HPLC波长切换法同时测定抗感颗粒中7个成分的含量

目的: 建立HPLC法同时测定抗感颗粒中7个成分的含量。方法: 采用Kromasil 100-5C₁₈色谱柱(250 mm×4.6 mm,5 μm),以乙腈(A)-0.4%磷酸溶液(B)为流动相,梯度洗脱(0~14 min,8%A→16%A;14~22 min,16%A;22~23 min,16%A→24%A;23~38 min,24%A;38~39 min,24%A→8%A),流速1.0 mL·min^-1,柱温10 ℃,波长切换[0~22 min,在327 nm波长下检测3-O-咖啡酰奎宁酸、绿原酸(5-O-咖啡酰奎宁酸)、4-O-咖啡酰奎宁酸;22~26 min,在230 nm波长下检测芍药苷;26~38 min,在327 nm波长下检测4,5-O-二咖啡酰奎宁酸、3,5-O-二咖啡酰奎宁酸、3,4-O-二咖啡酰奎宁酸]。结果: 3-O-咖啡酰奎宁酸、5-O-咖啡酰奎宁酸、4-O-咖啡酰奎宁酸、芍药苷、4,5-O-二咖啡酰奎宁酸、3,5-O-二咖啡酰奎宁酸、3,4-O-二咖啡酰奎宁酸进样量分别在0.010 7~0.214 6、0.044 9~0.898 0、0.010 0~0.199 6、0.099 9~1.998 6、0.010 3~0.206 2、0.010 6~0.211 4、0.010 1~0.201 4 μg范围内与色谱峰峰面积呈良好线性响应;平均回收率(n=6)分别为101.6%、102.0%、101.1%、101.6%、102.0%、100.6%、102.1%,RSD分别为0.9%、0.9%、0.6%、0.8%、1.5%、1.0%、0.5%;10批样品中上述7个成分质量分数分别为0.040~0.640、0.157~1.639、0.061~0.850、6.519~9.225、0.010~0.339、0.021~0.353、0.046~0.635 mg·g^-1。结论: 该方法对多种成分进行了同步测定,准确可靠,重复性好,可用于抗感颗粒的质量控制。

Objective: To develop an HPLC method for simultaneous determination of seven components in Kanggan granules.Methods: The determination was carried out with Kromasil 100-5C₁₈ column(250 mm×4.6 mm, 5 μm).The mobile phase consisted of acetonitrile(A)-0.4% phosphoric acid solution(B)with gradient elution(0-14 min, 8%A→16%A;14-22 min, 16%A;22-23 min, 16%A→24%A;23-38 min, 24%A;38–39 min, 24%A→8%A)at a flow rate of 1.0 mL·min^-1 at 10 ℃.The detection wavelengths were set at 327 nm in 0-22 min to determine 3-O-caffeoyl quinic acid, chlorogenic acid(5-O-caffeoyl quinic acid), and 4-O-caffeoyl quinic acid;230 nm in 22-26 min to determine paeoniflorin;and 327 nm in 26-38 min to determine 4, 5-O-dicaffeoyl quinic acid, 3, 5-O-dicaffeoyl quinic acid, and 3, 4-O-dicaffeoyl quinic acid.Results: The linear ranges of 3-O-caffeoyl quinic acid, 5-O-caffeoyl quinic acid, 4-O-caffeoyl quinic acid, paeoniflorin, 4, 5-O-dicaffeoyl quinic acid, 3, 5-O-dicaffeoyl quinic acid, and 3, 4-O-dicaffeoyl quinic acid fell within the ranges of 0.010 7-0.214 6μg, 0.044 9-0.898 0 μg, 0.010 0- 0.199 6 μg, 0.099 9-1.998 6 μg, 0.010 3-0.206 2 μg, 0.010 6-0.211 4 μg, and 0.010 1-0.201 4 μg, respectively.The average recoveries(n=6)were 101.6%, 102.0%, 101.1%, 101.6%, 102.0%, 100.6%, and 102.1%;the relative standard deviations were 0.9%, 0.9%, 0.6%, 0.8%, 1.5%, 1.0%, and 0.5%, respectively.The content ranges of seven aforementioned components in ten samples were 0.040-0.640 mg·g^-1, 0.157-1.639 mg·g^-1, 0.061-0.850 mg·g^-1, 6.519-9.225 mg·g^-1, 0.010-0.339 mg·g^-1, 0.021-0.353 mg·g^-1, 0.046-0.635 mg·g^-1, respectively.Conclusion: The established method for the determination of multi-components is accurate, reliable, and reproducible, which can be applied to the quality control of Kanggan granules.