Development and validation of a RPLC-MS/MS method for the quantification of ceramides in human serum
Development and validation of a RPLC-MS/MS method for the quantification of ceramides in human serum
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DOI：
10.1016/j.jchromb.2021.122734
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发表时间：
2021-05-12
影响因子：
3
通讯作者：
Gika, Helen G.
中科院分区：
文献类型：
--
作者：
Begou, Olga Angeliki;Deda, Olga;Gika, Helen G.
神经酰胺是参与许多中央细胞过程的关键作用脂质。大量研究表明，血液循环中的神经酰胺水平与不同的疾病状态有关，例如2型糖尿病、心血管疾病、卵巢癌、多发性硬化症等。本文建立并验证了人血清中神经酰胺Cer（d18：1/16：0）、Cer（d18：1/18：0）、Cer（d18：1/24：0）和Cer（d18：1/ 24：1）的RPLC-MS/MS快速定量方法。不同的样品制备策略，包括SLE，LLE和QuECheRS进行了测试，目的是实现有效，准确和重现性测定血清样品中的神经酰胺。所有神经酰胺的日内和日间准确度分别为80.0-111%和87.8- 106%，而日内和日间精密度分别为0.05%-10.2%RSD和2.2%-14.0%RSD。Cer（d18：1/ 16：0）和Cer（d18：1/18：0）的定量下限为2.3 ng/mL，Cer（d18：1/24：0）和Cer（d18：1/24：1）为1.4 ng/mL。根据生物分析方法指南对该方法进行了评价，并用于测定冠心病患者的血清神经酰胺，以评价其在临床分析中的实用性。
Ceramides are key-role lipids involved in numerous central cellular processes. A plethora of studies have demonstrated that the levels of ceramides in blood circulation are related to different disease states, such as type 2 diabetes, cardiovascular diseases, ovarian cancer, multiple sclerosis and others. Herein, a RPLC-MS/MS method for the rapid quantification of ceramides Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:0) and Cer(d18:1/ 24:1) in human blood serum was developed and validated. Different sample preparation strategies including SLE, LLE and QuECheRS were tested with the aim to attain effective, accurate and reproducible determination of ceramides in serum samples. Intra and inter-day accuracy were found to be between 80.0-111% and 87.8-106%, respectively, for all ceramides, while intra and inter-day precision were found to vary from 0.05% to 10.2% % RSD and 2.2% to 14.0% %RSD, respectively. The lower limits of quantification were 2.3 ng/mL for Cer(d18:1/ 16:0) and Cer(d18:1/18:0) and 1.4 ng/mL for Cer(d18:1/24:0) and Cer(d18:1/24:1). The method was evaluated in accordance to bioanalytical method guidelines and was used for the determination of serum ceramides of patients with coronary artery disease to evaluate its utility in clinical analyses.