Chromatographic analysis of dithiocarbamate residues and their metabolites in foods employed in dietary exposure studies-a review.

Dithiocarbamates (DTCs) belong to a group of compounds used as fungicides in food production and can be divided into three major groups. Since DTCs easily oxidise and hydrolyse in alkaline and acidic medium respectively, precautions have to be implemented during preparation/homogenisation and extraction of samples. As such, test samples are commonly prepared individually by cutting into small pieces just before the digestion of DTCs with a hot acid to give carbon disulphide (CS2) and the results are expressed as CS2 without any differentiation of individual DTCs. However, individual DTCs have different toxicological potencies whilst their metabolites are more toxic than the parent compound. Apart from the hot digestion method, chromatographic separation of three major groups of DTCs has been developed by a number of different researchers. This review provides a comprehensive examination of sample preparation, extraction, clean-up and chromatographic methods for the determination of individual DTCs and their more toxic metabolites in foodstuffs. Moreover, this review also studies on how dietary exposure of DTCs can be efficiently and effectively estimated using different methods of analysis.

Quantification of permitted synthetic colours in food by liquid chromatographic methods: A review on analytical methods and theirperformance.

Colours, natural and synthetic, are substances which add or restore colour to a food after processing or storage. They are widely used by food manufacturers but may pose a potential risk to human health. Most food safety authorities set up regulations to limit the use of synthetic colours, and monitor their levels and consumption by the general public. Therefore, validated analytical methods are needed to fulfil this requirement. This review presents a comprehensive overview of various liquid chromatographic methods used for quantification of permitted synthetic colours in foods. Available analytical methods have been assessed for their fitness for purpose in terms of extraction, clean-up, liquid chromatographic separation, quantification and method performance. The advantages and disadvantages are given of available analytical methods for analysing 24 synthetic colours, permitted for use by different jurisdictions. Gaps in the knowledge and levels of validation are identified and recommendations made on further research to develop suitable methods for routine monitoring of these permitted synthetic colours.

A critical review of analytical methods for ergot alkaloids in cereals and feed and in particular suitability of method performance for regulatory monitoring and epimer-specific quantification.

Cereals and feed contaminated with ergot alkaloids (EAs) have been of concern for several decades. Nowadays, analysis of EAs is focused on ergometrine, ergotamine, ergosine, ergocristine, ergocryptine (a mixture of α- and ß-isomers) and ergocornine and their related -inine epimers as listed in the European Commission Recommendation 2012/154/EU. Liquid chromatography with fluorescence detection has been used for quantification of EAs for decades whilst LC-MS has become the work-horse for quantification of EAs in the last decade. However, in LC-MS analysis matrix effects of different magnitudes exist for each EA epimer, especially ergometrine/ergometrinine, even after different clean-up procedures. This leads to an underestimation or overestimation of EAs levels. Moreover, isotopic labelled standards for EAs are still not available in the market. This review aims to provide background information on different analytical methods, discuss their advantages and disadvantages and possible advancement. Moreover, the method performance requirements to support forthcoming regulations are also discussed.

The development of isomer-specific analysis of branched 4-nonylphenol in food for dietary exposure - a critical review of analytical methods and occurrence in foodstuffs.

4-Nonylphenol (4-NP) is a para-substituted phenolic compound comprising a straight or branched carbon chain group while branched 4-NP consists of 211 possible structural isomers. NP is recognised as an environmental pollutant and exists ubiquitously in both the environment and in food. 4-NP, especially branched 4-NP, has been shown to have the potential role of endocrine disruptor and xeno-oestrogen. Moreover, different NP isomers also exhibit different oestrogen-like activities. Recently, it was reported that the isomer-specific profile of 4-NP in foodstuffs varies greatly between and within food groups. Hence, risk assessment based on total branched 4-NP cannot reflect dietary risk. This study reviews the analytical methods applicable to conduct an isomer-specific analysis of 4-NP and its occurrence in foodstuffs. Lastly, research gaps are identified for future research.

Occurrence of organotin compounds in seafood from Hong Kong market.

The degree of organotin compounds (OTCs), including dibutyltin, tributyltin, triphenyltin and dioctyltin, contamination in seafood purchased in 2017 and 2018 from Hong Kong market was studied. Edible portions of 341 seafood samples, including fish, crustaceans and molluscs, were used for analysis by gas chromatograph coupled to an inductively coupled plasma mass spectrometry (GC-ICP/MS). The method detection limits and quantification limits of OTCs were below or equal to 0.25 and 1.0 µg Sn kg-1 respectively. Triphenyltin accounted for the majority amongst other OTCs and was detected in 53% of samples. In general, mean total OTCs levels of fish (24 µg Sn kg-1) were higher than crustaceans (14 µg Sn kg-1) and molluscs (15 µg Sn kg-1). The highest detected levels of triphenyltin, tributyltin, dibutyltin and dioctyltin were found to be 480, 24, 4.5 and 0.89 µg Sn kg-1 in a mangrove snapper, noodle fish, coral clam and giant grouper respectively.

Development and validation of an analytical method for the analysis of Sterigmatocystin in roasted coffee beans and black pepper using liquid chromatography-tandem mass spectrometry.

Sterigmatocystin (STC) is a toxic and potentially carcinogenic fungal toxin found in a variety of food commodities. This study describes the development of an analytical method to determine STC in roasted coffee beans and black pepper using ultra performance liquid chromatography (UPLC) coupled with triple quadrupole tandem mass spectrometry (MS/MS). 13C18-STC was used as internal standard. STC was extracted with a mixture of acetonitrile/water, diluted with a buffer, followed by purification with a solid-phase extraction and an immunoaffinity column prior to the UPLC-MS/MS analysis. Two multiple reaction monitoring (MRM) transitions were employed, one for quantification and one for confirmation of STC. The UPLC-MS/MS analytical method was validated with respect to selectivity, linearity, sensitivity, accuracy, precision, recovery, and stability. Calibration curves were linear over a concentration range 25-2,500 pg mL-1 with correlation coefficients (r) > 0.998. The method limit of quantification for STC in roasted coffee beans and black pepper was 0.10 µg kg-1. The accuracy and precision of the analytical method were acceptable within 15% at all quality control levels. This method was suitable to determine STC levels because of its selectivity, precision, and accuracy. The method was successfully applied to roasted coffee beans and black pepper samples.

Development of an Analytical Method for Analyzing Pyrrolizidine Alkaloids in Different Groups of Food by UPLC-MS/MS.

Suspected nontargeted pyrrolizidine alkaloids (PAs), without analytical reference standard, were observed and interfered with the determination of targeted PAs in complex food matrices, especially for spices samples. Selectivity and applicability of multiple reaction monitoring (MRM) transitions, multistage fragmentation (MS3), and MRM with differential ion mobility spectrometry (DMS) for eliminating false positive identifications were evaluated. Afterward, a selective and sensitive LC-MS/MS method for the determination of 15 PAs and 13 PA N-oxides in foodstuffs was developed. The sample preparation and cleanup are applicable to a wide range of foodstuffs, including cereal products, dairy products, meat, eggs, honey, tea infusion, and spices. Freezing-out of the raw extract and the water/acetonitrile washing steps in a solid phase extraction was found to efficiently remove complex matrices. The method was validated at 0.05 µg kg-1 for general food and 0.5 µg kg-1 for spices, with reference to the Eurachem Guide. The estimated limit of quantifications of different PAs was in the range of 0.010-0.087 µg kg-1 for general food and 0.04-0.76 µg kg-1 for spices. Isotopically labeled PAs were used as internal standards to correct the variation of PAs/PANs performance in different food commodities. Matrix effects observed in complex food matrices could be reduced by solvent dilution. Recoveries of PAs and PA N-oxides were all seen within 50-120%.

Determination of butyltins, phenyltins and octyltins in foods with preservation of their moieties: A critical review on analytical methods.

Tributyltin and triphenyltin have been extensively applied in anti-fouling paints since the 1960s. Hence, organotin compounds (OTCs), especially butyltins and phenyltins, in seafood has been of concern for decades. Even though the "International Convention on the Control of Harmful Anti-Fouling Systems on Ships, 2001" entered into force internationally in 2008 and prohibited the use of OTCs in anti-fouling paints used on ships, the analysis of OTCs in seafood was not commonly included in routine monitoring programmes. Besides, species of triphenyltin, including triphenyltin hydroxide and acetate, have been used as pesticides and may accumulate in food. Moreover, the European Food Safety Agency established a group tolerable daily intake for tributyltin, dibutyltin, triphenyltin and dioctyltin in 2004 as they exert their immunotoxic effects by similar mode of action and potency. Therefore, suitable methods are needed to analyze butyl-, phenyl- and octyl-tins simultaneously in food without affecting their moieties. This review aims to provide background information in this area.

Investigation of pyrrolizidine alkaloids including their respective N-oxides in selected food products available in Hong Kong by liquid chromatography electrospray ionisation mass spectrometry.

This study determined the levels of pyrrolizidine alkaloids (PAs), including their respective N-oxides, in foodstuffs available in Hong Kong by liquid chromatography-electrospray ionisation tandem mass spectrometry. A total of 234 samples (48 food items) were collected randomly from a local market and analysed. About 50% of samples were found to contain detectable amount of PAs. Amongst the 48 food items, PAs were not detected in 11 food items, including barley flour, beef, cattle liver, pork, pig liver, chicken meat, chicken liver, milk, non-fermented tea, Melissa tea and linden tea. For those found to contain detectable PAs, the summed PA content ranged up to 11,000 µg kg-1. The highest sum of PA content among the 37 food items calculated with lower bound was cumin seed, then followed by oregano, tarragon and herbs de Provence with ranges of 2.5-11,000, 1.5-5100, 8.0-3300 and 18-1300 µg kg-1 respectively. Among the samples, the highest sum of PA content was detected in a cumin seed sample (11,000 µg kg-1), followed by an oregano (5100 µg kg-1), a tarragon (3300 µg kg-1) and a herbs de Provence (1300 µg kg-1). In general, the results of this study agreed well with other published results in peer-reviewed journals, except that the total PAs in honey and specific tea infusion in this study were comparatively lower.

Development of an ultraperformance liquid chromatography-tandem mass spectrometry method for the analysis of perfluorinated compounds in fish and Fatty food.

This paper reports the development of a method for the quantitative analysis of perfluorinated compounds (PFCs), including C6-C14 perfluorinated carboxylic acids (PFCAs) and C4-C12 perfluorinated sulfonates (PFSAs), in fish and fatty foods by ultraperformance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) in which the UPLC was equipped a PFC Analysis Kit to eliminate background contamination. Rapid baseline separation was achieved for 17 PFCs within 12 min, and PFCs were well-resolved from potential interferences from taurodeoxycholic acid and branched isomers of PFCs. The method was validated according to Commission Regulation 2002/657/EC of the European Commission with matrices including salmon, beef, egg, and butter. Average spiked recoveries, measured at concentration levels of 0.02 (method limit of quantification (MLOQ)), 0.2, and 2 µg/kg, were in the range of 68-117% with relative standard deviations below 20%. Matrix effects were evaluated and found to be correctable by internal standardization, especially for short- and long-chained PFCs. Trueness was verified against two certified reference materials. The method has also been successfully applied to the analysis of more than 200 food samples of a risk assessment study.

Dietary exposure of Hong Kong adults to pesticide residues: results of the first Hong Kong Total Diet Study.

The use of pesticides and other chemicals has become a common practice in modern agriculture to enhance and stabilise crop yield, protect the nutritional integrity of food, facilitate food storage to assure year-round supplies, and provide attractive and appealing food products. With the adoption of strict good agricultural practice (GAP), only minimal amounts of pesticide residues should remain on the crops or in connected foods of animal origin up the food chain. To assess their associated health risk to local people, the dietary exposure of Hong Kong adults to residues of four groups of pesticides or their metabolites - organophosphorus pesticides (OPPs), carbamates, pyrethrins and pyrethroids, and dithiocarbamate (DTC) metabolites - is estimated in the first Hong Kong Total Diet Study (TDS). A total of 150 commonly consumed food items were collected and prepared "as consumed". A total of 600 composite food samples were analysed for 85 pesticides or their metabolites by liquid chromatography-tandem mass spectrometry (LC-MS/MS). These pesticides were primarily found at low levels (highest mean = 350 µg kg⁻¹) in food samples of plant origin such as vegetables and fruits. Dietary exposures to pesticide residues were estimated based on the analytical results and the food consumption data of the local residents. The estimated dietary exposures of Hong Kong adults to all individual pesticides were well below their respective acceptable daily intakes (ADIs). The percentage contributions of the estimated mean and 95th percentile dietary exposures to the ADIs of individual pesticides were <6% and <24% for the OPPs, <1% for the carbamates and pyrethrins and pyrethroids, and <1% and <4% for the DTC metabolites, respectively. The findings indicate that dietary exposures to all the pesticide residues analysed in this study were unlikely to pose unacceptable health risks to the Hong Kong population.

Dietary exposure of Hong Kong adults to acrylamide: results of the first Hong Kong Total Diet Study.

Acrylamide is a processing contaminant in food formed during cooking at high temperature, such as frying and baking. To assess the associated health risk of the Hong Kong population, the dietary exposure of Hong Kong adults to acrylamide was estimated in the first Hong Kong Total Diet Study (TDS), where food samples were collected and prepared "as consumed". A total of 532 composite food samples were analysed for acrylamide using LC-MS/MS. Dietary exposures were estimated by combining the analytical results with the food consumption data of the Hong Kong adults. The mean and 95th percentile exposures to acrylamide of the Hong Kong population were 0.213 and 0.538 µg kg⁻¹ body weight (bw) day⁻¹, respectively, and their margins of exposure (MOEs) were all below 10,000. The main dietary source of acrylamide was "Vegetables and their products" (52.4% of the total exposure), particularly stir-fried vegetables (44.9%), followed by "Cereals and their products" (14.7%) and "Mixed dishes" (9.43%). The study findings suggest that the relatively low figures for MOE for a genotoxic carcinogen may indicate human health concern of the Hong Kong population. Efforts should continue to be made in the interest of reducing acrylamide levels in food locally.

Occurrence of bound 3-monochloropropan-1,2-diol content in commonly consumed foods in Hong Kong analysed by enzymatic hydrolysis and GC-MS detection.

The aim of this study was to determine the level of bound 3-monochloropropan-1,2-diol in foodstuffs commonly consumed in Hong Kong, China, by an enzymatic hydrolysis indirect method which proved to be free from interferences. A total of 290 samples were picked up randomly from the local market and analysed. About 73% of these samples were found to contain detectable amounts of bound 3-MCPD. Amongst the 73 food items, bound 3-MCPD was not detected in 13 food items, including extra virgin olive oil, beef ball/salami, beef flank, ham/Chinese ham, nuts, seeds, soy sauce, oyster sauce, butter, yoghurt, cream, cheese and milk. For those found to contain detectable bound 3-MCPD, the content ranged up to 2500 µg kg(-1). The highest mean bound 3-MCPD content among the 14 food groups was in biscuits (440 [50-860] µg kg(-1)), followed by fats and oils (390 [n.d.-2500] µg kg(-1)), snacks (270 [9-1000] µg kg(-1)), and Chinese pastry (270 [n.d.-1200] µg kg(-1)). Among the samples, the highest bound 3-MCPD content was in a grape seed oil (2500 µg kg(-1)), followed by a walnut flaky pastry (1200 µg kg(-1)) and a grilled corn (1000 µg kg(-1)). Basically, the results of this study agreed well with other published results in peer-reviewed journals, except for cheese, cream, ham, nuts and seeds.

Dietary exposure of Hong Kong adults to fatty acid esters of 3-monochloropropane-1,2-diol.

A total of 290 individual food samples were collected in Hong Kong, China, for 3-monochloropropane-1,2-diol (3-MCPD) fatty acid esters analysis. Most samples were processed food and in ready-to-eat form. The results show that the levels of 3-MCPD fatty acid esters were high in biscuits, fats and oils, snacks and Chinese pastry with mean bound 3-MCPD levels of 440, 390, 270 and 270 µg kg⁻¹, respectively. The dietary exposures to bound 3-MCPD of average and high adult consumers were estimated to be 0.20 and 0.53 µg kg bw⁻¹ day⁻¹, respectively. The primary toxicological concern of 3-MCPD fatty acid esters is its potential to release 3-MCPD in vivo during digestion in the gastrointestinal tract. 3-MCPD would affect the kidney, the central nervous system and the male reproductive system of rats. Assuming that 100% of the 3-MCPD was released from 3-MCPD fatty acid esters by hydrolysis in the digestive system, the dietary exposures to 3-MCPD for average and high adult consumers were only 10% and 26% of the provisional maximum tolerable daily intake (PMTDI) of 3-MCPD established by the Joint Food and Agriculture Organization/World Health Organization Expert Committee on Food Additives (JECFA) (2 µg kg bw⁻¹ day⁻¹), respectively. The results suggest that both average and high adult consumers are unlikely to experience major toxicological effects of 3-MCPD.

Dietary exposure to inorganic arsenic of the Hong Kong population: results of the first Hong Kong total diet study.

Inorganic arsenic, a human carcinogen, can be found in the environment and food. In the first Hong Kong Total Diet Study, the dietary exposure of the Hong Kong people, including various age-gender subgroups, to inorganic arsenic was estimated for assessing the associated health risk. Food samples, which represented the Hong Kong people's diet, were collected and prepared "as consumed" for analysis. Concentrations of inorganic arsenic, as sum of arsenite (As(III)) and arsenate (As(V)) were determined in 600 composite samples by using inductively coupled plasma mass spectrometry. The dietary exposures were estimated by combining the analytical results with the local food consumption data of the adult population. The mean and 95th percentile of inorganic arsenic exposures of the Hong Kong people were 0.22 and 0.38µg/kg body weight (bw)/day, respectively. Among the 12 age-gender subgroups, the respective exposures ranged from 0.19 to 0.26µg/kg bw/day and from 0.33 to 0.46µg/kg bw/day. The main food category that contributed inorganic arsenic was "cereals and their products" (53.5% of the total exposure), particularly rice. Having considered the carcinogenic risk of inorganic arsenic to humans, it is suggested that efforts should be made to reduce the inorganic arsenic exposure of the Hong Kong population.

Development and validation of a method for determination of residues of 15 pyrethroids and two metabolites of dithiocarbamates in foods by ultra-performance liquid chromatography-tandem mass spectrometry.

This paper reports a novel approach for the detection, confirmation, and quantification of 15 selected pyrethroid pesticides, including pyrethins, and two metabolites of dithiocarbamates in foods by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS). The proposed method makes use of a modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) procedure that combines isolation of the pesticides and sample cleanup in a single step. Analysis of pyrethroids and dithiocarbamate metabolites was performed by UPLC-MS-MS operated with electrospray and atmospheric pressure chemical ionization, respectively. Two specific precursor-product ion transitions were acquired per target compound in multiple reaction monitoring (MRM) mode. Such acquisition achieved the minimum number of identification points according to European Commission (EC) document no. SANCO/10684/2009, thus fulfilling the EC point system requirement for identification of contaminants in samples. The method was validated with a variety of food samples. Calibration curves were linear and covered from 1 to 800 µg kg(-1) in the sample for all target compounds. Average recoveries, measured at mass fractions of 10 and 100 µg kg(-1) for pyrethroids and 5 and 50 µg kg(-1) for dithiocarbamate metabolites, were in the range of 70-120% for all target compounds with relative standard deviations below 20%. Method limits of quantification (MLOQ) were 10 µg kg(-1) and 5 µg kg(-1) for pyrethroids and dithiocarbamate metabolites, respectively. The method has been successfully applied to the analysis of 600 food samples in the course of the first Hong Kong total diet study with pyrethroids and metabolites of dithiocarbamates being the pesticides determined.

Determination of organochlorine pesticide residues in fatty foods: a critical review on the analytical methods and their testing capabilities.

Organochlorine pesticide (OCP) residues in foods have been of concern for several decades. However, the analysis of some of the OCPs and their metabolites or derivatives, such as endrin aldehyde, endrin ketone, nonachlor, etc. in fatty foods (including foods of animal and plant origin), was not commonly included in routine monitoring programme. Recently, the Stockholm Convention introduced nine plus one new persistent organic pollutants (POPs) that included chlordecone and some other OCPs. Is there a method available that can analyze both traditional OCPs, together with their metabolites and derivatives in fatty foods? Furthermore, is there a suitable method that can monitor OCPs and the newly added POPs including chlordecone in fatty foods together in a pot? This review aims to provide some background information to answer these questions.

Nitrate and nitrite levels in commonly consumed vegetables in Hong Kong.

Levels of nitrate and nitrite in 73 different vegetables, a total of 708 individual samples grouped into leafy, legumes, root and tuber, and fruiting vegetables, which are traded mainly in Hong Kong, were measured. Where available, five samples of each vegetable type were purchased from different commercial outlets during the winter of 2008 and summer of 2009. Levels of nitrate and nitrite were determined by ion chromatography and flow injection analysis, respectively. Nitrate and nitrite levels of all samples ranged <4-6300 and <0.8-9.0 mg kg⁻¹, respectively. Nitrate concentrations for the different groups, in descending order, were leafy > root and tuber > fruiting and legume vegetables. More than 80% of vegetables had mean nitrate concentrations less than 2000 mg kg⁻¹, but mean nitrate concentrations of three types of leafy vegetables, namely Chinese spinach, Shanghai cabbage and Chinese white cabbage, were >3500 mg kg⁻¹. On the other hand, nitrite concentrations were generally low - <1 mg kg⁻¹ on average. Nitrate in vegetables (i.e. Chinese flowering cabbage, Chinese spinach and celery) can be reduced significantly (12-31%) after blanching for 1-3 min, but not after soaking.

Validation and use of a fast sample preparation method and liquid chromatography-tandem mass spectrometry in analysis of ultra-trace levels of 98 organophosphorus pesticide and carbamate residues in a total diet study involving diversified food types.

This paper reports a comprehensive sensitive multi-residue liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for detection, identification and quantitation of 73 pesticides and their related products, a total of 98 analytes, belonging to organophosphorus pesticides (OPPs) and carbamates, in foods. The proposed method makes use of a modified QuEChERS (quick, easy, cheap, effective, rigged, and safe) procedure that combines isolation of the pesticides and sample clean-up in a single step. Analysis is performed by liquid chromatography-electrospray ionization-tandem mass spectrometry operated in the multiple reaction monitoring (MRM) mode, acquiring two specific precursor-product ion transitions per target compound. Two main fragment ions for each pesticide were obtained to achieve the identification according to the SANCO guidelines 10684/2009. The method was validated with various food samples, including edible oil, meat, egg, cheese, chocolate, coffee, rice, tree nuts, citric fruits, vegetables, etc. No significant matrix effect was observed for tested pesticides, therefore, matrix-matched calibration was not necessary. Calibration curves were linear and covered from 1 to 20 microg L(-1) for all compounds studied. The average recoveries, measured at 10 microg kg(-1), were in the range 70-120% for all of the compounds tested with relative standard deviations below 20%, while a value of 10 microg kg(-1) has been established as the method limit of quantitation (MLOQ) for all target analytes. Similar trueness and precision results were also obtained for spiking at 200 microg kg(-1). Expanded uncertainty values were in the range 21-27% while the HorRat ratios were below 1. The method has been successfully applied to the analysis of 700 food samples in the course of a baseline monitoring study of OPPs and carbamates.

Dietary exposure to aluminium of the Hong Kong population.

A total of 256 individual food samples were collected in Hong Kong for aluminium testing. Most of food samples were analysed in ready-to-eat form. High aluminium levels were found in steamed bread/bun/cake (mean: 100-320 mg kg(-1)), some bakery products such as muffin, pancake/waffle, coconut tart and cake (mean: 250, 160, 120 and 91 mg kg(-1), respectively), and jellyfish (ready-to-eat form) (mean: 1200 mg kg(-1)). The results demonstrated that aluminium-containing food additives have been widely used in these food products. The average dietary exposure to aluminium for a 60 kg adult was estimated to be 0.60 mg kg(-1) bw week(-1), which amounted to 60% of the new PTWI established by JECFA. The main dietary source was "steamed bread/bun/cake", which contributed to 60% of the total exposure, followed by "bakery products" and "jellyfish", which contributed to 23 and 10% of the total exposure, respectively. However, the estimation did not include the intake of aluminium from natural food sources, food contact materials or other sources (e.g. drinking water). Although the results indicated that aluminium it is unlikely to cause adverse health effect for the general population, the risk to some populations who regularly consume foods with aluminium-containing food additives cannot be ruled out.