Simultaneous quantification of acylcarnitine isomers containing dicarboxylic acylcarnitines in human serum and urine by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry

Yasuhiro Maeda, Tetsuya Ito, Atsuko Suzuki, Yukihisa Kurono, Akihito Ueta, Kyoko Yokoi, Satoshi Sumi, Hajime Togari, Naruji Sugiyama

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Tandem mass spectrometry (MS/MS) has become a prominent method for screening newborns for diseases such as organic acidemia and fatty acid oxidation defects, although current methods cannot separate acylcarnitine isomers. Accurate determination of dicarboxylic acylcarnitines such as methylmalonylcarnitine and glutarylcarnitine has not been carried out, because obtaining standards of these acylcarnitines is difficult. We attempted the individual determinations of acylcarnitines with isomers and dicarboxylic acylcarnitines by applying high-performance liquid chromatography (HPLC). Chromatographic separation was performed by gradient elution using a mixture of 0.08% aqueous ion-pairing agent and acetonitrile as the mobile phase. Mass transitions of mlz 161.8→84.8 for carnitine and m/z 164.8→84.8 for deuterated carnitine were monitored in positive ion electrospray ionization mode. One carnitine and 16 acylcarnitines were quantified. The limit of quantitation (LOQ) was 0.1 μmol/L for methylmalonylcarnitine and 0.05 μmol/L for the other acylcarnitines. Intra-day and inter-day coefficients of variance (CVs) were <8.3% and <8.8%, respectively, for all acylcarnitines in serum, and both were <9.2% in urine. Mean recoveries were >90% for all acylcarnitines. Human samples were quantified by this method. After addition of deuterated acylcarnitines as internal standards, acylcarnitines in serum or urine were extracted using a solid-phase extraction cartridge. In healthy adult individuals, isobutyryl-, 2-methylbutyryl- and isovalerylcarnitine were detected in serum and urine. Dicarboxylic acylcarnitines were detected in urine. High concentrations of methylmalonylcarnitine and propionylcarnitine were found in both the serum and the urine of a patient with methylmalonic acidemia. The described HPLC/MS/MS method could separate most acylcarnitine isomers and quantify them, potentially allowing detailed diagnoses and follow-up treatment for those diseases.

Original languageEnglish
Pages (from-to)799-806
Number of pages8
JournalRapid Communications in Mass Spectrometry
Volume21
Issue number5
DOIs
Publication statusPublished - 07-03-2007
Externally publishedYes

Fingerprint

Electrospray ionization
High performance liquid chromatography
Isomers
Mass spectrometry
Carnitine
propionylcarnitine
acylcarnitine
Organic acids
Screening
Fatty Acids

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Spectroscopy
  • Organic Chemistry

Cite this

Maeda, Yasuhiro ; Ito, Tetsuya ; Suzuki, Atsuko ; Kurono, Yukihisa ; Ueta, Akihito ; Yokoi, Kyoko ; Sumi, Satoshi ; Togari, Hajime ; Sugiyama, Naruji. / Simultaneous quantification of acylcarnitine isomers containing dicarboxylic acylcarnitines in human serum and urine by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry. In: Rapid Communications in Mass Spectrometry. 2007 ; Vol. 21, No. 5. pp. 799-806.
@article{520fd0ba4c2d43e286a08ec24bd5c343,
title = "Simultaneous quantification of acylcarnitine isomers containing dicarboxylic acylcarnitines in human serum and urine by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry",
abstract = "Tandem mass spectrometry (MS/MS) has become a prominent method for screening newborns for diseases such as organic acidemia and fatty acid oxidation defects, although current methods cannot separate acylcarnitine isomers. Accurate determination of dicarboxylic acylcarnitines such as methylmalonylcarnitine and glutarylcarnitine has not been carried out, because obtaining standards of these acylcarnitines is difficult. We attempted the individual determinations of acylcarnitines with isomers and dicarboxylic acylcarnitines by applying high-performance liquid chromatography (HPLC). Chromatographic separation was performed by gradient elution using a mixture of 0.08{\%} aqueous ion-pairing agent and acetonitrile as the mobile phase. Mass transitions of mlz 161.8→84.8 for carnitine and m/z 164.8→84.8 for deuterated carnitine were monitored in positive ion electrospray ionization mode. One carnitine and 16 acylcarnitines were quantified. The limit of quantitation (LOQ) was 0.1 μmol/L for methylmalonylcarnitine and 0.05 μmol/L for the other acylcarnitines. Intra-day and inter-day coefficients of variance (CVs) were <8.3{\%} and <8.8{\%}, respectively, for all acylcarnitines in serum, and both were <9.2{\%} in urine. Mean recoveries were >90{\%} for all acylcarnitines. Human samples were quantified by this method. After addition of deuterated acylcarnitines as internal standards, acylcarnitines in serum or urine were extracted using a solid-phase extraction cartridge. In healthy adult individuals, isobutyryl-, 2-methylbutyryl- and isovalerylcarnitine were detected in serum and urine. Dicarboxylic acylcarnitines were detected in urine. High concentrations of methylmalonylcarnitine and propionylcarnitine were found in both the serum and the urine of a patient with methylmalonic acidemia. The described HPLC/MS/MS method could separate most acylcarnitine isomers and quantify them, potentially allowing detailed diagnoses and follow-up treatment for those diseases.",
author = "Yasuhiro Maeda and Tetsuya Ito and Atsuko Suzuki and Yukihisa Kurono and Akihito Ueta and Kyoko Yokoi and Satoshi Sumi and Hajime Togari and Naruji Sugiyama",
year = "2007",
month = "3",
day = "7",
doi = "10.1002/rcm.2905",
language = "English",
volume = "21",
pages = "799--806",
journal = "Rapid Communications in Mass Spectrometry",
issn = "0951-4198",
publisher = "John Wiley and Sons Ltd",
number = "5",

}

Simultaneous quantification of acylcarnitine isomers containing dicarboxylic acylcarnitines in human serum and urine by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry. / Maeda, Yasuhiro; Ito, Tetsuya; Suzuki, Atsuko; Kurono, Yukihisa; Ueta, Akihito; Yokoi, Kyoko; Sumi, Satoshi; Togari, Hajime; Sugiyama, Naruji.

In: Rapid Communications in Mass Spectrometry, Vol. 21, No. 5, 07.03.2007, p. 799-806.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Simultaneous quantification of acylcarnitine isomers containing dicarboxylic acylcarnitines in human serum and urine by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry

AU - Maeda, Yasuhiro

AU - Ito, Tetsuya

AU - Suzuki, Atsuko

AU - Kurono, Yukihisa

AU - Ueta, Akihito

AU - Yokoi, Kyoko

AU - Sumi, Satoshi

AU - Togari, Hajime

AU - Sugiyama, Naruji

PY - 2007/3/7

Y1 - 2007/3/7

N2 - Tandem mass spectrometry (MS/MS) has become a prominent method for screening newborns for diseases such as organic acidemia and fatty acid oxidation defects, although current methods cannot separate acylcarnitine isomers. Accurate determination of dicarboxylic acylcarnitines such as methylmalonylcarnitine and glutarylcarnitine has not been carried out, because obtaining standards of these acylcarnitines is difficult. We attempted the individual determinations of acylcarnitines with isomers and dicarboxylic acylcarnitines by applying high-performance liquid chromatography (HPLC). Chromatographic separation was performed by gradient elution using a mixture of 0.08% aqueous ion-pairing agent and acetonitrile as the mobile phase. Mass transitions of mlz 161.8→84.8 for carnitine and m/z 164.8→84.8 for deuterated carnitine were monitored in positive ion electrospray ionization mode. One carnitine and 16 acylcarnitines were quantified. The limit of quantitation (LOQ) was 0.1 μmol/L for methylmalonylcarnitine and 0.05 μmol/L for the other acylcarnitines. Intra-day and inter-day coefficients of variance (CVs) were <8.3% and <8.8%, respectively, for all acylcarnitines in serum, and both were <9.2% in urine. Mean recoveries were >90% for all acylcarnitines. Human samples were quantified by this method. After addition of deuterated acylcarnitines as internal standards, acylcarnitines in serum or urine were extracted using a solid-phase extraction cartridge. In healthy adult individuals, isobutyryl-, 2-methylbutyryl- and isovalerylcarnitine were detected in serum and urine. Dicarboxylic acylcarnitines were detected in urine. High concentrations of methylmalonylcarnitine and propionylcarnitine were found in both the serum and the urine of a patient with methylmalonic acidemia. The described HPLC/MS/MS method could separate most acylcarnitine isomers and quantify them, potentially allowing detailed diagnoses and follow-up treatment for those diseases.

AB - Tandem mass spectrometry (MS/MS) has become a prominent method for screening newborns for diseases such as organic acidemia and fatty acid oxidation defects, although current methods cannot separate acylcarnitine isomers. Accurate determination of dicarboxylic acylcarnitines such as methylmalonylcarnitine and glutarylcarnitine has not been carried out, because obtaining standards of these acylcarnitines is difficult. We attempted the individual determinations of acylcarnitines with isomers and dicarboxylic acylcarnitines by applying high-performance liquid chromatography (HPLC). Chromatographic separation was performed by gradient elution using a mixture of 0.08% aqueous ion-pairing agent and acetonitrile as the mobile phase. Mass transitions of mlz 161.8→84.8 for carnitine and m/z 164.8→84.8 for deuterated carnitine were monitored in positive ion electrospray ionization mode. One carnitine and 16 acylcarnitines were quantified. The limit of quantitation (LOQ) was 0.1 μmol/L for methylmalonylcarnitine and 0.05 μmol/L for the other acylcarnitines. Intra-day and inter-day coefficients of variance (CVs) were <8.3% and <8.8%, respectively, for all acylcarnitines in serum, and both were <9.2% in urine. Mean recoveries were >90% for all acylcarnitines. Human samples were quantified by this method. After addition of deuterated acylcarnitines as internal standards, acylcarnitines in serum or urine were extracted using a solid-phase extraction cartridge. In healthy adult individuals, isobutyryl-, 2-methylbutyryl- and isovalerylcarnitine were detected in serum and urine. Dicarboxylic acylcarnitines were detected in urine. High concentrations of methylmalonylcarnitine and propionylcarnitine were found in both the serum and the urine of a patient with methylmalonic acidemia. The described HPLC/MS/MS method could separate most acylcarnitine isomers and quantify them, potentially allowing detailed diagnoses and follow-up treatment for those diseases.

UR - http://www.scopus.com/inward/record.url?scp=33847391557&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33847391557&partnerID=8YFLogxK

U2 - 10.1002/rcm.2905

DO - 10.1002/rcm.2905

M3 - Article

VL - 21

SP - 799

EP - 806

JO - Rapid Communications in Mass Spectrometry

JF - Rapid Communications in Mass Spectrometry

SN - 0951-4198

IS - 5

ER -