苷和白藜芦醇的检测波长为291 nm，洋川芎内酯H、洋川芎内酯I、洋川芎内酯A、瑟丹酸内酯和藁本内酯的检测波长为280 nm。结果：5-O-咖啡酰莽草酸、落新妇苷、花旗松素、黄杞苷、白藜芦醇、洋川芎内酯H、洋川芎内酯I、洋川芎内酯A、瑟丹酸内酯和藁本内酯10 个成分的线性范围分别为9.13~182.60μg·mL^-1（r=0.999 4）、20.75~415.00 μg·mL^-1（r=0.999 9）、3.54~70.80 μg·mL^-1（r=0.999 5）、4.78~95.60μg·mL^-1（r=0.999 8）、2.42~48.40 μg·mL^-1（r=0.999 3）、3.38~67.60 μg·mL^-1（r=0.999 9）、4.94~98.80μg·mL^-1（r=0.999 6）、6.36~127.20 μg·mL^-1（r=0.999 8）、3.07~61.40 μg·mL^-1（r=0.999 7）、9.66~193.20μg·mL^-1（r=0.999 5）；平均加样回收率（n=6）分别为97.3%（RSD=1.1%）、99.2%（RSD=1.3%）、96.8%（RSD=0.44%）、98.8%（RSD=0.86%）、96.9%（RSD=1.1%）、98.3%（RSD=1.6%）、97.1%（RSD=1.2%）、100.0%（RSD=0.60%）、97.9%（RSD=1.3%）、99.2%（RSD=1.1%）。3 批样品中上述10 个成分的含量范围分别为0.542~0.579、1.519~1.663、0.105~0.117、0.314~0.331、0.072~0.081、0.101~0.113、0.164~0.183、0.406~0.437、0.163~0.182、0.585~0.607 mg·g^-1。结论：本文建立的HPLC 波长切换联合梯度洗脱法可为荆防败毒丸质量标准的提高提供依据。

Objective: To develop an HPLC method with wavelength switching and gradient elution for simultaneous determination of 5-O-caffeoylshikimic acid,astilbin,taxifolin,engeletin,resveratrol,senkyunolide H, senkyunolide I,senkyunolide A,sedanolide and ligustilide in Jingfang Baidu pills. Methods: The Shiseido C₁₈ column(4.6 mm×250 mm,5 μm)was adopted,the mobile phase was methanol-acetonitrile(3:1)(A)and 0.1% acetic acid glacial(B)with gradient elution at a flow rate of 1.2 mL·min^-1,the column temperature was set at 30℃. 5-O-caffeoylshikimic acid,astilbin,taxifolin,engeletin and resveratrol were detected at 291 nm; senkyunolide H,senkyunolide I,senkyunolide A,sedanolide and ligustilide were detected at 280 nm. Results: The linear ranges of 5-O-caffeoylshikimic acid,astilbin,taxifolin,engeletin,resveratrol,senkyunolide H, senkyunolide I,senkyunolide A,sedanolide and ligustilide were 9.13-182.60 μg·mL^-1(r=0.999 4),20.75-415.00 μg·mL^-1(r=0.999 9),3.54-70.80 μg·mL^-1(r=0.999 5),4.78-95.60 μg·mL^-1(r=0.999 8), 2.42-48.40 μg·mL^-1(r=0.999 3),3.38-67.60 μg·mL^-1(r=0.999 9),4.94-98.80 μg·mL^-1(r=0.999 6), 6.36-127.20 μg·mL^-1(r=0.999 8),3.07-61.40 μg·mL^-1(r=0.999 7),9.66-193.20 μg·mL^-1(r=0.999 5), respectively. The average recoveries(n=6)were 97.3%(RSD=1.1%),99.2%(RSD=1.3%),96.8% (RSD=0.44%),98.8%(RSD=0.86%),96.9%(RSD=1.1%),98.3%(RSD=1.6%),97.1%(RSD=1.2%), 100.0%(RSD=0.60%),97.9%(RSD=1.3%),99.2%(RSD=1.1%),respectively. The contents of ten components in 3 batches of samples were 0.542-0.579,1.519-1.663,0.105-0.117,0.314-0.331,0.072-0.081,0.101-0.113, 0.164-0.183,0.406-0.437,0.163-0.182,0.585-0.607 mg·g^-1,respectively. Conclusion: HPLC method with wavelength switching and gradient elution can be used for the quality control of Jingfang Baidu pills.

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