可靠的LC-MS/MS法测定干血斑中甲基丙二酸、甲基柠檬酸、丙二酸、乙基丙二酸和总同型半胱氨酸的建立和验证

IF 2.1 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinical biochemistry Pub Date : 2025-09-13 DOI:10.1016/j.clinbiochem.2025.111013

Chunyan Zhang , Nan Bai , Yanling Yang , Yuxuan Cheng , Xiyu He , Meng Wang , Honghui Zhou , Yaping Tian

{"title":"可靠的LC-MS/MS法测定干血斑中甲基丙二酸、甲基柠檬酸、丙二酸、乙基丙二酸和总同型半胱氨酸的建立和验证","authors":"Chunyan Zhang , Nan Bai , Yanling Yang , Yuxuan Cheng , Xiyu He , Meng Wang , Honghui Zhou , Yaping Tian","doi":"10.1016/j.clinbiochem.2025.111013","DOIUrl":null,"url":null,"abstract":"<div><h3>Objectives</h3><div>Metabolic disorders in newborns can cause significant morbidity and mortality if not diagnosed and managed early. In this study, we developed and validated a reliable LC-MS/MS method to simultaneously quantify methylmalonic acid, methylcitric acid, malonic acid, ethylmalonic acid, and total homocysteine associated with metabolic disorders.</div></div><div><h3>Design and methods</h3><div>According to Clinical Laboratory Standards Institute (CLSI) guidelines, various analytical performances (i.e., precision, linearity, accuracy and reference intervals) were rigorously evaluated. Meanwhile, sample preparation, chromatographic separation and mass spectrometric detection and incorporating stable isotope-labeled internal standards were optimized. Furthermore, calibrators and quality controls were developed to ensure standardization and traceability.</div></div><div><h3>Results</h3><div>The coefficient of variations for precision were less than 10.0 %. Meanwhile, the results showed high accuracy (recoveries: 94.57 %–109.60 %) and robust linearity (R<sup>2</sup> > 0.9935) over clinically relevant ranges. The limits of detection and limits of quantification for all analytes were established, enabling sensitive detection of pathological elevations. Reference intervals for pediatric populations (ages 0–1 month and 2 months to 18 years) were determined, providing essential baselines for clinical interpretation.</div></div><div><h3>Conclusions</h3><div>The newly developed method demonstrated good analytical performance and offers a standardized, high-throughput solution for clinical laboratories to improve early diagnosis and personalized management of metabolic diseases.</div></div>","PeriodicalId":10172,"journal":{"name":"Clinical biochemistry","volume":"140 ","pages":"Article 111013"},"PeriodicalIF":2.1000,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Reliable LC-MS/MS method development and validation for the determination of methylmalonic acid, methylcitric acid, malonic acid, ethylmalonic acid, and total homocysteine in dried blood spots\",\"authors\":\"Chunyan Zhang , Nan Bai , Yanling Yang , Yuxuan Cheng , Xiyu He , Meng Wang , Honghui Zhou , Yaping Tian\",\"doi\":\"10.1016/j.clinbiochem.2025.111013\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objectives</h3><div>Metabolic disorders in newborns can cause significant morbidity and mortality if not diagnosed and managed early. In this study, we developed and validated a reliable LC-MS/MS method to simultaneously quantify methylmalonic acid, methylcitric acid, malonic acid, ethylmalonic acid, and total homocysteine associated with metabolic disorders.</div></div><div><h3>Design and methods</h3><div>According to Clinical Laboratory Standards Institute (CLSI) guidelines, various analytical performances (i.e., precision, linearity, accuracy and reference intervals) were rigorously evaluated. Meanwhile, sample preparation, chromatographic separation and mass spectrometric detection and incorporating stable isotope-labeled internal standards were optimized. Furthermore, calibrators and quality controls were developed to ensure standardization and traceability.</div></div><div><h3>Results</h3><div>The coefficient of variations for precision were less than 10.0 %. Meanwhile, the results showed high accuracy (recoveries: 94.57 %–109.60 %) and robust linearity (R<sup>2</sup> > 0.9935) over clinically relevant ranges. The limits of detection and limits of quantification for all analytes were established, enabling sensitive detection of pathological elevations. Reference intervals for pediatric populations (ages 0–1 month and 2 months to 18 years) were determined, providing essential baselines for clinical interpretation.</div></div><div><h3>Conclusions</h3><div>The newly developed method demonstrated good analytical performance and offers a standardized, high-throughput solution for clinical laboratories to improve early diagnosis and personalized management of metabolic diseases.</div></div>\",\"PeriodicalId\":10172,\"journal\":{\"name\":\"Clinical biochemistry\",\"volume\":\"140 \",\"pages\":\"Article 111013\"},\"PeriodicalIF\":2.1000,\"publicationDate\":\"2025-09-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Clinical biochemistry\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0009912025001420\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MEDICAL LABORATORY TECHNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Clinical biochemistry","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0009912025001420","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MEDICAL LABORATORY TECHNOLOGY","Score":null,"Total":0}

引用次数: 0

摘要

目的新生儿代谢紊乱如不及早诊断和治疗，可导致严重的发病率和死亡率。在这项研究中，我们开发并验证了一种可靠的LC-MS/MS方法，可以同时定量与代谢紊乱相关的甲基丙二酸、甲基柠檬酸、丙二酸、乙基丙二酸和总同型半胱氨酸。设计和方法根据临床实验室标准协会（CLSI）的指南，严格评估各种分析性能（即精密度、线性度、准确度和参考区间）。同时，对样品制备、色谱分离、质谱检测及稳定同位素标记内标进行了优化。此外，校准器和质量控制的发展，以确保标准化和可追溯性。结果精密度的变异系数小于10.0%。同时，结果在临床相关范围内具有较高的准确度（回收率为94.57% ~ 109.60%）和良好的线性关系（R2 > 0.9935）。建立了所有分析物的检出限和定量限，使病理升高的敏感检测成为可能。确定了儿科人群（0-1个月和2个月至18岁）的参考区间，为临床解释提供了必要的基线。结论该方法具有良好的分析性能，为临床实验室提高代谢性疾病的早期诊断和个性化管理提供了标准化、高通量的解决方案。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

Reliable LC-MS/MS method development and validation for the determination of methylmalonic acid, methylcitric acid, malonic acid, ethylmalonic acid, and total homocysteine in dried blood spots

Objectives

Metabolic disorders in newborns can cause significant morbidity and mortality if not diagnosed and managed early. In this study, we developed and validated a reliable LC-MS/MS method to simultaneously quantify methylmalonic acid, methylcitric acid, malonic acid, ethylmalonic acid, and total homocysteine associated with metabolic disorders.

Design and methods

According to Clinical Laboratory Standards Institute (CLSI) guidelines, various analytical performances (i.e., precision, linearity, accuracy and reference intervals) were rigorously evaluated. Meanwhile, sample preparation, chromatographic separation and mass spectrometric detection and incorporating stable isotope-labeled internal standards were optimized. Furthermore, calibrators and quality controls were developed to ensure standardization and traceability.

Results

The coefficient of variations for precision were less than 10.0 %. Meanwhile, the results showed high accuracy (recoveries: 94.57 %–109.60 %) and robust linearity (R² > 0.9935) over clinically relevant ranges. The limits of detection and limits of quantification for all analytes were established, enabling sensitive detection of pathological elevations. Reference intervals for pediatric populations (ages 0–1 month and 2 months to 18 years) were determined, providing essential baselines for clinical interpretation.

Conclusions

The newly developed method demonstrated good analytical performance and offers a standardized, high-throughput solution for clinical laboratories to improve early diagnosis and personalized management of metabolic diseases.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Clinical biochemistry 医学-医学实验技术

CiteScore

5.10

自引率

0.00%

发文量

151

审稿时长

25 days

期刊介绍： Clinical Biochemistry publishes articles relating to clinical chemistry, molecular biology and genetics, therapeutic drug monitoring and toxicology, laboratory immunology and laboratory medicine in general, with the focus on analytical and clinical investigation of laboratory tests in humans used for diagnosis, prognosis, treatment and therapy, and monitoring of disease.