Food authentication, current issues, analytical techniques, and future challenges: A comprehensive review.

Food authentication and contamination are significant concerns, especially for consumers with unique nutritional, cultural, lifestyle, and religious needs. Food authenticity involves identifying food contamination for many purposes, such as adherence to religious beliefs, safeguarding health, and consuming sanitary and organic food products. This review article examines the issues related to food authentication and food fraud in recent periods. Furthermore, the development and innovations in analytical techniques employed to authenticate various food products are comprehensively focused. Food products derived from animals are susceptible to deceptive practices, which can undermine customer confidence and pose potential health hazards due to the transmission of diseases from animals to humans. Therefore, it is necessary to employ suitable and robust analytical techniques for complex and high-risk animal-derived goods, in which molecular biomarker-based (genomics, proteomics, and metabolomics) techniques are covered. Various analytical methods have been employed to ascertain the geographical provenance of food items that exhibit rapid response times, low cost, nondestructiveness, and condensability.

Application of carboxymethylcellulose in combination with essential oils nano-emulsions edible coating for the preservation of kiwifruit.

The present research investigates the effectiveness of nano-emulsified coatings (C-1, C-2, and C-3) in preserving the kiwifruit at a temperature of 10 ± 2 °C with 90-95 % relative humidity (RH) for 30 days. The nano-emulsions were prepared from varied carboxymethyl cellulose (CMC) concentrations with different combinations of essential oils such as thyme, clove, and cardamom. Dynamic light scattering investigation with Zeta Sizer revealed that C-1, C-2, and C-3 nano-emulsions have nano sizes of 81.3 ± 2.3, 115.3 ± 4.2, and 63.2 ± 3.2 nm, respectively. The scanning electron microscopy images showed that the nanoemulsion of C-1 had homogenous spherical globules, C-2 had voids, and C-3 showed a non-porous structure with uniform dispersion. The X-ray diffraction analysis indicated that C-1, C-2, and C-3 nano-emulsion exhibited distinct crystallinity and peaks. The nano-emulsion C-1 had reduced crystallinity, while C-2 had lower intensity peaks, and C-3 had increased crystallinity. The results documented that compared to control kiwifruit samples, the samples coated with C-3 nano-emulsion have decreased weight loss, decay incidence, soluble solids, maturity index activity, ethylene production, total bacterial count, and increased titratable acid, and firmness attributes. The results of current research are promising and would be applicable in utilization in industrial applications.

Physicochemical, Rheological and Sensory Evaluation of Herbal Bread Containing Turmeric, Ginger, and Black Cumin Powder.

The diversity in the global food market is expanding as thousands of new products enter the business every year, among which nutraceutical and functional foods hold important positions. The present research work aimed at the nutritional evaluation of three medicinal herbs, i.e., turmeric (Curcuma longa L.), ginger (Zingiber officinale), and black cumin (Nigella sativa). A bread formulation was enriched with the individual/combined supplementation (1-3%) of these herbs. Later, the bread was analyzed for nutritional, rheological, textural, and sensorial characteristics. The results revealed that the herbs improved the nutritional composition of bread, especially ash and fiber, as the maximum ash and fiber contents were noticed in T15 (2.0% dried powder of each plant) with values of 1.64 ± 0.04% and 4.63 ± 0.16%, respectively. The results regarding the rheological behavior showed minor variations in the rheological traits and a slight increase in dough development time up to 4.50 ± 0.20 min in T10 from 2.80 ± 0.13 min in T0. The sensorial attributes also indicated their marked suitability as external and internal characteristics were least affected by the addition of the herbs. Although some parameters like the crust and crumb colors were affected by the addition of black cumin, showing values of 6.25 ± 0.52 and 4.44 ± 0.19, respectively, in T15, and aroma characteristics were affected by the addition of ginger, supplementation with a combination of herbs at lower doses mitigated the adverse effects of other herbs. Moreover, shelf-life extension, especially with the addition of turmeric powder, was the hallmark of this research. This study concluded that medicinal herbs can be incorporated into baked products to improve the nutritional and sensorial attributes of functional herbal bread.

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).

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.

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.

Patulin Contamination of Citrus Fruits from Punjab and Northern Pakistan and Estimation of Associated Dietary Intake.

This research aims to assess the natural occurrence of patulin (PAT) in selected citrus fruits from central cities of Punjab and Pakistan's northern cities. A total of 2970 fruit samples from 12 citrus cultivars were examined using liquid chromatography fitted with a UV detector. The detection limit (LOD) and quantification limit were 0.04 and 0.12 µg/kg, respectively. About 56% of samples of citrus fruits from Punjab's central cities, Pakistan, were found to be contaminated with PAT, with values ranging from 0.12 to 1150 µg/kg in samples from central Punjab cities. Furthermore, 31.7% of samples of citrus fruits from northern cities of Pakistan were contaminated with PAT, with values ranging from 0.12 to 320 µg/kg. About 22.1% of citrus fruit samples had PAT levels greater than the suggested limits established by the European Union (EU). The dietary intake levels of PAT ranged from 0.10 to 1.11 µg/kg bw/day in the central cities of Punjab, Pakistan, and 0.13 to 1.93 µg/kg bw/day in the northern cities of Pakistan.

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).

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.

Assessment of Fumonisin B1 Concentrations in Wheat and Barley Products in the Punjab Region of Pakistan.

ABSTRACT: A total of 133 samples of whole wheat and barley grains and wheat and barley flour collected from retail markets in the main cities of Punjab, Pakistan, were analyzed for the mycotoxin fumonisin B1 (FB1) using reverse phase high-performance liquid chromatography with fluorescence detection. Of these samples, 120 (90%) were positive for FB1, and 75 (63%) of the 120 positive samples had FB1 concentrations higher than the European Union maximum (200 µg/kg). The limit of detection was 4 µg/kg. The highest mean (±SD) concentration of FB1 was found in whole wheat samples, 980.5 ± 211.4 µg/kg. The calculated dietary intakes of FB1 from wheat and barley flours were 4,456 and 503.7 ng/g of body weight per day, respectively.

Exploration of Carica papaya bioactive compounds as potential inhibitors of dengue NS2B, NS3 and NS5 protease.

Current outbreak of dengue has shown serious health concerns in Pakistan. The present study reports the anti-dengue potential of Carica papaya natural compounds. The leaves of C. papaya have previously shown promising results in cure of Dengue fever. The aim of this project is to find specific bioactive compounds by computational screening and biological activities of C. papaya against serine NS2B, NS3 and NS5 proteases of dengue virus. Docking study resulted in the screening of nine bioactive compounds having highest docking scores. However, three compounds namely epigallocatchin, catechin and protocatechuric acid had the strongest binding affinity with the active residues i.e., Ser135, His51 and Asp75 of dengue virus serine proteases. Results also indicated that the extract of C. papaya was a strong antimicrobial and antioxidant agent. It is concluded that the C. papaya compounds can be commercially applied for medical formulations against dengue virus.

Heavy Metals in Selected Vegetables from Markets of Faisalabad, Pakistan.

Two hundred ten samples of selected vegetables (okra, pumpkin, tomato, potato, eggplant, spinach, and cabbage) from Faisalabad, Pakistan, were analyzed for the analysis of heavy metals: cadmium (Cd), lead (Pb), arsenic (As), and mercury (Hg). Inductively coupled plasma optical emission spectrometry was used for the analysis of heavy metals. The mean levels of Cd, Pb, As, and Hg were 0.24, 2.23, 0.58, and 7.98 mg/kg, respectively. The samples with Cd (27%), Pb (50%), and Hg (63%) exceeded the maximum residual levels set by the European Commission. The mean levels of heavy metals found in the current study are high and may pose significant health concerns for consumers. Furthermore, considerable attention should be paid to implement comprehensive monitoring and regulations.

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.

Heterocyclic aromatic amines in deep fried lamb meat: The influence of spices marination and sensory quality.

The present study was focused to investigate the effect of selected spices (turmeric, torch ginger, lemongrass and curry leaves) on the formation of heterocyclic amines (HCAs, IQx, MeIQ, MeIQx, DiMeIQx, IQ, harman, norharman, and AαC) in deep fried lamb meat. Meat samples were marinated with optimized levels of turmeric (4 %), 10 % each of torch ginger, lemon grass, curry leaves at medium (70 °C) and well done (80 °C) doneness temperatures. The concentration of HCAs in deep fried meat samples were analysed using LC-MS/MS technique. The results revealed that torch ginger (10 %) has reduced 74.8 % of Me1Qx (1.39 to 0.35 ng/g) at medium doneness, followed by the 64.7 % reduction, using curry leaves and turmeric at medium degree of doneness. Torch ginger has reduced 86.6 % of AαC (2.59 to 0.40 ng/g) at well done doneness. The most prevalence level of HCAs was found in deep fried meat i.e. DiMeIQ (3.69 ng/g) at well done doneness. The sensory evaluation, using a 7 point hedonic test design for colour and texture in deep fried meat samples were resulted in a preferred color of golden brown and slightly tough texture. The use of local spices in marinating of deep fried lamb meat samples will certainly inhibit/reduce the level of these toxic and harmful HCAs.

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.

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.

Effect of Nigella sativa fixed and essential oils on antioxidant status, hepatic enzymes, and immunity in streptozotocin induced diabetes mellitus.

BACKGROUND: Nigella sativa fixed (NSFO) and essential (NSEO) oils have been used to treat diabetes mellitus and its complications. Present study was undertaken to explore and validate these folkloric uses. METHODS: Sprague dawley rats having streptozotocin (STZ) induced diabetes mellitus were used to assess the role of NSFO and NSEO in the management of diabetes complications. Parameters investigated were antioxidant potential, oxidative stress, and the immunity by in vivo experiments. RESULTS: The results indicated that STZ decreased the glutathione contents (25.72%), while NSFO and NSEO increased the trait significantly (P < 0.05). Experimental diets increased the tocopherol contents (P < 0.01) and enhanced the expression of hepatic enzymes (P < 0.01). Correlation matrix further indicated that antioxidant potential is positively associated (P < 0.05) responsible for the modulation of hepatic enzymes and the decrease of the nitric oxide production thus controlling the diabetes complications. CONCLUSIONS: Overall, results of present study supported the traditional use of N. sativa and its derived products as a treatment for hyperglycemia and allied abnormalities. Moreover, N. sativa fixed and essential oils significantly ameliorate free radicals and improve antioxidant capacity thus reducing the risk of diabetic complications.

Aflatoxin M1 in milk from urban and rural farmhouses of Punjab, Pakistan.

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 AFM1 from 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.

Assessment of aflatoxins, ochratoxin A and zearalenone in breakfast cereals.

Aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZEN) were analysed in 237 breakfast cereal samples collected from central areas of Punjab, Pakistan. According to the results, 41% of the samples were found contaminated with AFs, out of which 16% and 8% samples were found to be above the European Union (EU) maximum content for AFB1 and total AFs, respectively. About 48% samples were found contaminated with OTA and 30% samples were found to be above the EU maximum content. The results have shown that 53% samples of breakfast cereals were found contaminated with ZEN and 8% samples were found to be above the permissible limit of EU. The highest mean level of AFB1 and total AFs were found in semolina i.e. 3.60 and 4.55 µg/kg, respectively. Similarly, semolina was the highest contaminated breakfast cereal for OTA (3.90 µg/kg), while cornflakes (brand B) was found highest contaminated with ZEN (13.45 µg/kg).