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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.

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). [ABSTRACT FROM AUTHOR]

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). [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

In present study, proximate chemical composition and tocopherol content were evaluated for estimation of food quality. The maize genotypes with promising agronomic parameters were analyzed for their biochemical composition and further guidance for maize crop improvement with respect to quality traits. Twelve (12) genotypes (n= 60) of maize (Zea mays L.) cultivated at Maize and Millet Research Institute, Yousafwala, Sahiwal, Punjab, Pakistan were evaluated for diversity based on protein, crude fiber, moisture contents, oil contents and tocopherol. The results showed that the crude protein contents were ranged from 1.75 to 5.25%, crude fiber 2.12 to 2.55%, fat contents 2.83 to 5.20%, moisture contents 9.75 to 11.95% and total tocopherol 1971 to 9382 µg/g -tocopherol was found in maximum concentration and crude fiber in minimum percentage. These findings will be helpful to produce quality products from the maize and valuable to protect human beings against degenerative processes, such as cancer and cardiovascular diseases. [ABSTRACT FROM AUTHOR]

Abstract: A total of 320 samples of edible nuts (poppy seed, peanut, pistachio, cashew, almonds) and dry fruits (plum, raisins, figs, dates, apricot, watermelon seed, melon seeds, pomegranate seeds) collected from major cities of Punjab, and Khyber Pakhtunkhwa, Pakistan, were analyzed for the presence of aflatoxins (AFs) and ochratoxin A (OTA). Sample extracts were purified using immunoaffinity columns and analyzed by reversed phase HPLC with fluorescence detection. Aflatoxin B1 (AFB1) and total AFs (AFB1, AFB2, AFG1, AFG2) were found in 128 (40%) samples, and total AFs exceeded levels of 4 µg/kg and 10 µg/kg in 34 and 25% of samples. The highest mean levels of total AFs (7.30 ± 1.80 µg/kg) were found in peanuts without shell, the lowest mean levels (2.90 ± 1.50 µg/kg) were found in watermelon seeds with shell samples. OTA was found in 18 (5%) samples, the highest levels (5.60 ± 1.34 µg/kg) were found in raisins (maximum 18.5 µg/kg). Only four samples of edible nuts and dry fruits were contaminated with OTA at levels above the recommended limit of 10 µg/kg. The results have revealed the co‐occurrence of OTA and total AFs in 25% total samples of dry fruits and nuts with a mean level of OTA 3.58 ± 1.30 µg/kg and total AFs mean level of 4.13 ± 0.48 µg/kg, respectively. The high frequency especially of AFs in dry fruits and edible nuts is regarded as a health issue for consumers. Practical applications: The manuscript has provided the current status on the levels of AFs and OTA in dry fruits and nuts in Pakistan. The results of current finding revealed that the levels of AFs and OTA in those samples which have stored in shells are significantly less as compared to those samples of nuts and dry fruits which have been stored without shells. Therefore, to use good storage practices, farmers, venders, and traders can eliminate or minimize the levels of these toxins in their products. Furthermore, the study will provide the awareness and levels of AFs and OTA in nuts and dry fruits and help to implement strict regulations for these toxins by law enforcement agencies. [ABSTRACT FROM AUTHOR]

Salting-out-assisted liquid-liquid extraction (SALLE) was developed to extract thiacloprid (THI) from fruits and vegetables. SALLE conditions (NaCl/Na2SO4, pH, and solvent polarity) were investigated at various levels for the optimal recovery of THI. Meanwhile, reverse-phase high-performance liquid chromatographic (RP-HPLC) conditions were balanced over 1-100 µg/mL of THI. The optimized SALLE-RP-HPLC method offered 78.33-92.00% recovery of standard THI at an acceptable repeatability 1.81-4.30% and reproducibility 1.08-4.74%. The detection and quantification limits were found to be 0.03 and 0.05 µg/mL, respectively. The real-time analysis verifies its suitability and ease of use for the determination of THI in agricultural commodities. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the levels of the aflatoxins (AFs) and ochratoxin A (OTA) in 312 samples of whole chili, chili powder, crushed chili and chili sauce samples from suburbs, open market and food restaurants of Punjab, Pakistan. The analysis was carried out using HPLC with fluorescence detector, after immunoaffinity column clean-up. The results have shown that 176 out of 312 (56.4%) samples were to be positive with AFs and 126 out of 312 (40.4%) sample of chilies were found to be contaminated with OTA. Total mean level of AFB 1 and total AFs in chilies were 12.50 ± 1.91, 15.16 ± 2.22 μg/kg, respectively. The total mean level of OTA in chilies was found 16.68 ± 2.58 μg/kg, ranged from LOD to 120.9 μg/kg. Sample 39.5, 26.3 and 32.7% of chilies were found containing level of AFB 1 , total AFs and OTA, higher than the recommended limits for EU, respectively. The dietary levels of 3.26, 3.52, and 3.84 μg/kg were determined for AFB 1 , total AFs and OTA in chilies. The incidence and levels of AFs and OTA in chilies are higher and could pose serious health hazards for consumers. [ABSTRACT FROM AUTHOR]

Total 372 samples (169 sample from summer and 203 samples from winter) of milk and dairy products were collected from major cities of Punjab, Pakistan, during 2014–2015. The aflatoxin M 1 (AFM 1 ) levels were analyzed using HPLC, equipped with floresce n ce detector. The results have showed that AFM 1 was detected in 77 (45.5%) samples of milk and dairy products from summer at the range of LOD to 229.6 ng/L, and 114 (56.1%) samples from winter at a range of LOD to 345.8 ng/L. The highest mean of AFM 1 i.e. 94.9 ± 5.4 ng/L from summer and 129.6 ± 8.4 ng/L from winter was detected in raw milk samples. The results have shown that 37.5% of milk, 32% of UHT milk, 12.5% of powdered milk, 20% of flavored milk, 20% of yogurt and 16% of flavored milk samples from summer were found exceeded the recommended levels of AFM 1 i.e 50 ng/L (European Union permissible limit), as compared to 38.1%, 37.1%, 15.6%, 21.4%, 27.7% and 40% samples of milk, UHT milk, powdered milk, flavored milk, yogurt and flavored yogurt from winter, respectively. The mean level of AFM 1 in samples of milk, UHT milk and powdered milk samples from winter season has significantly (p˂ 0.05) higher as compared to the mean levels in samples from summer season. The dietary exposures of AFM 1 in milk and dairy products were calculated in five different age groups and the highest level of 3.42 and 3.33 ng/L/day in summer and 4.7 and 4.6 ng/L/day in winter were found in 1–5 years of male and female infants, respectively. The results of present study have shown significant levels of AFM 1 in milk and dairy products from Punjab, Pakistan and could pose considerable thereat to the health of local population. [ABSTRACT FROM AUTHOR]

Abstract: A total of 153 samples of dates and dates products (date cookies, date cake and date halva) from Punjab and Khyber Pakhtunkhwa, Pakistan were evaluated for aflatoxins (AFs) using HPLC equipped with fluorescence detection. Thirty eight out of 96 samples (39.6%) of different date varieties and 18 out of 57 (31.6%) samples of date products contained AFs. The total mean level of AFs ranged between 2.90 to 4.96 μg/kg and 2.76 to 4.80 μg/kg in dates and dates products, respectively. About 16 and 20 samples of dates were found above the permissible level for AFB1 and total AFs, respectively (i.e. 2 μg/kg, 4 μg/kg). Furthermore, two samples of date's cookies and one sample of date cake were found above the level of AFB1 and total AFs and three and five samples of date halva were found above the recommended limit for AFB1 and total AFs, respectively. The high occurrence of AFs may cause health hazards and limit exports. [Copyright &y& Elsevier]

Aflatoxin M1(AFM1) was determined in 107 milk samples collected from urban and rural farmhouses from Punjab, Pakistan, by HPLC with fluorescence detection. An incidence rate of 64% (38/59) in milk samples from urban farmhouses was found, with a mean concentration of 0.064 ± 0.023 μg L−1. In rural samples about 52% (25/48) of milk samples were contaminated with AFM1, with a mean of 0.04 ± 0.034 μg L−1. About 42% of milk samples from urban and 27% from rural farmhouses were well above the limit permitted by the European Union (EU). However, only 15% and 8% of milk samples from urban and rural farmhouses, respectively, exceeded the limit of USDA/Codex regulations. The results showed that the contamination of milk with AFM1from Punjab, Pakistan, when compared with Codex limit is present and needs continuous monitoring. The awareness and education among dairy farmers on the potential health risks associated with aflatoxins should be communicated. [ABSTRACT FROM AUTHOR]

Abstract: In present study the natural occurrence of aflatoxins (AFs) and zearalenone (ZEN) has determined in 147 samples of wheat derived products i.e. spaghetti (25), noodles (34), macaroni (29), lasagne (37), and bucatini (22), collected from major districts of Punjab, Pakistan. The mean of AFs was found 9.12 in spaghetti, 7.35 in noodles, 5.91 in macaroni, 8.39 in lasagne, and 9.61 μg/kg in bucatini and 28, 18, 17, 22 and 27% of samples were found above the European Union's legal limit (i.e. 4 μg/kg) for total AFs, respectively. However, mean value of ZEN was found 7.36 in spaghetti, 6.80 in noodles, 4.98 in macaroni, 6.90 in lasagne, and 8.89 μg/kg in bucatini samples and 16, 15, 21, 19 and 36% of samples were found to be above the recommended limit. The study urged the need to focus more comprehensive survey for these toxins in wheat derived products from Punjab, Pakistan. [Copyright &y& Elsevier]

Abstract: Total 221 samples of milk and milk products were collected during winter (November 2011–February 2012) and 212 samples were collected during summer (May–August 2012) from central areas of Punjab, Pakistan. The samples were analyzed for the presence of aflatoxin M1 (AFM1) with a validated HPLC method equipped with florescence detector. The results revealed that from winter season almost 45% samples of milk and milk products were found to be contaminated with AFM1 i.e. 40% of raw milk, 51% of UHT milk, 37% of yogurt, 60% of butter and 43% of ice cream samples and 27, 24, 25, 34 and 17% of samples were found above the recommended limit for AFM1, respectively. However, from summer season 32% samples of milk and milk products were found to be contaminated i.e. 36% of raw milk, 31% of UHT milk, 29% of yogurt, 40% of butter and 24% of ice cream and 23, 23, 18, 20 and 5% of samples were found above the permissible limit for AFM1, respectively. The levels of contamination in winter milk and milk product samples were significantly higher (α ≤ 0.05) than in summer season. The occurrence of AFM1 in milk and milk products were higher, demanding to implement strict regulations and also urged the need for continuous monitoring of milk and milk products in order to minimize the health hazards. [Copyright &y& Elsevier]

Abstract: A total 431 samples including halva (56), pistachio (71), almond (63), semolina (69), cardamom (34), raisins (46), halva puri (39) and wheat powder (53) were analyzed using HPLC equipped with florescence detector. The results have shown that 32 (57%) samples of halva, 45 (63%) pistachio, 43 (68%) almond, 46 (67%) semolina, 21 (62%) cardamom, 19 (41%) raisins, 21 (54%) halva puri and 22 (42%) of wheat powder samples were found contaminated with AFB1, and 11 (20%), 23 (32%), 34 (54%), 12 (17%), 11 (32%), 7 (15%), 9 (23%) and 11 (21%) samples, respectively were above the European Union permissible limit (2 μg/kg). The results have shown that 20 (59%) samples of halva that contained milk were found contaminated with AFM1 and 3 (9%) samples were found above the recommended limit for AFM1 i.e. 0.05 μg/kg. Limit of detection (LOD) and Limit of quantification (LOQ) for AFB1 and AFM1 were 0.04 μg/kg, 0.12 μg/kg, and 0.004 μg/L, 0.012 μg/L, respectively. [Copyright &y& Elsevier]

Abstract: A total 198 peanut and peanut products, commercially available in major cities of Punjab Pakistan were analyzed for aflatoxins (AFs) using HPLC equipped with florescence detector. The data revealed that 59% raw peanut with shell, 55% raw peanut without shell, 61% roasted peanut with shell, 68% roasted peanut without shell, 50% peanut butter, 42% peanut cookies and 20% peanut nimko were found contaminated with AFs. The mean concentration in raw peanut shell was (6.4 μg/kg), raw peanut without shell (9.6 μg/kg), roasted peanut with and without shell (10.4 and 12.3 μg/kg), peanut butter (2.4 μg/kg), peanut cookies (4.6 μg/kg) and peanut nimko (3.4 μg/kg). The present study was a first report on the occurrence of AFs contamination in peanut and peanut products from the markets of Punjab, Pakistan. The level of AFs in peanut and peanut products is high and poses a significant threat for the health of people. [Copyright &y& Elsevier]

Abstract: Total 170 samples of chillies (chilli sauce, crushed chilli and powdered chilli) were analyzed by HPLC coupled with fluorescence detector. AFs were detected in 39, 59 and 56% of chilli sauce, crushed chilli and chilli powder samples from open market compared with 29, 54 and 58% samples respectively, from food restaurants. OTA was found 5 (19%), 11 (38%) and 13 (38%), in chilli sauce, crushed chilli and chilli powder from open market compared to 4 (17%), 14 (50%), and 12 (41%) samples respectively, from food restaurants. About 12, 27 and 35% chilli sauce, crushed chilli and powdered chilli samples from open market were found above the European Union (EU) limit for total AFs compared to 8, 32, and 41% samples from food restaurants. The daily intakes of AFs and OTA were 3.3 and 2 μg/kg, respectively. [Copyright &y& Elsevier]

Abstract: The basic objective of this research was to investigate the incidence and occurrence of aflatoxin M1 (AFM1) in milk and dairy products produced in the province of Punjab, Pakistan. Milk samples (107) and dairy products including yogurt (96), white cheese (119), cheese cream (150) and butter (74) samples were analyzed using High performance liquid chromatography equipped with fluorescence detector. The results have shown that AFM1 was detected in 71% of milk samples with 58% samples were found above the permissible limit of European Union (EU). AFM1 was detected in 61% of yogurt, 78% white cheese, 59% cheese cream and 45% butter samples with 47, 15, 11 and 52% samples of yogurt, white cheese, cheese cream and butter, respectively were found above the recommended limit of EU. The data of the present study will be helpful for the implementation of regulatory limit for AFB1 in order to minimize or avoid AFM1 in milk and milk products from Pakistan and also gives insights that whether the occurrence of AFM1 in dairy products was considered as a possible risk for consumers health. [Copyright &y& Elsevier]

The effect of gamma radiation on moisture content, total mold counts, Aspergillus counts, and aflatoxins of three hot pepper hybrids (Sky Red, Maha, and Wonder King) was investigated. Whole dried peppers packed in polyethylene bags were gamma irradiated at 0 (control), 2, 4, and 6 kGy and stored at 25°C for 90 days. Gamma radiation proved to be effective in reducing total mold and Aspergillus counts in a dose-dependent relationship. Total mold counts in irradiated peppers immediately after treatments were significantly lowered compared with those in nonirradiated samples, achieving 90 and 99% reduction at 2- and 4-kGy doses, respectively. Aspergillus counts were significantly reduced, by 93 and 97%, immediately after irradiation at doses of 2 and 4 kGy, respectively. A radiation dose of 6 kGy completely eliminated the population of total molds and Aspergillus fungi. The evolution of total molds in control and irradiated samples indicated no further fungal proliferation during 3 months of storage at 25 °C. Aflatoxin levels were slightly affected by radiation doses of 2 and 4 kGy and showed a nonsignificant reduction of 6% at the highest radiation dose of 6 kGy. The distinct effectiveness of gamma radiation in molds and aflatoxins can be explained by the target theory of food irradiation, which states that the likelihood of a microorganism or a molecule being inactivated by gamma rays increases as its size increases. [ABSTRACT FROM AUTHOR]

Abstract: During October 2009 to September 2010, aflatoxin M1 (AFM1) levels were analyzed by HPLC-FLD in 356 milk samples of different lactating species (buffalo, cow, goat, sheep and camel) from Punjab (Pakistan). Recoveries of AFM1 ranged from 92 to 97% and the limit of detection (LOD) was 0.004 μg/L. For all lactating species the mean concentration of AFM1 was significantly higher in winter season than in summer (p < 0.05). The results showed that 55, 56, 32, 58 and 27% of winter milk samples of buffalo, cow, goat, sheep and camel exceeded the EU maximum limit (0.05 μg/kg), compared with 38, 33, 21, 36 and 14% of summer milk samples, respectively. For all lactating species the mean concentration of AFM1 was significantly higher in morning milks than in evening milks (p < 0.05). The percentage of morning milk samples exceeding the EU maximum limit was 72, 67, 69, 71 and 44% for buffalo, cow, goat, sheep and camel, while for evening milks percent non compliant rates were 39, 30, 18, 33 and 25%, respectively. The level of AFM1 tended to be higher in animal species fed mainly on concentrate mixtures (buffalo and cow) than in other species grazing on fresh greens. [Copyright &y& Elsevier]

Deferiprone (1, 2 dimethyl-3-hydroxypyrid-4-one) is considered to be the standard iron chelator. Pharmacokinetic studies of generic formulations are required in local condition before placed on the market. High performance liquid chromatographic (HPLC) method was used for quantification of deferiprone in human plasma using UV/VIS detector. Chromatographic separation was carried out on C18 column, with a mobile phase of methanol-buffer (18:82, v/v), pH 3.5, and caffeine was used as an internal standard. The calibration curve was linear over the range 0.25- 10 µg/mL in human plasma (R² = 0.9994). After oral administration of deferiprone (500 mg) to human, the plasma concentration-time curve of deferiprone was conformed to two-compartment open model. The deferiprone plasma concentration showed a rapid absorption and average area under the plasma concentration-time curve (AUC) of deferiprone was 17.0 ± 1.23 h.µg/mL. Average absorption and elimination half-life values of deferiprone of 24 volunteers were 0.62 ± 0.12 and 2.65 ± 0.43 hours. This study confirms the rapid absorption of deferiprone in humans. AUC was similar to that previously reported but Cmax was slightly lower than that stated in the literature. [ABSTRACT FROM AUTHOR]

Two different weight groups of farmed Labeo rohita were collected from fish hatchery, Satiana Road Faisalabad, Pakistan. Pesticide residues were determined by high-performance liquid chromatography (HPLC). HPLC analysis showed that seven pesticide residues (α-endosulfan, DDE, parathion methyl, isoproturon, atrazine, carbofuran, and deltamethrin) were detected in L. rohita in weight group W 1.Six pesticide residues (α-endosulfan, parathion methyl, isoproturon, atrazine, carbaryl, and carbofuran) were found in L. rohita weight group W 2. The overall comparison between two weight groups of L. rohita showed that maximal concentration of pesticide residues were observed in underweight group W 2 compared to W 1 weight group. [ABSTRACT FROM AUTHOR]

Aflatoxin contamination and mold proliferation in three hot pepper hybrids (Sky Red, Maha, and Wonder King) were studied during 5 months of storage at three temperatures (20, 25, and 30°C) and under different packaging conditions (low-density polyethylene bags and jute bags). The presence of aflatoxins in hot pepper samples was determined by high-performance liquid chromatography with a UV-Vis detector. Sampling for analysis of aflatoxins, total mold counts, and Aspergillus counts was carried out at 0, 50, 100, and 150 days of storage. Hot peppers packed in jute bags were more susceptible to aflatoxin contamination than those packed in polyethylene bags; aflatoxin concentrations were 75% higher in peppers stored in jute bags. The effect of storage temperature resulted in aflatoxin concentrations that were 61% higher in hot peppers stored at 25 and 30°C than in those stored at 20°C. Of the three pepper hybrids, Wonder King was more susceptible to aflatoxin contamination, with a maximum of 1.50 µg/kg when packed in jute bags and stored at 25°C for 150 days. However, no sample exceeded the maximum permitted level for total aflatoxins in spices established by European Union regulations (10 µg/kg). Total mold counts and Aspergillus counts increased with storage duration, but all counts were significantly lower in peppers stored in polyethylene bags. A gradual increase in temperature during prolonged storage of hot peppers in combination with aeration may be the main reasons for increases in fungal biomass and Aspergillus proliferation with the subsequent aflatoxin production. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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

Cirrhinus mrigala was exposed to sub-lethal concentrations of carbosulfan (carbamate) and parathion (organophosphate) to study the toxic effects on acetylcholinesterases (AChE) and butyrylcholinesterases (BuChE) activity in the brain, gills, liver, kidneys, blood and flesh, compared with control samples. The fingerlings of Cirrhinus mrigala were exposed to three sub-lethal concentrations of parathion (0.041, 0.020 and 0.013 mg/L) and carbosulfan (0.098, 0.049 and 0.033 mg/L) in three replicates for 60 days. The activity of cholinesterases (AChE and BuChE) was significantly inhibited, even at the lowest exposure levels of parathion and carbosulfan, in gills, liver, kidney and muscle of the fish. Findings from this study have demonstrated that inhibition of AChE and BuChE activity in C. mrigala is a useful biomarker in assessing the aquatic environment contaminated by anticholinesterases not only in the brain, but also in other vital organs; hence, it can be used as a biomarker for monitoring purposes. [ABSTRACT FROM AUTHOR]

The current study is focused to investigate the natural presence of Patulin (PAT) in various fruits, juices and smoothies marketed and consumed in Pakistan. A total of 237 samples of fruits, juices and smoothies were analysed using isocratic HPLC with UV detector. The results have shown that 136 out of 237 (57.4%) samples have found positive with PAT contamination with levels ranged from 0.04 to 1100 μg kg −1 . The highest mean level of PAT was found in red globe grapes i.e. 921.1 ± 22.4 μg kg −1 . About 33.8% samples have levels of PAT with concentration above the permissible regulations of European Union (EU) i.e. 50 μg kg −1 . The dietary intake of PAT from fruits, juices and smoothies were 0.0049, 0.0016 and 0.0014 μg kg −1 bw day −1 with hazard quotient (HQ) value of 1.22, 0.40 and 0.35%, respectively. The incidence and occurrence of PAT in fruits and fruits products have high and need urgent attention for comprehensive survey. The results of present study would be useful for farmers, traders, and consumers. [ABSTRACT FROM AUTHOR]

A total 307 samples of dry fruits and edible nuts from Northern areas and Khyber Pakhtunkhwa, Pakistan were evaluated for the presence of aflatoxins (AFs). The samples were analysed using a reversed phase HPLC, equipped with fluorescence detector. Results have revealed that 132 out of 307 samples of dry fruits and nuts were found positive with aflatoxin B 1 (AFB 1 ) and total AFs. The highest mean level of total AFs i.e. 7.89 ± 0.99 μg/kg was found in peanuts without shell and lowest mean level (2.45 ± 0.11 μg/kg) was found in watermelon seeds without shell samples. Samples 75 (24%) were found contaminated with AFB 1 , ranged from 8 to 10 μg/kg and 41 (13%) samples were found above the level of 10 μg/kg for total AFs. The high occurrence of AFs may cause health hazards for consumers and limit exports. [ABSTRACT FROM AUTHOR]

In the present study, a total of 380 samples of 5 rice varieties were investigated for the presence of aflatoxins (AFs) and tocopherols (Vitamin E) using HPLC, equipped with a fluorescence detector. The data revealed that super basmati rice has the highest mean level of total AFs (12.45 µg/kg) followed by KS-282 (11.2 µg/kg), basmati PK-385 (9.8 µg/kg), Irri-6 (9.7 µg/kg) and the lowest mean level was found in Irri-9 (8.9 µg/kg). Furthermore, the results have shown that the variety with lowest level of AFs has the highest level of tocopherol content; i.e. Irri-9 (53.2 mg/100 g), basmati PK-385 (45.9 mg/100 g), Irri-6 (45.3 mg/100 g), KS-282 (40.4 mg/100 g) and super basmati rice (40.2 mg/100 g). The data has shown correlation (r = -0.62, p < 0.05) between vitamin E content and AFs concentration in different rice varieties. The results are interesting and need further study to investigate the mechanistic background of vitamin E content and its effect on aflatoxins contamination. [ABSTRACT FROM AUTHOR]

Abstract: Aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZEN) were analyzed in 115 chicken meat and 80 eggs samples, collected from central areas of Punjab, Pakistan. The study was carried out using reverse phase HPLC, equipped with fluorescence detector. The results revealed that 35% samples of chicken and 28% samples of eggs were found contaminated with AFs, and maximum level of AFB1 and total AFs was found in the liver part of chicken (layer) 7.86 and 8.01 μg/kg, respectively. Furthermore, 41% samples of chicken and 35% sample of eggs were found contaminated with OTA and maximum level 4.70 μg/kg was found in the liver part of chicken meat. However, 52% samples of meat and 32% samples of eggs were found contaminated with ZEN and maximum level 5.10 μg/kg was found in the liver part of chicken meat. The occurrence and incidence of AFs, OTA and ZEN in chicken meat and eggs are alarming and it may produce health hazards and urged the need of continuous monitoring for these toxins in chicken meat and eggs. [Copyright &y& Elsevier]

Abstract: In present study, a total of 275 retail cereal samples including rice, corn and corn products was analyzed for the presence of aflatoxins and ochratoxin A (OTA) using HPLC equipped with a fluorescence detector. The data has shown that 38 out of 68 samples of rice, 37/105 of corn and 43/102 samples of corn products have been found contaminated with AFs, average level of AFB1 and total AFs, 8.23 and 19.54, 7.90 and 12.08, and 5.47 and 7.85 μg/kg, respectively. Furthermore, 34, 28 and 26 numbers of rice, corn and corn product samples were found to be contaminated with OTA with an average level of 12.94, 5.29 and 3.69 μg/kg, respectively. The samples of rice, corn and corn products found above the permissible European Union (EU) limit for AFB1 and total AFs were 18, 13 and 14 and 28, 14 and 20%, respectively; however, the samples of rice, corn and corn products above the EU limit were 40, 14 and 15%, respectively. [Copyright &y& Elsevier]

Abstract: Chillies are a very important cash crop of Pakistan. The effects of gamma irradiation on microbial load, aflatoxin B1 (AFB1) and total aflatoxins have been studied in chillies samples, collected from different districts of Punjab, Pakistan. Aflatoxins were analyzed using HPLC equipped with a fluorescence detector. The results revealed that among the Aspergillus species isolated, those belonging to section parasiticus were predominant. Gamma radiations of doses 2, 4 and 6kGy were employed on fungi and chilli samples. The results have demonstrated that the dose of 6kGy reduced the fungal load by 5logs. Furthermore, 6kGy reduced the level of AFB1 and total AFs in ground and whole chillies by 1–2logs (α < 0.05). [Copyright &y& Elsevier]

Leaf and internode explants from in vitro grown seedlings of Citrullus colocynthis (L.) Schard. were used to establish callus cultures in MS media supplemented with different concentrations (ranging from 1 to 3 mg/L) and combinations of plant growth regulators like BAP with NAA, and Kin with 2, 4-D. BAP 3 mg/L and NAA 2 mg/L supported green to yellowish green embryogenic callus from internode explant with callus index of 300 and callus initiation commenced in 3 days. Leaf explant produced green callus under influence of 2, 4-D 2 mg/L and Kin 1 mg/L with callus index of 200 and callus initiation duration was 15 d. Leaf and internode tissues from in vitro grown seedlings and leaf and internode calli were analyzed for secondary metabolites through GC-MS. Nineteen compounds were detected from leaf and internode tissues. From the comparative analysis of secondary metabolites it can be concluded that regardless of a few different compounds, most of the secondary metabolites from seedlings were present in their respective calluses. [ABSTRACT FROM AUTHOR]

The antioxidant potential of 80% methanolic extracts of some agro wastes (pomegranate peel, apple peel, banana peel, citrus peel, corncob, wheat husk, wheat bran, rice bran, and rice hull) was assessed. The yields of the extracts varied over a wide range (8.83 to 29.9 g/100g of dry weight). TPC, TFC, total flavonols (kaempeferol, quercetin, myricetin; HPLC method), DPPH radical scavenging and inhibition of linoleic acid peroxidation for the extracts varied significantly (P < 0.05) ranging from 0.16-36.40, 0.026-4.89 g/100g of dry matter, 1.06-2622.41 mg/Kg of dry matter, 11.2-52.6, and 64.8-96.4%, respectively. Pomegranate peel extract, offering the highest TPC, TFC, and total flavonols exhibited superior antioxidant activity followed by apple peel, citrus peel, banana peel, corncob, wheat bran, rice bran, wheat husk, and rice hull. Furthermore, the antioxidant effectiveness of the extracts was assessed using corn oil (CO) as the oxidation substrate. CO samples stabilized with extracts at a concentration of 600 ppm (w/w) were subjected to accelerated aging (60 °C, oven heating, 30 days; 8 h heating cycle per day), and analyzed periodically for the extent of oxidative alterations following the measurement of conjugated dienes (CD), conjugated trienes (CT), p-anisidine and peroxide values. The correlation between the results of different antioxidant assays and oxidation parameters investigated in the present study indicated that fruit peel extracts, exhibiting higher TPC, TFC, flavonol and scavenging power, were also more potent for enhancing the oxidative stability of corn oil. [ABSTRACT FROM AUTHOR]