目的: 建立多波长切换HPLC法同时测定五加皮多指标质量评价方法。方法: 采用Waters AtlantisTMT3色谱柱(250 mm×4.6 mm,5 μm),流动相为乙腈(A)-0.1%磷酸水溶液(B),梯度洗脱(0~15 min,10%A→20%A;15~25 min,20%A→40%A;25~30 min,40%A→85%A;30~45 min,85%A),流速1.0 mL·min-1,柱温35 ℃,检测波长为277 nm(0~30 min,检测新绿原酸、松柏苷、紫丁香苷、绿原酸、隐绿原酸、咖啡酸、1,3-二咖啡酰奎宁酸、异绿原酸A、异绿原酸C)、202 nm(30~45 min,检测异贝壳杉烯酸),进样量为20 μL。结果: 在优化的色谱条件下,新绿原酸、松柏苷、紫丁香苷、绿原酸、隐绿原酸、咖啡酸、1,3-二咖啡酰奎宁酸、异绿原酸A、异绿原酸C、异贝壳杉烯酸的质量浓度分别在1~100 μg·mL-1(r=1.000)、0.5~50 μg·mL-1(r=1.000)、0.25~25 μg·mL-1(r=1.000)、3.75~375 μg·mL-1(r=0.999 9)、0.25~25 μg·mL-1(r=0.999 9)、0.25~25 μg·mL-1(r=0.999 9)、0.25~25 μg·mL-1(r=1.000)、2~200 μg·mL-1(r=0.999 8)、0.5~50 μg·mL-1(r=0.999 9)、14.45~1 445 μg·mL-1(r=0.999 8)范围内均呈现良好的线性关系,精密度、重复性和稳定性的RSD均<3%,加样回收率(n=6)为90.4%~104.0%,RSD为0.4%~2.4%。11批样品中上述成分含量范围分别为0.03~0.19,0.06~0.87、0.02~0.48、0.16~4.51、0.04~0.20、0.04~0.13、0.02~0.45、0.04~3.03、0.22~1.03、5.26~21.18 mg·g-1。结论: 所建立的五加皮多指标质量评价方法快速、准确,有较好的重现性和稳定性,可用于五加皮的质量评价研究。
Objective: To establish a multi-index quality evaluation method for Acanthopanacis Cortex based on multi-wavelength switching HPLC. Methods: A Waters AtlantisTM T3(250 mm×4.6 mm, 5 μm)column was used in the HPLC assay. The mobile phase was acetonitrile(A)-0.1% phosphoric acid aqueous solution(B) by gradient elution(0-5 min, 10%A→20%A, 15-25 min, 20%A→40%A, 25-30 min, 40%A→85%A, 30-45 min, 85%A). The flow rate was 1.0 mL·min-1, the column temperature was maintained at 35 ℃ and the detection wavelengths were set at 270 nm (0-30 min) for neochlorogenic acid, abinoside, syringin, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 1,3-dicaffeioylquinic acid, isochlorogenic acid A, isochlorogenic acid C and 202 nm (30-45 min) for kaurenic acid. The injection volume was 20 μL. Results: Under the above conditions, neochlorogenic acid, abinoside, syringin, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 1, 3-dicaffeioylquinic acid, isochlorogenic acid A, isochlorogenic acid C and kaurenic acid showed good linearity in the ranges of 1-100 μg·mL-1(r=1.000), 0.5-50 μg·mL-1(r=1.000), 0.25-25 μg·mL-1(r=1.000), 3.75-375 μg·mL-1(r=0.999 9), 0.25-25 μg·mL-1(r=0.999 9), 0.25-25 μg·mL-1(r=0.999 9), 0.25-25 μg·mL-1(r=1.000), 2-200 μg·mL-1(r=0.999 8), 0.5-50 μg·mL-1(r=0.999 9), 14.45-1 445 μg·mL-1(r=0.999 8). The RSDs of precision, repeatability and stability were all less than 3.0%. The average recoveries (n=6) were 90.4%-104.0% with the RSD value of 0.4%-2.4%. The content ranges of the above-mentioned components in 11 samples were 0.03-0.19, 0.06-0.87, 0.02-0.48, 0.16-4.51, 0.04-0.20, 0.04-0.13, 0.02-0.45, 0.04-3.03, 0.22-1.03 and 5.26-21.18 mg·g-1, respectively. Conclusion: The multi-index quality evaluation method of Acanthopanacis Cortex is simple, accurate, reproducible and reliable, which can be used to evaluate the quality of it.
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