Semi-automated quantification of methylmalonic acid in human serum by LC-MS/MS
(2012) In Scandinavian Journal of Clinical & Laboratory Investigation 72(6). p.441-446- Abstract
- Background. Methylmalonic acid (MMA), a sensitive biomarker of functional vitamin B12 deficiency, is commonly determined by gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods using manual extraction and derivatization of MMA to reduce polarity prior to separation. Methods. In the present study we introduce a semi-automated extraction on a strong anion exchanger, HPLC separation on a BEH-amide column to separate serum MMA from its abundant isoform, succinic acid, followed by MS/MS detection and quantification. Results. The extraction of MMA plus internal standard provides full recovery and the method is linear between 0.03 mu mol/L and 20.0 mu mol/L (r(2) = 1.0) with intra-and... (More)
- Background. Methylmalonic acid (MMA), a sensitive biomarker of functional vitamin B12 deficiency, is commonly determined by gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods using manual extraction and derivatization of MMA to reduce polarity prior to separation. Methods. In the present study we introduce a semi-automated extraction on a strong anion exchanger, HPLC separation on a BEH-amide column to separate serum MMA from its abundant isoform, succinic acid, followed by MS/MS detection and quantification. Results. The extraction of MMA plus internal standard provides full recovery and the method is linear between 0.03 mu mol/L and 20.0 mu mol/L (r(2) = 1.0) with intra-and inter-assay imprecision of 2.2%. Agreement with other laboratories has been demonstrated in external proficiency testing. Compared to both conventional GC-MS and LC-MS/MS methods, the correlation is r(2) > 0.99. Conclusions. The use of robotic pipetting, elimination of derivatization and improved separation by the BEH-amide column combined with HILIC chromatographic conditions significantly improve sample throughput compared to conventional methods. Using a single pipetting robot and LC-MS/MS instrument, this method is currently performing 180 analyses per day from 10 regional hospitals and several additional distant sites. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/3190060
- author
- Nelson, Dick LU ; Xu, Ning LU and Carlson, Joyce LU
- organization
- publishing date
- 2012
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Chromatography, high performance liquid, tandem mass spectrometry, gas, chromatography-mass spectrometry, methylmalonic acid, vitamin B12
- in
- Scandinavian Journal of Clinical & Laboratory Investigation
- volume
- 72
- issue
- 6
- pages
- 441 - 446
- publisher
- Informa Healthcare
- external identifiers
-
- wos:000308941400001
- scopus:84866413491
- ISSN
- 1502-7686
- DOI
- 10.3109/00365513.2012.679963
- language
- English
- LU publication?
- yes
- id
- c3c52c62-c48e-4615-abd2-bf6850337e81 (old id 3190060)
- date added to LUP
- 2016-04-01 12:59:25
- date last changed
- 2022-01-27 08:40:30
@article{c3c52c62-c48e-4615-abd2-bf6850337e81,
abstract = {{Background. Methylmalonic acid (MMA), a sensitive biomarker of functional vitamin B12 deficiency, is commonly determined by gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods using manual extraction and derivatization of MMA to reduce polarity prior to separation. Methods. In the present study we introduce a semi-automated extraction on a strong anion exchanger, HPLC separation on a BEH-amide column to separate serum MMA from its abundant isoform, succinic acid, followed by MS/MS detection and quantification. Results. The extraction of MMA plus internal standard provides full recovery and the method is linear between 0.03 mu mol/L and 20.0 mu mol/L (r(2) = 1.0) with intra-and inter-assay imprecision of 2.2%. Agreement with other laboratories has been demonstrated in external proficiency testing. Compared to both conventional GC-MS and LC-MS/MS methods, the correlation is r(2) > 0.99. Conclusions. The use of robotic pipetting, elimination of derivatization and improved separation by the BEH-amide column combined with HILIC chromatographic conditions significantly improve sample throughput compared to conventional methods. Using a single pipetting robot and LC-MS/MS instrument, this method is currently performing 180 analyses per day from 10 regional hospitals and several additional distant sites.}},
author = {{Nelson, Dick and Xu, Ning and Carlson, Joyce}},
issn = {{1502-7686}},
keywords = {{Chromatography; high performance liquid; tandem mass spectrometry; gas; chromatography-mass spectrometry; methylmalonic acid; vitamin B12}},
language = {{eng}},
number = {{6}},
pages = {{441--446}},
publisher = {{Informa Healthcare}},
series = {{Scandinavian Journal of Clinical & Laboratory Investigation}},
title = {{Semi-automated quantification of methylmalonic acid in human serum by LC-MS/MS}},
url = {{https://lup.lub.lu.se/search/files/3091363/3737314.pdf}},
doi = {{10.3109/00365513.2012.679963}},
volume = {{72}},
year = {{2012}},
}