UPE-HILIC-DAD-ESI-TOF/MS法测定山东产区丹参中丹酚酸类成分

立超高压提取-亲水色谱-二极管阵列检测器-电喷雾飞行时间质谱法（UPE-HILIC-DAD-ESI-TOF/MS）测定丹参中丹酚酸类成分的方法，并用于山东产区丹参中丹酚酸类成分的测定。采用单因素试验对超高压提取条件进行了系统优化；采用Waters XBridge Amide色谱柱进行分析，以水溶液（0.8%甲酸、20 mmol/L甲酸铵、30%乙腈、10%正丁醇）-乙腈（0.8%甲酸）为流动相体系，梯度洗脱，流速为0.8 mL/min，柱温为25 ℃，检测波长为280 nm；采用DAD-ESI-TOF/MS对各化合物进行鉴别，采用HILIC-DAD对各化合物进行定量测定。在优化的提取条件下，超高压提取法的提取率明显优于超声波辅助提取法和水浴回流提取法；采用HILIC法可以实现丹参中丹酚酸类成分的较好分离；通过ESI-TOF/MS获得的精确分子量信息和DAD获得的紫外吸收信息结合标准对照品可以对各化合物进行准确鉴别；对7种丹酚酸类成分（丹酚酸A、迷迭香酸、原儿茶醛、原儿茶酸、熊果酸、紫草酸、丹酚酸B）进行了含量测定，结果表明各化合物的峰面积与其质量浓度呈良好的线性关系（r=0.999 0～1.000 0），各化合物的浓度范围分别为0.311~0.438、1.218~1.517、0.104~0.147、0.375~0.527、0.303~0.401、4.025~4.935、35.628~44.255 mg/g。该方法提取效率高、分离度好，为丹参中丹酚酸类成分提取、测定和质量评价研究提供了技术支持。

Determination of salvianolic acids in Salvia miltiorrhiza Bge. from Shandong region by UPE-HILIC-DAD-ESI-TOF/MS

A method involving ultrahigh pressure extraction-hydrophilic interaction chromatography-diode array detector-electrospray time-of-flight mass spectrometry (UPE-HILIC-DAD-ESI-TOF/MS) was developed in this study for determining the content of salvianolic acids in Salvia miltiorrhiza Bge.(Danshen) from Shandong region. The UPE conditions were optimized in detail through a single-factor experiment; analysis was performed based on a Waters XBridge Amide column with water solution (0.8% formic acid, 20 mmol/L ammonium formate, 30% acetonitrile, and 10% n-butanol)-acetonitrile (0.8% formic acid) as mobile phase, gradient elution, flow rate 0.8 mL/min, column temperature at 25°C, and detection wavelength 280 nm; various compounds were identified by DAD-ESI-TOF/MS and quantified by HILIC-DAD. Under optimized extraction conditions, the extraction rate of UPE method was significantly higher than that of ultrasonic assisted extraction and reflux extraction method; desirable separation effect could be achieved by HILIC method for the salvianolic acids in Salvia miltiorrhiza Bge. In combination with standard reference materials, compounds could be accurately identified using precise molecular weight information obtained by ESI-TOF/MS and ultraviolet absorption information acquired by DAD. Seven types of salvianolic acids (including salvianolic acid A, rosmarinic acid, protocatechuic aldehyde acid, ursolic acid, purple oxalic acid, and salvianolic acid B) were determined. The results showed that there was a good linear relationship between peak area and mass concentration of each compound(r=0.999 0~1.000 0), the concentration range of each compound were 0.311~0.438, 1.218~1.517, 0.104~0.147, 0.375~0.527, 0.303~0.401, 4.025~4.935, and 35.628~44.255 mg/g. This method features high extraction efficiency and good degree of separation, providing technical evidence for extraction, determination, and quality evaluation of salvianolic acids from Danshen.