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目的:利用DNA条形码技术对中药紫草进行基源鉴定,并建立紫草的功效多组分含量测定方法,为全面评价紫草药材质量提供依据。方法:对不同产地紫草进行DNA条形码基原鉴定;采用高效液相色谱法测定紫草中萘醌类成分与含量,Thermo BDS Hypersil C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相0.2%冰乙酸-乙腈梯度洗脱,流速0.8 m L·min~(-1),检测波长275 nm,柱温35℃。结果:经DNA条形码鉴定了11批药材,均为紫草科植物新疆紫草Arnebia euchroma的干燥根;测定了11批新疆紫草的8种萘醌类功效组分含量,各组分均显示良好分离;不同成分在线性范围内线性关系良好,平均回收率在95.26%~101.40%,RSD均在0.7%~1.8%(n=6)。结论:DNA条形码鉴定确保了新疆紫草药材基源的准确性,组分含量测定方法准确可靠、重复性好,二者结合可作为紫草的质量控制方法。
OBJECTIVE: To identify the origin of Lithospermum bungei using DNA barcoding technique and to establish the multi-component method for the determination of Lithospermum phaeoides. It provides a basis for the comprehensive evaluation of Lithospermum herbs. Methods: The DNA of barnyard grass was identified by DNA barcoding. The components and contents of naphthoquinones in Lithospermum officinale were determined by high performance liquid chromatography (HPLC) on a Thermo BDS Hypersil C 18 column (4.6 mm × 250 mm, 5 μm) The mobile phase was eluted with a gradient of 0.2% glacial acetic acid - acetonitrile at a flow rate of 0.8 mL · L -1. The detection wavelength was set at 275 nm and the column temperature was 35 ℃. Results: 11 batches of medicinal materials were identified by DNA barcoding, all of them were the dried roots of Arnebia euchroma. The contents of functional components of 8 kinds of naphthoquinones in 11 batches of Xinjiang Lithospermum were determined, and all the components showed good The average recoveries ranged from 95.26% to 101.40% with RSDs ranging from 0.7% to 1.8% (n = 6). Conclusion: The identification of DNA barcodes ensures the accuracy of Radix Gentii Medicinal Herbs, the determination of components is accurate and reproducible, and the combination of the two can be used as the quality control method of Lithospermum.