Quantification of lactose using ion-pair RP-HPLC during enzymatic lactose hydrolysis of skim milk

The correct labelling of dairy foods as “lactose-free” requires a suitably sensitive and valid analytical method for the quantification of lactose in complex food matrices. Thus, an ion-pair RP-HPLC method for the simultaneous determination of lactose, glucose and galactose in original skim milk was investigated. The samples derived from an enzymatic lactose hydrolysis approach (0.5 L) using the commercial β-galactosidase Godo-YNL2. After derivatisation with p-aminobenzoic acid and sodium cyanoborohydride, the samples were injected on a RP-C₁₈ column. Tetrabutylammonium hydrogen sulphate was used as the ion-pair reagent in the eluent system. The sugars were quantified using photometric- (UV; 303 nm) and fluorescence-detection (λ_ex 313 nm, λ_em 358 nm). The overall run time was 27 min. The limits of detection (LOD) were estimated at 2 mg L⁻¹ (UV detection) and at 0.13 mg L⁻¹ (fluorescence detection). The limits of quantification were 6 mg L⁻¹ (UV detection) and 0.45 mg L⁻¹ (fluorescence detection). Thus, this analytical method is suitable for sensitive lactose quantification in milk systems of less than 10 mg L⁻¹.     

5-Hydroxymethylfurfural content in foodstuffs determined by micellar electrokinetic chromatography

Micellar electrokinetic chromatography (MEKC) has been applied for the determination of 5-hydroxymethylfurfural in several foodstuffs. A 75 mM phosphate buffer solution at pH 8.0 containing 100 mM sodium dodecylsulphate was used as background electrolyte (BGE), and the separation was performed by applying +25 kV in a 50 μm I.D. uncoated fused-silica capillary. Good linearity over the range 2.5–250 mg kg⁻¹ (r² ? 0.999) and run-to-run and day-to-day precisions at low and medium concentration levels were obtained. Sample limit of detection (0.7 mg kg⁻¹) and limit of quantification (2.5 mg kg⁻¹) were established by preparing the standards in blank matrix. The procedure was validated by comparing the results with those obtained with liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS). Levels of HMF in 45 different foodstuffs such as breakfast cereals, toasts, honey, orange juice, apple juice, jam, coffee, chocolate and biscuits were determined.     

Validation of a method for simultaneous quantification of total carotenes and tocols in vegetable oils by HPLC

A normal-phase HPLC method for analysis of carotenes, tocopherols and tocotrienols has been developed and validated. In this work we presented a modification to the official AOCS method for analysis of tocols which allowed simultaneous quantification of the three groups of compounds, including carotenes. Analytes were separated using a gradient mobile phase (hexane and isopropanol) and with a gradient flow rate (1–2 mL min⁻¹). The column effluent was monitored by Photo Diode Array detector (PDA) set at 292 nm (tocols) and 455 nm (β-carotene) and by fluorescence detector set at an excitation wavelength of 290 and 330 nm emission. Inter- and intra-run accuracies and precision of the analytical method were better than ±15%. The lower limit of quantification was 5.0 mg L⁻¹ for the tocols and 0.1 mg L⁻¹ for carotenes. The method has been applied for the quantification of these compounds in Amazon oils.     

Development of a fast MECK method for determination of 5-HMF in honey samples

In this study, 5-hydroxymethylfurfural (5-HMF) determination was carried out by a micellar electrokinetic capillary chromatography (MEKC) methodology, using caffeine as the internal standard (IS). The optimisation of the electrolyte composition was approached using a 3² full factorial design with a central point to study the MEKC electrolyte components. Inspection of the response surface indicated that the optimal electrolyte composition was 5 mmol L⁻¹ sodium tetraborate (STB, pH 9.3) containing 120 mmol L⁻¹ sodium dodecyl sulphate (SDS). Under optimal CE conditions, separation of the investigated substance was achieved in less than 0.7 min. Quality parameters, such as linearity (R² > 0.99), precision (RSD < 5.41%), detection and quantification limits (3.37 and 11.24 mg kg⁻¹ for honey samples) and recovery (96.37–99.56%). The proposed methodology was successfully applied to the analysis of 5-HMF in honey samples. The analytical performance of this method makes it suitable for implementation in food laboratories for the routine determination of 5-HMF in honey samples.     

Pervaporation flow injection analysis for the determination of sulphite in food samples utilising potassium permanganate–rhodamine B chemiluminescence detection

A simple pervaporation flow injection chemiluminescence (PFI-CL) procedure was utilised as an on-line separation for the analysis of contaminated sulphite in food samples. The method involves the injection of standard and/or sulphite sample solutions into a 0.20 M sulphuric acid donor stream. Sulphite is converted to sulphur dioxide and transported to the donor chamber of a pervaporation module. The sulphur dioxide gas then evaporates into the headspace and diffuses across a semi-permeable PTFE membrane into an acceptor stream containing 0.75% (m/v) sodium hexametaphosphate and 1.0 mg L⁻¹ rhodamine B in 0.02 M H₃PO₄, which functions as a carrier solution for the chemiluminescence detection. The sulphur dioxide in the acceptor stream merges at a T-piece with a reagent stream consisting of potassium permanganate (8.0 × 10⁻⁵ M) prepared in the acidic sodium hexametaphosphate carrier solution. The elicited chemiluminescence intensity of the resulting reaction mixture was measured at a red sensitive photomultiplier tube operated at a voltage of 1.00 kV. Optimal experimental conditions for an on-line determination of sulphite were investigated. The second-order polynomial calibration curve was developed over the concentration range of 0.5–10.0 mg L⁻¹ sulphite with a resulting equation of I = −0.239C² + 4.846C − 1.64, r² = 0.9997. The detection limit was found to be 0.2 mg L⁻¹ with a sampling frequency of 30 h⁻¹. The effects of common anionic and cationic interferences were also investigated. The proposed PFI procedure was successfully applied to the determination of sulphite in different food samples. The PFI data was validated versus standard differential pulse polarography.     

Simultaneous kinetic-spectrophotometric determination of maltol and ethyl maltol in food samples by using chemometrics

A fast and accurate procedure has been researched and developed for the simultaneous determination of maltol and ethyl maltol, based on their reaction with iron(III) in the presence of o-phenanthroline in sulfuric acid medium. This reaction was the basis for an indirect kinetic spectrophotometric method, which followed the development of the pink ferroin product (λ_max = 524 nm). The kinetic data were collected in the 370–900 nm range over 0–30 s. The optimized method indicates that individual analytes followed Beer’s law in the concentration range of 4.0–76.0 mg L⁻¹ for both maltol and ethyl maltol. The LOD values of 1.6 mg L⁻¹ for maltol and 1.4 mg L⁻¹ for ethyl maltol agree well with those obtained by the alternative high performance liquid chromatography with ultraviolet detection (HPLC-UV). Three chemometrics methods, principal component regression (PCR), partial least squares (PLS) and principal component analysis–radial basis function–artificial neural networks (PC–RBF–ANN), were used to resolve the measured data with small kinetic differences between the two analytes as reflected by the development of the pink ferroin product. All three performed satisfactorily in the case of the synthetic verification samples, and in their application for the prediction of the analytes in several food products. The figures of merit for the analytes based on the multivariate models agreed well with those from the alternative HPLC-UV method involving the same samples.     

Evaluation of QuEChERS method for the determination of organochlorine pesticide residues in selected groups of fruits

This paper reports a method for organochlorine pesticide determination in selected fruit species where pesticide residues were extracted and cleaned using a buffered QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method, followed by GC–MS analysis. The method results showed the matrix-matched calibration curve linearity was >0.99 for all target analytes. With pesticide recovery rates (spiked at 0.008 mg kg⁻¹) ranging from 70% to 120%, and RSD values <17% for most compounds, the limit of quantification ranged from 0.001–0.013 mg kg⁻¹. Finally, the method ruggedness was further demonstrated by analysis of actual commercial fruits and baby food samples.     

A rapid liquid chromatography method for determination of glufosinate residue in maize after derivatisation

A rapid liquid chromatographic method for glufosinate analysis in maize samples after derivatisation has been developed. The labelled glufosinate was separated on a Kromasil C₁₈ column (250 mm × 4.6 mm, 5 μm) and UV detection was applied at 360 nm. The optimisation of derivatisation conditions and the influence of different ion-pair reagents on the separation were discussed. The method linearity correlation coefficient was 0.9998 in concentrations ranging from 0.1 to 20 mg L⁻¹. The level of quantification was set to 0.02 mg kg⁻¹, and reached pesticide EU-MRLs for glufosinate in the maize samples. The proposed method was applied to the quantitative determination of glufosinate in samples with recoveries of 98.0–100.5% and RSDs of 2.13–4.13%.     

Chemical composition of cultivated seaweed Ulva clathrata (Roth) C. Agardh

Samples of cultivated Ulva clathrata were collected from a medium scale system (MSS, 1.5 × 1.5 m tank), or from a large scale system (LSS, 0.8 ha earthen pond). MSS samples were dried directly while the LSS sample was washed in freshwater and pressed before drying. Crude protein content ranged 20–26%, essential amino acids accounting for 32–36% of crude protein. The main analysed monosaccharides were rhamnose (36–40%), uronic acids (27–29%), xylose (10–13%) and glucose (10–16%). Some notable variations between MSS and LSS samples were observed for total dietary fibre (26% vs 41%), saturated fatty acids (31% vs 51%), PUFAS (33% vs 13%), carotenoids (358 vs 169 mg kg⁻¹ dw) and for Ca (9 vs 19 g kg⁻¹), Fe (0.6 vs 4.2 g kg⁻¹), Cu (44 vs 14 mg kg⁻¹), Zn (93 vs 17 mg kg⁻¹) and As (2 vs 9 mg kg⁻¹). The chemical composition of U. clathrata indicates that it has a good potential for its use in human and animal food.     

Multiresidue method for determination of 88 pesticides in berry fruits using solid-phase extraction and gas chromatography–mass spectrometry: Determination of 88 pesticides in berries using SPE and GC–MS

A method using solid phase extraction (SPE) cleanup followed by gas chromatography–mass spectrometry (GC–MS) has been established for quantitative determination of 88 pesticide residues in berry fruits including raspberry, strawberry, blueberry and grape. Based on an appraisal of the characteristics of GC–MS, validation experiments were conducted for 88 pesticides. In the method, solid-phase extraction was carried out using Envi-Carb cartridge coupled with NH₂-LC cartridge with acetonitrile–toluene (3:1, v/v) as the eluted solvent. In the linear range of each pesticide, the correlation coefficient was R² ? 0.99. At the low, medium and high three fortification levels of 0.05–0.5 mg kg⁻¹, recoveries fell within 63–137%. The relative standard deviation was between 1% and 19% for all 88 pesticides. Low limits of detection (0.006–0.05 mg kg⁻¹) and quantification (0.02–0.15 mg kg⁻¹) were readily achieved with this method for all tested pesticides.     

Comparison of positive and negative ion detection of tea catechins using tandem mass spectrometry and ultra high performance liquid chromatography

Tea catechins are an important group of natural compounds associated with health promoting effects and desired commodities for the growing market of dietary supplements and functional foods. Consequently these compounds attract more interest of research groups worldwide. A reliable quantitative analysis of tea catechins is essential for human intervention studies, manufacturers of dietary supplements and quality control by authorities. UHPLC–ESI-MS/MS analytical method was chosen due to rapid runtime, high sensitivity and selectivity. The chromatographic separation of eight tea catechins was achieved within 2.5 min on C₁₈ BEH analytical column (100 mm × 2.1 mm i.d.; 1.7 μm), whilst the gradient elution mode was employed using water:methanol mobile phase with addition of volatile organic acid. The concentration of organic acids in the mobile phase was optimised within the range of 0.01–0.1% (v/v). High sensitivities were achieved in positive (10.2-16.8 fmol/inj.) and negative ion detection mode (102.1-168.1 fmol/inj.), through accurate and complex tuning of MS parameters. The UHPLC-ESI-MS/MS method was validated in terms of linearity (>0.9997; >0.9990), range (0.02–2.40 mg L⁻¹; 0.15-24.00 mg L⁻¹), LOD (3.0–4.8 μg L⁻¹; 30.1–48.0 μg L⁻¹), LOQ (9.9–15.8 μg L⁻¹; 150.5-240.0 μg L⁻¹), intra-day precision (4.4-7.1% RSD; 3.3-5.1% RSD), accuracy (94.06-113.7%; 89.5-108.4%), retention time repeatability (0.0-0.5% RSD; 0.0-0.6% RSD), and peak area repeatability (1.2-4.0% RSD; 2.4-3.5% RSD) for positive and negative ion detection modes, respectively. The statistical comparison of the quantitative results obtained in positive and negative ion detection mode was performed.     

Multi-residue method for determination of seven neonicotinoid insecticides in grains using dispersive solid-phase extraction and dispersive liquid–liquid micro-extraction by high performance liquid chromatography

A method using dispersive solid-phase extraction and dispersive liquid–liquid micro-extraction cleanup followed by high performance liquid chromatography (HPLC) has been established for determination of seven neonicotinoid insecticides residues in grains including brown rice, maize, millet and oat. Based on an appraisal of the characteristics of HPLC, validation experiments were conducted for seven neonicotinoid insecticides. In the method, dispersive solid-phase extraction was carried out using PSA and bonded C₁₈ coupled with graphitised carbon black with acetonitrile as the eluted solvent. In the linear range of each pesticide, the correlation coefficient was R² ? 0.99. At the low, medium and high three fortification levels of 0.05–0.8 mg kg⁻¹, recoveries fell within 76–123%. The relative standard deviation was between 0.9% and 12.6% for seven neonicotinoid pesticides. Low limits of detection (0.002–0.005 mg kg⁻¹) and quantification (0.007–0.018 mg kg⁻¹) were readily achieved with this method for all tested pesticides.     

Development of polyclonal antibody-based indirect competitive enzyme-linked immunosorbent assay for sodium saccharin residue in food samples

A sensitive and specific polyclonal antibody (PcAb)-based indirect competitive enzyme-linked immunosorbent assay (icELISA) for sodium saccharin is described. 6-Amino saccharin was coupled to carrier protein for artificial antigen by diazotisation. New Zealand white rabbits were immunised to obtain anti-sodium saccharin PcAb and then icELISA was developed. The assay showed high sensitivity and specificity to sodium saccharin, with the 50% inhibition value (IC₅₀) of 0.243 μg mL⁻¹, workable range (IC₃₀–IC₇₀) of 0.050–12.8 μg mL⁻¹ and limit of detection (LOD, IC₂₀) of 0.021 μg mL⁻¹. The average recoveries of sodium saccharin in spiked food samples were estimated ranging from 70.7% to 98.8%. A statistically significant correlation of results was obtained between this new ELISA and previously established HPLC approaches with the food-relevant sodium saccharin concentration range 0–320 μg mL⁻¹ (R² = 0.9887–0.9975). These results indicated that the established ELISA was a potential and useful analytical tool for rapid determination of sodium saccharin residue in food samples.     

Effect of cross-linking on the resistance to enzymatic hydrolysis of waxy maize starch and low-methoxy pectin

Gelatinised waxy maize starch and low-methoxy pectin mixtures were solubilised/dispersed in water and cross-linked with sodium trimetaphosphate (STMP). The polysaccharides were subjected to α-amylase, β-amylase or amyloglucosidase (AMG) hydrolysis for different times, and at two starch to pectin combination ratios (3:2 and 2:3). The extent of hydrolysis by porcine pancreatic α-amylase of the cross-linked (gelatinised) starch was 54.8–58.9% in comparison with gelatinised starch (for different incubation times), corresponding to 52.3–58.9% and 51.3–55.3% of the starch in the uncross-linked (UL) 3:2 and 2:3 starch to pectin ratios (for the same hydrolysis times). Blends of individually cross-linked starch to pectin ratios (3:2 and 2:3) were hydrolysed to 66.2–67.0% and 65.4–71.8%, respectively, compared with the corresponding UL counterparts. When the gelatinised starch was incubated for 0.5–36 h with β-amylase, hydrolysis ranged from 9.2% to 26.2%, and from 5.4% to 12.2% when the starch was cross-linked (corresponding to 40.0–58.7% of the gelatinised starch). For starch to pectin ratios of 3:2 or 2:3 blended after cross-linking, or by simultaneous cross-linking, hydrolysis represented 2.3–3.4% and 0.3–0.6% for the 3:2 ratio but only 1.2–2.0% and 0–0.3% for the 2:3 ratio. Hydrolysis with AMG using a 0.1 mg ml⁻¹ enzyme concentration caused 51.8% hydrolysis of gelatinised starch, which was lowered to 35.2% (i.e. by 68%) after cross-linking. At a higher enzyme concentration (1 mg ml⁻¹), the comparable figures were 91.7% and 71.9% (a reduction of 78.4%). For the UL 3:2 starch to pectin ratio and 0.1 and 1 mg ml⁻¹ enzyme concentrations, there was 27.8% and 56.5% hydrolysis of the polysaccharide which translated to 24.3% (87.4%) and 45.8% (81.1%), respectively, after cross-linking. Comparable figures for the 2:3 ratio (for the 0.1 and 1 mg ml⁻¹ enzyme concentrations) were 20.2% and 36.5% hydrolysis of the UL samples and 18.2% (90.1%) and 32.5% (89.0%) hydrolysis, respectively, after cross-linking.     

Effect of modifiers for As,Cu and Pb determinations in sugar-cane spirits by GF AAS

Palladium plus magnesium nitrates with and without Ir, Ru and W were evaluated for the simultaneous determination of As, Cu and Pb in cachaça by graphite furnace atomic absorption spectrometry. For 20 μL of sample, 5 μL Pd(NO₃)₂ and 3 μL Mg(NO₃)₂ dispensed together onto the Ir-coated platform of the THGA, analytical curves in the 0–30.0 μg L⁻¹ As, 0–1.50 mg L⁻¹ Cu and 0–60.0 μg L⁻¹ Pb were built up and typical linear correlation coefficients were always better than 0.999. The limit of detection was 1.30 μg L⁻¹ As, 140 μg L⁻¹ Cu and 0.90 μg L⁻¹ Pb. As, Cu and Pb contents in 10 cachaça samples agreed with those obtained by ICP-MS. Recoveries of spiked samples varied from 96% to 106% (As), 97% to 112% (Cu) and 92% to 108% (Pb). The relative standard deviation (n = 12) was typically 2.7%, 3.3% and 1.9%.     

Spectrophotometric determination of iodate in iodised salt by flow injection analysis

A simple and sensitive flow injection (FI) method for the determination of iodate is proposed. The method is based on the reaction of iodate with hydroxylamine in acidic solution. Sulfanilamide is diazotised by the nascent nitrite and the diazonium ion produced is then coupled with N-(1-naphthyl)ethyenediamine in hydrochloric acid medium to form an azo dye which is measured spectrophotometrically. The calibration graph for iodate is linear in the range of 0.1–30 mg L⁻¹ with a correlation coefficient of 0.9992. The limit of detection and relative standard deviation are 0.02 mg L⁻¹ and 1.2% (5 mg L⁻¹, n = 8). The method has been applied to the determination of iodate in table salts and accuracy was assessed through recovery experiments and independent analysis by a conventional titrimetric method.     

Analysis of potential adulteration in herbal medicines and dietary supplements for the weight control by capillary electrophoresis

Four different phytopharmaceutical dosage forms for use in weight control programs were analyzed. Two different ground herbal blends and their correspondent infusions, a capsule and a tincture were investigated for the presence of compounds used as adulterants in these products. A capillary electrophoresis (CE) method was developed and validated. The optimized experimental conditions were: BGE, sodium tetraborate buffer 20 mM, pH 9.2, voltage applied 30 kV, capillary temperature 25 °C, injection sample at 0.5 Psi during 5 s. Ephedrine, norephedrine, caffeine and furosemide were baseline separated in less than 7 min; the migration times were found to be 2.65, 2.90, 3.75 and 6.58 min, respectively. The analysis showed in sample 3 concentrations of 0.45 ± 0.03 mg g⁻¹ (ephedrine), 0.33 ± 0.02 mg g⁻¹ (norephedrine), 1.09 ± 0.41 mg g⁻¹ (caffeine) and 0.80 ± 0.17 mg g⁻¹ (furosemide). Caffeine content in samples 1, 2 and 4 was 0.61 ± 0.06 mg g⁻¹, 15.66 ± 1.05 mg g⁻¹ and 2.27 ± 0.13 mg ml⁻¹, respectively. Linearity was obtained in the concentration range of 1–1000 μg ml⁻¹. Limits of detection (LOD) and quantification (LOQ) were determined as 0.42 μg ml⁻¹ and 1.40 μg ml⁻¹ (ephedrine), 0.47 μg ml⁻¹ and 1.40 μg ml⁻¹ (norephedrine), 0.12 μg ml⁻¹ and 0.48 μg ml⁻¹ (caffeine), 0.22 μg ml⁻¹ and 0.73 μg ml⁻¹ (furosemide).     

Microwave-assisted extraction and determination of cyanuric acid residue in infant formula samples by liquid chromatography–tandem mass spectrometry

In the present work, microwave-assisted extraction method in combining with liquid chromatography–tandem mass spectrometry (LC–MS/MS) was proposed for the determination of cyanuric acid (CYA) in infant formula samples. The separation was performed on a MERCK ZIC HILIC column (150 × 2.1 mm i.d., 5 μm) with gradient elution of 20 mM ammonium acetate solution – acetonitrile. The method could respond linearly with cyanuric acid at concentrations from 1.0 to 50 ng mL⁻¹ with a quantification limit of 0.25 mg kg⁻¹. The intra- and inter-day precision was less than 4% and the recovery of the assay was in the range of 86.7–93.1%. In the analysis of practical spiked infant formula samples, the new method yielded satisfactory results. Due to its simplicity and accuracy the straightforward method is particularly suitable for routine cyanuric acid detection.     

Simultaneous determination of calcium,magnesium and zinc in different foodstuffs and pharmaceutical samples with continuous wavelet transforms

This work presents novel and very simple spectrophotometric methods by ratio spectra–continuous wavelet transformation for the simultaneous determination of ternary mixtures of calcium, magnesium and zinc without prior separation steps. The methods are based on the complexation reaction of these elements with bromopyrogallo red (BPR) at pH 9.4. The results showed that calcium, magnesium and zinc could be determined simultaneously in the range of 0.1–3.5 mg L⁻¹, 0.2–3.2 mg L⁻¹ and 0.5–3.8 mg L⁻¹, respectively. Mexican hat and Morlet from the family of continuous wavelet transforms were selected and applied under the optimal conditions for multi-component determinations. In this study for improving the sensitivity the normalized spectra of divisors were used instead of standard spectrum of divisor. The method was tested by analyzing various synthetic ternary mixtures of Ca²⁺, Mg²⁺ and Zn²⁺. Under the working conditions, the proposed methods were successfully applied to simultaneous determination of elements in cows’ milk, powder milk, pharmaceutical product and tea samples.     

Improved sample treatment and chromatographic method for the determination of isoflavones in supplemented foods

Isoflavones are a group of substances that belong to the family of phytoestrogens. These natural substances may offer several benefits to human health. One of the most important sources for human isoflavone intake is soy and soybean food derivatives. An improved sample treatment followed by a high performance liquid chromatographic method for the determination of isoflavones in supplemented milk and juices is proposed and compared to the AOAC official method. Detection limits found were between 0.2 and 0.3 mg L⁻¹ for daidzein and genistein respectively. Quantification limits found were between 0.7 mg L⁻¹ for daidzein to 1.0 mg L⁻¹ for genistin, while inter and intra-day variability was under 10% in all cases. Recoveries for spiked samples were over 90% and under 110%. The method was validated by comparison with the AOAC method and by recovery assay methodology.