北京生物醫(yī)學(xué)工程

基于UPLC-MS/MS的人血漿膽固醇酯分子檢測方法及其在冠心病患者中的應(yīng)用

UPLC-MS/MS based measurement for human plasma cholesteryl esters quantification and its application in coronary heart disease

作者：王宇欣李曉璐陳旭謝佳宏蔣宏峰

單位：北京市心肺血管疾病研究所（北京 100029）<br />首都醫(yī)科大學(xué)附屬北京安貞醫(yī)院（北京 100029）<br />通信作者：蔣宏峰。E-mail：[email protected]

關(guān)鍵詞：質(zhì)譜檢測膽固醇酯冠心病急性心梗生物樣本

分類號：R318 

出版年·卷·期（頁碼）：2023·42·2（138-143）

摘要：

目的使用高通量超高效液相色譜-串聯(lián)質(zhì)譜（Ultra-high performance liquid chromatography-tandem mass spectrometry，UPLC-MS/MS）建立人血漿中各膽固醇酯分子的測量方法，并對患者血漿樣本實(shí)現(xiàn)定量分析。方法使用UPLC-MS/MS，從基質(zhì)效應(yīng)、檢測限、回收率、精確度、準(zhǔn)確度等方法學(xué)評價(jià)指標(biāo)對13種膽固醇酯標(biāo)準(zhǔn)品進(jìn)行方法學(xué)驗(yàn)證，同時(shí)收集來自首都醫(yī)科大學(xué)附屬北京安貞醫(yī)院的穩(wěn)定型冠心病患者、急性心梗患者與體檢中心對照人群血樣各50例，檢測血漿中13種膽固醇酯的的分子水平，比較各組結(jié)果差異。結(jié)果通過對膽固醇酯標(biāo)準(zhǔn)品的方法學(xué)驗(yàn)證，本檢測方案達(dá)到定量檢測方法學(xué)驗(yàn)證要求。對人血樣檢測結(jié)果顯示，膽固醇酯CE 22:0、CE 22:5、CE 20:5、CE 18:0、CE 18:3等分子與冠心病的發(fā)生發(fā)展相關(guān)。結(jié)論基于UPLC-MS/MS高通量、高靈敏度和高特異性的檢測特性，本文建立的膽固醇酯分子檢測方法能夠?qū)ρ獫{中多種膽固醇酯分子實(shí)現(xiàn)良好的分離和檢測，為臨床精準(zhǔn)測量不同膽固醇酯分子提供了方法學(xué)參考。

Objective Using high throughput ultra high performance liquid chromatography-tandem mass spectrometry (UPLC/MS/MS) to establish and quantify the cholesteryl ester species in human plasma. Methods Using UPLC/MS/MS to verify method's detection and quantification limitation, matrix effect, recovery rate, accuracy and precision. We collected serum from the patients with stable coronary artery disease or acute myocardial infarction, and from control group from health screening center visitors. Plasma cholesteryl ester levels were measured and compared. Results The method was successfully validated and tested for plasma cholesteryl esters determination. Plasma cholesterol esters CE 22:0, CE 22:5, CE 20:5, CE 18:0 and CE 18:3 were associated with disease occurrence and development in the patient. Conclusions By using a UPLC-MS/MS approach of high throughput, sensitivity and specificity, we set up a method for fine separation and detection of plasma cholesteryl esters, which potentially provides a quantification protocol of plasma cholesteryl ester species for clinical lab test.

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