Analysis of catecholamines in urine by unique LC/MS suitable ion-pairing chromatography

Marianne L. Bergmann*, Seyed Sadjadi, Anne Schmedes

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Resumé

The catecholamines, epinephrine (E) and norepinephrine (NE) are small polar, hydrophilic molecules, posing significant challenges to liquid chromatography − tandem mass spectrometry (LC–MS/MS) method development. Specifically, these compounds show little retention on conventional reversed-phase liquid chromatography columns. This work presents development and validation of an LC–MS/MS method for determining catecholamines in urine, based on a new approach to ion-pairing chromatography (IPC), in which the ion-pairing reagent (IPR), 1-Heptane Sulfonic Acid (HSA), is added to the extracted samples instead of the mobile phases. A Hamilton STARlet workstation carried out the solid phase extraction of urine samples. The extracted samples were diluted with 60 mmol/L HSA and injected on a Kinetex core-shell biphenyl column with conventional LC–MS/MS suitable mobile phases. Chromatographic separation of E and NE was achieved successfully with very stable retention times (RT). In 484 injections, the RTs were steady with a CV of less than ±4%. Furthermore, HSA was separated from E and NE, allowing HSA to be diverted to waste instead of entering the mass spectrometer ion chamber. The method was validated with good analytical performance, and even though the analysis for urinary catecholamines is increasingly being replaced by plasma free metanephrines in diagnosing pheochromocytomas, this work represents the application of a new analytical technique that can be transferred to other small polar molecules, that are difficult to chromatograph on traditional reversed phase columns.

OriginalsprogEngelsk
TidsskriftJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Vol/bind1057
Sider (fra-til)118-123
ISSN1570-0232
DOI
StatusUdgivet - 2017

Fingeraftryk

Heptanes
Ion chromatography
Sulfonic Acids
Catecholamines
Norepinephrine
Liquid chromatography
Metanephrine
Ionization chambers
Molecules
Mass spectrometers
Epinephrine
Mass spectrometry
Ions
Plasmas

Citer dette

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title = "Analysis of catecholamines in urine by unique LC/MS suitable ion-pairing chromatography",
abstract = "The catecholamines, epinephrine (E) and norepinephrine (NE) are small polar, hydrophilic molecules, posing significant challenges to liquid chromatography − tandem mass spectrometry (LC–MS/MS) method development. Specifically, these compounds show little retention on conventional reversed-phase liquid chromatography columns. This work presents development and validation of an LC–MS/MS method for determining catecholamines in urine, based on a new approach to ion-pairing chromatography (IPC), in which the ion-pairing reagent (IPR), 1-Heptane Sulfonic Acid (HSA), is added to the extracted samples instead of the mobile phases. A Hamilton STARlet workstation carried out the solid phase extraction of urine samples. The extracted samples were diluted with 60 mmol/L HSA and injected on a Kinetex core-shell biphenyl column with conventional LC–MS/MS suitable mobile phases. Chromatographic separation of E and NE was achieved successfully with very stable retention times (RT). In 484 injections, the RTs were steady with a CV of less than ±4{\%}. Furthermore, HSA was separated from E and NE, allowing HSA to be diverted to waste instead of entering the mass spectrometer ion chamber. The method was validated with good analytical performance, and even though the analysis for urinary catecholamines is increasingly being replaced by plasma free metanephrines in diagnosing pheochromocytomas, this work represents the application of a new analytical technique that can be transferred to other small polar molecules, that are difficult to chromatograph on traditional reversed phase columns.",
keywords = "Catecholamine, Epinephrine, Ion-pairing chromatography, Mass spectrometry, Norepinephrine, Urine",
author = "Bergmann, {Marianne L.} and Seyed Sadjadi and Anne Schmedes",
year = "2017",
doi = "10.1016/j.jchromb.2017.04.011",
language = "English",
volume = "1057",
pages = "118--123",
journal = "Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences",
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Analysis of catecholamines in urine by unique LC/MS suitable ion-pairing chromatography. / Bergmann, Marianne L.; Sadjadi, Seyed; Schmedes, Anne.

I: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Bind 1057, 2017, s. 118-123.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Analysis of catecholamines in urine by unique LC/MS suitable ion-pairing chromatography

AU - Bergmann, Marianne L.

AU - Sadjadi, Seyed

AU - Schmedes, Anne

PY - 2017

Y1 - 2017

N2 - The catecholamines, epinephrine (E) and norepinephrine (NE) are small polar, hydrophilic molecules, posing significant challenges to liquid chromatography − tandem mass spectrometry (LC–MS/MS) method development. Specifically, these compounds show little retention on conventional reversed-phase liquid chromatography columns. This work presents development and validation of an LC–MS/MS method for determining catecholamines in urine, based on a new approach to ion-pairing chromatography (IPC), in which the ion-pairing reagent (IPR), 1-Heptane Sulfonic Acid (HSA), is added to the extracted samples instead of the mobile phases. A Hamilton STARlet workstation carried out the solid phase extraction of urine samples. The extracted samples were diluted with 60 mmol/L HSA and injected on a Kinetex core-shell biphenyl column with conventional LC–MS/MS suitable mobile phases. Chromatographic separation of E and NE was achieved successfully with very stable retention times (RT). In 484 injections, the RTs were steady with a CV of less than ±4%. Furthermore, HSA was separated from E and NE, allowing HSA to be diverted to waste instead of entering the mass spectrometer ion chamber. The method was validated with good analytical performance, and even though the analysis for urinary catecholamines is increasingly being replaced by plasma free metanephrines in diagnosing pheochromocytomas, this work represents the application of a new analytical technique that can be transferred to other small polar molecules, that are difficult to chromatograph on traditional reversed phase columns.

AB - The catecholamines, epinephrine (E) and norepinephrine (NE) are small polar, hydrophilic molecules, posing significant challenges to liquid chromatography − tandem mass spectrometry (LC–MS/MS) method development. Specifically, these compounds show little retention on conventional reversed-phase liquid chromatography columns. This work presents development and validation of an LC–MS/MS method for determining catecholamines in urine, based on a new approach to ion-pairing chromatography (IPC), in which the ion-pairing reagent (IPR), 1-Heptane Sulfonic Acid (HSA), is added to the extracted samples instead of the mobile phases. A Hamilton STARlet workstation carried out the solid phase extraction of urine samples. The extracted samples were diluted with 60 mmol/L HSA and injected on a Kinetex core-shell biphenyl column with conventional LC–MS/MS suitable mobile phases. Chromatographic separation of E and NE was achieved successfully with very stable retention times (RT). In 484 injections, the RTs were steady with a CV of less than ±4%. Furthermore, HSA was separated from E and NE, allowing HSA to be diverted to waste instead of entering the mass spectrometer ion chamber. The method was validated with good analytical performance, and even though the analysis for urinary catecholamines is increasingly being replaced by plasma free metanephrines in diagnosing pheochromocytomas, this work represents the application of a new analytical technique that can be transferred to other small polar molecules, that are difficult to chromatograph on traditional reversed phase columns.

KW - Catecholamine

KW - Epinephrine

KW - Ion-pairing chromatography

KW - Mass spectrometry

KW - Norepinephrine

KW - Urine

U2 - 10.1016/j.jchromb.2017.04.011

DO - 10.1016/j.jchromb.2017.04.011

M3 - Journal article

VL - 1057

SP - 118

EP - 123

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

SN - 1570-0232

ER -