基于UHPLC-MS/MS法测定青海产黑枸杞中六种绿原酸单体含量

Determination of the Content of Six Types of Chlorogenic Acid Compounds in Lycium ruthenicum Murr from Qinghai Using UHPLC-MS/MS

摘要:

黑枸杞中含有丰富的多酚成分，绿原酸类成分是一类重要的酚酸，为测定青海黑枸杞中绿原酸类物质的含量，本研究采用Folin-Ciocalteu法测定黑枸杞中总多酚含量，运用pH示差法测定其总花青素含量，并利用超高效液相色谱串联质谱（UHPLC-MS/MS）质谱多反应检测方法（MRM）对黑枸杞中结构相似的六种绿原酸单体含量进行了测定。青海黑枸杞中总多酚含量为(4906.5±60.6)?10-2 mg GAE/g DW；总花色苷含量为(87.6±34.1)?10-2 mg CYG/g DW。通过UHPLC-MS/MS测得绿原酸含量最高，为902.97 μg/g、新绿原酸含量3.72 μg/g、隐绿原酸含量为81.78 μg/g、异绿原酸A含量为13.71 μg/g、异绿原酸B含量为1.10 μg/g、异绿原酸C含量为1.80 μg/g。实验结果表明黑枸杞中绿原酸成分种类较多且含量丰富，UHPLC-MS/MS法分析速度快、重复性好、结果准确，具有较好的线性关系（0.9989~0.9994），平均回收率为83.00%~103.89%，此方法可用于黑枸杞中绿原酸含量的测定及黑枸杞的品质控制。

Abstract:

Lycium ruthenicum Murr is rich in polyphenols, and chlorogenic acids are important phenolic acids. In order to determine the content of chlorogenic acids in Lycium ruthenicum Murr grown in Qinghai province, the Folin-Ciocalteu method was used to determine the total polyphenolic content, and the pH differential method was applied to determine the total anthocyanin content. The content of six types of chlorogenic acids with similar structure was determined with ultra-high performance liquid chromatography-tandem mass-spectrometry (UHPLC-MS/MS) by multiple reaction monitoring. The total polyphenolic content and the total anthocyanin content of Lycium ruthenicum Murr were (4906.5±60.6)?10-2 mg GAE/g DW and (87.6±34.1)?10-2 mg CYG/g DW, respectively. The UHPLC-MS/MS results showed that the highest chlorogenic acid content was 902.97 μg/g, and the content of neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A, isochlorogenic acid B, and isochlorogenic acid C was 3.72 μg/g, 81.78 μg/g, 13.71 μg/g, 1.10 μg/g, and 1.80 μg/g, respectively. The experimental results indicated that Lycium ruthenicum Murr contained numerous types of chlorogenic acids at high concentrations. The UHPLC-MS/MS method provided rapid measurement, good reproducibility, accurate results, and a good linear relationship (R2=0.9989~0.9994), and the average recovery was in the range of 83.00%~103.89%. This method can be applied in the determination of chlorogenic acid content and the quality control of Lycium ruthenicum Murr.