High levels of blood glutamic acid and ornithine in children with intellectual disability.

Objectives: Aminoacidopathies are inborn errors of metabolism (IEMs) that cause intellectual disability in children. Luckily, aminoacidopathies are potentially treatable, if diagnosed earlier in life. The focus of this study was the screening of aminoacidopathies in a cohort of patients suspected for IEMs. Methods: Blood samples from healthy (IQ > 90; n = 391) and intellectually disabled (IQ < 70; n = 409) children (suspected for IEMs) were collected from different areas of Northern Punjab, Pakistan. An analytical HPLC assay was used for the screening of plasma amino acids. Results: All the samples (n = 800) were analyzed on HPLC and forty-three out of 409 patient samples showed abnormal amino acid profiles mainly in the levels of glutamic acid, ornithine and methionine. Plasma concentration (Mean ± SD ng/mL) were significantly high in 40 patients for glutamic acid (patients: 165 ± 38 vs. controls: 57 ± 8, p < 0.00001) and ornithine (patients: 3177 ± 937 vs. controls: 1361 ± 91, p < 0.0001). Moreover, 3 patients showed abnormally high (53.3 ± 8.6 ng/mL) plasma levels of methionine. Conclusion: In conclusion, biochemical analysis of samples from such patients at the metabolites level could reveal the underlying diseases which could be confirmed through advanced biochemical and genetic analyses. Thus, treatment to some of such patients could be offered. Thus burden of intellectual disability caused by such rare metabolic diseases could be reduced from the target populations.

Seasonal Variation of Aflatoxin Levels in Selected Spices Available in Retail Markets: Estimation of Exposure and Risk Assessment.

A total of 603 samples of selected spices from different seasons (winter and summer) were analyzed for the occurrence of aflatoxin B1 (AFB1), total AFs, and tocopherols. The findings revealed that 120 (38.7%) samples from the summer and 136 (46.4%) samples from the winter season were observed to be infected with AFB1 and a large amount of AFs. The highest means of both AFB1 and total Afs were observed in red pepper, i.e., 15.5 ± 3.90 µg/kg and 22.90 ± 4.10 µg/kg, respectively. The minimum averages of AFB1 and total AFs were observed in cloves of 6.32 ± 1.8 and 8.40 ± 1.60 µg/kg, respectively (from the winter season). The seasonal variations in the levels of the total AFs in selected spices were observed to be nonsignificant (p ≥ 0.05), except for the levels in red pepper and ginger samples, which showed significant differences (p ≤ 0.05). The maximum average of the dietary intake of Afs, 4.80 µg/day/kg, was found in ginger from the winter season in individual females. Furthermore, the findings document that the maximum level of total tocopherol, i.e., 44.8 ± 9.3 mg/100 g, was observed in black pepper from the winter season. A significant difference in the concentration of total tocopherols was observed in selected spices from the summer and the winter seasons (p ≤ 0.05).

Variation of Aflatoxin Levels in Stored Edible Seed and Oil Samples and Risk Assessment in the Local Population.

Five hundred and twenty samples of edible seeds and oilseeds (sunflower, palm, peanut, sesame, cotton, and grapeseed) were purchased from markets, farmers, and superstores in the central cities of Punjab, Pakistan. A total of 125 (48.1%) edible seed samples from a 6 ≤ months storage period, and 127 (48.8%) from a 2 ≥ years storage period were found to be infested with AFs. The average elevated amount of AFB1 and total AFs was observed in a 2 ≥ years storage period, i.e., 28.6 ± 4.5 and 51.3 ± 10.4 µg/kg, respectively, in sesame seeds. The minimum amount of AFB1 and total AFs was observed in palm seed samples with a storage period of 6 ≤ months, i.e., 9.96 ± 2.4, and 11.7 ± 1.90 µg/kg, respectively. The maximum amount of AFB1 and total AFs were observed in peanut oil samples, i.e., 21.43 ± 2.60 and 25.96 ± 4.30 µg/kg, respectively, with a storage period of 2 ≥ years. Therefore, the maximum dietary intake of 59.60 ng/kg/day was observed in oil samples stored at a ≥ 2 years storage period. The results of the present study concluded that a significant difference was found in the amounts of total AFs in edible seed samples stored at 6 ≤ months and 2 ≥ years storage periods (p < 0.05).

Impact of sonolytic ozonation (O3/US) on degradation of pesticide residues in fresh vegetables and fruits: Case study of Faisalabad, Pakistan.

High pesticide residues in fresh produce is a serious food safety issue. This study was aimed at assessing the pesticides residues in some important vegetables and fruits marketed in Faisalabad, Pakistan and the impact of sonolytic ozonation (O3/US) treatment in removing these contaminants. From a short grower's survey, five registered and mostly used pesticides (acetamiprid, carbendazim, imidacloprid, thiacloprid and thiamethoxam) were identified. A time optimization trial of O3/US application (05, 10 and 15 min) on okra, showed that 10 min treatment significantly reduced three identified chemicals (thiamethoxam 100 %, imidacloprid and thiacloprid 97.17 %), without any adverse effect on its quality. In follow up trial, five fresh vegetables (cauliflower, chillies, cucumber, spinach and tomato) three fresh fruits (grapes, guava and peach) collected from three markets of Faisalabad, were pooled together to have uniform samples. Vegetables and fruits were treated with O3/US for 10 and 6 min, respectively, along with control (simple tap wash) for determining the impacts on pesticides degradation. Samples were processed for extraction, clean up and analysis using HPLC-UV-Vis in isocratic mode. The data revealed the presence of five mentioned chemicals, with an accumulative mean residue of 9.006 and 1.921 µg/g in tested vegetables and fruits, respectively. After subjecting to O3/US, the accumulative chemical residues were reduced to 3.214 µg/g (64.313 %) and 1.064 (44.6 %) in treated vegetables and fruits respectively. Irrespective of fresh produce, the mean residues of thiamethoxam, imidachloprid, acetamiprid and thiachloprid and carbendazim were reduced by 99.3 %, 52.6 %, 65.2 %, 87.3 % and 72% respectively. It was concluded that sonolytic ozonation treatment was effective in significant reduction of pesticide residues from vegetables and fruits and thus can be employed as a good food safety practice at culinary level to reduce the associated health hazardous risks.

Bioherbicidal Activity and Metabolic Profiling of Potent Allelopathic Plant Fractions Against Major Weeds of Wheat-Way Forward to Lower the Risk of Synthetic Herbicides.

The productivity of major field crops is highly compromised due to weed infestation. Inefficient weed management practices and undue and excessive use of chemical herbicides have drastically contaminated the environment and human health, in addition to resistance development in weed species. Therefore, utilization of allelopathic plants to explore phytochemicals as potent organic alternatives to such chemical herbicides has become indispensable. The current study evaluates the comparative bio-herbicidal potential of methanolic extracts of castor (Ricinus communis), artemisia (Artemisia santolinifolia), wheat (Triticum aestivum), and sorghum (Sorghum bicolor) to suppress growth of major weeds, i.e., wild mustard (Sinapis arvensis), Italian ryegrass (Lolium multiflorum), and carrot grass (Parthenium hysterophorus). The results demonstrated a concentration-dependent effect on weeds' growth. Overall, in vitro seed germination was reduced from 60 to 100% in response to 5% (w/v) extract concentration. Significant reduction in radicle length, hypocotyl length, and fresh biomass of the weeds was also observed. A strong inhibitory effect was seen in in vivo pot experiments, revealing that application of 10-20% methanolic extracts induced permanent wilting and substantial reduction in the chlorophyll content of weeds along with 20-80% increase in oxidative stress. Artemisia showed the most significant allelopathic effect, on account of highest phenolic and flavonoid contents, followed by castor, wheat, and sorghum, against S. arvensis, L. multiflorum, and P. hysterophorus, respectively. Phytochemical analysis, through high-performance liquid chromatography (HPLC), also exhibited a correlation between extract's phytotoxicity and their antioxidant potential due to their major constituents (rutin, quercetin, catechin, gallic acid, vanillic acid, syringic acid, ferulic acid, p-hydroxy benzoic acid, p-coumaric acid, and sinapic acid), among the total of 13 identified in methanolic fractions. Comprehensive profiling of allelochemicals with liquid chromatography-mass spectrometry (LC-MS) determined 120, 113, 90, and 50 derivates of phenolic acids, flavonoids, and alkaloids, reported for the first time through this study, demonstrating significant allelopathic potential of the targeted plant fractions, which can be explored further to develop a sustainable bio-herbicidal formulation.

Occurrence of Aflatoxins in Edible Vegetable Seeds and Oil Samples Available in Pakistani Retail Markets and Estimation of Dietary Intake in Consumers.

Aflatoxins (AFs) are secondary metabolites toxic to humans as well as animals. The environmental conditions, conventional agricultural practices, and illiteracy are the main factors which favor the production of AFs in food and feed. In the current study 744 samples of vegetable seeds and oils (soybean, sunflower, canola, olive, corn, and mustard) were collected and tested for the presence of aflatoxin B1 (AFB1) and total AFs. Liquid-liquid extraction was employed for the extraction of AFs from seeds and oil samples. Reverse phase high performance liquid chromatography equipped with fluorescence detection was used for the analysis. The results have shown that 92 (56.7%) samples of imported and 108 (57.0%) samples of local edible seeds were observed to be contaminated with AFs. All samples of edible seeds have AFB1 levels greater than the proposed limit set by the European Union (EU, 2 µg/kg) and 12 (7.40%) samples of imported seeds and 14 (7.40%) samples of local seeds were found in the range ≥ 50 µg/kg. About 78 (43.3%) samples of imported edible oil and 103 (48.3%) sample of local edible oil were observed to be positive for AFs. Furthermore, 16 (8.88%) and six (3.33%) samples of imported vegetable oil have levels of total AFs in a range (21-50 µg/kg) and greater than 50 µg/kg, respectively. The findings indicate significant differences in AFs levels between imported and local vegetable oil samples (t = 22.27 and p = 0.009) at α = 0.05 and a significant difference in AFs levels were found between vegetable seeds and oil samples (t = -17.75, p = 0.009) at α = 0.05. The highest dietary intake was found for a local sunflower oil sample (0.90 µg/kg/day) in female individuals (16-22 age group). The results have shown considerably high levels of AFB1 and total AFs in seeds and oil samples and emphasise the need to monitor carefully the levels of these toxic substances in food and feed on regular basis.

Sonolytic-ozonation technology for sanitizing microbial contaminants and pesticide residues from spinach (Spinacia oleracea L.) leaves, at household level.

Increasing foodborne illnesses linked with the consumption of contaminated food pose serious health risks. Fresh fruits and vegetables can potentially be contaminated (microbes/chemicals) throughout the supply chain. Various chemical and thermal approaches have been used in the past to decontaminate fresh produce, which have had a negative impact on commodities and health hazardous. Henceforth, this conducted study was aim to test an ecofriendly/green decontamination technique, for impact on food safety and quality of spinach. Freshly harvested spinach leaves were treated with a combined application of ozone and ultrasound (O3+US) for different times (5, 10, and 15 min) in an aqueous medium. Different food safety (microbes and pesticide residues) and quality (VC content, TSS, TA, and weight loss) parameters were studied in comparison with tap washed samples. Total plate count method was used to evaluate microbial contamination and pesticide residues were determined by HPLC. There was gradual decrease in contaminants with increase in O3+US treatment time. Sonolytic-ozonation 10 and 15 min treatments were found optimal in reduction of microbial counts (TPC, E. coli, Salmonella, and Listeria) and pesticide residues (10 min 94.04% and 15 min 99.77% residue reduction). However, chlorophyll degradation was observed in 15 min treated sample under 1-week household storage. There was no significant detrimental impact on quality parameters of spinach during 1 week of storage under refrigerated conditions. So, it was concluded that O3+US treatment for 10 min can be employed for microbial and chemical decontamination in spinach leaves at household level without any negative effect on its quality up to 1 week under refrigerator storage (5 ± 2 ○C).

Variation of Deoxynivalenol Levels in Corn and Its Products Available in Retail Markets of Punjab, Pakistan, and Estimation of Risk Assessment.

This study investigated the natural incidence of deoxynivalenol (DON) in corn and corn products from corn-producing districts of Punjab, Pakistan. The analysis was carried out using high performance liquid chromatography (HPLC) with UV detector and immunoaffinity cleanup columns. The detection limit (LOD) and limit of quantification were 25 and 50 µg/kg, respectively. A total of 1220 samples of corn and corn products were analyzed to detect DON, and 539 (44.2%) samples were observed to be contaminated with DON (n ≥ LOD). Furthermore, 92 (7.5%) samples of corn and corn products had DON levels that were higher than the proposed limits of the EU. The data are significantly different from a normal distribution of DON in samples of corn and corn products from different locations (p < 0.05) for Shapiro-Wilk and Kolmogorov-Smirnov values. However, a significant difference in DON levels was found between corn and corn-derived products (p ≤ 0.05). The lowest and highest exposures, and hazard quotient (HQ) values of 0.92 and 9.68 µg/kg bw/day, were documented in corn flour samples.

Seasonal Variation in Aflatoxin Levels in Edible Seeds, Estimation of Its Dietary Intake and Vitamin E Levels in Southern Areas of Punjab, Pakistan.

A total of 779 samples of edible nuts (melon seeds, watermelon seeds, pumpkin seeds, and cantaloupe seeds) from Southern Punjab (Pakistan), were collected during the summer and the winter seasons. The natural occurrence of aflatoxins (AFs) and vitamin E (tocopherols) levels were investigated using HPLC. The results have shown that 180 (43.4%) of samples from the winter season and 122 (33.4%) samples from the summer season were found positive for AFs. Elevated average levels of total AFs (20.9 ± 3.10 µg/kg, dry weight) were observed in watermelon seeds without shell, and the lowest average amount (15.9 ± 3.60 µg/kg) were documented in melon seeds without shell samples from the winter season. An elevated average amount of total AFs 17.3 ± 1.50 µg/kg was found in pumpkin seeds available without a shell. The results have documented a significant difference in total AFs levels in edible seeds available with shells versus without shells (α = 0.05 & 0.01). The highest dietary intake of 6.30 µg/kg/day was found in female individuals from consuming pumpkin seeds (without shell) in the winter season. A value of 3.00 µg/kg/day was found in pumpkin seed without shell in the summer season in female individuals. The highest total tocopherol levels were 22.2 ± 7.70 ng/100 g in pumpkin seeds samples from the winter season and 14.5 ± 5.50 mg/100 g in melon seed samples from the summer season. The variation of total tocopherol levels in edible seeds among the winter and summer seasons showed a significant difference (p ≤ 0.0054), except watermelon seeds samples with non-significant differences (p ≥ 0.183).

Surveillance of Patulin in Apple, Grapes, Juices and Value-Added Products for Sale in Pakistan.

The prime objective of the current study was to investigate the occurrence of mycotoxin patulin (PAT) in apples, grapes and their value added products. PAT was determined by a validated method based on HPLC with UV detector. A total of 381 samples comprising apple and grape fruits (n = 133 each), apple-based products (n = 76, juice, puree, jam) and grape juice (n = 39) were analyzed. PAT was found in 58.9% samples of apple and apple-based products, with a mean of 49.8 µg/kg (maximum 396 µg/kg), while 27.3% samples contained PAT beyond the maximum regulatory limit of 50 µg/kg. The average levels of PAT contamination in apple-derived products was higher in apple juice concentrate, followed by apple puree, apple juice and apple jam. The incidence of PAT in table grapes was 65.1%, with a mean of 53.9 µg/kg (maximum 505 µg/kg), whereas 23.8% exceeded the maximum level. Among the fruit samples, there were differences in PAT contents due to apple variety (6 types) or grape variety (8 types), as well as for sampling location. Our investigations showed the wide PAT occurrence in fruits and derived value-added products affecting consumer product safety, so that the population is chronically exposed to this toxin.

Effect of Postharvest Application of Aloe Vera Gel on Shelf Life, Activities of Anti-Oxidative Enzymes, and Quality of 'Gola' Guava Fruit.

Guava is an important climacteric fruits in terms of taste and aroma, which contains various vital nutrients such as minerals, carotenoids, ascorbic acid, and polyphenols. At ambient conditions, it exhibits a short shelf life, which makes it difficult for marketing and subsequent storage. Therefore, it is necessary to develop procedures to extend its shelf life and conserve quality. For this purpose, an aloe vera (AV) gel coating was assessed for its potential to enhance the shelf life of guava fruits. Guava fruits coated with AV gels (0, 20, 40, 60 and 80%, v/v) were evaluated for postharvest shelf life extension, changes in quality attributes, anti-oxidative activities, and flavonoid content when stored at ambient conditions (23 ± 2 °C and 70-75% relative humidity) for 12 days. The AV gel-treated fruits showed reduced increments in total sugar, malondialdehyde, and total carotene contents compared to untreated controls. AV gel-treated fruits exhibited higher contents of ascorbic acid, flavonoids (quercetin and rutin), and total phenolics in comparison to control fruits. Moreover, AV gel-treated fruits displayed greater activities of superoxide dismutase, catalase, and ascorbate peroxidase, along with higher antioxidant capacity and higher levels of total soluble solids, than untreated fruits. These results demonstrate that AV gel coating, especially at high concentrations, can be considered an eco-friendly and non-chemical substitute treatment for maintaining the postharvest quality of guava fruit.

Assessment of Deoxynivalenol in Wheat, Corn and Its Products and Estimation of Dietary Intake.

The main goal of the present research was to explore the seasonal variation of deoxynivalenol (DON) in wheat, corn, and their products, collected during 2018-2019. Samples of 449 of wheat and products and 270 samples of corn and their products were examined using reverse-phase liquid chromatography with a UV detector. The findings of the present work showed that 104 (44.8%) samples of wheat and products from the summer season, and 91 (41.9%) samples from winter season were contaminated with DON (concentration limit of detections (LOD) to 2145 µg/kg and LOD to 2050 µg/kg), from summer and winter seasons, respectively. In corn and products, 87 (61.2%) samples from summer and 57 (44.5%) samples from winter season were polluted with DON with levels ranging from LOD to 2967 µg/kg and LOD to 2490 µg/kg, from the summer and winter season, respectively. The highest dietary intake of DON was determined in wheat flour 8.84 µg/kg body weight/day from the summer season, and 7.21 µg/kg body weight/day from the winter season. The findings of the work argued the need to implement stringent guidelines and create awareness among farmers, stakeholders, and traders of the harmful effect of DON. It is mostly observed that cereal crops are transported and stockpiled in jute bags, which may absorb moisture from the environment and produce favorable conditions for fungal growth. Therefore, these crops must store in polyethylene bags during transportation and storage, and moisture should be controlled. It is highly desirable to use those varieties that are more resistant to fungi attack. Humidity and moisture levels need to be controlled during storage and transportation.

Comparative efficacy of mitigation techniques for the detoxification of Prunus persica (L.) from selected pesticide residues.

Fruits are the valuable and important components of human diet. Among them, Prunus persica is a rich source of different minerals and dietary fibers. In Pakistan, the total annual production of P. persica is approximately 837,000 tons. In order to enhance agricultural yield and quality, the plant protection agents are employed during fruit production. Ultimately, this in turn leads to the incorporation of pesticide residues in fruits. In present study, an effort has been made for the determination of three selected pesticide residues, i.e., chlorpyrifos (CPF), difenoconazole (DFN), and carbendazim (CRB) in samples of P. persica collected from Swat territory. Samples were analyzed through high performance liquid chromatography (HPLC). Results revealed the occurrence of all three pesticides in studied samples; however, levels of CPF and DFN were found to be higher than MRLs. Moreover, the effects of different mitigation techniques revealed that highest reduction of CPF, DFN, and CRB (86%, 97%, 89%) residues was obtained by treatment with 10% CH3COOH followed by 10% NaCl (74%, 78%, 84%). The lowest reduction was obtained by treatment with 10% solution of NaOH (52%, 55%, 63%).

Patulin Mycotoxin in Mango and Orange Fruits, Juices, Pulps, and Jams Marketed in Pakistan.

The objective of the study was to explore the incidence of patulin (PAT) mycotoxin in mango and orange fruits and derived products marketed in Pakistan. A total of 274 samples, including 70 mango fruits, 63 mango-based products (juices, pulp, and jam), 77 orange fruits, and 64 orange-based products, were collected. PAT was determined by reverse-phase high-performance liquid chromatography (HPLC) with UV-Vis detector (276 nm). Linear detector response was observed (R2 > 0.99), the limit of detection (LOD) was 5 µg/kg and recovery percentage was 97.4%. The incidence of PAT in mango samples was 61.7%, and the concentration ranged from

In vitro anticancer activities of Withania coagulans against HeLa, MCF-7, RD, RG2, and INS-1 cancer cells and phytochemical analysis.

BACKGROUND: The Pakistani Salt Range has a rich floral diversity including Withania coagulans from the Solanaceae family. METHODS: The crude methanolic extracts of the root, leaf, leaf stalk, and fruit of this plant were screened for their cytotoxic activity against human (HeLa, MCF-7, RD) and rat (RG2 and INS-1) cancer cell lines at 20 µg/mL and compared to methotrexate. The IC50 values indicated that leaf stalk and fruit extracts exert an 80% or higher cytotoxic activity against all cell lines at 24 hours. RESULTS: The leaf stalk extract showed the highest cytotoxic efficacy against all tested cell lines, with IC50 values ranging from 0.96 ± 0.01 µg/mL to 4.73 ± 0.05 µg/mL followed by the fruit extract with IC50 values of 0.69 ± 0.01-6.69 ± 0.06 µg/mL after 48-72 hours incubation. The leaf stalk and seed extracts were analyzed for polyphenols and flavonoids using RP-HPLC. The total flavonoid content (TFC) was calculated for all tested samples, and the highest TFC was recorded for the root extract (394.34 ± 1.26 µg/g). The total phenolic content (TPC) was found in the seed extract (307.86 ± 9.42 µg/g) of W. coagulans. The highest contents of myricetin (358.46 ± 2.91 µg/g) were noted in the leaf extract, and highest quercetin was recorded in the seed extract (21.43 ± 0.13 µg/g). The highest gallic acid concentration (83.62 ± 0.71 µg/g) was recorded in leaf stalk extract and p-hydroxybenzoic acid in the seed extract (157.46 ± 1.43 µg/g). CONCLUSION: The present study gives a scientific insight and comparative analysis of various plant parts in this medicinally important plant species from the Salt Range of Pakistan against both human and rat cancer cells.

Analysis of Nutritional Traits and Aflatoxin Contamination in Selected Maize Varieties from Pakistan.

Recently, cultivation of high-yielding hybrid maize varieties has revolutionized maize production in Pakistan. Analyses of nutritional traits and aflatoxin (AF) contamination in these varieties can aid in the identification of susceptible and resistant varieties, particularly for cultivation in the Pakistani agro-climatic environment. Five spring maize varieties-Pioneer, Neelam, DK-919, Desi, and Hi-maize-were selected for analyses of their nutritional, tocopherol, and AF contents. Protein, carbohydrate, oil, ash, fiber, and moisture contents ranged between 8.7 and 10.8%, 68 and 71%, 3.72 and 5.56%, 1.09 and 1.81%, 1.1 and 3.1%, and 11.7 and 14.2%, respectively. Tocopherol levels in selected varieties were in the range of 461 to 1,430 µg/g. Hi-maize exhibited significantly higher protein and tocopherol contents than the other varieties, indicating its better suitability for feed and silage applications. The highest mean level of total AFs, 14.5 ± 0.12 µg/kg, was found in Desi, and results showed that the most dominant AF found in the maize varieties was AFB1. Furthermore, the results showed that the higher the level of tocopherol, the lower the concentration of total AFs and vice versa in maize varieties. The results can be used to investigate additional susceptible maize varieties that are resistant to fungal attack.

LC-MS/MS based method development for the analysis of florfenicol and its application to estimate relative distribution in various tissues of broiler chicken.

Florfenicol, a broad spectrum bacteriostatic antibiotic belonging to amphenicol class, is widely used in poultry and livestock for the treatment of various infections. The major metabolite of florfenicol in different animal species is florfenicol amine which is exploited as the marker residue for the determination of florfenicol. Analysis of florfenicol merely by solvent extraction cannot determine the accurate amount of the drug present in incurred tissues (muscle, liver and kidney) of treated birds, as indicated by this study. Thus the methods solely based on solvent extraction may lead to false negative results. A reliable LC-MS/MS based confirmatory method for the analysis of florfenicol and its metabolites in chicken muscle was developed and validated according to the European Union Commission Decision 2002/657/EC. The method was based on acid hydrolysis to liberate non-extractable residues having presumably been covalently bound to tissues, and to convert all the florfenicol residues as well as its metabolites into florfenicol amine. The amine was subsequently recovered with ethyl acetate at pH 10.5, defatted and further cleaned up with dispersive solid phase extraction (dSPE). The LC separation was achieved on reverse phase C-18 column with isocratic elution using acetonitrile/water mobile phase and the analysis was performed on linear ion trap mass spectrometer. Calibration curve was obtained over a concentration range of 25-600µg/kg for chicken muscles. The accuracy values ranged from 84 to 101.4% and the precision values for within day and between days ranged from 1.2-11.7%, respectively. Limit of detection (LOD), limit of quantification (LOD), CCα and CCß values were 0.98, 3.2, 113 and 126µg/kg, respectively. The developed method was highly robust and was further applied to estimate the relative distribution of solvent-extractable against solvent-non-extractable florfenicol drug residues in muscle, liver and kidney samples of broiler chicken after 5days of oral dosing.

The presence of aflatoxins and ochratoxin A in rice and rice products; and evaluation of dietary intake.

In present study aflatoxins (AFs) and ochratoxin A (OTA) were analysed in 208 samples of rice and products collected from central areas of Punjab, Pakistan. The analysis was carried out using HPLC equipped with fluorescence detector. The results have shown that 35% of the samples were found contaminated with AFs, out of which 19% and 24% samples were found to be above the European Union (EU) maximum content for AFB1 and total AFs, respectively. About 19% samples were found contaminated with OTA and 14% samples were found to be above the EU maximum content. The highest mean level of AFB1 and total AFs were found in brown rice samples i.e. 8.91 and 12.4µg/kg, respectively. However, white rice samples have shown the highest mean level of OTA (8.50µg/kg) with highest level of 24.9µg/kg. The high mean dietary exposure 22.2 and 24.2ngkg(-1)bwday(-1) to AFB1 and OTA, respectively poses significant health hazard for local population.

Irradiation Maintains Functional Components of Dry Hot Peppers (Capsicum annuum L.) under Ambient Storage.

Hot peppers used as natural flavoring and coloring agents are usually irradiated in prepacked form for decontamination. The effects of gamma radiation on the stability of functional components such as capsaicinoids and antioxidant compounds (carotenoids, ascorbic acid and total phenolics) were investigated in hot peppers (Capsicum annuum). Whole dried peppers packed in polyethylene bags were gamma irradiated at 0 (control), 2, 4, and 6 kGy and subsequently stored at 25 °C for 90 days. The irradiation dose did not substantially affect the initial contents of capsaicinoids, ascorbic acid and total phenolics, though the concentration of carotenoids declined by 8% from the control (76.9 mg/100 g) to 6 kGy radiation dose (70.7 mg/100 g). Similarly, during storage for 90 days at ambient temperature the concentrations of capsaicinoids and total phenolics remained fairly stable with mean percent reductions from 3.3% to 4.2%, while the levels of total carotenoids and ascorbic acid significantly (p < 0.05) declined by 12% and 14%, respectively. Overall, neither irradiation nor subsequent ambient storage could appreciably influence the contents of functional components in hot peppers. These results revealed that gamma irradiation up to 6 kGy can be safely used for decontamination to meet the needs for overseas markets without compromising product quality.

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.