Occurrence of Alternariol and Alternariolmonomethyl ether in edible oils: Their thermal stability and intake assessment in state of Uttar Pradesh, India.

Alternariol (AOH) and Alternariol monomethyl ether (AME) mycotoxins are found to be present naturally in various food commodities, such as barley, oats, pepper, rye, sorghum, sunflower seeds, tomatoes, and wheat. A few epidemiological studies have correlated the consumption of Alternaria-contaminated cereal grains with higher occurrence of esophageal cancer in Chinese populations. In addition, several studies have reported the toxicological properties of Alternaria mycotoxins. However, surveillance data on AOH and AME occurrence are still limited. Therefore, the goal of this study was to determine the presence of AOH and AME in various commonly consumed, edible oils using HPLC-FLD method. Thirty four percent of samples were found positive for AOH and 35% for AME. Moreover, AOH retained 80% stability, while AME retained 84% stability, after deep frying for 25 min, which is an important factor with respect to Indian cooking style. To the best of our knowledge, this is the first report on the presence of Alternaria mycotoxins in edible oils and their probable dietary intake in Indian population. This surveillance study may help in formulating guidelines for Alternaria mycotoxin levels in India, which are not yet implemented by Food Safety and Standards Authority of India. PRACTICAL APPLICATIONS: At present, no safety guidelines exist for Alternaria mycotoxins in any part of the world. This study will help the regulatory bodies to set permissible levels of Alternaria mycotoxins to safeguard the health of consumers. This study shows that Alternaria mycotoxins are heat stable even after deep frying for 25 min. The data will also help to issue guidelines against exposure of these mycotoxins, keeping in the mind the heat stability factor.

Fungal mediated biotransformation reduces toxicity of arsenic to soil dwelling microorganism and plant.

Rhizospheric and plant root associated microbes generally play a protective role against arsenic toxicity in rhizosphere. Rhizospheric microbial interaction influences arsenic (As) detoxification/mobilization into crop plants and its level of toxicity and burden. In the present investigation, we have reported a rhizospheric fungi Aspergillus flavus from an As contaminated rice field, which has capability to grow at high As concentration and convert soluble As into As particles. These As particles showed a reduced toxicity to soil dwelling bacteria, fungi, plant and slime mold. It does not disrupt membrane potential, inner/outer membrane integrity and survival of the free N2 fixating bacteria. In arbuscular mycorrhiza like endophytic fungi Piriformospora indica, these As particles does not influence mycelial growth and plant beneficial parameters such as phosphate solubilizing enzyme rAPase secretion and plant root colonization. Similarly, it does not affect plant growth and chlorophyll content negatively in rice plant. However, these As particles showed a poor absorption and mobilization in plant. These As particle also does not affect attachment process and survival of amoeboid cells in slime mold, Dictyostelium discoideum. This study suggests that the process of conversion of physical and chemical properties of arsenic during transformation, decides the toxicity of arsenic particles in the rhizospheric environment. This phenomenon is of environmental significance, not only in reducing arsenic toxicity but also in the survival of healthy living organism in arsenic-contaminated rhizospheric environment.

Methylenecyclopropyl glycine, not pesticide exposure as the primary etiological factor underlying hypoglycemic encephalopathy in Muzaffarpur, India.

Some districts of Bihar, especially Muzzaffarpur district, have been known to be affected by annual outbreak, called locally as Acute Encephalitis Syndrome (AES) which became one of the major health concerns in Bihar, due to its high fatality and complications. Several hypotheses like bat virus, heat stroke, pesticide exposure and the presence of a compound - methylenecyclopropyl glycine (MCPG) in Litchi have been proposed by different investigators for AES. When the investigators examined the symptoms, signs and the epidemiological data, bat virus and heat stroke hypothesis were ruled out. Two major hypotheses which remain in question were the exposure to pesticides or MCPG present in litchi. Therefore, this study was designed to check the presence of both in the Muzzaffarpur samples of ripe and semi ripe litchi fruits. The fruit cover of ripe and semi ripe litchi showed the traces of Malathion (0.18-0.19 µg/g) and p'-p'-DDT (0.022-0.023 µg/g), while no pesticide residues were detected in the pulp of ripe or semi ripe litchi thereby ruling out the possibility of pesticide exposure in children of Muzzaffarpur. However, MCPG was detected in the pulp of semi ripe (0.57 µg/g) and ripe litchi fruits (0.19 µg/g). Further, when the human condition was simulated in animals, there was deprivation in body weight and glucose levels in starved litchi seed dosed rats, causing hypoglycemia. These results suggest that the cause of hypoglycemic encephalopathy in Muzzaffarpur is related to the consumption of semi ripe and ripe litchi fruits by undernourished children.

Presence of Zearalenone in Cereal Grains and Its Exposure Risk Assessment in Indian Population.

Zearalenone (ZEA) is a toxic metabolite of Fusarium genera that frequently contaminates cereal grains. India being a tropical country provides suitable conditions for fungal invasion to the cereals. In the absence of any regulatory limits for ZEA in India, the present study was carried out to analyze the contamination levels of ZEA in different cereal samples consumed by Indian population and its exposure assessment through intake. Out of 117 cereal samples comprising of wheat, rice, corn, and oats, 70 (84%) were found to be positive for ZEA contamination, among which 24 (33%) samples exceeded the permissible limits proposed by European Union when analyzed by high-performance liquid chromatography. The positive samples were further validated by Liquid Chromatography-Mass Spectroscopy (LC-MS) analysis. Based on the quantitative estimation of ZEA contamination in cereals and their daily consumption values, the probable daily intake of ZEA was found to be 16.9- and 7.9-fold higher in rice and wheat samples, respectively, than the tolerable daily intake prescribed by European Food Safety Authority. The presence of ZEA at high levels indicates a higher exposure risk for Indian population as wheat and rice are staple foods in India. Thus, there is an immediate need to set the permissible levels of ZEA in India to safeguard the health of 1.34 billion people. PRACTICAL APPLICATION: High levels of ZEA contaminated wheat and rice samples suggest that the consumers are at a greater exposure risk. The study will help the Indian regulatory bodies to set the permissible level of ZEA in different cereal grains so as to safeguard the health of common masses. This can happen by simply adopting to European Food Safety Authority standards or depending on the consumption pattern of food and its occurrence, the new safe limit can be prescribed in India like in other Asian countries.

Simultaneous Determination of Acetaminophen and Synthetic Color(s) by Derivative Spectroscopy in Syrup Formulations and Validation by HPLC: Exposure Risk of Colors to Children.

Color additives are used in pediatric syrup formulations as an excipient; though not pre-requisite, but pediatric syrup formulations are normally colored. An attempt has been made to measure simultaneously the single drug, acetaminophen (AT), along with the colors, carmoisine (CA), erythrosine (ET), and sunset yellow FCF (SSY) added in it by three derivative spectroscopy methods namely, 1st order, ratio, and differential derivative methods. Moreover, evaluation has been made for the exposure assessment of the colors added as excipient because some colors have been reported to cause allergic reactions and hypersensitivity in children. The present methods provide simple, accurate, and reproducible quantitative determination of the drug, AT, along with the color in synthetic mixtures and commercial drug formulations without any interference. The limit of detection varied from 0.0001-0.31 µg/ml while limit of quantification ranged from 0.002-1.04 µg/ml in all the three methods. The calibration curve of all the three derivative methods exhibited good linear relationship with excellent regression coefficients (0.9986-1.000). Both intra-day and inter-day precisions showed %RSD value less than 2% while the percentage recovery was found between 96.8-103.8%. The sensitivity of the proposed methods is almost comparable to HPLC and thus, can be used for determination of drug AT, and color simultaneously in pharmaceutical formulation on routine basis. The present methods also showed that colors like SSY and ET are saturating more than 50% of acceptable daily intake (ADI) value which is alarming and needs to be considered for modification by regulatory authorities to safeguard the health of children.

Influence of temperature and pH on the degradation of deoxynivalenol (DON) in aqueous medium: comparative cytotoxicity of DON and degraded product.

Deoxynivalenol (DON), a toxic fungal metabolite, is stable under different processing conditions; however, its stability in aqueous medium at different temperatures and low pH (1-2) (present in the gastrointestinal tract) has not been investigated. In the present study, DON standard was used to study the influence of temperature and pH on DON stability in aqueous medium, the characterisation of the degraded product, and the comparative toxicity profile of the degraded and the parent compound. The results suggest that standard DON was unstable at 125-250°C showing 16-100% degradation whereas DON at pH 1-3 had 30-66% degradation, with a concomitant increase in the formation of a degraded product. Further ESI-MS characterisation of the dominant precursor ion of the HPLC eluate of the DON-degraded product was found to be m/z 279, resembling the known metabolite DOM-1. The degraded product of DON was reconfirmed as DOM-1 by comparison with standard DOM-1 and both gave a similar λmax at 208 nm. Comparative studies of both standard DOM-1 and the degraded product of DON showed no cytotoxicity up to 6400 ng ml(-1) while significant cytotoxicity was observed for DON (400 ng ml(-1)). The results suggest that a highly acidic environment (pH 1-2) could be responsible for the de-epoxydation of DON leading to the formation of DOM-1.

All India survey for analyses of colors in sweets and savories: exposure risk in Indian population.

In the present study, an attempt has been made to understand the exposure assessment of food colors through 2 major groups, sweets and savories, at a national level so as to evolve a scientific yardstick to fix levels of colors in commodities based on technological and safety requirement. A vast majority of colored food commodities (83.6%) were found to employ permitted colors and confirmed a marked decline in the trend of use of nonpermitted colors (NPCs). Of the 4 zones of India, East zone showed the maximum adulteration (80.3%) both by exceeding the prescribed limits of permitted colors (72.3%) and the use of NPCs (28.7%). Tartrazine was the most popular color among the permitted list, which ranged from 12.5 to 1091 mg/kg. Rhodamine B was the most prevalent dye in the NPCs group. On the basis of average consumption of food commodities and average levels of detected colors, the intake of Sunset Yellow FCF saturates the acceptable daily intake limit to a maximum of 47.8% in children, which is a cause of concern. The uniform maximum permissible limit of synthetic colors at 100 mg/kg under the Indian rules thus needs to be reviewed and should rather be governed by the technological necessity and the consumption profiles of food commodities so that the vulnerable population should not unnecessary be exposed to excessive amounts of synthetic colors to pose health risks.

Fatty acid composition including trans-fatty acids in edible oils and fats: probable intake in Indian population.

UNLABELLED: The susceptibility of trans-fat to the human health risk prompted the Food and Agriculture Organization (FAO) and World Health Organization (WHO) to prepare regulations or compulsory claims for trans-fatty acids (TFA) in edible oils and fats. In this study, analysis of fatty acid composition and TFA content in edible oils and fats along with the possible intake of trans-fat in Indian population was carried out. The analysis was carried out as per the Assn. of Official Analytical Chemists (AOAC) methodology and the results were statistically analyzed. The average TFA content in nonrefined mustard and refined soybean oils exceeded by 1.16- to 1.64-fold as compared to the Denmark limit of 2% TFA in fats and oils destined for human consumption. In branded/nonbranded butter and butter oil samples, average TFA limit exceeded by 4.2- to 9.5-fold whereas hydrogenated vegetable oil (HVO) samples exceeded the limit by 9.8-fold, when compared to Denmark standards. The probable TFA intake per day through different oils in Indian population were found to be less than WHO recommendation. However Punjab having highest consumption of HVO (-15 g/d) showed 1.09-fold higher TFA intake than the WHO recommendation, which is alarming and may be one of the factors for high cardiovascular disease mortality rate that needs further elucidation. Thus there is a need to prescribe TFA limit for edible oil, butter, and butter oil in India and to reduce the already proposed TFA levels in HVO to safeguard the health of consumers. PRACTICAL APPLICATION: The probable daily intake of trans-fatty acid (TFA) especially through hydrogenated vegetable oil (HVO) was assessed. In absence of any specification for TFA and fatty acid composition for edible oils, butter, and butter samples, a pressing need was felt to prescribe TFA limit in India. The study indicates that TFA intake through HVO consumption is higher in States like Punjab than the recommended daily intake prescribed by WHO. Hence, strategies should be adopted to either decrease the consumption of HVO or to modify the industrial processing method of HVO with less content of TFA to safeguard the health of consumers.

Usage pattern of synthetic food colours in different states of India and exposure assessment through commodities preferentially consumed by children.

Exposure studies in children are emphasized nowadays given children's higher consumption vulnerability. The present study generated national-level data covering 16 major states of India on the usage pattern of colours and it identified food commodities through which a particular colour has the scope to exceed ADI limits. Out of the total analysed samples, 87.8% contained permitted colours, of which only 48% adhered to the prescribed limit of 100 mg kg(-1). The majority of candyfloss, sugar toys, beverages, mouth fresheners, ice candy and bakery product samples exceeded the prescribed limit. Non-permitted colours were mostly prevalent in candyfloss and sugar toy samples. Though sunset yellow FCF (SSYFCF) and tartrazine were the two most popular colours, many samples used a blend of two or more colours. The blend of SSYFCF and tartrazine exceeded the prescribed limit by a factor of 37 in one sample, and the median and 95th percentile levels of this blend were 4.5- and 25.7-fold, respectively. The exposure assessment showed that the intake of erythrosine exceeded the ADI limits by two to six times at average levels of detected colours, whereas at the 95th percentile level both SSYFCF and erythrosine exceeded the respective ADI limits by three- to 12-fold in all five age groups. Thus, the uniform prescribed limit of synthetic colours at 100 mg kg(-1) under Indian rules needs to be reviewed and should be governed by consumption profiles of the food commodities to check the unnecessary exposure of excessive colours to those vulnerable in the population that may pose a health risk.

A simple method for simultaneous determination of basic dyes encountered in food preparations by reversed-phase HPLC.

The present method utilizes a simple pretreatment step, cleanup on polyamide SPE cartridges, and HPLC resolution on reversed-phase C18 for the detection of the three basic nonpermitted dyes encountered in food matrixes. Polyamide cartridges were chosen because both acidic and basic dyes can be cleaned up due to their amphoteric nature. Analysis was performed on a reversed-phase C18 micro-Bondapak column using the isocratic mixture of acetonitrile-sodium acetate with a flow rate of 1.5 mL/min and a programmable lambda(max) specific visible detection to monitor colors, achieving higher sensitivity and expanded scope to test multicolor blends. All the colors showed linearity with the regression coefficient, from 0.9983 to 0.9995. The LOD and LOQ ranged between 0.107 and 0.754 mg/L and 0.371 and 2.27 mg/L or mg/kg, respectively. The intraday and interday precision gave good RSDs, and percentage recoveries in different food matrixes ranged from 75 to 96.5%. The study demonstrates that the use of a combination of a simple SPE cleanup and HPLC resolution with UV-Vis end point detection was successful in screening the presence of these three basic nonpermitted dyes individually or in blend, in a variety of food matrixes.

Simultaneous determination of eight synthetic permitted and five commonly encountered nonpermitted food colors in various food matrixes by high-performance liquid chromatography.

A simple and sensitive HPLC method has been developed for the simultaneous determination of eight permitted food colors and five commonly encountered nonpermitted colors in various food commodities, including sugar-, fat-, and starch-based food matrixes. The method uses a specific food category-based cleanup/treatment procedure before color extraction to avoid the interference of food matrixes, and to obtain the optimal color extraction. Analysis was performed on a reversed-phase C18 micro-Bondapak column with ammonium acetate and acetonitrile gradient elution as the mobile phase; a programmable lamda max-specific visible detection was used to monitor colors to obtain the higher sensitivity and expanded scope needed for multicolor blends having diverse absorption maxima. All colors showed good linearity, with regression coefficients of 0.9974-0.9999. The LOD and LOQ values ranged from 0.01 to 0.12 mg/L, and from 0.04 to 0.83 mg/L or mg/kg, respectively. The intraday and interday precision tests produced good RSD values, and the recoveries from different food matrixes ranged from 82 to 104%. The method offers high sensitivity for analysis of a wide variety of food matrixes containing a broad scope of multicolor blends. Two nonpermitted colors, orange II and metanil yellow, were found. Also, a number of samples contained permitted colors at levels two- to seven-fold higher than those prescribed.

A simple 2-directional high-performance thin-layer chromatographic method for the simultaneous determination of curcumin, metanil yellow, and sudan dyes in turmeric, chili, and curry powders.

A method using simple extraction and 2-directional high-performance thin-layer chromatography (HPTLC) was developed for the simultaneous determination of curcumin, metanil yellow, and sudan dyes in turmeric, chili, and various mixed curry powder formulations. The method offers resolution (Rf) of turmeric pigments, namely, curcumin (0.77), demethoxycurcumin (0.69), bis(demethoxy)curcumin (0.61), and the synthetic dye metanil yellow (0.05) by the first-directional mobile phase, chloroform-methanol (9 + 1, v/v). The resolution (Rf) of sudan I (0.30) and sudan IV (0.23) was achieved by the second-directional mobile phase, toluene-hexane-acetic acid (50 + 50 + 1, v/v/v). Natural pigments of both turmeric and chili showed no interference in the detection and quantification of synthetic colors. The limit of detection and limit of quantification values for curcumin, metanil yellow, sudan I, and sudan IV were 17.39, 42.90, 15.45, and 7.01 and 52.71,130.0, 46.80, and 21.24 ng/spot, respectively. Analysis of a few market samples showed the presence of metanil yellow (1.5-4.6 mg/g), sudan I (4.8-12.1 mg/g), and sudan IV (0.9-2.0 mg/g) in loose turmeric and chili samples, whereas the curcumin content in turmeric and mixed curry powder samples ranged from 6.5 to 36.4 and from 0.3 to 1.9 mg/g, respectively. The method is relatively simple, offers reasonable sensitivity, and can be used to screen a large number of samples.