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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 C_(18)色谱柱(250 mm×4.6 mm,5μm),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1.0mL·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4.512~1.444×10~2μg·mL~(-1)(r=0.999 9)、14.162~4.532×10~2μg·mL~(-1)(r=1.0000)、2.538~0.812×10~2μg·mL~(-1)(r=0.999 9)、3.275~1.048×10~2μg·mL~(-1)(r=0.999 9)、2.638~0.844×10~2μg·mL~(-1)(r=0.999 9)、5.912~1.892×10~2μg·mL~(-1)(r=0.999 9)、8.162~2.612×10~2μg·mL~(-1)(r=0.999 9)、30.288~9.692×10~2μg·mL~(-1)(r=0.999 9)、3.288~1.052×10~2μg·mL~(-1)(r=0.999 9)、5.812~1.860×10~2μg·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。
OBJECTIVE: To establish a method for the simultaneous determination of neo-chlorogenic acid, chlorogenic acid, cryptocerbic acid, caffeic acid, iso-chlorogenic acid B, and iso-chlorogenic acid in Huabaytang by HPLC-DAD with wavelength- The original acid A, chlorogenic acid C, baicalin, forsythin and arctiin active ingredients of 10 quantitative analysis methods. Methods: A SHIMADZU Inserts Cain 18 column (250 mm × 4.6 mm, 5 μm) was used. The mobile phase consisted of gradient elution of acetonitrile and 0.2% phosphoric acid. The detection wavelength was 327 nm (0 ~ 37 min, Chlorogenic acid, chlorogenic acid, caffeic acid, isooctanoic acid B, iso-chlorogenic acid A and iso-chlorogenic acid C) at 278 nm (37-41.3 min for the detection of baicalin), 228 nm (41.3 ~ 50 min, detection of forsythin and arctiin), the flow rate of 1.0mL · min -1, the column temperature was 35 ℃. Results: All the constituents were detected effectively and the separation was good. Chlorogenic acid, chlorogenic acid, cryptocholic acid, caffeic acid, chlorogenic acid B, isocrochloric acid A, isocrochloric acid C, baicalin , The concentrations of forsythin and arctiin were 4.512-1.444 × 10-2 μg · mL -1 (r = 0.999 9), 14.162 ~ 4.532 × 10-2 μg · mL -1 (r = 0.999 9), 2.638 ~ 0.812 × 10 ~ 2μg · mL -1 (r = 0.999 9), 3.275 ~ 1.048 × 10 ~ 2μg · mL -1 ~ 0.844 × 10 ~ 2μg · mL -1 (r = 0.999 9), 5.912 ~ 1.892 × 10 ~ 2μg · mL -1 (r = 0.999 9), 8.162 ~ 2.612 × 10 ~ 2μg · mL (R = 0.999 9), 30.288 ~ 9.692 × 10 ~ 2μg · mL -1 (r = 0.999 9), 3.288 ~ 1.052 × 10 ~ 2μg · mL -1 (r = 0.999 9), 5.812 ~ 1.860 × 10 ~ 2μg · mL -1 (r = 0.999 9), and the peak area was good. The recoveries of n = 6 were 99.1% 99.5% (RSD = 1.4%), 100.3% (RSD = 1.0%), 100.0% (RSD = (RSD = 2.0%), 99.7% (RSD = 1.5%), 99.6% (RSD = 1.5%) and 99.7% (RSD = 1.9%). Conclusion: HPLC-DAD wavelength-switching method can be used to determine the contents of 10 active ingredients in Huabian Decoction simultaneously. The method is simple, reliable and reproducible. It provides a scientific basis for the quality control and evaluation of Huabian Tang.