Determination of histamine in tomatoes by liquid chromatography/mass spectrometry

The histamine level in tomato fruits and pastes was determined by 2 orthogonal techniques as a means of comparing accuracy. Close statistical agreement was found between assays for free histamine by capillary electrophoresis (with UV absorbance detection), and for the dansyl derivative by reversed-phase liquid chromatography (LC). Both techniques have adequate sensitivity for the analysis of endogenous histamine in tomatoes, but LC/electrospray tandem mass spectrometry was more sensitive by at least an order of magnitude, down to a level of 0.05 mg/kg.     

A Rapid Qualitative and Quantitative Method of Assaying Histamine in Small Plasma Volume

Abstract

A modified method for a qualitative and quantitative determination of histamine in small plasma volume (≤300 μl) was developed. According to this method, blood samples containing methylhistamine, the internal standard, are centrifuged to collect plasma. These plasma samples which contain underivatized histamine are injected into a Dionex BioLC System coupled with a pulsed amperometric detector. Histamine and methylhistamine are separated through a C-18 Zorbax ODS 4.5mm ID ± 25cm (5 microns) column. Histamine is quantitated by comparing histamine peak height with that of known quantity of the internal standard. The sensitivity of the method is 0.03 pmols. The peak heights were found to be linearly related to histamine concentrations providing a quantitative means of assaying histamine in biological samples. The retention time of histamine was 6 min in contrast to that of methlyhistamine which was 10 min.     

Determination of Histamine in Rat Plasma and Tissue Extracts by Intramolecular Excimer-Forming Derivatization and LC with Fluorescence Detection

A highly sensitive and selective method for determination of histamine in rat plasma and tissue extracts by liquid chromatography with fluorescence detection is described. The method is based on precolumn derivatization of amino groups of histamine with two molecules of 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester that allows generation of intramolecular excimer fluorescence. The detection limit for histamine was 0.183 nM. This sensitivity allowed determination of histamine in 10 μL of rat plasma or in the extracts from less than 1 mg of tissue.     

A sensitive and selective determination method of histamine by HPLC with intramolecular excimer-forming derivatization and fluorescence detection

A highly sensitive, selective and simple method is described for the determination of histamine by high-performance liquid chromatography (HPLC) with fluorescence detection. The method is based on an intramolecular excimer-forming fluorescence derivatization of histamine with 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester (PSE), followed by reversed-phase HPLC. Histamine, having two amino moieties in a molecule, was converted to the dipyrene-labeled derivative by reaction with PSE. The derivative afforded intramolecular excimer fluorescence (450-540 nm), which can clearly be discriminated from the monomer fluorescence (370-420 nm) emitted from PSE. Typically, a 10 micro L sample solution was mixed with 100 micro L of derivatization reagent solution, which was a mixture of 0.5 mm PSE in acetonitrile and 0.5 mm potassium carbonate in water (8:2, v/v). The derivatization was carried out at 100 degrees C for 90 min. The PSE derivative of histamine could be separated by reversed-phase ODS column with isocratic elution using acetonitrile:water (82:18, v/v) containing 0.03% triethylamine. The detection limit (singnal-to-noise ratio = 3) of histamine was 0.5 fmol for a 30 micro L injection. The method was successfully applied to the determination of histamine in human urine, and had enough selectivity and sensitivity for urinary histamine quantification.     

Homogeneous time resolved fluorescence assay to measure histamine release

Histamine is a biogenic amine synthesized by the enzymatic decarboxylation of histidine. Implication of histamine in allergy is well described but histamine is also found in some specific neurones, functions as a neurotransmitter and regulates sleep/wake cycles, hormonal secretion, cardiovascular control and thermo-regulation. We have developed a TR-FRET histamine assay, based on the competition between sample histamine and allophycocyanine (XL665) labelled histamine for binding to a Europium cryptate (EuK) labelled antibody. As histamine is a small monoamine molecule, high affinity antibodies have been raised against carrier protein conjugated histamine. Therefore, sample histamine needs to be derivatized in the same way as the conjugated histamine, so that the antibody will have a similar affinity for both molecules. This acylation step is performed directly in wells and does not need to be done in separate vials, making handling easier for large numbers of samples. The incubation takes place at room temperature for 3 hours. The assay covers a measurement range of 1.56 to 400 nM and shows an analytical sensitivity of 1.3nM. We have shown that miniaturization of sample and reagents volumes down to 20 micro l does not alter these performances. This histamine release assay provides a particularly well adapted procedure for HTS and secondary screening compared to current heterogeneous methods.     

Field-amplified sample injection and in-capillary derivatization for sensitivity improvement of the electrophoretic determination of histamine

The feasibility of the combination of field-amplified sample injection (FASI) and in-capillary derivatization was explored for improving sensitivity of histamine in capillary electrophoresis (CE). Naphthalene-2,3-dicarboxaldehyde (NDA) was used as derivatization reagent. The reagent and sample was introduced by tandem mode. The derivatization was accomplished by at-inlet mode with standing time of 1.5 min. The combination of FASI and in-capillary derivatization was successfully achieved with about 400-fold concentration sensitivity enhancement compared to pre-capillary derivatization at the same set-up. The detection limit of concentration for histamine reached 1.25 x 10(-11) M by CE and fluorescence detection with S/N = 3. Parameters affecting FASI and in-capillary derivatization process including sample matrix, buffer concentration and reagent injection amount, were investigated.     

s-BLM修饰电极对组胺电化学传感行为的研究

The electrochemical biosensor properties of the molecular devices based on solid supported bilayer lipid membranes were investigated. The results showed that the system embedded with antihistaminic agent had strong response for the histamine. The response currents for the histamine will rise following the increasing concentrations of the histamine,as a result of the special opposing effect between histamine and antihistamine. Near the normal physiological condition（pH=7. 5）,a good linearity occurred for the current response against histamine concentration ranging from 3. 0 to 60. 0 μg / L,and the relation coefficient R is 0. 9878. All the experiments mentioned above showed good sensitivity,selectivity and stability. This system can measure the concentration of histamine under -200 mV by voltammetry and be expected to form the electrochemical biosensor with high stability and low detection limit.     

Development of an automated on-line pre-column derivatization procedure for sensitive determination of histamine in food with high-performance liquid chromatography-fluorescence detection

An improved sensitive method was developed and validated for the determination of histamine in food samples by using automated on-line pre-column derivatization coupled with high performance liquid chromatography and fluorescence detection (HPLC-FLD). o-Phthaldialdehyde (OPA) was adopted as derivatization reagent, and a "sandwich" (OPA+histamine+OPA) aspiration mode for the automated on-line pre-column derivatization was found to efficiently enhance the method sensitivity and precision. Histamine in food samples was efficiently extracted with a methanol-phosphate buffer solution (50:50, v/v) at 60 degrees C for 30 min, and purified with Waters Oasis MCX solid-phase extraction (SPE) cartridge. The limit of quantification for this method is 0.2 mg/kg, which is very sensitive for histamine determination. With the "sandwich" injection program, 3.7% of relative standard deviation (RSD) was achieved by five replicative determinations of a sample blank spiked with 0.25 mg/kg histamine standard. Histamine in food samples such as fumitory skipjack and mackerel was analyzed with relative recoveries over 95% at spiking level of 150 mg/kg, as well as canned tuna fish and cheese with relative recoveries up to 98% at spiking levels of 0.50 and 5.0 mg/kg, respectively. The proposed method was validated with a sample from the Food Analysis Performance Assessment Scheme (FAPAS) as a standard certified material; and the results (140+/-6 mg/kg) agreed well with the assigned value (139 mg/kg).     

In vitro determination of diamine oxidase activity in food matrices by an enzymatic assay coupled to UHPLC-FL

Intestinal diamine oxidase (DAO) acts as a protective barrier against exogenous histamine. A deficit of DAO activity can lead to the appearance of histamine intolerance, a clinical condition that may be treated by a low-histamine diet and oral DAO supplementation to enhance intestinal histamine degradation. As sources of DAO, porcine kidneys and certain legume seedlings are suitable components for the formulation of a DAO supplement. The aim of this work was to develop a rapid and reliable methodology for the in vitro determination of DAO activity in food matrices based on an enzymatic assay coupled to UHPLC-FL. The proposed method showed a satisfactory linearity and sensitivity and provided a relative standard deviation lower than 3%, guaranteeing method precision, and a mean recovery greater than 99% both for lyophilized pea sprouts and porcine kidney protein extracts. A high specificity is a key attribute of this method due to the use of histamine as the reaction substrate and the direct quantification of its degradation. Moreover, the lack of interference of catalase and hydrogen peroxide is another advantage in comparison with previously published methods. Lyophilized pea sprouts showed the greatest histamine-degrading activity (0.40 ± 0.01 mU/mg), followed by porcine kidney protein extracts (0.23 ± 0.01 mU/mg) and commercial DAO supplements (0.09 ± 0.06 mU/mg). This technique could be used as a tool to validate the DAO activity of food matrices of potential interest for the treatment of histamine intolerance.

     

Use of 2,6-dinitro-4-trifluoromethylbenzenesulfonic acid as a novel derivatizing reagent for the analysis of catecholamines, histamines and related amines by gas chromatography with electron-capture detection

We have found that 2,6-dinitro-4-trifluoromethylbenzenesulfonic acid reacts rapidly and specifically with primary amines at room temperature. We have used this reagent for derivatizing phenylethylamines, including catecholamines, and histamine and 1-methylhistamine. After the reaction, hydroxyl groups were derivatized to form the corresponding trimethylsilyl ethers, and the final derivatives were analyzed by gas chromatography with electron-capture detection. These derivatives are stable, possess excellent gas chromatographic properties and are detected with high sensitivity. We have applied this method to the analysis of histamine and 1-methylhistamine in human urine.     

An easy-to-use excimer fluorescence derivatization reagent, 2-chloro-4-methoxy-6-(4-(pyren-4-yl)butoxy)-1,3,5-triazine,for use in the highly sensitive and selective liquid chromatography analysis of histamine in Japanese soy sauces

In this study, a novel pre-column excimer fluorescence derivatization reagent, 2-chloro-4-methoxy-6-(4-(pyren-4-yl)butoxy)-1,3,5-triazine (CMPT), was developed for polyamines, specifically histamine. By CMPT derivatization, the polyamines, histamine and tyramine were converted to polypyrene derivatives, and emitted intra-molecular excimer fluorescence at 475 nm. This could clearly be distinguished from the normal fluorescence emitted from reagent blanks at 375 nm. Unlike conventional excimer fluorescence derivatization reagents, CMPT is chemically stable and its reactivity sustained over at least 36 days even in solution state. We successfully applied this reagent to the sensitive and selective analysis of histamine in different kinds of Japanese commercial soy sauces. The detection and quantification limits of histamine were 15 and 50 μg L⁻¹, respectively, equating to 1.35 pmol and 4.5 pmol for a 6 μL injection. This sensitivity helped the direct analysis of soy sauce samples only treated by one-step liquid–liquid extraction without concentration. The histamine levels of commercial soy sauce samples (koikuchi, usukuchi and saishikomi) investigated were 1.24–768.5 mg L⁻¹.     

Cis-UROCANIC ACID SYNERGIZES WITH HISTAMINE FOR INCREASED PGE2 PRODUCTION BY HUMAN KERATINOCYTES: LINK TO INDOMETHACIN-INHIBITABLE UVB-INDUCED IMMUNOSUPPRESSION

Abstract— There is considerable evidence that suppression of the immune system by UVB (280–320 nm UV) irradiation is initiated by UVB-dependent isomerization of a specific skin photoreceptor, urocanic acid (UCA), from the trans to the cis form. Previous studies have confirmed that cis -UCA administration to mice 3–5 days prior to hapten sensitization at a distant site, suppresses the contact hypersensitivity (CHS) response upon challenge. This study demonstrates in mice that cis -UCA, like UVB, suppresses CHS to trinitrochlorobenzene by a mechanism partly dependent on prostanoid production. In vitro experimentation showed that human keratinocytes, isolated from neonatal foreskin, increased prostaglandin E₂ (PGE₂) production in response to histamine but not UCA alone. However, cis -UCA synergized with histamine for increased PGE₂ production by keratinocytes. cis -urocanic acid also increased the sensitivity of keratinocytes for PGE₂ production in response to histamine. Prostaglandin E₂ from keratinocytes exposed to cis -UCA and histamine may contribute directly, or indirectly, to the regulation of CHS responses by UVB irradiation.     

The affinity of histamine to serum albumin: Capillary electrophoresis-frontal analysis and in-silico molecular docking approaches

Histamine is a biogenic amine found in various body tissues and responsible for many critical vital activities. It is also responsible for allergic reactions in the body. Ingestion of foods containing high amounts of histamine can cause fatal allergic reactions. Albumin in plasma controls drugs and free concentrations of bioactive constituents taken to the body with food. Hence, this study aimed to characterise the interactions of histamine with bovine serum albumin. Capillary electrophoresis in the frontal analysis mode was employed in this study as a practical approach for assessing histamine-bovine serum albumin affinity. The plateau-shaped free histamine peak was well separated from the bovine serum albumin (BSA)-histamine complex peak. The free histamine concentration was obtained by following the height of the free histamine peak. Whereas the bound histamine concentrations were obtained by calculating the difference between the height of total and free histamine peaks. Histamine bound to BSA at one independent site with a K_b value of 2.50 × 10³ L/mol. Moreover, an in-silico molecular docking method was performed, and it was revealed that the binding site of histamine was located closer to Lysine-131 in subdomain IIA of bovine serum albumin.     

Determination of histamine in biological fluids by capillary isotachophoresis and fluorescence

Analytical isotachophoresis was used for the determination of histamine in biological fluids. For biological fluids with a very low content of histamine a method was developed that combines the fluorescing condensation product of histamine and o-phthaldialdehyde with the high concentration effect of isotachophoresis. This method permits the determination of histamine in serum and other biological fluids down to less than 3 ng/ml.     

Determination of histamine, 1-methylhistamine and N-methylhistamine by capillary electrophoresis with micelles

Urinary histamine and Ngamma-methylhistamine (1-MH), a histamine metabolite, are highly correlated with histamine in plasma. Therefore, allergic reactions can be examined by determination of histamine and 1-MH in urine. We separated histamine, 1-MH and Nalpha-methylhistamine (N-MH) by capillary electrophoresis with UV detection at 210nm, using borate buffer (pH 9) containing 100 mM SDS. The absolute detection limits were 200, 100 and 50 pg for histamine, 1-MH and N-MH, respectively. To purify histamine 1-MH and N-MH in urine, a silica cartridge was used. Recovery rates of histamine, 1-MH and N-MH in physiological saline were 90.0, 91.4 and 95.4%, respectively. We measured histamine and 1-MH levels in urine from a normal female volunteer before and after a meal, and a male bronchial asthma patient. The results showed clearly that the concentrations of histamine and its metabolite rose after eating or asthma attack. N-MH was not detected in the urine.     

Flow-through chemiluminescence sensor using immobilized histamine oxidase from Arthrobacter crystallopoietes KAIT-B-007 and peroxidase for selective determination of histamine.

A flow sensor with immobilized oxidases is proposed for the determination of histamine in fish meat. Chemiluminometric measurement of histamine was based on the luminol reaction with hydrogen peroxide produced by immobilized histamine oxidase (EC 1.4.3.-.) and peroxidase (EC 1.11.1.7.) within a flow cell. Histamine oxidase was found in cells of Arthrobacter crystallopoietes KAIT-B-007 isolated from soil. The oxidase and peroxidase were coimmobilized covalently on tresylated hydrophilic vinyl polymer beads and packed into transparent PTFE; the tubing was used as the flow cell. One assay for histamine was done at intervals of 2 min without carryover. The calibration curve for histamine was linear from 0.1 microM to 50 microM. The response was reproducible within 1.25% of the relative standard deviation for 115-replicate injections of 50 microM histamine. The sensor system was applied to the determination of histamine in fish meat extracts.     

Amine oxidase amperometric biosensor coupled to liquid chromatography for biogenic amines determination

A selective and sensitive method is presented for biogenic amines (BA) determination. The novelty consists in coupling a highly selective electrochemical biosensor to a weak acid cation-exchange column for online detection of amines. A bienzyme design, based on a recently isolated amine oxidase from grass pea and commercial horseradish peroxidase, was used for the biosensor construction. The enzymes were co-immobilized on the surface of a graphite electrode together with the electrochemical mediator (Os-redox polymer). The electrochemical detection was performed at a low applied potential (?50 mV vs. Ag/AgCl, KCl_0.1 M), where biases from interferences are minimal. The separation and determination of six BA, with relevance in food analysis (tyramine, putrescine, cadaverine, histamine, agmatine and spermidine), were investigated. Irrespective of the BA nature, the amine oxidase-based biosensor showed a linear response up to 5 mM, and its sensitivity decreases in the following order: cadaverine, putrescine, spermidine, agmatine, histamine and tyramine. The approach was used to estimate the BA content in fish samples, after their extraction with methanesulfonic acid.     

Separation of histamine and its metabolites using glass capillary gas chromatography and a nitrogen detector

Complete separation of the TMS derivatives of histamine, 1-methyl histamine and 3-methyl histamine was achieved in two minutes in a 7 m × 0.25 mm column coated with SP 2250. Using NPD, histamine was detectable at levels as low as 20 pg. This offers a rapid, sensitive procedure with potential for clinical diagnosis and food analysis.     

Improved detection of the histamine receptor on human peripheral blood mononuclear cells by crosslinking using tritiated as compared with radioiodinated histamine

In order to detect histamine receptors on the surface of human peripheral blood mononuclear cells, the cells were incubated in the presence of radiolabelled histamine and then the bifunctional crosslinker disuccimidyl suberate was added in various concentrations. They were then solubilized with sodium dodecyl sulphate, boiled, reduced and the lysate separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Both 3H and 125I-radiolabelled ligands bound to a 16 kDa band, to be defined although a much clearer and obviously unequivocal signal was obtained with 3H-labelled histamine. This molecule migrated with the same mass on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a 16 kDa subunit which had been purified on a histamine affinity column from Triton X-100 solubilized mononuclear cells, indicating it to be the ligand-binding subunit for the histamine receptor on these cells. For 3H, fluorography with Entensify was required to obtain an autoradiographic signal. Although 3H took much longer to give a signal than 125I, the considerable background, artefacts and heavy lane trailing seen with [125I] histamine were completely abrogated when [3H]histamine was used. In addition, the distinction between specific and nonspecific binding was more clearly seen using [3H]histamine. The modifications reported here which improve signal detection for 3H should encourage the use of tritiated ligands in radioreceptor crosslinking, particularly those of low molecular weight which might otherwise undergo steric modification due to iodination, this having the potential for interfering with receptor ligand binding.     

Electrochemical determination of histamine derivatized with o-phthalaldehyde and 2-mercaptoethanol

High-performance liquid chromatography coupled with electrochemical detection was used to determine histamine following precolumn derivatization with o-phthalaldehyde (OPA) and 2-mercaptoethanol. The isoindole derivative which is obtained as reaction product was electrochemically active at a moderate potential (peak potential +0.4 V). Direct oxidation of histamine required a much higher potential (peak potential +1.05 V) and was of no practical use. No electrochemical signal was observed for the reaction product of histamine with OPA. Changing the pH of the mobile phase had little effect on the electrochemical response of the isoindole derivative of histamine, which was well separated from analogous derivatives of methylated histamines, mono- and polyamines and amino acids by isocratic elution from a reversed-phase column. An example of a practical application of the method to the estimation of histamine in rat brain is presented.