Pallerla Pavankumar, Ragi Nagarjunachary, Gari Aravind Reddy Babi Reddy, Bhumireddy Sudarshana Reddy, Addipilli Ramunaidu, Rodda Ramesh, Yadla Manjusha, Sripadi Prabhakar
Centre for Mass Spectrometry, Department of Analytical & Structural Chemistry, CSIR-Indian Institute of Chemical Technology, Hyderabad, 500007, Telangana, India.
Academy of Scientific and Innovative Research, Ghaziabad, 201002, India.
Anal Bioanal Chem. 2023 Nov;415(26):6491-6509. doi: 10.1007/s00216-023-04926-x. Epub 2023 Sep 27.
End-stage renal disease (ESRD) is a rapidly increasing health problem, and every year, about 2 million ESRD cases are reported worldwide. Hemodialysis (HD) is the vital renal reinstatement therapy for ESRD, and HD patterns play a crucial role in patients' health. Plasma metabolomics is the potential approach to understanding the HD process, effectiveness, and patterns. The lack of protein vitality is a primary problem for HD patients, and the quantities of amino acids intracellularly and in the blood are considered to be a symbolic index of protein metabolism and nutrition conditions. In the current study, LC/MS/MS and GC/MS methods were developed for 29 targeted plasma metabolites and validated as per ICH bioanalytical method validation M10 guidelines. The 29 metabolites included 20 proteinogenic amino acids and nine other related metabolites. The methods were employed to measure the absolute quantities (µM) of the targeted metabolites in HD patients (n=60) before and after dialysis (PRE-HD and POST-HD), and compared with the healthy control (HC) group (n=60). Phenylacetylglutamine was found to be higher in both PRE-HD (72.88±14.5 µM) and POST-HD (26.62±7.9 µM), when compared to HC (1.61±0.6 µM). On the other hand, glutamic acid was lower in PRE-HD (14.90±6.5 µM), and POST-HD (13.6±6.1 µM) than that of HC (245.4±37.8 µM). The dialytic loss was found to be 52-45% for arginine, lysine, and histidine, while it was 38-26% for glycine, cysteine, proline, alanine, threonine, glutamine, valine, and methionine. The dialytic loss was low (≤12%) for aspartic acid, glutamic acid, asparagine, leucine, tyrosine, tryptophan, and isoleucine. Graphical abstract adapted from mass spectrometry templates by Biorender.com retrieved from https://app.biorender.com/biorender-templates .
终末期肾病（ESRD）是一个迅速加剧的健康问题，全球每年约报告200万例ESRD病例。血液透析（HD）是ESRD至关重要的肾脏恢复治疗方法，HD模式对患者健康起着关键作用。血浆代谢组学是了解HD过程、效果和模式的潜在方法。蛋白质活力不足是HD患者的一个主要问题，细胞内和血液中的氨基酸数量被视为蛋白质代谢和营养状况的标志性指标。在本研究中，针对29种靶向血浆代谢物开发了液相色谱/串联质谱（LC/MS/MS）和气相色谱/质谱（GC/MS）方法，并根据国际人用药品注册技术协调会（ICH）生物分析方法验证M10指南进行了验证。这29种代谢物包括20种蛋白质氨基酸和9种其他相关代谢物。这些方法用于测量HD患者（n = 60）透析前（HD前）和透析后（HD后）靶向代谢物的绝对量（µM），并与健康对照组（HC，n = 60）进行比较。发现苯乙酰谷氨酰胺在HD前（72.88±14.5 µM）和HD后（26.62±7.9 µM）均高于HC组（1.61±0.6 µM）。另一方面，谷氨酸在HD前（14.90±6.5 µM）和HD后（13.6±6.1 µM）均低于HC组（245.4±37.8 µM）。精氨酸、赖氨酸和组氨酸的透析损失为52 - 45%，而甘氨酸、半胱氨酸、脯氨酸、丙氨酸、苏氨酸、谷氨酰胺、缬氨酸和蛋氨酸的透析损失为38 - 26%。天冬氨酸、谷氨酸、天冬酰胺、亮氨酸、酪氨酸、色氨酸和异亮氨酸的透析损失较低（≤12%）。图形摘要改编自Biorender.com的质谱模板，从https://app.biorender.com/biorender-templates获取。
