基于UPLC-Q-Exactive-MS及多种技术研究威廉环毛蚓提取物化学成分组成

目的 对威廉环毛蚓 Pheretima guillelmi 提取物中化学成分和蛋白质、游离氨基酸、脂质、总糖、多种元素、水分、灰分及硫酸铵等成分进行质量分数测定。方法 采用 UPLC-Q-Exactive-MS 技术探究威廉环毛蚓提取物化学组成；采用仲裁法——凯氏定氮法对蛋白质质量分数进行测定；采用索氏抽提法测定脂质的质量分数，采用皂化及甲基化法测定各脂肪酸的质量分数；采用 ICP-MS 及 ICP-OES 技术测定 23 种元素的质量分数；采用《中国药典》2020 年版中烘干法和总灰分测定法测定水分和总灰分的质量分数；采用离子色谱法测定硫酸铵的质量分数。结果 共鉴定出 54 种化学物质，主要为氨基酸类、核苷类及有机酸类等成分；蛋白质的质量分数为 53.01%，游离氨基酸为 4.18%；脂质为 5.62%，其中总脂肪酸质量分数为 1.23%；Mo、Se、As、V、Mn、Co、Ni、Pb、Sn、Cd、Cr、Cu、Li、B、Fe、Zn、Al、K、Ca、Mg、Na、Hg、Si 等元 素 质 量 分 数 分 别 为 2.65、 4.88、 2.31、 1.12、 25.2、 8.32、 2.39、 1.59、 0.0572、 9.27、 43.7、 27.7、 0.642、 1.33、 759、185、245、614、3.28×10³、970、1.32×10³、0.191、433 mg?kg^-1；总糖质量分数为 11.42%；水分 4.05%、总灰分 2.36%；硫酸铵 9.89%。结论 基本确定威廉环毛蚓提取物中大部成分组成。

Probing chemical composition of Pheretima guillelmi extracts based on UPLCQ-Exactive-MS and various techniques

Objective To explore and study the chemical composition and the content of protein, free amino acid, lipid, total sugar, many elements, moisture, ash and ammonium sulphate in the extract of Pheretima guillelmi. Clarify its main material composition and promote the development and application of its preparations. Methods UPLC-Q-Exactive-MS technique was used to investigate the chemical composition of the extract of P. guillelmi; Kjeldahl nitrogen determination was used as an arbitration method for the determination of proteins, Soxhlet extraction was used to determine the content of lipids, and saponification and methylation were used to determine the content of each fatty acid; ICP MS and ICP-OES techniques for the determination of 23 elements; Moisture and total ash content were determined using the drying method and total ash determination method of the Chinese Pharmacopoeia (2020 edition); And ion chromatography for the determination of ammonium sulfate.Results A total of 54 chemical substances were identified, which were categorized into amino acid, nucleoside and organic acid components, of which the content of protein was 53.01%, the content of lipids was 5.62%, the content of free amino acid was 4.18%, the total fatty acid content was 1.23%. Mo, Se, As, V, Mn, Co, Ni, Pb, Sn, Cd, Cr, Cu, Li, B, Fe, Zn, Al, K, Ca, Mg, Na, Hg, and Si were 2.65, 4.88, 2.31, 1.12, 25.2, 8.32, 2.39, 1.59, 0.057 2, 9.27, 43.7, 27.7, 0.642, 1.33, 759, 185, 245, 614, 3.28×10³, 970, 1.32×10³, 0.191, and 433 mg·kg^-1. Total sugar content was 11.42%, moisture was 4.05%, total ash was 2.36%, and ammonium sulfate content was 9.89%. Conclusion The contents of most of the components in the extract of William''s ring worm were basically determined.