Aflatoxins in cotton seeds and cotton seed cake from Pakistan.

A study was conducted to evaluate the natural occurrence of aflatoxins (AFB1, AFB2, AFG1, AFG2) in cotton seeds (n = 110) and cotton seed cake (CSC; n = 110) from Pakistan. All samples were screened by Enzyme-Linked ImmunoSorbent Assay (ELISA). Positive samples were further quantified by IAC-HPLC-FLD. Total contamination frequency and aflatoxins mean levels were 80% and 69 µg/kg in cotton seeds and the corresponding values for cotton seed cake 88% and 89 µg/kg, respectively. Aflatoxin B1was found in all positive samples and co-occurred with AFB2, AFG1, and AFG2. Sixty-four cotton seeds and 71 CSC samples contained aflatoxins levels higher than the ML set for animal feed (20 µg/kg). The results of the present study will help the regulatory authorities to formulate strategies for monitoring aflatoxins in animal feeds.

Quantitative structure-activity relationship between antioxidant capacity of phenolic compounds and the plasmonic properties of silver nanoparticles.

Plasmonic response of silver nanoparticles (AgNPs) reduced with phenolic compounds were evaluated and correlated with the antioxidant capacities of corresponding phenolic compounds and their relative chemical structures. The reference methods including DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)) radical scavenging assays were used for the measurement of antioxidant capacity of phenolic compounds which was positively correlated with redox characteristics of these compounds against the formation of AgNPs. It is found that the higher plasmonic response of AgNPs corresponds to the highest antioxidant capacity of phenolic acids, which in turn depends on chemical structures and degree of hydroxylation. The higher degree of hydroxylation in chemical structures of phenolic compounds demonstrated the higher radical scavenging capacity and higher tendency to reduce Ag+ to AgNPs. The influence of reaction time and temperature on reducing efficiency of the tested phenolic compounds is found to be different. Some phenolic compounds such as quercetin, rutin and gallic acid reacted fast (< 1.0 min) while others were found slow reacting. This study establishes the relationship between the antioxidant capacity of phenolic acids and corresponding optical response by means of plamonics, which can be used as an innovative antioxidant detection assay for samples rich in phenolic compounds.

A simplified FTIR chemometric method for simultaneous determination of four oxidation parameters of frying canola oil.

Transmission Fourier transform infrared (FTIR) spectroscopic method using 100 µm KCl cell was applied for the determination of total polar compounds (TPC), carbonyl value (CV), conjugated diene (CD) and conjugated triene (CT) in canola oil (CLO) during potato chips frying at 180 °C. The calibration models were developed for TPC, CV, CD and CT using partial least square (PLS) chemometric technique. Excellent regression coefficients (R(2)) and root mean square error of prediction values for TPC, CV, CD and CT were found to be 0.999, 0.992, 0.998 and 0.999 and 0.809, 0.690, 1.26 and 0.735, respectively. The developed calibration models were applied on samples of canola oil drawn during potato chips frying process. A linear relationship was obtained between CD and TPC with a good correlation of coefficient (R(2)=0.9816). Results of the study clearly indicated that transmission FTIR-PLS method could be used for quick and precise evaluation of oxidative changes during the frying process without using any organic solvent.

Occurrence of ochratoxin A in poultry feeds and feed ingredients from Pakistan.

A study was conducted to evaluate the occurrence of ochratoxin A (OTA) in complete poultry feeds (n=80) and poultry feed ingredients (n=286) from Pakistan. All samples were first analyzed by indirect enzyme linked immunosorbent assay (ELISA), samples with OTA concentrations above the European Union maximum regulatory limit (MRL, 100 µg/kg) were further confirmed by HPLC-FLD. Contamination frequency and mean OTA levels were 31% and 51 µg/kg in feed ingredients, and the corresponding values for complete feeds were 38% and 75 µg/kg. Ten samples of complete poultry feed and 19 samples of feed ingredients contained OTA at levels higher than the MRL. The results of the present study indicate that there is a strong need for a more intense monitoring programs for OTA in poultry feed.

Application of multivariate chemometric techniques for simultaneous determination of five parameters of cottonseed oil by single bounce attenuated total reflectance Fourier transform infrared spectroscopy.

Single bounce attenuated total reflectance (SB-ATR) Fourier transform infrared (FTIR) spectroscopy in conjunction with chemometrics was used for accurate determination of free fatty acid (FFA), peroxide value (PV), iodine value (IV), conjugated diene (CD) and conjugated triene (CT) of cottonseed oil (CSO) during potato chips frying. Partial least square (PLS), stepwise multiple linear regression (SMLR), principal component regression (PCR) and simple Beer׳s law (SBL) were applied to develop the calibrations for simultaneous evaluation of five stated parameters of cottonseed oil (CSO) during frying of French frozen potato chips at 170°C. Good regression coefficients (R(2)) were achieved for FFA, PV, IV, CD and CT with value of >0.992 by PLS, SMLR, PCR, and SBL. Root mean square error of prediction (RMSEP) was found to be less than 1.95% for all determinations. Result of the study indicated that SB-ATR FTIR in combination with multivariate chemometrics could be used for accurate and simultaneous determination of different parameters during the frying process without using any toxic organic solvent.

Natural co-occurrence of aflatoxins and deoxynivalenol in poultry feed in Pakistan.

Two hundred and fifteen broiler poultry feed samples were analysed over the time period of one year for the co-occurrence of aflatoxins and deoxynivalenol (DON). These were determined by GC-MS and ELISA, respectively. LOD values for aflatoxins and DON were 0.5 and 5 µg/kg, respectively. From all investigated 215 poultry feed samples, aflatoxins and DON co-occurred in 100 samples (46%). DON was detected in 114 samples while 100 samples also were positive for aflatoxins. Mean concentrations of positive samples for aflatoxins and DON were 18 and 807 µg/kg, respectively. Twenty-one DON-positive and 42 aflatoxin positive samples were contaminated above the EU maximum legal limits of 1000 µg/kg and 20 µg/kg, respectively. The present study provided useful data on aflatoxin and DON contamination, which may be helpful for future strategies to control contamination and to formulate standards in poultry feeds.

Application of attenuated total reflectance Fourier transform infrared spectroscopy for determination of cefixime in oral pharmaceutical formulations.

A quick and reliable analytical method for the quantitative assessment of cefixime in orally administered pharmaceutical formulations is developed by using diamond cell attenuated total reflectance (ATR) Fourier transform infrared (FT-IR) spectroscopy as an easy procedure for quality control laboratories. The standards for calibration were prepared in aqueous medium ranging from 350 to 6000mg/kg. The calibration model was developed based on partial least square (PLS) using finger print region of FT-IR spectrum in the range from 1485 to 887cm(-1). Excellent coefficient of determination (R(2)) was achieved as high as 0.99976 with root mean square error of 44.8 for calibration. The application of diamond cell (smart accessory) ATR FT-IR proves a reliable determination of cefixime in pharmaceutical formulations to assess the quality of the final product.

Rapid detection of melamine adulteration in dairy milk by SB-ATR-Fourier transform infrared spectroscopy.

Melamine is a nitrogenous chemical substance used principally as a starting material for the manufacture of synthetic resins. Due to its very high proportion of nitrogen melamine has been added illegitimately to foods and feeds to increase the measured protein content, which determines the value of the product. These issues prompted private as well as governmental laboratories to develop methods for the analysis of melamine in a wide variety of food products and ingredients. Owing to this fact present study is aimed to use single bounce attenuated total reflectance (SB-ATR) Fourier transform infrared spectroscopy (FTIR) method as an effective rapid tool for the detection and quantification of melamine in milk (liquid and powder). Partial least-squares (PLS) models were established for correlating spectral data to melamine concentration with R(2)>0.99, and RMSEC 0.370. Linear calibration curves were obtained over the calibration range of 25-0.0625%. The LOD and LOQ of the method was 0.00025% (2.5 ppm) and 0.0015% (15 ppm) respectively. Proposed SB-ATR-FTIR method requires little or no sample preparation with an assay time of 1-2 min.

Estimation of ibuprofen in urine and tablet formulations by transmission Fourier Transform Infrared spectroscopy by partial least square.

A rapid, reliable and cost effective analytical procedure for the estimation of ibuprofen in pharmaceutical formulations and human urine samples was developed using transmission Fourier Transform Infrared (FT-IR) spectroscopy. For the determination of ibuprofen, a KBr window with 500 µm spacer was used to acquire the FT-IR spectra of standards, pharmaceuticals as well as urine samples. Partial least square (PLS) calibration model was developed based on region from 1807 to 1,461 cm(-1) using ibuprofen standards ranging from 10 to 100 µg ml(-1). The developed model was evaluated by cross-validation to determine standard error of the models such as root mean square error of calibration (RMSEC), root mean square error of cross validation (RMSECV) and root mean square error of prediction (RMSEP). The coefficient of determination (R(2)) achieved was 0.998 with minimum errors in RMSEC, RMSECV and RMSEP with the value of 1.89%, 1.63% and 4.07%, respectively. The method was successfully applied to urine and pharmaceutical samples and obtained good recovery (98-102%).

A novel approach for determination of free fatty acids in vegetable oils by a flow injection system with manual injection.

A non-aqueous flow injection method for determining free fatty acid (FFA) content in corn and sunflower oil samples was developed. A single-line manifold system was built by modification of an HPLC for flow injection analysis (FIA). Without pre-treatment, oil samples were injected into a n-propanol solution containing KOH and phenolphthalein (PHP). The main parameters, such as flow rate of carrier phase, length, geometry, inner diameters of the coils and reagent concentration were all optimized. The proposed FIA method was validated for precision, accuracy, linear region, limit of detection (LOD) and limit of quantification (LOQ). The intra- and inter-day measurements of the precision of the method were found to be within the limits of acceptance criteria (RSD < 1%), and were rugged when the method was performed by a different analyst. The linear concentration range was calculated as 0.09-1.50 and 0.07-1.40 FFA% for corn and sunflower oils, correspondingly. The LOD and LOQ were found to be 7.53 × 10(-4)-2.28 × 10(-3) oleic acid % and 7.11 × 10(-4)-2.23 × 10(-3) oleic acid % for corn and sunflower oils, respectively. The results were compared with those obtained by the AOCS (Ca-5a-40) method using statistical t and F tests, and a significant difference was not observed between the methods at a 95% confidence level. The proposed method is suitable for quality control of routine applications due to its simplicity, high sample throughput, and economy of solvents and sample, offering considerable promise as a low cost analytical system that needs minimum human intervention over long periods of time.

Assessment of pesticide residues in commonly used vegetables in Hyderabad, Pakistan.

The aim of present study was to assess pesticide residues in vegetables in the Hyderabad region of Pakistan. The concentrations of six pesticides were determined by gas chromatography coupled with mass selective detector (GC-MSD) in locally produced vegetables purchased from wholesale markets. A total of 200 samples of eight vegetables viz. cauliflower, green chilli, eggplant, tomato, peas, bitter gourd, spinach and apple gourd were analyzed for pesticide residues. The results indicated that almost all samples were contained pesticides, only 39% contained pesticide residues at or below maximum residue limits (MRLs), and 61% contained pesticide residues above MRLs. From the six analyzed pesticides, carbofuran and chlorpyrifos were found above to MRLs with concentrations ranging from 0.01-0.39 and 0.05-0.96 mg/kg, respectively. The results provided important information on the current pesticide contamination status of some commonly used vegetables and pointed an urgent need to control the use of some excessively applied and potentially persistent pesticides, such as carbofuran and chlorpyrifos.

Synthesis and application of p-tert-butylcalix[8]arene immobilized material for the removal of azo dyes.

The present study describes synthesis of a new resin through immobilization of p-tert-butylcalix[8]arene onto silica and its application for the removal of azo dyes from aqueous media as well as from textile effluents. The newly synthesized material 4 is characterized by FT-IR spectroscopy, scanning electron microscope (SEM) and thermogravimetric analysis (TGA). Reactive Black-5 (RB-5) and Reactive Red-45 (RR-45) azo dyes were used as sorbate. Batch wise sorption experiments were conducted to optimize various experimental parameters such as the effect of sorbent dosage, electrolyte, pH, dye concentration, and contact time. The optimized pH for the effective removal of RB-5 and RR-45 dyes was 9 and 3, respectively. The increase in material 4 dosage increased the percent sorption. Both Langmuir and Freundlich isotherm models were applied to experimental data and Langmuir isotherm model found to be best fit. The results revealed that material 4 was potentially more effective sorbent for the sorption of selected azo dyes as compared to pure silica and p-tert-butylcalix[8]arene. The field studies also supported the effectiveness of material 4, which could be useful for the removal of both the dyes and also for the normalization of pH, TDS, conductivity and salinity near to the drinking water.

Simultaneous assessment of zinc, cadmium, lead and copper in poultry feeds by differential pulse anodic stripping voltammetry.

In the present work four metals (Zn, Cd, Pb and Cu) were determined simultaneously in 28 commercial broiler poultry feeds by differential pulse anodic stripping voltammetry (DPASV) using hanging mercury drop electrode (HMDE). The digestion of poultry feeds was carried out with concentrated nitric acid and hydrogen peroxide (2:1) with the help of microwave heating. Acetate buffer of pH 5 was used as a supporting electrolyte. The limit of detection for Zn, Cd, Pb and Cu was 0.69, 0.35, 0.68 and 0.24 microg/kg, respectively. The amount of Zn, Cd, Pb and Cu in the analyzed poultry feeds was ranged between 54.3-482.2, 3.8-33.6, 23.2-32.6 and 12.3-65.8 mg/kg, respectively. In most of analyzed poultry feed samples, the amount of Cd and Pb was found to be greater than the maximum tolerable level (MTL) which could be harmful for the poultry.

A simplified UV spectrometric method for determination of peroxide value in thermally oxidized canola oil.

The aim of present study was to develop a simple method on UV spectrometer for the determination of peroxide value (PV) of the frying oil. The basis of the PV determination was the stoichiometric reaction of triphenylphosphine (TPP) with the hydroperoxides present in frying oil to produce triphenylphosphine oxide (TPPO), which exhibits a readily measurable absorption band at 240 nm by ultraviolet region. The PV ranged between 0.15 and 11.66 meq. of active oxygen per kilogram of oil as the canola oil was heated from 0 to 12h in the fryer at 180 degrees C. The proposed method was correlated with official AOCS titration method and best correlation coefficient (R(2)=0.99525) was achieved, proving that there is no significant difference in the results. Therefore, developed method could serve as an alternative to the titration method, for the determination of PV in frying oils.

Main fatty acid classes in vegetable oils by SB-ATR-Fourier transform infrared (FTIR) spectroscopy.

The prospect of using single bounce attenuated total reflectance (SB-ATR)-Fourier transform infrared (FTIR) spectroscopy as a rapid quantitative tool to determine the main fatty acid groups present in different edible oils was investigated. Partial least squares (PLS) calibrations were developed using SB-ATR-FTIR spectra which were associated with fatty acid groups (saturated, trans, mono- and polyunsaturated) using quantitative data obtained by gas chromatography (GC). Good calibrations were obtained for all main four fat groups (saturated, mono, trans and polyunsaturated) with excellent precision. The coefficient of determination (R(2)), root mean square error of prediction (RMSEP) and bias for validation set were obtained as 0.999, 2.43 and 0.998 for saturated; 0.999, 1.850 and 0.003 for mono; 0.999, 0.625 and -0.001 for trans while for poly the values were 0.999, 1.170 and 0.003, respectively. The results of 13 validation samples for total saturated, mono, trans and polyunsaturated fats by FI-IR were found in the range of 8.16-55.16, 37.62-74.75, 0.20-18.16 and 1.36-62.35%, respectively. The present study shows that it may well be possible to expand the utility of SB-ATR-FTIR spectroscopy not only to provide isolated trans data, but also serve as a simple, rapid and quantitative means of categorizing the main groups present in the edible oils. The information obtained would be useful for meeting the new lipid nutritional labeling requirements.

Synthesis and application of calix[4]arene based resin for the removal of azo dyes.

The present study describes a novel synthetic method for the immobilization of calix[4]arene (II) onto the surface of modified Amberlite XAD-4 resin (4), which does not require the derivatization of calixarene moiety. The novel calix[4]arene based resin (C4 resin) 5 was used as sorbent for the removal of azo dyes. Batch-wise sorption study was carried out and observed that the C4 resin (5) is more effective as compared to compound II as well as pure Amberlite XAD-4 resin (1) to remove the selected dyes [i.e. Reactive Black-5 (RB-5), Reactive Red-45 (RR-45) and Congo Red (CR)]. The effect of sorbent dosage and pH on % sorption was studied. During the extraction process, various kinds of interactions such as electrostatic repulsion, deprotonation of the hydroxyl groups of C4 resin, dissociation of reactive dyes into anions/cations and structural variations were monitored and found that they are highly pH dependent.

Application of microwave heating for the fast extraction of fat content from the poultry feeds.

A rapid method has been developed to extract and quantitatively measure the total oil content in poultry feeds using a domestic microwave oven. The optimized extraction procedure involves the replicate (6x) extraction of 5 g of ground feed with 12 ml of hexane for 20 s in a 900 W oven. Each replicate involves the collection of the resulting miscella and its replacement with fresh solvent for re-extraction. The collected extracts were centrifuged and transferred to a vial. The solvent was evaporated to a constant weight and the residual lipid weighed. In comparison to conventional Soxhlet extraction method, lipid contents obtained using the optimized microwave procedure was not significantly different. However, FTIR analysis indicated that the microwave procedure was superior in minimizing the formation of free fatty acids (FFA) relative to the Soxhlet procedure if the temperature of the sample was kept within the range of 45-50 degrees C. This simple, sequential extraction procedure is rapid, highly efficient and provides a simple mean of quantitating the lipid content of poultry feed in less than 40 min without the need for specialized microwave oven.

Correcting for underlying absorption interferences in Fourier transform infrared trans analysis of edible oils using two-dimensional correlation techniques.

Substantive improvement in the sensitivity of the AOAC/AOCS spectral ratioing method for the determination of isolated trans isomers in edible oils was recently achieved by the application of a new spectral reconstitution (SR) technique that facilitates the FTIR analysis of edible oils in the transmission mode. However, the general applicability of the spectral ratioing method is still severely limited by the requirement to know the provenance of the oil to be analyzed and have on hand its trans-free counterpart so that the underlying triacylglycerol absorptions in the trans measurement region (990-945 cm(-1)), henceforth referred to as UAt , may be ratioed out. To eliminate the need for a trans-free reference oil, we have employed two-dimensional (2D) correlation spectroscopy to search for other spectral features that might correlate with and serve to estimate the UAt . The three-dimensional contour maps obtained by 2D correlation analysis of the spectra of 10 trans-free oils of different oil types, recorded using the SR procedure, revealed such correlations in two spectral regions, 1700-1600 and 4500-4300 cm(-1), exhibiting one maximum and two maxima, respectively, with wavenumber coordinates of (968, 4407), (968, 4299), and (968, 1650). The latter two correlations, when optimized, produced excellent linear regression relationships (r>0.95) with the UAt . The spectra of five sets of trielaidin-spiked oils were corrected for the UA t using these relationships, and their trans contents were predicted from the calibration equation generated for the spectral ratioing procedure. Linear regression of predicted versus added trans over the range of 0-1.6% trans, which is below the limit of quantitation of the AOAC/AOCS spectral ratioing method, yielded r=0.88-0.90 with an SD of approximately 0.2% trans. These results indicate that the combination of the SR technique with the UA t correction approach may provide a simple and accurate FTIR method for the analysis of the trans content of fats and oils that would be competitive with GC.

Intramuscular fatty acid profile of longissimus dorsi and semitendinosus muscle from Pateri goats fed under traditional feeding system of Sindh, Pakistan.

The present study was undertaken to assess the intramuscular fatty acid composition of different muscles taken from male Pateri (n=15) goats, reared on naturally grown grasses, leaves and pods of Acacia nilotica and crushed cereal, under traditional way of feeding. Goats were slaughtered with an average weight of 68±7kg and age 12±1 month. The samples were taken from longissimus dorsi region (between the 12th and 13th rib) and distal region of semitendinosus muscle. Results of total fat content and fatty acids composition does not show significant (P>0.05) variation among muscles investigated. The fatty acid composition of muscles studied were primarily composed of oleic (31.50-33.38%), followed by palmitic acid (19.84-22.05%) and stearic acid (22.25-24.91%) respectively. Muscle tissue in general contained an average 51.13% of saturated fatty acids and 48.87% of unsaturated fatty acids. The mean conjugated linoleic acid was found 0.41%, 0.43% and 0.47% in ribeye and loin portion of longissimus dorsi muscle and distal region of semitendinosus muscle, respectively.

GC-MS quantification of fatty acid profile including trans FA in the locally manufactured margarines of Pakistan.

Ten margarine brands of Pakistan were analyzed for their fatty acid composition with emphasis on trans fatty acids (TFA) using GC-MS. Saturated, cis-monounsaturated and polyunsaturated fatty acids were present at 24.2-58.1, 5.7-35.4 and 3.8-37.4% of total fatty acids, respectively. Among the saturated fatty acids, palmitic acid (16.9-33.8%) was dominant in all analyzed margarine brands and its higher amount indicates that palm oil was a major contributor in the margarine manufacturing. Among samples tested only one contained a low level of TFA (2.2%) while the rest contained very high amounts of TFA (11.5-34.8%) which clearly shows that hydrogenated oils were used in the formulation of margarines. Fatty acid profiles demonstrated that all samples belong to the hard margarine category containing high amounts of trans and saturated fatty acids which is an alarming issue for the health of consumers.