忍冬藤中咖啡酰奎宁酸葡萄糖苷的分离及含量测定

Isolation and Content Determination of Caffeoylquinic Acid Glucoside in Lonicerae Japonicae Caulis

目的：忍冬藤中5-O-[4＇-O-（β-D-吡喃葡萄糖基）咖啡酰基]奎宁酸的分离鉴定及HPLC含量测定方法的建立。方法：采用色谱分离方法从忍冬藤50%甲醇提取物中分离得到5-O-[4＇-O-（β-D-吡喃葡萄糖基）咖啡酰基]奎宁酸,并通过核磁共振波谱和质谱数据确定其结构。含量测定色谱条件：Dikma Technologies PLATISILTMODS C18色谱柱（4.6 mm×250 mm,5μm）;流动相乙腈（A）-0.4%冰乙酸溶液（B）梯度洗脱（0~20 min,8%~15%A）;流速1.0 m L·min^-1;检测波长327 nm;柱温35℃。结果：5-O-[4＇-O-（β-D-吡喃葡萄糖基）咖啡酰基]奎宁酸在0.15~3.07μg与峰面积呈良好的线性关系,其线性回归方程为Y=1.521 3X-3.777 7（r=1.000 0）。供试品溶液在24 h内稳定性良好,平均加样回收率（n=6）为95.95%,RSD1.5%。对收集的8批忍冬藤药材进行测定,5-O-[4＇-O-（β-D-吡喃葡萄糖基）咖啡酰基]奎宁酸质量分数1.737~7.390 mg·g-1。结论：首次从忍冬藤中分离鉴定了5-O-[4＇-O-（β-D-吡喃葡萄糖基）咖啡酰基]奎宁酸,报道了其完整核磁数据,建立了忍冬藤中该成分的含量测定方法,并设立了合理的含量限度值。建立的含量测定方法准确、可靠,可用于忍冬藤药材的质量控制。

Objective： To isolate and identify 5-O-（4＇-O-glucosylcaffeoyl） quinic acid from Lonicerae Japonicae Caulis, and develop a high performance liquid chromatography（HPLC） method for its content determination. Method： 5-O-（4＇-O-glucosylcaffeoyl） quinic acid was isolated from 50% methanol extract of Lonicerae Japonicae Caulis by chromatographic methods, and its structure was identified by nuclear magnetic resonance（NMR） spectrum and mass spectrum（MS） data. Dikma Technologies PLATISILTMODS C18 column（4. 6 mm × 250 mm,5 μm） was used with a mobile phase consisting of acetonitrile（A）-0. 4% acetic acid solution（B） for gradient elution（0-20 min,8%-15% A） at a flow rate of 1. 0 m L·min^-1. The detection wavelength was 327 nm and the column temperature was 35 ℃. Result： 5-O-（4＇-O-glucosylcaffeoyl） quinic acid showed a good linear relationship with peak area in the range of 0. 15-3. 07 μg,with the linear regression equation of Y = 1. 521 3 X-3. 777 7（r = 1. 000 0）; the test sample solution was stable within 24 h; the average recovery was 95. 95%（RSD 1. 5%,n = 6）. The content of 5-O-（4＇-O-glucosylcaffeoyl） quinic acid in eight batches of samples was in the range of 1. 737-7. 390 mg·g-1. Conclusion： The 5-O-（4＇-O-glucosylcaffeoyl） quinic acid was obtained from Lonicerae Japonicae Caulis for the first time. The complete NMR data of 5-O-（4＇-Oglucosylcaffeoyl） quinic acid was presented in this paper for the first time, and the method for its content determination and content limits in Lonicerae Japonicae Caulis were established. The established method is accurate and reliable,which could be applied for the quality control of Lonicerae Japonicae Caulis.