Review on development of assigned value microbiological reference materials used in food testing.

Evolving testing methods in food Microbiology have resulted in the need for different types of microbiological reference materials. Based on the research articles available in this area, it is evident that development has been quite substantial in chemical testing compared to Microbiology. The primary reason could be the ease of spiking, and recovery in chemical RM as compared to microbiological RM. A significant challenge faced in recovery and stability during the development of quantitative microbiological RM depends on temperature, type of microbiological media used, method of analysis including reconstitution method, interference due to antimicrobial agents in food matrices, and competitor microorganisms present in higher numbers then the target microorganisms. Most of the research papers published on microbiological reference materials are contributed by developed economies were in the information related to complex food matrices are limited. Further analysis of different International Depository Agencies under the Budapest treaty indicates that there are only three institutes based in Europe providing quantitative or assigned value RM. This, in turn, highlights the scarcity in the availability of quantitative RM in Microbiology. This article provides a global overview of the availability of microbiological RM, microbial preservation techniques, protective agents, and elaboration on developing different formats of microbiological RM used in food testing.

Modifications of the U.S. food and drug administration validated method for detection of Cyclospora cayetanensis oocysts in prepared dishes: Mexican-style salsas and guacamole.

Although multiple outbreak clusters of Cyclospora cayetanensis have been traced back to consumption of dishes in Mexican-style restaurants, the FDA Bacteriological Analytical Manual (BAM) does not currently provide methods to detect C. cayetanensis in dishes that contain multiple produce ingredients, such as salsas and guacamole. These complex food matrices also may contain high levels of fats, which can interfere with the detection. Several modifications to the BAM Chapter 19b method (washing produce, DNA extraction, and a TaqMan real-time PCR assay targeting the 18S rRNA gene of C. cayetanensis) were assessed with the goal to detect as few as 5 oocysts of C. cayetanensis in 25 g samples of commercial salsa/pico de gallo, guacamole, and salsa verde. Both freshly prepared and frozen versions of these foods were seeded with 5, 10 and 200 oocysts. For salsa samples, using a gentler washing step than recommended by BAM, we achieved detection of 5 oocysts in the samples (81.8%, n = 11). Increasing the amount of Alconox® in the wash solution to 1%, rather than the 0.1% used in BAM, and adjusting the DNA extraction protocol to process large wash pellets, enabled detection of 5 oocysts in guacamole. To reach the desired level of detection in salsa verde, two types of modifications were necessary: gentler washing and DNA extraction modifications. The use of these same method modifications on previously frozen food samples, provided levels of detection similar to those achieved with fresh dishes. Our modifications enabled robust and reproducible detection of C. cayetanensis in multi-ingredient Mexican dishes, detecting as few as 5 oocysts in 25 g samples. Validating and deploying effective methods to detect C. cayetanensis in high risk fresh produce and prepared dishes are critically important for prevalence studies and outbreak investigations of this parasite.

Detection of Cyclospora cayetanensis on bagged pre-cut salad mixes within their shelf-life and after sell by date by the U.S. food and drug administration validated method.

Recently, outbreaks of Cyclospora cayetanensis in the U.S. were linked to the consumption of a variety of salads containing romaine and/or iceberg lettuce, carrots and/or red cabbage. The Bacteriological Analytical Manual (BAM) Chapter 19b method was validated for the detection of C. cayetanensis in carrots, cabbage and romaine lettuce, but has not been previously evaluated in ready-to-eat (RTE) salad mixes. In addition, the only samples available for traceback investigations are sometimes leftovers in bad conditions. This study evaluated the validated BAM method for detection of C. cayetanensis in two different RTE mixed salads (mix 1: romaine and iceberg lettuces, carrots, and red cabbage and mix 2: romaine and iceberg lettuces, carrots, red cabbage, radish, and pea pods) in good condition and after their sell by date. Individual samples (25 g) were seeded with five and 200 C. cayetanensis oocysts. Unseeded produce was used as negative control. The method included washing of the produce, concentration and extraction of C. cayetanensis DNA and molecular detection of C. cayetanensis 18 S rRNA gene. As few as five oocysts were detected in both fresh and after sell by date mix salads. All unseeded samples were negative, and all samples of both salad types seeded with 200 oocysts were positive. In samples seeded with 200 oocysts, average 18 S rRNA C. cayetanensis CT values were significantly higher in fresh salad mix 1 compared to fresh salad mix 2; CT values were significantly higher in the after sell by date salads compared to their respective fresh mixes (p < 0.05). In conclusion, the BAM method was able to detect as few as five oocysts even in after sell by date RTE mix salads. However, the differences in detection observed, highlight the importance of evaluating the performance of the validated C. cayetanensis detection method in different food matrices and conditions, in advance for future outbreak investigations.

Standardized Extraction Techniques for Meat Analysis with the Electronic Tongue: A Case Study of Poultry and Red Meat Adulteration.

The electronic tongue (e-tongue) is an advanced sensor-based device capable of detecting low concentration differences in solutions. It could have unparalleled advantages for meat quality control, but the challenges of standardized meat extraction methods represent a backdrop that has led to its scanty application in the meat industry. This study aimed to determine the optimal dilution level of meat extract for e-tongue evaluations and also to develop three standardized meat extraction methods. For practicality, the developed methods were applied to detect low levels of meat adulteration using beef and pork mixtures and turkey and chicken mixtures as case studies. Dilution factor of 1% w/v of liquid meat extract was determined to be the optimum for discriminating 1% w/w, 3% w/w, 5% w/w, 10% w/w, and 20% w/w chicken in turkey and pork in beef with linear discriminant analysis accuracies (LDA) of 78.13% (recognition) and 64.73% (validation). Even higher LDA accuracies of 89.62% (recognition) and 68.77% (validation) were achieved for discriminating 1% w/w, 3% w/w, 5% w/w, 10% w/w, and 20% w/w of pork in beef. Partial least square models could predict both sets of meat mixtures with good accuracies. Extraction by cooking was the best method for discriminating meat mixtures and can be applied for meat quality evaluations with the e-tongue.

Intra-Laboratory Validation of Alpha-Galactosidase Activity Measurement in Dietary Supplements.

INTRODUCTION: Alpha-galactosidase (α-Gal) is an enzyme responsible for the hydrolyzation of glycolipids and glycoprotein commonly found in dietary sources. More than 20% of the general population suffers from abdominal pain or discomfort caused by intestinal gas and by indigested or partially digested food residuals. Therefore, α-Gal is used in dietary supplements to reduce intestinal gases and help complex food digestion. Marketed enzyme-containing dietary supplements must be produced in accordance with the Food and Drug Administration (FDA) regulations for Current Good Manufacturing Practice (cGMPs). AIM: in this work we illustrated the process used to develop and validate a spectrophotometric enzymatic assay for α-Gal activity quantification in dietary supplements. METHODS: The validation workflow included an initial statistical-phase optimization of materials, reagents, and conditions, and subsequently a comparative study with another fluorimetric assay. A final validation of method performance in terms of specificity, linearity, accuracy, intermediate-precision repeatability, and system precision was then executed. RESULTS AND CONCLUSIONS: The proven method achieved good performance in the quantitative determination of α-Gal activity in commercial food supplements in accordance with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals (ICH) guidelines and is suitable as a rapid in-house quality control test.

Determination of aquacultured whiteleg shrimp (Litopanaeus vannemei) quality using a sensory method with chemical standard references.

BACKGROUND: Fresh shrimp are highly perishable seafood and a reliable spoilage assessment method is necessary to ensure sufficient quality control. The current quality evaluation method employed by the US Food and Drug Administration (FDA)/National Oceanic and Atmospheric Administration (NOAA) uses subjective terms 'odor of decomposition' to reject shrimp shipments, which lacks reference standards to anchor the concept and can cause ambiguity. The present study aimed to develop chemical reference standards to assist in a more objective and consistent sensory evaluation of shrimp quality. RESULTS: Chemical references were developed and used by the descriptive panel to demonstrate the aroma quality indicators of shrimp. The most important aroma attributes describing shrimp quality changes were 'salty water-like', 'natto water-like' and 'sour milk-like' based on the results of multiple linear regression analysis. The overall rating consistency of the key quality indicators was confirmed by trained a descriptive panel such that the intensity scores in two separated evaluation sessions (30 days apart) were not significant different (P > 0.05). The sensory ratings also corresponded well with presumed FDA/NOAA grades of shrimp. An untrained panel also confirmed 'salty water-like' as the main indicator of freshness and 'natto water-like' as the main indicator of spoilage, whereas the discriminative capacity was lower compared to the trained panel. CONCLUSION: The developed chemical references of key aroma quality indicators allowed the trained and untrained panels to distinguish shrimp of different freshness levels. The results indicate the potential of using chemical references as a new evaluation tool for on-site quality inspection or industrial quality assurance/quality control of shrimp with improved objectivity and consistency. © 2021 Society of Chemical Industry.

Development of certified reference materials for the determination of cadmium and acrylamide in cocoa.

Since 1 January 2019 a maximum content of 0.6 mg kg-1 cadmium (Cd) in cocoa powder sold to the final consumer or as an ingredient in sweetened cocoa powder sold to the final consumer (drinking chocolate) is set by the Commission Regulation (EU) No. 488/2014. Monitoring compliance with the specified limit value requires analytical measuring methods and reference materials for quality control. However, suitable certified reference materials intended for quality assurance and quality control purposes are still lacking. Therefore, three cocoa reference materials (ERM®-BD513, ERM®-514 and ERM®-515) were developed according to the requirements of ISO 17034 and the recommendations of ISO Guide 35. The whole process of reference material development, including material preparation, assessment of homogeneity and stability, characterisation and value assignment is presented. The assignment of the certified mass fractions was based upon an interlaboratory comparison study involving 19 expert laboratories for Cd and 12 laboratories for acrylamide. The certified mass fractions and expanded uncertainties (k = 2) of the reference materials were (0.181 ± 0.009) mg kg-1 Cd (ERM®-BD513), (0.541 ± 0.024) mg kg-1 Cd (ERM®-BD514) and (0.690 ± 0.029) mg kg-1 Cd (ERM®-BD515). Acrylamide contents are given for information.

New matrix certified reference material for accurate measurement of ciprofloxacin residue in egg.

Matrix certified reference materials (CRMs) are an indispensable part of method validation and have played an important role in ensuring reliable analytical results. To retain similarity to real samples, a new matrix CRM for the mass fraction of ciprofloxacin in whole liquid egg was developed by use of incurred materials with a target value corresponding to residue levels in real sample. The source materials were collected from laying hens following oral administration of ciprofloxacin. An optimized homogenization method and a strict bottling process were applied to bulk whole egg materials to prepare the CRM candidate. The mass fraction of ciprofloxacin in whole liquid egg was certified by a collaborative characterization program with eight accredited participating laboratories. Liquid chromatography coupled with isotope dilution mass spectrometry was studied as a reliable reference method for value assignment and was used by all participating laboratories. The certified value and expanded uncertainty (k = 2, at a confidence level of 95%) was 39.7 ± 5.2 µg/kg for ciprofloxacin in whole liquid egg. Homogeneity, long-term stability at -70 °C for 12 months, and short-term stability at -18 °C, 4 °C, and room temperature were assessed for 9 days. Additionally, uncertainties arising from inhomogeneity, instability, and characterization were analyzed in detail and fully estimated. This CRM would be a useful tool for validation of analytical methods and proficiency testing in ciprofloxacin residue analysis of egg. Graphical Abstract.

Comparative study of multiplex real-time recombinase polymerase amplification and ISO 11290-1 methods for the detection of Listeria monocytogenes in dairy products.

Dairy products have been implicated in foodborne infections caused by different bacterial pathogens. Among them, Listeria monocytogenes is of particular concern due to its ubiquity, resistance to sanitation processes and high mortality rates resulting from infection. These issues make the development of novel methods for the rapid detection of this bacterium of high interest. The evaluation of a novel multiplex real-time Recombinase Polymerase Amplification method including an internal amplification control is reported in the present work. The method performance was compared to that of the European reference method (ISO 11290-1) for the detection of the species in samples from 40 commercial products, including 14 UHT milk samples, 16 hard cheese samples, 6 infant dairy preparation samples and 4 fresh cheese samples. A limit of detection below 10 cfu/25 g or mL sample was achieved, and values higher than 90% were obtained for relative sensitivity, specificity, accuracy, positive and negative predictive values and the index (kappa) of concordance. Analysis was achieved within one working day, compared to the six days required using the ISO method. Moreover, slight modification of the ISO 11290-1 method to include secondary enrichment in half Fraser broth resulted in the confirmation of all positive samples.

Validation of the 2H-SNIF NMR and IRMS Methods for Vinegar and Vinegar Analysis: An International Collaborative Study.

This paper presents the results of a collaborative study involving seven laboratories and concerning two samples of wine vinegar, one of apple vinegar and four of balsamic vinegar. The aim of the study was to define standard deviations of repeatability (sr) and reproducibility (sR) for vinegar and balsamic vinegar stable isotope ratios of H (D/H), C (δ13C) and O (δ18O), in order to establish them as fully recognized official standards. Acetic acid was extracted and subjected to (D/H)CH3 and δ13C analysis. δ18O analysis was performed on whole samples. The grape must solution remained after distillation of balsamic vinegar was fermented and the resulting ethanol was subjected to (D/H)I, (D/H)II, R and δ13C analysis. The sr and sR were 0.6 ppm and 1.1 ppm for (D/H)CH3, 0.14‱ and 0.25‱ for δ13C of acetic acid, 0.1‱ and 0.17‱ for δ18O of water, 0.19 ppm and 0.64 ppm for ethanol (D/H)I, 1.14 and 1.31 ppm for (D/H)II, 0.09 and 0.11‱ for δ13C of ethanol. These data are in line with those in the literature or reported in corresponding official methods, and sr and sR of balsamic vinegar are in line with those of vinegar and must.

[Proficiency Testing Schemes for Food Nutrition Analysis in Japan (2017-2018)].

This paper deals with proficiency testing schemes for food nutrition analysis in Japan. In schemes in 2017 and 2018, 65 and 73 organizations participated, respectively, and more than 70% of the participants were public organizations responsible for a nutrition-labeling compliance test. The food matrices were pork and chicken sausages, and analytes were protein, fat, ash, moisture, carbohydrate, energy, sodium, salt equivalent, calcium (2018 only), and iron (2018 only). The organizations reporting inadequate laboratory values in one or more nutrients for mandatory declaration (energy, protein, fat, carbohydrate, or salt equivalent) were 11 and 15% of all organizations and 9 and 13% of public organizations in the 2017 and 2018 schemes, respectively. The approximate relative standard deviations for proficiency assessment (RSDr) were as follows: protein, 2%; fat, 3%; ash, 2%; moisture, 0.5%; carbohydrate, 9%; energy, 1%; sodium (salt equivalent), 4%; calcium, 7%; and iron, 7%. Notably, the large RSDr value for carbohydrate may cause inconsistency among laboratories in compliance tests for foods containing several grams or less of carbohydrate per 100 grams.

[Validation Study on Developed Methods for Total Organic Carbon in Some Kinds of Mineral Water].

Total organic carbon (TOC) was measured in some kinds of mineral water, and the method was validated. In mineral water, there are many kinds of elements such as carbon dioxide and a wide range of hardness. The official method for amount of TOC in tap water was validated in non-carbonated mineral water regardless of the degree of hardness. However, the amount of TOC was not accurately measured in two kinds of carbonated mineral water with medium or high degree of hardness. In our method of this study, the removal of carbon dioxide from the two kinds of mineral water was achieved by making bubbling time longer and additive rate of HCl upper than the official condition of tap water. And then, the method we developed was validated in the two kinds of mineral water. Our results suggested that the method we developed could be useful to measure the amount of TOC in many kinds of mineral water on the market.

[Method Verification for Vitamin D Analysis in Processed Foods Based on the Analytical Manual for the Standard Tables of Food Composition in Japan 2015 (Seventh Revised Edition)].

Considerable amounts of processed foods contain vitamin D (ergocalciferol (D2) and cholecalciferol (D3)) as food additives. For field surveys on food additives, the analytical method for vitamin D should be well-validated. However, the current official method in Japan cannot separately determine the concentrations of D2 and D3, whereas the method for the Standard Tables of Food Composition in Japan 2015 (STFC method) can. Therefore, in this study, we verified the applicability of the STFC method to processed foods. During the course of this research, we added some improvements to the original method. Spike and recovery experiments using vegetable juice, soymilk, and corn flakes as food matrices showed that the recovery rates (relative standard deviation) of D2 and D3 were 103-112% (4.7-12.6%) and 102-109% (2.4-21.8%), respectively, at the estimated method limit of quantification (EMLOQ) level; and 100-110% (4.0-7.4%) and 102-105% (3.8-4.8%), respectively, at 10 times the EMLOQ level. These results indicated that accuracy and precision of the modified STFC method were enough to determine dietary D2 and D3 as endogenous nutrients and/or food additives, and suggested that this method is appropriate for analyzing vitamin D concentrations in processed foods.

[Comparison of Validation Results Obtained by GC-MS/MS and LC-MS/MS for the Analysis of Residual Pesticides in Agricultural Products].

In the field of food analysis and regulation, different instruments are used to determine the accuracy of quantification values. This is essential, as inconsistencies in values are commonly encountered. To visualize the degree of these discrepancies in each food matrix, we compiled a validation study based on a routine method developed in our laboratory, for 121 pesticides in six agricultural products, namely the grapefruit, potato, paprika, cabbage, spinach, and brown rice. These were analyzed by GC-MS/MS and LC-MS/MS, and the results were compared mainly on the basis of trueness. According to the results of the validation study when using GC-MS/MS, of the 121 pesticides tested in each product class, the number of analytes that satisfied the criteria of the Japanese validation guidelines was 97 in grapefruit, 111 in potato, 110 in paprika, 118 in cabbage, 111 in spinach, and 63 in brown rice. In contrast, in the analysis of the same samples by using LC-MS/MS, the number of analytes that satisfied the criteria of the validation guidelines was 50 in grapefruit, 114 in potato, 103 in paprika, 112 in cabbage, 100 in spinach, and 103 in brown rice. Inconsistences in the differences of trueness were mainly attributed to matrix effects of each instrument, as well as to food matrices, of which the most diverged matrix was that of brown rice (over 20%).

What's for lunch? The content and quality of lunches consumed by Dutch primary schoolchildren and the differences between lunches consumed at home and at school.

BACKGROUND: Lunch is an important part of a healthy diet, which is essential for the development, growth and academic performance of school-aged children. Currently there is an increasing number of Dutch primary schoolchildren who are transitioning from eating lunch at home to school. There is limited knowledge about the current quality of the lunches consumed by primary schoolchildren in the Netherlands and whether there are any differences between lunches consumed at home or at school. To investigate differences in content and quality of lunches consumed by Dutch primary schoolchildren at home and at school. METHODS: Cross-sectional study among 363 Dutch primary schoolchildren aged 4-12 years based on the first two years of the 2012-2016 Dutch National Food Consumption Survey. Demographic characteristics were obtained through a questionnaire. Diet was assessed with two non-consecutive 24-h dietary recalls. Quality of lunches was assessed on their nutritional quality whether they fitted the nutritional guidelines. 'Nonparametric tests were used to examine the content and quality of the lunches between place of consumption and parental educational position. RESULTS: The most consumed lunch products among primary schoolchildren were bread, dairy products and sugar-sweetened beverages. Fruit and vegetable consumption was very low. Consumption of milk and other dairy products was higher among children who eat lunch at home than children who eat lunch at school (p < 0.01). Consumption of sugar-sweetened beverages was higher among children who eat lunch at school than children who eat lunch at home (p < 0.01), and at school a higher proportion of the drinks did not fit within the Dutch dietary recommendations (p < 0.01). CONCLUSIONS: The current content of the lunches consumed by Dutch primary schoolchildren leaves room for improvement, especially regarding fruit and vegetables. The statistically significantly higher consumption of sugar-sweetened beverages and lower consumption of milk and dairy products at school vs. home is worrisome, as currently more children in the Netherlands are transitioning to having lunch at school.

Co-Extraction and Co-Purification Coupled with HPLC-DAD for Simultaneous Detection of Acrylamide and 5-hydroxymethyl-2-furfural in Thermally Processed Foods.

Acrylamide and 5-hydroxymethyl-2-furfural (5-HMF) are two of the most abundant compounds generated during thermal processing. A simple method for the simultaneous quantitation of acrylamide and 5-HMF was developed and successfully applied in thermally processed foods. Acrylamide and 5-HMF were co-extracted with methanol and then purified and enriched by an Oasis HLB solid-phase extraction cartridge, simultaneously analyzed by high-performance liquid chromatography and detected with a diode array detector, respectively, at their optimal wavelength. The linear concentration range was found to be 25-5000 µg/L with high linear correlation coefficients (R > 0.999). The limit of detection and the limit of quantitation for acrylamide and 5-HMF were 6.90 µg/L and 4.66 µg/L, and 20.90 µg/L and 14.12 µg/L, respectively. The recovery of acrylamide and 5-HMF in biscuits, bread, Chinese doughnuts, breakfast cereals, and milk-based baby foods was achieved at 87.72-96.70% and 85.68-96.17% with RSD at 0.78-3.35% and 0.55-2.81%, respectively. The established method presents simplicity, accuracy and good repeatability, and can be used for the rapid simultaneous quantitation of acrylamide and 5-HMF in thermally processed foods.

Comparing Friedman versus Mack-Skillings data analyses on duplicated rank data: a case of visual color intensity.

BACKGROUND: In duplicated ranking tests, panelists either rank duplicates separately (2SS) or jointly in a single session (1SS). This study compared data analyses of duplicated yellow color intensity rank data using Friedman versus Mack-Skillings (M-S) tests. Panelists (n = 75) ranked an orange juice set twice - a similar-samples set (100%, 95% versus 90%); samples other than the 100% juice were prepared by dilution with water. Rank sum data were obtained from (a) intermediate ranks from jointly re-ranked scores of 2SS for each panelist, and (b) joint rank data of all panelists from the two replications in 1SS. Both (a) and (b) were analyzed using the M-S test. The median rank data (c) for each panelist from two replications were analyzed using the Friedman test. RESULTS: Comparing M-S with the Friedman tests, the former generally produced higher test statistics and lower P-values than the latter. However, when considering the pattern of post hoc pairwise significant differences, both tests yielded similar conclusions. CONCLUSION: This study demonstrated that, in a duplicated ranking test with three samples that were very similar in color, separating the two replications into two complete individual ranking tests or serving sessions (2SS) may prevent sensitivity loss due to fatigue that is otherwise experienced when evaluating all samples together in a single session (1SS). We expected to find the M-S test to be more sensitive than the Friedman test; however, this hypothesis was not supported by the post hoc (Tukey's honest significant difference (HSD)) multiple comparison test results under the specific test conditions in this study. © 2019 Society of Chemical Industry.

Evaluation of the impact of matrix effects in LC/MS measurement on the accurate quantification of neonicotinoid pesticides in food by isotope-dilution mass spectrometry.

The use of isotope-labeled internal standards is the most widely accepted approach to overcome the matrix effects on quantification of pesticides in food by LC/MS. We evaluated the impact of the matrix effects on quantification of six neonicotinoid pesticides, acetamiprid, clothianidin, dinotefuran, imidacloprid, thiacloprid, and thiamethoxam, in food by using deuterated internal standards. The calibration curves for each pesticide were obtained by using matrix-free and matrix-matched calibration solutions with blank brown rice, carrot, and green onion extracts. For brown rice and carrot, the matrix effects were not observed. In contrast, the slopes of calibration curves for each pesticide were influenced by presence of green onion extracts in calibration solutions (variability of the slopes was 4-9%), because the ratios of peak area for native pesticide to those for internal standards were influenced by matrix. The spike-and-recovery test with green onion was also performed. The analytical values obtained by using matrix-free calibration solution were biased from the spiked concentration, whereas those obtained by using matrix-matched calibration solution were comparable to the spiked concentration. These results indicate that matrix-matched calibration solution should be used for accurate quantification of neonicotinoid pesticides in food by LC/MS using deuterated internal standards.

A free sugars daily value (DV) identifies more "less healthy" prepackaged foods and beverages than a total sugars DV.

Regulatory changes in Canada will require food labels to have a benchmark [% Daily Value, %DV] for total sugars, based on 100 g/day, while US labels will require a %DV for added sugars, based on 50 g/day. The objective of this study was to compare two labelling policies, a total sugars DV (100 g/day) and a free sugars DV (50 g/day) on food labels. This cross-sectional analysis of the Food Label Information Program database focussed on top sources of total sugars intake in Canada (n = 6924 foods). Products were categorized as "less healthy" using two sets of criteria: a) free sugars levels exceeding the WHO guidelines (≥10% energy from free sugars); and b) exceeding healthfulness cut-offs of the Food Standards Australia New Zealand Nutrient Profiling Scoring Criterion (FSANZ-NPSC). The proportion of "less healthy" products with ≥15%DV (defined as "a lot" of sugars i.e. high in sugars, based on Health Canada's %DV labelling footnote and educational message for dietary guidance) were compared for each sugar labelling scenario. The free sugars DV showed better alignment with both methods for assessing "healthfulness" than the total sugars DV. The free sugars DV identified a greater proportion of "less healthy" foods with ≥15%DV, based on both the FSANZ-NPSC (70% vs. 45%, p < .0001) and WHO guidelines (82% vs. 55%, p < .0001); particularly in sweet baked goods, sugars and preserves, chocolate bars, confectionery, and frozen desserts categories. Compared to total sugars DV labelling, using a free sugars DV identified more "less healthy" foods. Findings support the adoption of free sugars labelling.

Stable oxygen isotopes in water of concentrated liquid foodstuffs: Are the commonly determined values accurate?

RATIONALE: Oxygen isotope analysis of water molecules of liquid foodstuffs is commonly performed under isotopic equilibrium between water in the solution and the vapour water, assuming that the liquid water activity is equal to unity and that liquid water is an ideal mixture of H2 O isotopologues. A priori this behaviour is not realistic for all foodstuffs, which frequently are very concentrated solutions. In this paper we mainly consider "balsamic vinegar" with the aim of defining an appropriate procedure of oxygen isotope ratio analysis of water molecules in these concentrated solutions. METHODS: Isotope ratio mass spectrometry (IRMS) measurements of the oxygen isotope ratios (δ18 O values) were carried out on CO2 equilibrated with water molecules at 22 ± 0.1°C. Three independently calibrated, very low salinity waters were used as standards. RESULTS: For grape must and wine vinegar (density < 1.15 g/cm3 ) the δ18 O values for water determined directly on these solutions are "true" values. On the contrary, for balsamic vinegar with density higher than 1.15-1.20 g/cm3 , the δ18 O values obtained directly on the solutions are systematically different from those obtained on water produced by distillation of the same samples at 70°C under vacuum. CONCLUSIONS: In the case of balsamic vinegar with density higher than 1.15-1.20 g/cm3 , to avoid severe systematic errors, the isotopic analyses must be carried out on water obtained by distillation under stirring.