Development and validation of a highly sensitive LC–MS/MS method for in vitro measurement of histamine concentration

Kwang Youl Kim, Hyun Jung Kwon, Sang Heon Cho, Moonsuk Nam, Cheol Woo Kim

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The basophil histamine release test (HRT) is an important in vitro diagnostic assay to evaluate immunoglobin E (IgE)-mediated allergic responses. In this study, a bioanalytical LC–MS/MS method was developed and validated to quantify histamine in the leukocyte suspension from human peripheral blood. The method used pre-column derivatization with phenylisothiocyanate (PITC) and the resulting phenylthiocarbamyl (PTC) histamine was analyzed by positive-ion electrospray ionization using the multiple reaction monitoring mode. Chromatographic separation was achieved using an Imtakt-HT C18 column (2.1 mm × 50 mm, 3.0 μm), with a flow rate of 0.35 mL/min, 2 μL injection, and gradient elution with a mixture of acetonitrile-2 mM ammonium acetate buffer (both containing 0.1% formic acid). The total runtime of the method was 3.0 min including equilibration time. The method had a lower limit of detection of 0.1 ng/mL, and the quantifiable range was 0.1–100 ng/mL in the leukocyte suspension. The intra-day and inter-day precision and accuracy results were within the acceptable limits. It was established that histamine quantification should be performed within 2 h of preparing the leukocyte suspension, and freezing and thawing should be avoided. This method was successfully applied to the diagnosis and evaluation of the pathophysiologic mechanism of respiratory or cutaneous allergic diseases.

Original languageEnglish
Pages (from-to)33-41
Number of pages9
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume172
DOIs
StatePublished - 5 Aug 2019

Fingerprint

Histamine
Suspensions
formic acid
Leukocytes
Electrospray ionization
Thawing
Freezing
Basophils
Histamine Release
Assays
Buffers
Blood
Skin Diseases
Positive ions
Flow rate
Limit of Detection
In Vitro Techniques
Monitoring
Ions
Injections

Keywords

  • Basophil histamine release test
  • Leukocyte suspension
  • Multiple reaction monitoring
  • Phenylisothiocyanate

Cite this

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title = "Development and validation of a highly sensitive LC–MS/MS method for in vitro measurement of histamine concentration",
abstract = "The basophil histamine release test (HRT) is an important in vitro diagnostic assay to evaluate immunoglobin E (IgE)-mediated allergic responses. In this study, a bioanalytical LC–MS/MS method was developed and validated to quantify histamine in the leukocyte suspension from human peripheral blood. The method used pre-column derivatization with phenylisothiocyanate (PITC) and the resulting phenylthiocarbamyl (PTC) histamine was analyzed by positive-ion electrospray ionization using the multiple reaction monitoring mode. Chromatographic separation was achieved using an Imtakt-HT C18 column (2.1 mm × 50 mm, 3.0 μm), with a flow rate of 0.35 mL/min, 2 μL injection, and gradient elution with a mixture of acetonitrile-2 mM ammonium acetate buffer (both containing 0.1{\%} formic acid). The total runtime of the method was 3.0 min including equilibration time. The method had a lower limit of detection of 0.1 ng/mL, and the quantifiable range was 0.1–100 ng/mL in the leukocyte suspension. The intra-day and inter-day precision and accuracy results were within the acceptable limits. It was established that histamine quantification should be performed within 2 h of preparing the leukocyte suspension, and freezing and thawing should be avoided. This method was successfully applied to the diagnosis and evaluation of the pathophysiologic mechanism of respiratory or cutaneous allergic diseases.",
keywords = "Basophil histamine release test, Leukocyte suspension, Multiple reaction monitoring, Phenylisothiocyanate",
author = "Kim, {Kwang Youl} and Kwon, {Hyun Jung} and Cho, {Sang Heon} and Moonsuk Nam and Kim, {Cheol Woo}",
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Development and validation of a highly sensitive LC–MS/MS method for in vitro measurement of histamine concentration. / Kim, Kwang Youl; Kwon, Hyun Jung; Cho, Sang Heon; Nam, Moonsuk; Kim, Cheol Woo.

In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 172, 05.08.2019, p. 33-41.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Development and validation of a highly sensitive LC–MS/MS method for in vitro measurement of histamine concentration

AU - Kim, Kwang Youl

AU - Kwon, Hyun Jung

AU - Cho, Sang Heon

AU - Nam, Moonsuk

AU - Kim, Cheol Woo

N1 - Copyright © 2019 Elsevier B.V. All rights reserved.

PY - 2019/8/5

Y1 - 2019/8/5

N2 - The basophil histamine release test (HRT) is an important in vitro diagnostic assay to evaluate immunoglobin E (IgE)-mediated allergic responses. In this study, a bioanalytical LC–MS/MS method was developed and validated to quantify histamine in the leukocyte suspension from human peripheral blood. The method used pre-column derivatization with phenylisothiocyanate (PITC) and the resulting phenylthiocarbamyl (PTC) histamine was analyzed by positive-ion electrospray ionization using the multiple reaction monitoring mode. Chromatographic separation was achieved using an Imtakt-HT C18 column (2.1 mm × 50 mm, 3.0 μm), with a flow rate of 0.35 mL/min, 2 μL injection, and gradient elution with a mixture of acetonitrile-2 mM ammonium acetate buffer (both containing 0.1% formic acid). The total runtime of the method was 3.0 min including equilibration time. The method had a lower limit of detection of 0.1 ng/mL, and the quantifiable range was 0.1–100 ng/mL in the leukocyte suspension. The intra-day and inter-day precision and accuracy results were within the acceptable limits. It was established that histamine quantification should be performed within 2 h of preparing the leukocyte suspension, and freezing and thawing should be avoided. This method was successfully applied to the diagnosis and evaluation of the pathophysiologic mechanism of respiratory or cutaneous allergic diseases.

AB - The basophil histamine release test (HRT) is an important in vitro diagnostic assay to evaluate immunoglobin E (IgE)-mediated allergic responses. In this study, a bioanalytical LC–MS/MS method was developed and validated to quantify histamine in the leukocyte suspension from human peripheral blood. The method used pre-column derivatization with phenylisothiocyanate (PITC) and the resulting phenylthiocarbamyl (PTC) histamine was analyzed by positive-ion electrospray ionization using the multiple reaction monitoring mode. Chromatographic separation was achieved using an Imtakt-HT C18 column (2.1 mm × 50 mm, 3.0 μm), with a flow rate of 0.35 mL/min, 2 μL injection, and gradient elution with a mixture of acetonitrile-2 mM ammonium acetate buffer (both containing 0.1% formic acid). The total runtime of the method was 3.0 min including equilibration time. The method had a lower limit of detection of 0.1 ng/mL, and the quantifiable range was 0.1–100 ng/mL in the leukocyte suspension. The intra-day and inter-day precision and accuracy results were within the acceptable limits. It was established that histamine quantification should be performed within 2 h of preparing the leukocyte suspension, and freezing and thawing should be avoided. This method was successfully applied to the diagnosis and evaluation of the pathophysiologic mechanism of respiratory or cutaneous allergic diseases.

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