Analysis of Non-volatile Organic Acids in Fermented and Dried Cocoa Beans by High Performance Liquid Chromatography

A high peiformance liquid chromatographic methodfor analysis of non-volatile acids (oxalic, citric, tartaric, succinic, malic and lactic) in fermented and dried cocoa beans is described. Bean samples were pulverized in dionized water using a Polytron Homogenizer (Brinkman)for 20 sec and centrifuged at 14000 rpm for 45 min at 25°C. The extract was alkalized to pH between 8-9 and passed through intermediate base anion exchange resin; the acidic fraction was eluted after adding 10lt/o sulphuric acid to the column. Polyphenols in the fraction were then eliminated by passing the acidic fraction through a reverse phase SEP-PAK that had been pre-wet with methanol. The eluate was analyzedfor non-volatile acids using Organic Acid Column (Bio-Rad) with O.lN H SO as a mobile phase at 65°C. The acids were detected at 214nm and quantified by comparing peak 2hezght of sample to those of standards. The method demonstrated excellent reproducibility and recoveries of the added acids

Validation of the procedure for the simultaneous determination of aflatoxins ochratoxin A and zearalenone in cereals using HPLC-FLD

Method validation for quantitative analysis of aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZEA) in cereals using HPLC with fluorescence detector (FLD) is described. Mycotoxins were extracted with methanol : water (80 : 20) and purified with a multifunctional AOZ immunoaffinity column before HPLC analysis. The validation of the analytical method was performed to establish the following parameters: specificity, selectivity, linearity, limits of detection (LOD) and quantification (LOQ), accuracy, precision (within- and between-day variability), stability, robustness, measurement of performance, and measurement of uncertainty. Calibration curves were linear (r > 0.999) over the concentration range, from the LOQ to 26, 40 and 400 ng/g for AFs, OTA and ZEA, respectively. LOD and LOQ were 0.0125 and 0.05 ng/g for aflatoxin B1 (AFB1) and G1 (AFG1), 0.0037 and 0.015 ng/g for aflatoxin B2 (AFB2) and G2 (AFG2), as well as 0.05 and 0.2 ng/g for OTA and 0.5 and 2 ng/g for ZEA, respectively. The mean recovery values were 77-104% for different concentrations of AFs, OTA and ZEA in spiked cereal samples. Both intra- and inter-day accuracy and precision were within acceptable limits. This method was successfully applied for the simultaneous determination of mycotoxins for 60 cereal samples collected from Malaysian markets. Fifty per cent of the cereal samples were contaminated with at least one of these mycotoxins, at a level greater than the LOD. Only one wheat sample and two rice samples were contaminated with levels greater than the European Union regulatory limits for AFs and OTA (4 and 5 ng/g). The means and ranges of mycotoxins obtained for the cereal samples were 0.4 ng/g and 0.01-5.9 ng/g for total AFs; 0.18 ng/g and 0.03-5.3 ng/g for OTA; and 2.8 ng/g and 2.4-73.1 ng/g for ZEA, respectively. The results indicate that the method is suitable for the simultaneous determination of AFs, OTA and ZEA in cereals and is suitable for routine analysis

Suitable coating material for microencapsulation of spray-dried fish oil

This study was conducted to screen the most suitable coating material for the production of microencapsulated fish oil powder using ternary blends of maltodextrin (15, 25 % w/w), Arabic gum (2.5, 7.5 % w/w), and methylcellulose (0.5, 1.5 % w/w). The physical properties of fish oil emulsion and encapsulated powders were evaluated. Arabic gum (5 % w/w) showed the most significant (p < 0.05) effect on the surface mean diameter of the droplets in the emulsion. Maltodextrin had the most significant (p < 0.05) effect on the centrifuge stability of the emulsion and the amount of surface oil of the powder at 15 and 20 % (w/w) respectively, whereas methylcellulose (0.5 % w/w) had the most significant (p < 0.05) effect on the width distribution of the droplets in the emulsion. The total optimal area leading to the formation of coating material with desirable physical properties was expected to be obtained by the combination of 16 % (w/w) maltodextrin, 6.5 % (w/w) Arabic gum, and 0.88 % (w/w) methylcellulose respectively

Optimization and validation of a method for extraction and quantification of ochratoxin A in black pepper

Abstract The extraction method for the determination of ochratoxin A (OTA) in black pepper was optimized. The influence of three variables, i.e., type of solvent, solvent-volume-to-samplesize ratio (v/w) and amount of sodium chloride (NaCl) (g), on OTA recovery was evaluated. Analysis of variance was used to compare recovery values obtained from different solvents, and response surface methodology (RSM) was used to determine the optimum amount of NaCl and the solvent-volume-to-sample-size ratio. The concentration of OTA was determined by high-performance liquid chromatography with fluorescence detection. The highest recovery (95.2 %) was obtained when methanol/water (80:20, v/v) was used as the solvent. The RSM results showed that the experimental data could be adequately fitted to a second-order polynomial model with multiple regression coefficients (R 2 ) of 0.962. The optimum amount of NaCl was determined to be 3 g, whereas the optimum solvent-volume-to-sample-size ratio (v/w) was found to be 4. The proposed method was applied to 20 samples, and the presence of OTA was found in 8 (40%) samples ranging from 0.11 to 3.16 ng g -1

Acrylamide formation in vegetable oils and animal fats during heat treatment

The method of liquid chromatographic tandem mass spectrometry was utilized and modified to confirm and quantify acrylamide in heating cooking oil and animal fat. Heating asparagine with various cooking oils and animal fat at 180 °C produced varying amounts of acrylamide. The acrylamide in the different cooking oils and animal fat using a constant amount of asparagine was measured. Cooking oils were also examined for peroxide, anisidine and iodine values (or oxidation values). A direct correlation was observed between oxidation values and acrylamide formation in different cooking oils. Significantly less acrylamide was produced in saturated animal fat than in unsaturated cooking oil, with 366 ng/g in lard and 211 ng/g in ghee versus 2447 ng/g in soy oil, followed by palm olein with 1442 ng/g

A limited survey of aflatoxins and zearalenone in feed and feed ingredients from Pakistan

This work presents current information on the presence of aflatoxins (AFs) and zearalenone (ZEN) in feed and feed ingredients from Punjab, Pakistan. The 105 samples tested were concentrated feed, i.e., cotton seed meal (18 samples) and soybean meal (14), and feed ingredients, i.e., crushed corn (17), crushed wheat (15), barley (17). and poultry feed (24). Samples were analyzed using high-performance liquid chromatography equipped with a fluorescence detector. Analysis revealed that 69 of 105 samples were contaminated with AFs, and the highest mean concentrations of AFB1 (6.20 μg/kg) and total AFs (9.30 μg/kg) were found in poultry feed samples. The mean total AF concentrations ranged from the limit of quantification to 165.5 μg/kg. However, 75 of the 105 samples were positive for ZEN. The highest mean concentration (19.45 μg/kg) was found in poultry feed samples. The mean ZEN concentrations were 0.15 to 145.30 μg/kg. The prevalence of AFs and ZEN was high in feed and feed ingredients and needs urgent attention

Multivarate calibration of fourier transform spectra in determining the malonaldehyde as a TBA reactive substance (TBARS) in palm olein

Fourier transform infrared (FTIR) spectra of palm oil samples between 2900 and 2800 cm–1 and 1800 and 1600 cm–1 were used to compare different multivariate calibration techniques for quantitative determination of their thiobarbituric acid-reactive substance (TBARS) content. Fifty spectra (in duplicate) of palm oil with TBARS values between 0 and 0.25 were used to calibrate models based on partial least squares (PLS) and principal components regression (PCR) analyses with different baselines. The methods were compared for the number of factors, coefficients of determination (R2), and accuracy of estimation. The standard errors of prediction (SEP) were calculated to compare their predictive ability. The calibrated models generated three to eight factors, R2 of 0.9414 to 0.9803, standard error of estimation (SEE) of 0.0063 to 0.0680, and SEP of 1.20 to 6.67