HPLC-DAD波长切换法同时测定化扁汤中10种有效成分的含量

目的：采用高效液相色谱-二极管阵列检测（HPLC-DAD）波长切换法建立同时测定化扁汤中新绿原酸、绿原酸、隐绿原酸、咖啡酸、异绿原酸B、异绿原酸A、异绿原酸C、黄芩苷、连翘苷和牛蒡苷10种有效成分的定量分析方法。方法：采用SHIMADZU Insertsustain C₁₈色谱柱（250 mm×4.6 mm，5 μm），以流动相乙腈-0.2%磷酸溶液梯度洗脱，DAD检测器，检测波长为327 nm（0~37 min，检测新绿原酸、绿原酸、隐绿原酸、咖啡酸、异绿原酸B、异绿原酸A、异绿原酸C）、278 nm（37~41.3 min，检测黄芩苷）、228 nm（41.3~50 min，检测连翘苷和牛蒡苷），流速1.0 mL·min^-1，柱温为35℃。结果：各成分均能有效检出且分离度良好；新绿原酸、绿原酸、隐绿原酸、咖啡酸、异绿原酸B、异绿原酸A、异绿原酸C、黄芩苷、连翘苷、牛蒡苷10种成分的进样浓度分别在4.512~1.444×10² μg·mL^-1（r=0.999 9）、14.162~4.532×10² μg·mL^-1（r=1.000 0）、2.538~0.812×10² μg·mL^-1（r=0.999 9）、3.275~1.048×10² μg·mL^-1（r=0.999 9）、2.638~0.844×10² μg·mL^-1（r=0.999 9）、5.912~1.892×10² μg·mL^-1（r=0.999 9）、8.162~2.612×10² μg·mL^-1（r=0.999 9）、30.288~9.692×10² μg·mL^-1（r=0.999 9）、3.288~1.052×10² μg·mL^-1（r=0.999 9）、5.812~1.860×10² μg·mL^-1（r=0.999 9）与峰面积呈良好的线性关系；加样回收率（n=6）分别为99.1%（RSD=1.1%）、99.6%（RSD=0.9%）、100.2%（RSD=1.5%）、99.8%（RSD=1.6%）、99.5%（RSD=1.4%）、100.3%（RSD=1.0%）、100.0%（RSD=2.0%）、99.7%（RSD=1.5%）、99.6%（RSD=1.5%）、99.7%（RSD=1.9%）。结论：利用HPLC-DAD波长切换法可同时测定化扁汤中10种有效成分的含量，该法简便可靠，重现性好，为化扁汤的质量控制与评价提供科学依据。

Simultaneous determination of ten active components in Huabian decotion by HPLC-DAD method

OBJECTIVE To establish a simultaneous quantitative analysis method to determine 10 active components (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlomgenic acid B, isochlomgenic acid A, isochlomgenic acid C, baicalin, phillyrin, arctiin) in Huabian decotion by HPLC-DAD method.METHODS SHIMADZU Insertsustain C₁₈ (250 mm×4.6 mm,5 μm) was chosen, gradient elution was conducted with acetonitrile-0.2% phosphoric acid as mobile phase, DAD detection wavelength at 327 nm (0-37 min for neochlorogenic acid, chlorogenic acid, 4-dicaffeoylquinic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C), 278 nm (37-41.3 min for baicalin), 228 nm (41.3-50 min for phillyrin and arctiin), flow rate at 1.0 mL·min^-1, column temperature at 35℃.RESULTS The ingredients could be effectively detected and separated. For neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlomgenic acid B, isochlomgenic acid A, isochlomgenic acid C, baicalin, phillyrin and arctiin, the linear ranges were 4.512-1.444×10² μg·mL^-1(r=0.999 9), 14.162-4.532×10² μg·mL^-1 (r=1.000 0), 2.538-0.812×10² μg·mL^-1 (r=0.999 9), 3.275-1.048×10² μg·mL^-1 (r=0.999 9), 2.638-0.844×10² μg·mL^-1 (r=0.999 9), 5.912-1.892×10² μg·mL^-1 (r=0.999 9), 8.162-2.612×10² μg·mL^-1 (r=0.999 9), 30.288-9.692×10² μg·mL^-1 (r=0.999 9), 3.288-1.052×10² μg·mL^-1 (r=0.999 9), 5.812-1.860×10² μg·mL^-1 (r=0.999 9), respectively. The average recoveries (n=6) of the ten components were 99.1% (RSD=1.1%), 99.6% (RSD=0.9%), 100.2% (RSD=1.5%), 100.3% (RSD=1.0%), 99.7% (RSD=1.5%), 99.6% (RSD=1.5%), 99.7% (RSD=1.9%).CONCLUSION This method determines 10 active components in Huabian decotion by switching HPLC-DAD wavelength, which is simple and reproducible, and can provide a scientific basis for quality control and evaluation of Huabian decotion.