Rapid determination of 5-hydroxymethylfurfural by DART ionization with time-of-flight mass spectrometry.

DART (direct analysis in real time), a novel technique with wide potential for rapid screening analysis, coupled with high-resolution time-of-flight mass spectrometry (TOF-MS) has been used for quantitative analysis of 5-hydroxymethylfurfural (5-HMF), a typical temperature marker of food. The DART/TOF-MS method was optimised and validated. Quantification of 5-HMF was achieved by use of a stable isotope-labelled 5-HMF standard prepared from glucose. Formation of 5-HMF from saccharides, a potential source of overestimation of results, was evaluated. Forty-four real samples (honey and caramelised condensed sweetened milk) and 50 model samples of heated honey were analysed. The possibility of using DART for analysis of heated samples of honey was confirmed. HPLC and DART/TOF-MS methods for determination of 5-HMF were compared. The correlation equation between these methods was DART = 1.0287HPLC + 0.21340, R(2) = 0.9557. The DART/TOF-MS method has been proved to enable efficient and rapid determination of 5-HMF in a variety of food matrices, for example honey and caramel.

Determination of phytic acid and inositolphosphates in barley.

Phytic acid (PA) and lower inositolphosphates (InsP(n) ) is the main storage form of phosphorus in grains or seeds. The content of PA and InsP(n) in different varieties of barley was analyzed by capillary isotachophoresis and online-coupled capillary isotachophoresis with CZE. The electrolytes (in demineralized water) for the isotachophoretic analysis consisted of 10 mM HCl, 14 mM glycylglycine, and 0.1% 2-hydroxyethylcellulose (leading) and 10 mM citric acid (terminating). The optimized electrolytes for the online coupling isotachophoresis with zone electrophoresis analysis were mixtures of 5 mM HCl, 7 mM glycylglycine, and 0.1% 2-hydroxyethylcellulose (leading), 20 mM citric acid, 10 mM glycylglycine, and 0.1% 2-hydroxyethylcellulose (background) and 10 mM citric acid (terminating). PA and all studied InsP(n) were separated within 25 min and detected by a conductivity detector. Simple sample preparation (acidic extraction), sufficient sensitivity, speed of analysis, and low running cost are important attributes of the electrophoretic methods. The method was used for the determination of PA and InsP(n) in barley varieties within an ongoing research project.

Electrophoretic determination of calystegines A3 and B2 in potato.

Potatoes, members of the Solanaceae plant family, contain calystegines, water-soluble nortropane alkaloids, which are biologically active as glycosidase inhibitors. The content of calystegines A(3) and B(2) in different varieties of potato and in various parts of the tubers (whole potato, peel, flesh, and sprouts) were analysed by new capillary zone electrophoresis and capillary isotachophoresis methods and by the routine GC method. The optimized background electrolyte for capillary zone electrophoretic analysis was mixture of 20 mM histidine, 20 mM N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid and 20% (v/v) methanol in demineralized water. Calystegines were detected by indirect UV detection at 210 nm. A clear separation of calystegines from other components of the methanolic sample extract was achieved within 4 min. The electrolytes for isotachophoretic analysis consisted of 5 mM NH(4)OH, 10 mM N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid, 0.1% hydroxyethylcellulose and 20% (v/v) methanol in demineralized water (leading) and 5 mM histidine+10 mM acetic acid+20% (v/v) methanol in demineralized water (terminating). Calystegines were separated within 20 min and detected by a conductimeter. Method characteristics of both zone electrophoresis and isotachophoresis, i.e., linearity (10-100 ng/microl and 1-10 ng/microl), accuracy (recovery 96+/-5% and 98+/-4%), intra-assay repeatability (4.2% and 3.5%), and detection limit (3 and 0.4 ng/microl) were evaluated. Simple sample preparation, sufficient sensitivity, speed of analysis, and low running cost are important attributes of the electrophoretic methods. The overall results of electrophoretic methods were comparable with GC.

Determination of glycyrrhizin in liqueurs by on-line coupled capillary isotachophoresis with capillary zone electrophoresis.

An on-line coupled capillary isotachophoresis-capillary zone electrophoresis method for the determination of glycyrrhizin in liqueurs is described. The optimised electrolyte system was 5 mM HCl+11 mM epsilon-aminocaproic acid+0.05% hydroxyethylcellulose+30% methanol (leading electrolyte), 5 mM caproic acid+30% methanol (terminating electrolyte) and 20 mM caproic acid+10 mM histidine+0.1% hydroxyethylcellulose+30% methanol (background electrolyte). Method characteristics, i.e., linearity (20-500 ng/ml), accuracy (recovery 99+/-4%), intra-assay repeatability (2%), intermediate repeatability (3.8%) and detection limit (8 ng/ml) were determined. Speed of analysis, low laboriousness, high sensitivity and low-running cost are the typical attributes of the capillary isotachophoresis-capillary zone electrophoresis method. Developed method was successfully applied to analysis of liqueurs with liquorice extract and some foods (sweets and food supplements) containing liquorice. Found levels of glycyrrhizin in liqueurs, sweets and food supplements varied between 1-16 mg/l, 850-1050 mg/kg and 1.6-1.8 g/kg, respectively.

Determination of biogenic amines by capillary zone electrophoresis with conductometric detection.

A capillary zone electrophoresis (CZE) method with conductometric detection of biogenic amines (cadaverine, putrescine, agmatine, histamine, tryptamine and tyramine) is described. The optimised background electrolyte was the following: 15 mM histidine + 5 mM adipic acid + 1.5 mM sulphuric acid + 0.1 mM ethylenediaminotetraacetic acid + 0.1% hydroxyethylcellulose + 50% methanol. A clear separation of six biogenic amines from other components of acidic sample extract was achieved within 10 min. Method characteristics, i.e., linearity (0-100 micromol/ml), accuracy (recovery 86-107%), intra-assay repeatability (2-4%), and detection limit (2-5 micromol/l) were evaluated. Low laboriousness, sufficient sensitivity, speed of analysis, and low running cost are important attributes of this method. The developed method was successfully applied on the determination of biogenic amines in selected food samples.

Determination of domoic acid by on-line coupled capillary isotachophoresis with capillary zone electrophoresis.

An on-line coupled capillary isotachophoresis--capillary zone electrophoresis (cITP-CZE) method for the determination of domoic acid in shellfish and algae is described. The optimised cITP-CZE electrolyte system was 10 mM HCl + 20 mM beta-alanine (BALA) + 0.05% hydroxyethylcellulose (leading electrolyte), 5 mM caproic acid (terminating electrolyte) and 20 mM caproic acid + 20 mM BALA + 0.1% HPMC (background electrolyte). A clear separation of the domoic acid from the other components of methanolic sample extract was achieved within 25 min. Method characteristics, i.e., linearity (0-200 microg/l), accuracy (recovery 101+/-3%), intra-assay repeatability (2.4%) and detection limit (1.5 microg/l) were determined. Speed of analysis, low laboriousness, high sensitivity and low running cost are the typical attributes of the cITP-CZE method. Developed method was successfully applied to analysis of shellfish samples and food supplements containing algae extract.

Capillary electrophoresis determination of carnitine in food supplements.

L-Carnitine is a substance natural for human body which transfers fatty acids to the place of burning-mitochondria and aids the transformation of fats into energy and this way supports overweight reduction and immediate physical performance, increases resistance from physical load and protect heart from overload. In this study are described newly developed electrophoretic methods (ITP, CZE with direct and/or indirect UV detection) for carnitine determination in various samples. The results were compared with results obtained by validated HPLC method. All of these methods gave comparable results. The detection limits of the electrophoretic methods were between 2.4 and 4.7 microg/ml, reproducibility (relative standard deviation, RSD%) was between 1.2 and 4.4% and recoveries were between 91 and 113% in different samples. The shorter analysis and low running cost are the main advantages of CE methods.

Separation of enantiomers of butorphanol and cycloamine by capillary zone electrophoresis.

Butorphanol tartrate is a synthetic opioid agonist-antagonist used as analgesic, possessing three chiral centres in the basic part of the molecule. Its chiral purity is routinely controlled only by optical rotation. A new capillary zone electrophoresis method, capable to separate the enantiomers of butorphanol and intermediate of its synthesis, cycloamine, was developed. Different electrolyte composition (type and concentration of carrier ion, pH, and organic solvent addition), and type and concentration of several chiral selectors (natural and modified cyclodextrins) were tested. Using the optimized conditions (acidic electrolyte with the addition of highly sulphated gamma-cyclodextrin) as low as 0.05% of undesirable enantiomers can be detected. Selected method characteristics, i.e., linearity (0-50 mg/l), precision (2.5% at 20 mg/l), and accuracy (101 +/- 2% at 20 mg/l) were evaluated. The optimized method was applied for the analysis of real batches of butorphanol and cycloamine. It was found that butorphanol tartrate manufactured by IVAX Pharmaceuticals contains less than 0.05% of undesirable enantiomer.