超高效液相色谱法同时测定丹参酮提取物中隐丹参酮、丹参酮Ⅰ和丹参酮Ⅱ_A

目的：建立超高效液相色谱法同时测定丹参酮提取物中特征性的3个脂溶性成分（隐丹参酮、丹参酮Ⅰ和丹参酮Ⅱ_A）含量。方法：采用Waters Acquity UPLC BEH Shield RP18色谱柱（1.7 μm，2.1 mm×100 mm），预柱为Phenomenex SecurityGuard C₁₈ Cartridges Polar-RP（4 mm×3.0 mm），以乙腈（A）-5%乙腈水溶液（含0.1%甲酸，pH 2.5）（B）为流动相，梯度洗脱（0~5 min，52%A→70%A；5~7 min，70%A→80%A；7~10 min，80%A→52%A；10~12 min，52%A），流速0.25 mL·min^-1，柱温35 ℃，进样量5 μL，检测波长269 nm。结果：隐丹参酮、丹参酮Ⅰ和丹参酮Ⅱ_A 3个被测成分在线性范围内均具有良好的线性关系（r>0.999）；平均回收率（n=6）分别为96.7%（RSD=1.3%），96.4%（RSD=0.94%），98.5%（RSD=1.2%）；丹参酮提取物中主峰与其他共存组分达到基线分离。结论：与常规液相色谱法比较，本方法分离效率高，重复性好，可作为实验室方法改进的参考依据，为丹参酮提取物的质量标准的提升做准备。

Objective: To establish an ultra-performance liquid chromatography method for simultaneous determination of three representative tanshinones (cryptotanshinone,tanshinone Ⅰ and tanshinone Ⅱ_A) in ethanol extract of Salviae Miltiorrhizae Radix et Rhizoma. Methods: The method was performed on a Waters Acquity UPLC BEH Shield RP18 (1.7 μm,2.1 mm×100 mm) analytical column employed with a guard column of Phenomenex SecurityGuard C₁₈ Cartridges Polar-RP (4 mm×3.0 mm) at the column temperature of 35 ℃.The gradient mobile phase consisted of acetonitrile(A)-5% acetonitrile aqueous solution containing 0.1% formic acid (pH 2.5) (B) with a flow rate of 0.25 mL·min^-1(0-5 min,52%A→70%A;5-7 min,70%A→80%A;7-10 min,80%A→52%A;10-12 min,52%A).The injection volume was 5 μL.The UV detection wavelength was set at 269 nm. Results: The relationship between the concentration and the peak area of cryptotanshinone,tanshinone Ⅰ and tanshinone Ⅱ_A was all in good linearity (r>0.999) over the linear ranges.The average recoveries (n=6) of the investigated compounds were 96.7% (RSD=1.3%),96.4% (RSD=0.94%) and 98.5% (RSD=1.2%);the chromatographic peaks of investigated compounds and coexisting components were well isolated. Conclusion: Compared with routine liquid chromatography,this method with high separation efficiency and good repeatability can be used as a reference for improvement of laboratory methods and make preparation for improvement of the quality standard of tanshinone extract.