US EPA

3860327 

Journal Article 

Histamine quantification in human plasma using high resolution accurate mass LC-MS technology 

Laurichesse, M; Gicquel, T; Moreau, C; Tribut, O; Tarte, K; Morel, I; Bendavid, C; Amé-Thomas, P 

2016 

Yes 

Clinical Biochemistry
ISSN: 0009-9120
EISSN: 1873-2933 

PERGAMON-ELSEVIER SCIENCE LTD 

OXFORD 

49 

1-2 

111-116 

English 

26282720 

10.1016/j.clinbiochem.2015.08.012 

WOS:000368752700020 

BACKGROUND: Histamine (HA) is a small amine playing an important role in anaphylactic reactions. In order to identify and quantify HA in plasma matrix, different methods have been developed but present several disadvantages. Here, we developed an alternative method using liquid chromatography coupled with an ultra-high resolution and accurate mass instrument, Q Exactive™ (Thermo Fisher) (LCHRMS).

METHODS: The method includes a protein precipitation of plasma samples spiked with HA-d4 as internal standard (IS). LC separation was performed on a C18 Accucore column (100∗2.1mm, 2.6μm) using a mobile phase containing nonafluoropentanoic acid (3nM) and acetonitrile with 0.1% (v/v) formic acid on gradient mode. Separation of analytes was obtained within 10min. Analysis was performed from full scan mode and targeted MS2 mode using a 5ppm mass window. Ion transitions monitored for targeted MS2 mode were 112.0869>95.0607m/z for HA and 116.1120>99.0855m/z for HA-d4. Calibration curves were obtained by adding standard calibration dilution at 1 to 180nM in TrisBSA.

RESULTS: Elution of HA and IS occurred at 4.1min. The method was validated over a range of concentrations from 1nM to 100nM. The intra- and inter-run precisions were <15% for quality controls. Human plasma samples from 30 patients were analyzed by LCHRMS, and the results were highly correlated with those obtained using the gold standard radioimmunoassay (RIA) method.

CONCLUSION: Overall, we demonstrate here that LCHRMS is a sensitive method for histamine quantification in biological human plasmas, suitable for routine use in medical laboratories. In addition, LCHRMS is less time-consuming than RIA, avoids the use of radioactivity, and could then be considered as an alternative quantitative method.