枸杞子芦丁微波提取-高效液相含量测定法研究

The HPLC Assay for Content Determination of Rutoside in the Extracts of Wolfberry from Microwave Extraction

目的建立一种微波提取-反相高效液相测定枸杞子芦丁含量的方法。方法色谱条件:Dikma Kromasil C18色谱柱(4.6mm×200mm,5μm),以甲醇-0.3%磷酸(37∶63)为流动相,采用等度洗脱的方式,流速为1ml.min-1,检测波长为257nm,柱温为25℃,进样量为5μl;用回流提取、超声提取、微波提取三种方法前处理样品,考察它们对枸杞芦丁含量测定的影响;对枸杞芦丁微波提取-反相高效液相色谱含量测定方法进行了方法学考察。结果芦丁的线性范围为0.004～0.11mg/ml(相关系数为0.9998),平均回收率(n=5)为101.48%,不同产地枸杞子样品中的芦丁含量存在着较大的差异,真伪枸杞子样品中的芦丁含量存在着极大的差异。结论该方法准确可靠,重现性好,适用于测定枸杞子中芦丁的含量,为枸杞子的质量标准研究提供了实验数据。

Objective To develop a HPLC assay for the content determination of rutoside in the extracts of wolfberry from mi crowave extraction. Methods HPLC method： Column： Dikma Kromasil C 18 （4.6 mm ×200 mm, 5μm）. The column temperature was mainrained at 25 ℃. Mobile phase： methanol-0.3% phosphoric acid （37 ： 63）, with flow rate 1 ml ·min^-1. UV detector wavelength： 257 nm. Inject volume： 5μL. The established HPLC assay for the quantitative determination ofrutoside was fully evaluated. Then, three different extraction methods including reflux extraction, ultrasonic extraction, and microwave extraction were applied to evaluate the impaction of different extraction methods on the content determination of rutoside. Results The HPLC assay for the rutoside was linear within 0.004 - 0.11 mg/ml （RZ=-0.9998）. The average recovery was 101.48 % （n=5）. Significant difference in the rutoside content was identified among the wolfberry from different producing areas. Extremely high difference in the rutoside content was identified between the real and fake wolfberry. Conclusion In current research, an accurate and reproducible HPLC method is developed and evaluated to determine the content of rutoside in the extracts of wolfberry. Current research provides valuable data for the establishment of the wolfberry quality standard.