Application of Bismuth(III)-Entrapped XO Biosensing System for Xanthine Determination in Beverages

A xanthine biosensor was prepared by electrochemical immobilization of xanthine oxidize enzyme onto carbon paste electrode via entrapment of Bi³⁺. After the optimization of experimental parameters, analytical characteristics were investigated. Two linear ranges between 0.02 and 0.06 and 1–7.5 μM with the equation y?=?93.00x?+?0.12 and y?=?1.07x?+?18.03 with the correlation coefficients of R ²?=?0.9951 and R ²?=?0.9931, respectively, were obtained for this biosensing system. RSD value was calculated for 0.04 μM xanthine (n?=?5) and found as 3.84%. LOD and LOQ values were also calculated and revealed as 1.30?×?10^?8 and 4.3?×?10^?8 M, respectively. Then, this biosensor was applied for xanthine detection in real samples. As a sample treatment, only necessary dilutions were made. Four types of beverages including wine, energy drink, peach, and sour cherry juice were used for this purpose. Obtained recovery values demonstrate that this system is applicable for xanthine detection in real samples without needing any laborious sample pretreatment procedures.     

Simultaneous Detection of Azodicarbonamide and the Metabolic Product Semicarbazide in Flour by Capillary Electrophoresis

In the present work, capillary electrophoresis (CE) was used for the first time for the simultaneous analysis of azodicarbonamide (ADA) and semicarbazide (SEM), and the capillary electrophoresis separation conditions, extraction agents, and derivatization conditions were investigated. In 20 mmol L^?1 sodium tetraborate, 30 mmol L^?1 β-cyclodextrin (β-CD), 17 % isopropanol (v/v), and 25 mmol L^?1 sodium dodecyl sulfate (SDS) running buffer, ADA and SEM previously derivatized with 9-fluorenylmethyl chloroformate (FMOC) were separated in less than 25 min with good sensitivity. The linear ranges were 8.3?×?10^?4～6.6?×?10^?2 mmol L^?1 and 1.9?×?10^?3～3.4?×?10^?2 mmol L^?1, and detection limits (S/N?=?10) were 0.5 and 0.15 mg kg^?1 for ADA and SEM, respectively. The proposed method was successfully applied for the simultaneous analysis of ADA and SEM in five flour samples with satisfactory recovery data from 88.0 to 93.0 % for ADA and 98.0 to 106.0 % for SEM, indicating the valuable potential application of this method for food analysis.     

Optimization of the QuEChERS Method for Determination of Pesticide Residues in Chicken Liver Samples by Gas Chromatography-Mass Spectrometry

The goal of this research was to evaluate the application of Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method for the determination of organochlorine, organophosphate, and carbamate pesticides in fatty animal matrices such as liver of chicken obtained from National Research Institute of Animal Production in Balice (Poland). Pesticides extraction effectiveness was evaluated at two different spiking levels (0.010 and 0.020 mg kg^?1) and efficiency of the dispersive solid-phase extraction (d-SPE) clean-up step was evaluated by comparison adding different d-SPE sorbent combinations (PSA?+?GCB, PSA?+?C₁₈, PSA?+?SAX, and PSA?+?NH₂). The analysis of pesticide residues was performed by gas chromatography ion trap mass spectrometry (GC/IT-MS). The linear relation was observed from 0 to 400 ng mL^?1 and the determination coefficient R ²?>?0.997 in all instances for all target analytes. Better recoveries were obtained in samples at 0.020 mg kg^?1 spiking level. The recoveries were in the range 70–120 %, with relative standard deviation (RSD) values lower than 15 % at 0.020 mg kg^?1 spiking level for most pesticides. Similar recovery ratios were obtained with the four different combinations of sorbents tested in the clean-up step, with better precision when the (PSA?+?SAX) combination was tested. Limits of detection (LODs) ranged from 0.001 to 0.005 mg kg^?1 and limits of quantification (LOQs) ranged from 0.003 to 0.015 mg kg^?1. The proposed method was successfully applied analyzing pesticide residues in real chicken liver samples; detectable pesticide residues were observed, but in all of the cases, the contamination level was lower than the default maximum residue levels (MRLs) set by European Union (EU), Regulation (EC) N 396/2005.     

Validated RP-HPLC Method for Simultaneous Determination of Tocopherols and Tocotrienols in Whole Grain Barley Using Matrix Solid-Phase Dispersion

The vitamers of vitamin E such as α-, β-, γ-, and δ-tocotrienol and α-, β-, γ-, and δ- tocopherol are important phytochemical compounds with antioxidant activity and with potential benefits for human health. A high-performance liquid chromatography with fluorescence detection (HPLC-FLD) method was validated for their determination in whole grain barley samples. Tocol extraction was performed by an optimized matrix solid-phase dispersion (MSPD) protocol with neutral alumina (0.5 g) as the dispersion agent and methanol (5 mL) as the elution solvent. The analytical column was an Eclipse XDB C18 column (150?×?4.6 mm, 5 μm) and it was operated at room temperature. Mobile phase was consisted of methanol/acetonitrile/i-propanol (55:40:5?v/v?%) and the elution was isocratic at a flow rate of 0.8 mL/min. Total analysis time was 12 min, and the detection of the tocols was performed with a fluorimetric detector where the excitation and emission wavelengths were set at 295 and 335 nm, respectively. Method validation was performed by means of intra-day (n?=?5) and inter-day accuracy and precision (n?=?8), sensitivity, and linearity. The linear regression coefficient (R ²) was higher than 0.99. The recoveries of the tocols from barley samples with the proposed extraction method were in an acceptable level (74–91 %) where the relative standard deviation ranged from 4.2 to 15.0 %. Limits of detection (LODs) and limits of quantification (LOQs) varied from 0.03 to 0.11 mg kg^?1 and 0.11 to 0.34 mg kg^?1, respectively.     

Development of an Aqueous Polyethylene Glycol-Based Extraction and Recovery Method for Almond (<Emphasis Type="Italic">Prunus armeniaca L</Emphasis>.) Protein

A novel method for protein extraction from sweet almonds with aqueous polyethylene glycol (PEG) as solvent and recovery from the extraction solution was developed. The extraction yields of different solvents, such as sodium hydroxide, sodium chloride, PEG 200, PEG 400, and PEG 600 aqueous solutions, were investigated and PEG 200 showed the highest extraction efficiency. The PEG-based microwave-assisted extraction (MAE) parameters were then optimized using response surface methodology. Under optimum condition, PEG 200 concentration of 25 % (w/w), liquid to solid ratio of 22 mL g^?1, microwave power of 120 W, extraction temperature of 45 °C, and extraction time of 4 min, the average extraction yield was 93.75?±?3.15 %. Subsequently, the almond protein was recovered from the extraction solution containing PEG with an isoelectric point-ethanol synergy precipitation protocol. The combined technique integrated the speed of isoelectric point precipitation with the completeness of alcohol precipitation. The recovery of almond protein was 98.81 % with a time of 3–5 min. The proposed PEG-based MAE and synergy precipitation protocol provide a rapid and effective method for almond protein extraction and recovery and have the potential to be used for other plant proteins.     

Determination of Hormones Residues in Milk by Gas Chromatography-Mass Spectrometry

The article presents the use of gas chromatography-mass spectrometry (GC-MS) technique in a method for the determination of 18 anabolic hormones from synthetic stilbenes, steroids and resorcylic acid lactones (RALs) groups in raw milk and milk powder. Sample preparation consisted of liquid-liquid extraction with diethyl ether and purification by solid phase extraction (SPE). Prior to instrumental analysis, the reaction of derivatisation with the heptafluorobutyric anhydride or N-methyl-N-trimethylsilyltrifluoroacetamide was performed. Method validation was carried out according to the required performance criteria of the Commission Decision 2002/657/EC. The apparent recovery of all analytes at 1 μg L^?1 (kg^?1) level was ranged between 70.4 and 119.4 % with the coefficients of variation values less than 30 %. The decision limits (CCα) and the detection capabilities (CCβ) were in the range of from 0.11 to 0.44 μg L^?1 (kg^?1) and from 0.19 to 0.75 μg L^?1 (kg^?1), respectively. The procedure has been accredited and successfully applied as a screening method for the presence of hormone residues in the study of commercial samples of milk.     

Ion-Pair-Based Air-Assisted Liquid–Liquid Microextraction for the Extraction and Preconcentration of Phthalic Acids from Aqueous Samples

In the present study, a rapid, simple, and highly efficient sample preparation method based on ion-pair air-assisted liquid–liquid microextraction using a low-density extraction solvent followed by high performance liquid chromatography–diode array detection has been developed for the extraction, preconcentration, and determination of three phthalic acids (phthalic acid, iso-phthalic acid, and terephthalic acid) in aqueous samples. In this method, a mixture of tri-butyl amine (as an ion-pair reagent) and toluene (as an extraction solvent) is transferred into an aqueous sample solution. Fine organic solvent droplets are formed by aspirating and dispersing of the mixture via syringe needle. After that, the formed ion-pairs are extracted into toluene, and after centrifuging, the obtained collected phase is transferred into a microtube and is evaporated to dryness under a stream of nitrogen at room temperature. The residue is re-dissolved in mobile phase and injected into the separation system for analysis. Under the optimum extraction conditions, the method showed low limits of detection and quantification between 0.09–0.24 and 0.29–0.78 ng mL^?1, respectively. Extraction recoveries and enrichment factors were from 88 to 98 % and 443 to 491, respectively. Relative standard deviations for the extraction of 5 ng mL^?1 of each analyte were less than 8.4 % for intra-day (n?=?6) and inter-days (n?=?5) precisions. Finally, different aqueous samples were successfully analyzed using the proposed method, and the target analytes were determined in some of them at ng mL^?1 level.     

Steady- and Unsteady-State Determination of the Water Vapor Permeance (<Emphasis Type="Italic">WVP</Emphasis>) of Polyethylene Film to Estimate the Moisture Gain of Packed Dry Mango

The water vapor permeance (WVP; g m^?2 d^?1 Pa^?1) of packaging films quantifying the water vapor transfer rate between foods and its surroundings is usually determined in units operating under steady-state conditions that do not necessarily reflect food handling scenarios. This study evaluated the determination of the WVP of a polyethylene (PE) film by steady-state method ASTM F1249-06 using a permeability cell and unsteady-state method ASTM E96/E96M in which 102 vacuum-sealed PE bags containing silica gel were stored (37.8 °C, 75% relative humidity) and weighed over 25 days. Average steady-state WVP (2.935 ± 0.365 × 10^?3, n = 4) fell within the 95% quantiles of unsteady-state WVP values (1.818–3.183 × 10^?3, n = 2142). Moisture uptake of dehydrated mango stored at 37.8 °C and 75% relative humidity was predicted with WVP values obtained by both methods. Predictions were validated by monitoring over 25 days the weight gain of 100 PE bags with dry mango. Experimental moisture averages during storage fell within one standard deviation of predictions using the unsteady-state WVP (R ² = 0.974). The same was observed only until day 15 for predictions obtained with the steady-state WVP. Calculations for days 20–25 overestimated the moisture uptake by 6.0–7.2%, resulting in registered R ² = 0.924. The unsteady-state WVP determination is low-cost, uses large numbers of film samples, and allowed more accurate predictions of dry mango moisture uptake. Knowledge of the moisture uptake controlled by the film WVP is essential when predicting the safety and quality changes limiting the shelf-life of foods.     

An approach for extraction,purification, characterization and quantitation of acylated-anthocyanins from <Emphasis Type="Italic">Nitraria tangutorun</Emphasis> Bobr. fruit

In the present study, a systematic approach for extraction, purification and analysis of acylated-anthocyanins from Nitraria tangutorun Bobr. fruit was explored. Six acylated-anthocyanins in N. tangutorun fruit were identified by HPLC-MS/MS, and a rapid and efficient HPLC-DAD method was developed to analyze the acylated-anthocyanins. Ultrasonic-assisted extraction conditions of acylated-anthocyanins were optimized using response surface methodology, extraction at 70 °C for 32 min using 70% methanol solution (0.1% HCl, v/v) rendered an extract with 80.37?±?2.66 mg/100 g of cyanidin-3-O-(trans-p-coumaroyl)-diglucoside and 97.88?±?4.06 mg/100 g of total acylated-anthocyanins. Nine macroporous resins were investigated for preliminary purification of acylated-anthocyanins. According to the static/dynamic adsorption and desorption tests, XDA-6 macroporous resin exhibited the maximum potential for preparing acylated-anthocyanins. The purity of cyanidin-3-O-(trans-p-coumaroyl)-diglucoside (43.30 mg/g) in purified acylated-anthocyanins was 201.89 times of that of the extract (0.21 mg/g), and the purity of total acylated-anthocyanins increased from 0.36 to 56.44 mg/g. Besides, the stability (t _1/2) of cyanidin-3-O-(trans-p-coumaroyl)-diglucoside and total acylated-anthocyanins increased by more than five-fold after purification using XDA-6. The established methods of analysis, extraction and purification of acylated-anthocyanins were hopefully utilized in food industry.     

Determination of Fatty Acids in Beef by Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry

A method was developed to separate, identify, and quantify 28 fatty acids of potential health relevance using liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS). Optimization of the experimental factors enabled baseline separation of the fatty acids including three pairs of closely related fatty acid isomers (C18:3n-3 and C18:3n-6; C18:1 and C18:1 t; and C20:3n-3 and C20:3n-6) that are challenging to separate. The limits of detection ranged from 0.01 to 0.26 mg L^?1 for the 28 fatty acids, and average recovery (mean, n?=?4) was found to be 102?±?12 %. In addition, the proposed method was validated using a quality control standard mix of fatty acids which yielded acceptable precision and accuracy. Fatty acid concentrations in conventional grain-fed and organic grass-fed beef were determined, and the results show that grass-fed beef have a lower omega-6 to omega-3 ratio (1.6–2.8) compared to grain-fed beef (9.3–13.5). Principal component analysis (PCA) was applied on the resulting data to find correlations between significant fatty acid composition and the diet of beef samples.     

Validation of a HS-SPME-GC Method for Determining Higher Fatty Esters and Oak Lactones in White Rums

Higher fatty esters and oak lactones are the main components of white rum aroma and furthermore, they have an important sensorial impact in these distilled alcoholic beverages. A method for analyzing these volatile compounds was validated. It involves a separation and concentration step using headspace solid-phase microextraction (HS-SPME) and determination by capillary gas chromatography using flame ionization detection. The method showed a good within-day (RSD?<?3 %) and between-day precision (RSD?<?5 %). The calibration curves were linear at the tested ranges (R?>?0.99) and the limits of detection and quantification were 0.001–0.018 and 0.003–0.054 mg L^?1 (12 %?v/v alcohol), respectively. Good recoveries were obtained (98.6–100.3 %). The method is suitable for the quality control of higher fatty esters and oak lactones in white rums.     

A New Ultrasonic Thermostatic-Assisted Cloud Point Extraction/Spectrophotometric Method for the Preconcentration and Determination of Bisphenol A in Food,Milk, and Water Samples in Contact with Plastic Products

Bisphenol A (BPA) contamination in foods and beverages usually occurs as a result of migration from the packages that contain it. In this context, a simple, easy-to-use, and efficient method was developed for the spectrophotometric determination of BPA in food, milk, and water samples in contact with plastic products after preconcentration by ultrasonic-thermostatic-assisted cloud point extraction (UTA-CPE). The method is based on the charge transfer-sensitive complexation of BPA with 3-methylamino-7-dimethylaminophenothiazin-5-ium chloride (AzB) in the presence of cetyltrimethylammonium bromide (CTAB) at pH 8.5 and then extraction of the formed complex into the micellar phase of polyethylene glycol dodecyl ether (Brij 35). The effects of the analytical variables affecting complex formation and extraction efficiency were systematically studied and optimized. Under optimized conditions, a good linear relationship was obtained in the range of 1.2–160 μg L^?1 with a detection limit of 0.35 μg L^?1. After preconcentration of a sample of 20 mL, a sensitivity enhancement factor was found to be 180. The accuracy and reliability of the method were evaluated by recovery studies from the spiked quality control samples and intraday and interday precision studies. From the studies conducted, the extraction efficiency (E%) was in the range of 94–103% with a relative standard deviation lower than 5.2% (as RSD%, n = 5). The method was successfully applied to the preconcentration and determination of BPA from the selected sample matrices.

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Graphical Abstract Migration of bisphenol A into the foodstuffs

     

Dispersive Liquid–Liquid Microextraction Combined With Micellar Electrokinetic Chromatography for the Determination of Some Photoinitiators in Fruit Juice

A fast and simple technique composed of dispersive liquid–liquid microextraction (DLLME) and micellar electrokinetic chromatography (MEKC) with diode array detector (DAD) was developed for the determination of multi-photoinitiators in fruit juice. Seven photoinitiators were separated in MEKC using a 25 mM borate buffer of pH 8.0, containing 24 mM sodium dodecyl sulfate (SDS), 10 mM β-cyclodextrin (β-CD), and 12.5 % acetonitrile (v/v). A CD-modified MEKC made this method more suitable for the determination of isopropylthioxanthone (ITX) isomers including 2-IXT and 4-ITX than the recently prescribed methods. A DLLME procedure was used as an offline preconcentration strategy. The satisfactory recoveries obtained by DLLME spiked at two spiked levels ranged from 85.6 to 124.7 % with relative standard deviations (RSDs) below 14 %. The limits of quantification (LOQs) ranged from 2.1 to 6.0 μg kg^?1.     

Determination of Ultraviolet Absorbers and Light Stabilizers in Food Packaging Bags by Magnetic Solid Phase Extraction Followed by High-Performance Liquid Chromatography

A sensitive and effective extraction technique based on magnetic solid phase extraction (MSPE) was developed for separating and detecting trace amounts of ultraviolet (UV) absorbers and light stabilizers in plastic packaging prior to high-performance liquid chromatography (HPLC). The magnetic Fe₃O₄/Au composites were synthesized by chemical co-precipitation, and a HAuCl₄ solution was added to the Fe₃O₄ NPs, followed by the addition of hydroxylamine hydrochloride as a reducing agent for Au shell formation. The main factors affecting the adsorbability of UV absorbers, such as the effects of pH of the sample solution, the amount of adsorbent, extraction time, and desorption conditions on the magnetic nanoparticles (MNPs), were investigated and optimized. The proposed method showed good linearity in the range from 0.75 to 100 μg/mL with good presenting regression coefficients (R ²) ≥0.9999. Low limits of detection (LODs) were achieved at 0.05, 0.14, and 0.04 μg mL^?1 for 2-hydroxy-4-n-octoxy-benzophenone (UV-531), 2-tert-butyl-6-(5-chloro-2H-benzotriazol-2-yl)-4-methylphenol (UV-326), and 2-(2H-benzotriazol-2-yl)-4,6-di-tert-pentylphenol (UV-328), respectively. The mean recoveries were in the range from 88.06 to 100.29% at 2.5, 5, and 10 μg mL^?1 spiked levels, and the relative standard deviations (RSDs) were in the range from 0.98 to 3.56%.     

Optimization of Ultrasound-Assisted Extraction of Lycopene from Papaya Processing Waste by Response Surface Methodology

Lycopene possesses strong antioxidant ability, which may provide protection against cancer and other degenerative diseases. An ultrasound-assisted extraction method has been developed for the extraction of lycopene from papaya (Carica papaya L.) processing waste. Optimization conditions were firstly determined using single factor experiment, and then response surface methodology was used. Analysis showed that second-order polynomial models produced a satisfactory fitting of the experimental data with regard to lycopene (R ²?=?0.9147, P?<?0.001). The optimal conditions were 42.28 % ethanol in ethyl acetate as a solvent and extraction for 26.09 min at 50.12 °C. Under these conditions, the maximum yield of lycopene was 189.8?±?4.5 μg/g fresh weight (FW), which was higher than that obtained using the traditional extracting method (153.9?±?7.8 μg/g FW) and the Soxhlet extraction method (68.3?±?4.1 μg/g FW). The crude extract obtained could be used as either food additive or for further isolation and purification of lycopene. The results obtained are helpful for full utilization of papaya.     

Optimizing the Supercritical Fluid Extraction of Lutein from Corn Gluten Meal

Supercritical CO₂ was used to extract xanthophylls from corn gluten meal (CGM). Data from a Box-Behnken experimental design was used to model optimal lutein extraction based on extraction temperature (40–80 °C), pressure (5500–7500 psi), and fraction of ethanol co-solvent added (5–15% by volume of total solvent). Lutein extraction was also strongly correlated with zeaxanthin extraction with a correlation coefficient (r) of 0.995. The response surface model for lutein extraction indicated that the amount of co-solvent had the largest impact (p?<?0.001) on lutein extraction yield. Influence of temperature and pressure were limited to quadratic or interaction effects (p?<?0.15). The optimal lutein extraction conditions predicted with the model were a temperature of 40 °C, pressure of 6820 psi, and co-solvent (ethanol) addition of 15% by volume. At these conditions, lutein recovery from CGM was 2.6 times higher than the amount recovered with a quintuple extraction using ethanol and chloroform/dichloromethane (2:1). The strong linear effect of co-solvent addition suggests the possibility of further increasing lutein extraction with the addition of more co-solvent. CGM protein loss during extraction was also calculated and determined to be negligible.     

Determination of Eugenol in Aquatic Products by Dispersive Solid-Phase Extraction and Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry

A novel procedure, dispersive solid-phase extraction coupled with ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), was developed for the determination of eugenol in aquatic products (shrimp, crab, and carp). Aquatic products were extracted with acetonitrile and primarily purified by dispersive solid-phase extraction with graphitized carbon black as absorbent. The pretreated acetonitrile extract was detected by UHPLC-MS/MS. UHPLC was carried out on Dikma Endeavorsil C₁₈ (30 mm × 2.1 mm, 1.8 μm) column eluted by methanol and water (80:20 v/v) at a rate of 0.30 mL min^?1. Tandem mass spectrometry was performed by electrospray ionization in negative ion mode to identify and quantify eugenol during multiple reaction monitoring. Under optimized analytical conditions, the matrix-matched spiked calibration sample demonstrates good linearity between 5.0 and 500.0 μg kg^?1 with a linear regression coefficient of 0.9996. The average recovery of eugenol from aquatic products is 95.3–103.4% at spiked levels between 5 and 50 μg kg^?1 with a relative standard deviation (n = 6) less than 5.4%. The limits of detection and quantification for eugenol were calculated to be 1.47 and 4.91 μg kg^?1, respectively. In comparison with those reported, the proposed method has advantages in low detection limit, high recovery, and short analysis time, meeting the requirements for the determination of trace eugenol residue in aquatic products.     

Reusable 2-(Dimethylamino)ethyl Methacrylate Polymers On-line Solid-Phase Extraction Coupled to Liquid Chromatography Tandem Mass Spectrometry for Rapid Determination of Acidic Pesticides in Foods

A rapid and easy method was developed for the sensitive determination of six acidic pesticides (2,4,5-trichlorophenoxyacetic acid, acifluorfen, methyl-4-chlorophenoxyacetic acid, fluroxypyr, haloxyfop, and bispyribac-sodium) in pear, apple, cucumber, and rice samples. This method was a full automatic platform using a novel polymeric monolith-based on-line solid-phase extraction (SPE) coupled to liquid chromatography-tandem mass spectrometry (LC-MS/MS). A poly(2-(dimethylamino)ethyl methacrylate-co-ethylene dimethacrylate) monolith was fabricated as the sorbent, and the morphology, surface area, and extraction performance of the polymers were characterized. With the addition of 10.0 mL of acetonitrile and 1.0 to 5.0 g of anhydrous NaCl, the food samples were homogenized and centrifuged. The extracted samples were directly loaded and cleaned on the polymers using a 5 mmol L^?1 ammonium acetate solution and subsequently eluted into the analytical column for next separation with an acetonitrile-1.5% formic acid solution as eluting solvent. The detection limit (S/N?=?3) was 0.2 to 2.0 μg kg^?1 for the analytes, and the intra- and inter-day recoveries in pear, apple, cucumber, and rice samples ranged from 80 to 104%, and 83 to 115% with relative standard deviations of less than 13.5%. The developed on-line SPE LC-MS/MS protocol permits an automated and high-throughput determination in only 15 min. Moreover, the intra-batch (n?=?5) and inter-batch precisions (n?=?3) for the preparation of the polymers were lower than 11.6% and the sorbent was sufficiently stable for 500 extraction cycles without a significant loss in the extraction efficiency. The developed method was successfully applied to monitor six acidic pesticides in vegetable, fruit, and cereal samples.     

A Short Extraction Time of Polysaccharides from Fenugreek (<Emphasis Type="Italic">Trigonella foencem graecum</Emphasis>) Seed Using Continuous Ultrasound Acoustic Cavitation: Process Optimization,Characterization and Biological Activities

In this study, the effective variables involved in ultrasound-assisted extraction (UAE) of fenugreek (Trigonella foenum graecum) seeds polysaccharides (FSPs) were optimized through Box–Behnken response surface design. The maximum yield of FSPs was 33.49% under the modified optimum conditions of an ultrasound power of 120 W, an irradiation time of 22 min and a liquid-to-solid ratio (L/S) of 30:1 mL g^?1. The predicted yield was in a very good agreement with the experimental yield of 33.41%. This value was higher than the FSPs yield obtained using conventional extraction (CE) method for 180 min at the same extraction temperature and L/S ratio. The primary chemical and structural characteristics were investigated by UV, FT-IR, and GC-MS. FSPs found to be a heteropolysaccharide consisted of galactose (38.18%), glucose (3.71%), mannose (46.13%), rhamnose (1.02%), and arabinose (0.83%). Furthermore, the FSPs exhibited considerable scavenging activity against 1,1-Diphenyl-2-Picrylhydrazyl free radicals and ferric reducing antioxidant power and reducing power, in a concentration-dependent mode in vitro. Our results suggested that UAE technique gave a higher yield of FSPs with a shorter extraction time than the CE and FSPs can also be used as promising resources of natural agent for functional foods and medicinal industries.     

TLC-Digital Image-Based Fluorometric Analysis of Ergosterol and Chitin Content in Food Grains Artificially Infested with <Emphasis Type="Italic">Aspergillus flavus</Emphasis> and <Emphasis Type="Italic">Fusarium verticillioides</Emphasis>

Novel thin-layer chromatography-digital image-based analytical methods were developed for the quantitation of ergosterol and chitin content in six food matrices (rice, wheat, maize, sorghum, groundnut, and sunflower), artificially infested with Aspergillus flavus (MTCC 6513)/Fusarium verticillioides (MRC 826). For ergosterol, single-step method, based on liquid/liquid extraction, was followed by thin-layer chromatography (TLC). Chitin was solubilized using lithium chloride (5%) in dimethyl acetamide and converted to chitosan using 5 N NaOH and subsequently complexed with calcofluor white dye. The absorption and emission maxima of chitosan-calcofluor complex were recorded at λ 350/230 and 430 nm, respectively. The sensitivity based on the limit of detection (LOD) was found to be 100 ng both for ergosterol and chitin analysis. Based on ergosterol and chitin analysis, groundnut and maize were found to be suitable substrates for A. flavus (p?<?0.013 and p?<?0.01), while sorghum followed by groundnut and sunflower were found to be ideal for F. verticillioides (p?<?0.01 and p?<?0.0001) and rice was established as poor substrate as there was no growth on it up to 12 days of incubation. A strong correlation was found between ergosterol and chitin contents with regression (r ²) values of 0.974 and 0.997 in food grains inoculated with A. flavus and F. verticillioides, respectively, during the period of infection. The authenticity of the two methods developed was further confirmed by applying them to commercial food grains and flours. Thus, ergosterol in combination with chitin analysis could be successfully used as an index of fungal contamination employing TLC-digital-based analytical methods.