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


CONCLUSIONS
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.
CHU Rennes, Laboratoire d'Immunologie, Thérapie Cellulaire et Hématopoïèse, F-35033 Rennes, France; CHU Rennes, Laboratoire de Biochimie, F-35033 Rennes, France. Electronic address: m_laurichesse@hotmail.com.
Mathieu
M
Laurichesse
CHU Rennes, Laboratoire de Toxicologie Biologique et Médico-Légale, F-35033 Rennes, France; Université de Rennes 1, Faculté de Pharmacie, F-35043 Rennes, France; INSERM Unité Mixte de Recherche 991, F-35043 Rennes, France.
Thomas
T
Gicquel
CHU Rennes, Laboratoire de Biochimie, F-35033 Rennes, France; Université de Rennes 1, Faculté de Médecine, F-35043 Rennes, France; INSERM Unité Mixte de Recherche 991, F-35043 Rennes, France.
Caroline
C
Moreau
CHU Rennes, UF Biomarqueurs, F-35033 Rennes, France.
Olivier
O
Tribut
CHU Rennes, Laboratoire d'Immunologie, Thérapie Cellulaire et Hématopoïèse, F-35033 Rennes, France; Université de Rennes 1, Faculté de Médecine, F-35043 Rennes, France; INSERM Unité Mixte de Recherche 917, F-35043 Rennes, France.
Karin
K
Tarte
CHU Rennes, Laboratoire de Toxicologie Biologique et Médico-Légale, F-35033 Rennes, France; Université de Rennes 1, Faculté de Pharmacie, F-35043 Rennes, France; INSERM Unité Mixte de Recherche 991, F-35043 Rennes, France.
Isabelle
I
Morel
CHU Rennes, Laboratoire de Biochimie, F-35033 Rennes, France; Université de Rennes 1, Faculté de Pharmacie, F-35043 Rennes, France; INSERM Unité Mixte de Recherche 991, F-35043 Rennes, France.
Claude
C
Bendavid
CHU Rennes, Laboratoire d'Immunologie, Thérapie Cellulaire et Hématopoïèse, F-35033 Rennes, France; Université de Rennes 1, Faculté de Médecine, F-35043 Rennes, France; INSERM Unité Mixte de Recherche 917, F-35043 Rennes, France. Electronic address: patricia.ame@univ-rennes1.fr.
Patricia
P
Amé-Thomas
D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D006632:Histamine / Q000097:blood*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D011786:Quality Control; D011863:Radioimmunoassay; D012015:Reference Standards
D006632:Histamine
Accurate mass; Anaphylactic reaction; Histamine; LC–high resolution-MS