Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept Validation
Gas chromatography-mass spectrometry has been widely used to analyze hundreds of organic acids in urine to provide a diagnostic basis for organic acidemia. However, it is difficult to operate in clinical laboratories on a daily basis due to sample pretreatment processing. Therefore, we aimed to deve...
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MDPI AG
2021-11-01
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| Series: | Diagnostics |
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| author | Yasushi Ueyanagi Daiki Setoyama Daisuke Kawakami Yuichi Mushimoto Shinya Matsumoto Taeko Hotta Dongchon Kang |
| author_facet | Yasushi Ueyanagi Daiki Setoyama Daisuke Kawakami Yuichi Mushimoto Shinya Matsumoto Taeko Hotta Dongchon Kang |
| author_sort | Yasushi Ueyanagi |
| collection | DOAJ |
| description | Gas chromatography-mass spectrometry has been widely used to analyze hundreds of organic acids in urine to provide a diagnostic basis for organic acidemia. However, it is difficult to operate in clinical laboratories on a daily basis due to sample pretreatment processing. Therefore, we aimed to develop a fully automated system for quantifying serum organic acids using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The pretreatment CLAM-2030 device was connected to an LC-MS/MS system for processing serum under optimized conditions, which included derivatizing serum organic acids using 3-Nitrophenylhydrazine. The derivatized organic acids were separated on a reverse-phase Sceptor HD-C column and detected using negative-ion electrospray ionization multiple reaction monitoring MS. The automated pretreatment-LC-MS/MS system processed serum in less than 1 h and analyzed 19 serum organic acids, which are used to detect organic acidemias. The system exhibited high quantitative sensitivity ranging from approximately 2 to 100 µM with a measurement reproducibility of 10.4% CV. Moreover, a proof-of-concept validation of the system was performed using sera from patients with propionic acidemia (<i>n</i> = 5), methylmalonic acidemia (<i>n</i> = 2), and 3-methylcrotonylglycinuria (<i>n</i> = 1). The levels of marker organic acids specific to each disease were significantly elevated in the sera of the patients compared to those in control samples. The automated pretreatment-LC-MS/MS system can be used as a rapid in-hospital system to measure organic acid levels in serum for the diagnosis of organic acidemias. |
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| last_indexed | 2024-03-10T04:20:26Z |
| publishDate | 2021-11-01 |
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| series | Diagnostics |
| spelling | doaj.art-e66379ba4e364341934bd0129d7b5ebe2023-11-23T07:52:43ZengMDPI AGDiagnostics2075-44182021-11-011112219510.3390/diagnostics11122195Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept ValidationYasushi Ueyanagi0Daiki Setoyama1Daisuke Kawakami2Yuichi Mushimoto3Shinya Matsumoto4Taeko Hotta5Dongchon Kang6Department of Clinical Chemistry and Laboratory Medicine, Kyushu University Hospital, Fukuoka 812-8582, JapanDepartment of Clinical Chemistry and Laboratory Medicine, Kyushu University Hospital, Fukuoka 812-8582, JapanShimadzu Corporation, Kyoto 604-8511, JapanDepartment of Pediatrics, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, JapanDepartment of Clinical Chemistry and Laboratory Medicine, Kyushu University Hospital, Fukuoka 812-8582, JapanDepartment of Clinical Chemistry and Laboratory Medicine, Kyushu University Hospital, Fukuoka 812-8582, JapanDepartment of Clinical Chemistry and Laboratory Medicine, Kyushu University Hospital, Fukuoka 812-8582, JapanGas chromatography-mass spectrometry has been widely used to analyze hundreds of organic acids in urine to provide a diagnostic basis for organic acidemia. However, it is difficult to operate in clinical laboratories on a daily basis due to sample pretreatment processing. Therefore, we aimed to develop a fully automated system for quantifying serum organic acids using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The pretreatment CLAM-2030 device was connected to an LC-MS/MS system for processing serum under optimized conditions, which included derivatizing serum organic acids using 3-Nitrophenylhydrazine. The derivatized organic acids were separated on a reverse-phase Sceptor HD-C column and detected using negative-ion electrospray ionization multiple reaction monitoring MS. The automated pretreatment-LC-MS/MS system processed serum in less than 1 h and analyzed 19 serum organic acids, which are used to detect organic acidemias. The system exhibited high quantitative sensitivity ranging from approximately 2 to 100 µM with a measurement reproducibility of 10.4% CV. Moreover, a proof-of-concept validation of the system was performed using sera from patients with propionic acidemia (<i>n</i> = 5), methylmalonic acidemia (<i>n</i> = 2), and 3-methylcrotonylglycinuria (<i>n</i> = 1). The levels of marker organic acids specific to each disease were significantly elevated in the sera of the patients compared to those in control samples. The automated pretreatment-LC-MS/MS system can be used as a rapid in-hospital system to measure organic acid levels in serum for the diagnosis of organic acidemias.https://www.mdpi.com/2075-4418/11/12/2195liquid chromatography-mass spectrometryautomated sample preparationorganic acid analysis3-Nitrophenylhydrazineorganic acid disorders |
| spellingShingle | Yasushi Ueyanagi Daiki Setoyama Daisuke Kawakami Yuichi Mushimoto Shinya Matsumoto Taeko Hotta Dongchon Kang Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept Validation Diagnostics liquid chromatography-mass spectrometry automated sample preparation organic acid analysis 3-Nitrophenylhydrazine organic acid disorders |
| title | Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept Validation |
| title_full | Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept Validation |
| title_fullStr | Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept Validation |
| title_full_unstemmed | Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept Validation |
| title_short | Fully Automated Quantitative Measurement of Serum Organic Acids via LC-MS/MS for the Diagnosis of Organic Acidemias: Establishment of an Automation System and a Proof-of-Concept Validation |
| title_sort | fully automated quantitative measurement of serum organic acids via lc ms ms for the diagnosis of organic acidemias establishment of an automation system and a proof of concept validation |
| topic | liquid chromatography-mass spectrometry automated sample preparation organic acid analysis 3-Nitrophenylhydrazine organic acid disorders |
| url | https://www.mdpi.com/2075-4418/11/12/2195 |
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