Optimized Dispersive Liquid–Liquid Microextraction and Determination of Sorbic Acid and Benzoic Acid in Beverage Samples by Gas Chromatography

A new rapid method for direct determination of trace levels of sorbic and benzoic acids was developed by dispersive liquid–liquid microextraction and gas chromatography with flame ionization detection. In the proposed approach, the separation procedure of sorbic and benzoic acids was performed on a general chromatographic column without any prior derivatization processes. Some effective parameters on the microextraction recovery were studied and optimized utilizing multilevel factorial and central composite experimental designs. The best concurrent extraction efficiency acquired using ethanol and chloroform as dispersive and extraction solvents. Central composite design (CCD) resulted in the optimized values of microextraction parameters as follows: 1.0 mL of dispersive and 0.1 mL of extraction solvents, ionic salt concentration of 50 g?L^?1 at pH 4. Under optimum conditions, the calibration curve was linear over the range 0.5–20 mg L^?1. Relative standard deviation was 11% and 13% for five repeated determinations for sorbic and benzoic acids, respectively. Limits of detection were acquired as 0.2 mg L^?1 for sorbic acid and 0.5 mg L^?1 for benzoic acid. The average recoveries were 31% and 39% for sorbic and benzoic acids, respectively. The method was successfully applied to the determination of sorbic and benzoic acids as preservatives in beverage samples.     

Inhibitory effect of organic acids on arcobacters in culture and their use for control of Arcobacter butzleri on chicken skin

The inhibitory effects of 17 organic acids (C₂-C₁₆ fatty acids, sorbic, benzoic, phenylacetic, fumaric, succinic, lactic, malic and citric) on Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii were investigated by determining their IC₅₀ values, defined as the concentration of acid at which the target DNA sequence was expressed at 50% of the positive control level in cultures incubated at 30 °C for 24 h. DNA was analysed by real-time PCR. The Arcobacter strains tested were inhibited by all the organic acids, with the sensitivities in the order A. skirrowii > A. cryaerophilus > A. butzleri. Eight acids with IC₅₀ values of < 1 mg/mL against A. butzleri were tested for their effects on A. butzleri inoculated on chicken carcasses at a concentration of 5 log CFU/g of skin. Inoculated halved carcasses were immersed in solutions of the acids at 5 mg/mL for 1 min. Samples of skin were collected from carcass halves after storage at 4 °C for 0, 1, 2 or 3 days for enumeration of arcobacters on Muller-Hinton agar. All eight tested acids suppressed bacterial proliferation. The highest inhibitory activities were observed for benzoic, citric, malic and sorbic acids. Subsequent sensory analysis revealed benzoic acid to be the most suitable organic acid for chicken skin treatment.     

Effect of pre- and postharvest salicylic acid treatment on ethylene production,fungal decay and overall quality of Selva strawberry fruit

The effect of salicylic acid (SA) treatment at different concentrations and growth stages of strawberry (Fragaria ananassa cv. Selva) fruit on postharvest ethylene production, fungal decay and overall quality index was studied. SA at all concentrations effectively reduced fruit ethylene production and fungal decay and retained overall quality. Treatment of plants at vegetative stage and fruit development stage followed by postharvest treatment of fruits with 1 and 2 mmol L⁻¹ was the most effective strategy, whilst with decrease in treatment time the effects of treatment decreased. Single stage treatment strategy of fruits with 2 mmol L⁻¹ SA at postharvest stage was most effective. Postharvest treatment with 4 mmol L⁻¹ SA slightly damaged the fruits and was less effective than 2 mmol L⁻¹ in retaining fruit quality.     

Occurrence of caffeine,saccharin, benzoic acid and sorbic acid in soft drinks and nectars in Portugal and subsequent exposure assessment

A reliable method for the determination of benzoic and sorbic acids, caffeine and saccharin in soft drink and nectars using high performance liquid chromatography and UV detection was validated. The chromatographic separation was achieved with a C18 column (250 × 4.6 mm) and one buffered mobile phase, KH₂PO₄ 0.02 M/ACN (90:10)/phosphoric acid at pH 4.2. The effluent was monitored at 220 nm.     

Efficacy of sodium chlorite as an inhibitor of enzymatic browning in apple slices

Sodium chlorite (SC) is an effective sanitizer for inhibiting microbial growth. This investigation was conducted to determine the efficacy of SC as a browning control agent for use on fresh-cut apple slices, applied alone, or in conjunction with organic acids. Additionally, the authors compared the efficacy of SC to that of acidified sodium chlorite (ASC) and to several other salts and examined the effect of pH and several different organic acids on efficacy of SC. The fresh-cut apple slices were dipped in treatment solutions for 1 min, then drained and placed in plastic containers at 20 °C for 24 h, and finally stored in polyethylene bags at 5 °C for 2 weeks. Color was measured periodically during storage. Lightness (L) values for all treated and control samples measured at 4 h, 24 h, and 2 weeks of storage were compared to L value for untreated samples measured immediately after cutting. Percent decrease in L-values was calculated for each sample at each time interval. Apple slices treated in ASC or SC solution had a significantly smaller decrease in L value indicating less browning than those treated in citric acid or water control at 4 h (P < 0.01), and with the exception of 1 g L⁻¹ ASC and 0.1 g L⁻¹ SC, all other ASC and SC treated slices still had significantly less browning than those for the water control (P < 0.01) at 24 h. After 2 weeks of storage, only SC (0.5–1.0 g L⁻¹), sodium bisulfite (0.5 g L⁻¹) and calcium l-ascorbate (10 g L⁻¹) continued to inhibit browning. Treatment with 0.5 g L⁻¹ SC and pH adjusted in the range from 3.9 to 6.2 using citric acid (CA) reduced browning more effectively than 0.5 g L⁻¹ SC without pH adjustment. Two organic acids, salicylic acid and cinnamic acid, when added to SC solution, were found to achieve even better inhibition of browning than CA at the same pH value.     

Sorbic,benzoic and propionic acids in fishery products: a survey of the South Korean market

This study was conducted to provide basic data as part of a project to distinguish naturally occurring organic acids from added preservatives. Accordingly, we investigated naturally occurring levels of sorbic, benzoic and propionic acids in fish and their processed commodities. The levels of sorbic, benzoic and propionic acids in 265 fish and their processed commodities were determined by high-performance liquid chromatography–photodiode detection array (HPLC-PDA) of sorbic and benzoic acids and gas chromatography-mass spectrometry (GC/MS) of propionic acid. For propionic acid, GC-MS was used because of its high sensitivity and selectivity in complicated matrix samples. Propionic acid was detected in 36.6% of fish samples and 50.4% of processed fish commodities. In contrast, benzoic acid was detected in 5.6% of fish samples, and sorbic acid was not detected in any sample. According to the Korean Food and Drug Administration (KFDA), fishery products and salted fish may only contain sorbic acid in amounts up to 2.0 g kg^?1 and 1.0 g kg^?1, respectively. The results of the monitoring in this study can be considered violations of KFDA regulations (total 124; benzoic acid 8, propionic acid 116). However, it is difficult to distinguish naturally generated organic acids and artificially added preservatives in fishery products. Therefore, further studies are needed to extend the database for distinction of naturally generated organic acids and added preservatives.     

Capillary zone electrophoresis of organic acids in beverages

Capillary zone electrophoresis (CZE) with photodiode array detection (PDA) was applied to simultaneously determine eleven organic acids in wine, beer, and fruit and vegetable juices after derivatisation with 2-nitrophenylhydrazine (2-NPH) in the presence of N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC·HCl). The parameters affecting CZE separation included type and concentration of buffer, pH, organic additive and applied voltage. Optimum conditions at 25 °C were: 30 mmol l⁻¹ borate buffer, pH 10.0, containing 100 ml l⁻¹ acetonitrile, at 20 kV, sample injection at 0.5 psi for 5 s, with direct detection at 230 nm. Separation of eleven organic acids was achieved within 12 min. Linear calibration curves with good fit were obtained in the range 10.0-100.0 mg l⁻¹. Limits of detection ranged from 2.0 to 10.0 mg l⁻¹. Intra-day precision with RSD?4.0% for migration time and ?5.0% for peak area, and inter-day precision with RSD?6.0% and ?9.0% for migration time and peak area, were obtained. Recovery for all beverage samples was 97±15%.     

Potentiometric sensor for sorbic acid determination in food products

The construction, evaluation and application of a new potentiometric sensor, namely, Pt|Hg|Hg₂(SOB)₂ |Graphite, where SOB stands for sorbate ion, are described. This electrode has a wide linear dynamic range between 5.0 × 10⁻⁷ and 1.0 × 10⁻² mol L⁻¹ with a near-Nernstian slope of (−58.6 ± 1.3) mV decade⁻¹ and a detection limit of 4.3 × 10⁻⁷ mol L⁻¹. The potentiometric response is independent of the pH of the solution in the pH range 6.0–9.0. The electrode is easily constructed at a relatively low-cost with fast response time (within 15–30 s) and can be used for a period of 4 months without significant change in its performance characteristics. The proposed sensor displayed good selectivities over a variety of other anions (carboxylates and inorganic anions). The potentiometric sensor was successfully applied to the determination of sorbic acid in real food samples, that is, soft drinks, skim yogurts, jams and sauces.     

Validation of a HPLC method for simultaneous determination of main organic acids in fruits and juices

A liquid chromatographic method for fast and simultaneous determination of tartaric, malic, ascorbic and citric acids was validated for further application to fruits and juices. Moreover, the organic acids content of commercial samples of fruits and juices were evaluated, as well as the ascorbic acid stability during the storage. Determination of organic acids was carried out using a liquid chromatograph coupled to a diode array detector, with reversed phase (C₁₈ column) and isocratic elution with 0.01 mol L⁻¹ KH₂PO₄ (pH = 2.60) mobile phase. The validation parameters showed efficiency, adequate linearity, relative standard deviation values between 0.4% and 2.3% (n = 10) for repeatability and from 1.2% to 5.0% (n = 18) for reproducibility, limits of detection (LD) were between 0.03 and 3.31 μg mL⁻¹ and quantification (LQ) were between 0.10 and 11.03 μg mL⁻¹, recovery rates were between 82% and 110%, for two levels. In addition, the method is fast (10 min) and generates low and non-toxic residues. The values found for vitamin C were about 10 times above the values declared at the package. Ready to drink juices have a composition similar to the fruit, concerning to organic acids, except for the powder juice, in which only ascorbic and citric acids were found, for all tastes. After opening the package, a decrease of 14.0% and 27.0% in ascorbic acid content was observed for orange powder and ready to drink juices, respectively.     

Effects of skin and grilling method on formation of heterocyclic amines in chicken pectoralis superficialis muscle

Mutagenic heterocyclic amines (HAs) originate in processed proteinaceous foods. The effects of the presence of skin (with vs. without) and of grilling method (two plate vs. infrared) on the content of HAs in grilled chicken pectoralis superficialis muscle (temperature, 220 °C) were investigated. HA precursors (creatine, creatinine, free amino acids and carbohydrates) and HAs of these raw and grilled breast muscles were determined. The muscles originated from 24 birds of either sex (provenance Ross; aged 40-45 days). The HA content was determined in homogenates of the upper and lower surface slices of the grilled muscles (T_i = 82 °C). A higher content of total free amino acids was seen for the muscle (27.1 mmol kg⁻¹ raw meat) than for the skin (21.7 mmol kg⁻¹ raw meat). The creatine, creatinine and carbohydrate levels in the skin were below the limits of detection. The contents of creatine (31.8-38.7 mmol kg⁻¹) and creatinine (0.24-0.33 mmol kg⁻¹) in the breast muscle were determined. Relatively high levels were seen for glucose (23 mmol kg⁻¹ raw meat) and fructose (10 mmol kg⁻¹ raw meat) in the muscle, with other sugars present at low levels (<2 mmol kg⁻¹ raw meat). For the chicken muscle grilled on a two-plate grill, the contents of total HAs (PhIP, MeIQx, DiMeIQx, Harman and Norharman) were lower with the skin in place than in the muscle grilled without the skin (3.5 μg kg⁻¹vs. 4.8 μg kg⁻¹). Also, during infrared grilling with the skin, lower amounts of HAs were formed than with grilling on the two-plate grill (2.4 μg kg⁻¹vs. 3.5 μg kg⁻¹). On average, the infrared-grilled samples with skin contained 3-fold more total HAs than similar samples without the skin (2.4 μg kg⁻¹vs. 0.8 μg kg⁻¹), with the highest levels seen for PhIP and MeIQx.     

Partition of selected food preservatives in fish oil–water systems

The partition coefficients (K_ow) of benzoic acid and sorbic acid in systems of fish oil (sand eel)–water, fish oil–buffer solution, rape oil–water and olive oil–water were experimentally determined in a temperature range from 5 to 43 °C and pH from 4.5 to 6.5 °C. The dimerization of benzoic acid in fish oil–water system was observed at 25 °C. Two modifications have been made to the Nordic Food Analysis Standard for the determination of sorbic acid by HPLC. The experimental results show that the K_ow of benzoic acid and sorbic acid in fish oil–buffer system is ca. 100 times lower than that in fish oil–water system. The K_ow values of benzoic acid and sorbic acid in fish oil and water system decrease with increasing system pH values. The partition coefficients of plant origin and fish origin oils are in the same order of magnitude even though their molecular structures are very different.     

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.     

Determination of selenium in garlic (Allium sativum) and onion (Allium cepa) by electro thermal atomic absorption spectrometry

A separation/enrichment procedure has been developed for the determination of selenium in garlic and onion samples by electrothermal atomic absorption spectrometry (ET-AAS) as a slurry sampling after preconcentration with 3,3-diaminobenzidine (DAB) reagent on the activated carbon. The influences of pH, time, amount of carbon and complexing reagent were outlined. The effect of acids used in the digestion of samples was also studied and compared. Selenium level was found to be 0.024 μg g⁻¹ for onion (n = 5; LOD – 0.5 μg L⁻¹; LOQ – 1.7 μg L⁻¹) and 0.015 μg g⁻¹ for garlic (n = 5; LOD – 1.3 μg L⁻¹; LOQ – 3.3 μg L⁻¹). Three different samples of garlic were analyzed by k₀-instrumental neutron activation analysis (k₀-INAA) at the Jozef Stefan Institute (JSI). The data obtained by k₀-INAA show that the content of selenium overlapped the results obtained by ET-AAS.     

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.     

Estimated intake of benzoic and sorbic acids in Denmark

The monitoring of food additives and recent dietary surveys carried out in Denmark have earlier been used to estimate the intake of sweeteners and nitrite in relation to acceptable daily intakes. The ubiquitous use of the preservatives benzoic and sorbic acids raises the question of the magnitude of the intake of these preservatives in relation to acceptable daily intakes. This area is explored in this paper. The content of benzoic and sorbic acids in all food groups, where they are allowed, was monitored in Denmark 17 times between 2001 and 2006 with a total of 1526 samples. Transgressions of maximum limits, illegal use or declaration faults were found in about 3% of samples. From repeated investigations on fat-based foods (salads and dressings), marmalade and stewed fruit, it is concluded that the amounts used in industry have been relatively stable throughout the whole period, although limited data for marmalade show some variation. Most foods in the categories soft drinks, dressings, fat-based salads, pickled herrings, and marmalade contain benzoic and sorbic acid, and sliced bread also contains in some cases sorbic acid. The median daily intake and intake distribution of benzoic and sorbic acids were calculated with data from the Danish National Survey of Dietary Habits and Physical Activity (age from 4 to 75 years) conducted in 2000–2004 with 5785 participants. The median intakes of both benzoic acid and sorbic acid are well below the acceptable daily intakes of 0–5 and 0–25 mg kg^?1 body weight (bw) day^?1 for benzoic and sorbic acid, respectively. However, the 90th percentile based on the average of the samples with a content of benzoic acid is higher than the acceptable daily intake for both men and women, with the highest value of 16 mg kg^?1 bw day^?1 for both boys and girls in the 4–6-year-old age group. Based on the average of all samples, the 95th percentile is over the acceptable daily intake for men up to 34 years and for women up to 24 years, and the 90th percentile for men up to 18 years and for women up to 10 years. Soft drinks, salads and dressings are the main contributors to benzoic acid intake. The sorbic acid intake based on the average of all samples is well below the acceptable daily intake. However, for the intake based on the average of samples with content, the 95th percentile exceeds the acceptable daily intake. This is caused by the dominating contribution to the intake of sorbic acid from sliced bread, but since only seven out of 42 samples have added sorbic acid, the calculation based on the average of samples with content will exaggerate the intake. With a built-in safety factor of 100 in the acceptable daily intakes and judging from the literature, the high intakes of benzoic acid should not cause any concern for ill-effects. However, there must be a reason to reconsider the maximum limits especially for benzoic acid in soft drinks, dressings and salads and for sorbic acid in sliced bread.     

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%.     

Determination of nitrofurans residues in animal feeds by flow injection chemiluminescence procedure

A simple flow injection chemiluminescence (FI-CL) method was developed for the rapid and sensitive determination of nitrofurans, including furazolidone, nitrofurantoin and nitrofurazone, in animal feeds based on its chemiluminescence induced by potassium permanganate in sulphuric acid medium. The method involves the injection of nitrofuran samples or standards into H₂SO₄ carrier stream, which then merges at a T-piece with a reagent stream consisting of KMnO₄ in the H₂SO₄ carrier solution. The elicited chemiluminescence intensity of the resulting reaction mixture was measured by photomultiplier tube operated at a voltage of 950 V. Optimum CL signals were given using 2.5 × 10⁻⁵ mol L⁻¹ potassium permanganate in 0.1 mol L⁻¹ sulphuric acid as an oxidant stream and a carrier stream of 0.1 mol L⁻¹ sulphuric acid with a total flow rate of 7.0 mL min⁻¹. Results detailing the optimisation of the analytical signal, calibration, and common interferences of animal feeds were also discussed. The proposed FI-CL method was successfully applied to the determination of nitrofurans in animal feeds, with excellent recoveries, as the determination is free from interference. The method validation has been compared versus HPLC method for animal feed samples.     

Electrochemical determination of glucose using polyaniline electrode modified by glucose oxidase

Polyaniline (PANI) enzyme electrode was formed by immobilisation of Glucose oxidase (GOx) via glutaraldehyde into electrochemically polymerised PANI on graphite electrode. Electrochemical polymerisation of PANI on graphite was performed from aqueous solution of 1.0 mol dm⁻³ HCl and 0.25 mol dm⁻³ aniline at constant current density of 2.0 mA cm⁻². Hronopotentiometric curves of the PANI enzyme electrode obtained at current density of 10 μA cm⁻² were recorded in different glucose concentrations. The linearity response range was between 1.0 and 5.0 mmol dm⁻³ of glucose concentration. The estimated apparent Michaelis–Menten constant, was K_m^′ = 0.30 mmol dm⁻³, which is significantly lower than that of free enzyme.     

Optimisation of a CE method for caffeine analysis in decaffeinated coffee

Multiple response simultaneous optimisation was used to develop a micellar electrokinetic chromatography method for caffeine determination in decaffeinated coffees. Buffer composition and voltage were optimised using a central composite design. Six responses were evaluated and each set of response values was regressed on the factor levels of the experimental design using linear and quadratic models. The regression models, correlation coefficients and a principal component analysis (PCA) were used to determine the optimum conditions. Successful results were obtained using a buffer containing 10 mmol L⁻¹ sodium carbonate and 50 mmol L⁻¹ sodium dodecylsulfate, 15 kV voltage, 25 °C temperature, 48 cm × 50 μm fused-silica capillary, hydrodynamic injection of 50 mBar during 7 s and detection at 206 nm. This optimised method was applied for caffeine analysis in 45 samples of commercial decaffeinated coffee.