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In Japan, existing food additives are those included in the List of Existing Food Additives specified in the Supplementary Provisions to the Law Concerning Amendments to the Food Sanitation Law and Nutrition Improvement Law. Most of the currently available food additives are natural extracts containing various ingredients. However, the characteristic and active components of existing food additives are not always properly defined due to poor characterization of the constituents of the respective raw materials. For that reason, the characteristic components of existing food additives from natural extracts have been evaluated using various methods and reported. Here we review examples of our research on the characterization of marker constituents of existing food additives from natural products.

In order to understand the actual state of residual solvents contained in commercial supplements, we performed a simultaneous analysis of residual solvents by headspace (HS)-GC-MS with reference to the Japanese Pharmacopoeia's "Residual Solvents", for 29 products selected from among commercial supplements (e.g., revitalizers, weight loss pills) that are deeply colored or contain coating agents and extract powder. As a result, benzene (class 1) was detected in eight black-colored supplements, and hexane (class 2B) was also detected in one of those products. On the other hand, methanol (class 2A) was detected in four products containing coating agents and extract powders, such as citrus peel extract. None of these residual solvents exceeded the concentration limits set by the Japanese Pharmacopoeia. Benzene was detected at 1.7 μg/g, which was near the concentration limit, in some products. As raw materials used for the manufacture of the black-colored supplements from which benzene was detected commonly included activated carbon, we analyzed the residual solvents contained in activated carbon commercially available for use as food additive and in food production and medicine. As a result, benzene was detected at high concentrations in activated carbon made from hemp (approximately 29 μg/g) and bamboo (approximately 140 μg/g).

Mogroside V is one of the characteristic and effective components of luohanguo extract, a food additive used as a sweetener in Japan as per Japan's Standards and Specifications for Food Additives (JSFA; 9th ed.). JSFA stipulates that the quantitative determination for mogroside V content in luohanguo extract applies HPLC using analytical standard mogroside V. However, no mogroside V reagents with proven purities are commercially available. Therefore the current JSFA determination method is not particularly suited for daily quality control operations involving luohanguo extract. In this study, we applied an alternative quantitative method using a single reference with relative molar sensitivity (RMS). It was possible to calculate the accurate RMS by an offline combination of 1 H-quantitative NMR spectroscopy ( 1 H-qNMR) and an HPLC/variable-wavelength detector (VWD). Using the RMS of mogroside V to a commercial certified reference material grade caffeine, the mogroside V contents in luohanguo extracts could be determined using HPLC/VWD without analytical standard mogroside V. There was no significant difference between the mogroside V contents in luohanguo extracts determined using the method employing single-reference caffeine with the RMS and using the JSFA method. The absolute calibration curve for the latter was prepared using an analytical standard mogroside V whose purity was determined by 1 H-qNMR. These results demonstrate that our proposed method using a single reference with RMS is suitable for quantitative determination of mogroside V in luohanguo extract and can be used as an alternative method to the current assay method in JSFA.

Violation of the Food Sanitation Act regarding detection of Patent Blue V, which is one of the non-permitted dyes for food in Japan, in imported food occurs every year. With respect to the identification of dyes of Patent Blue group, in some cases, each dye has several different names, and in other cases, different dyes have the same name. Thus, there is a risk that the detected dye is misidentified with other dyes of Patent Blue group. In this study, nine commercial available dyes of Patent Blue group, including a reagent with unclear product information, were analyzed by TLC, HPLC and LC-MS/MS. The result showed that with all three methods, the dyes could be clearly identified into one of four types of blue dyes, i.e. Patent Blue V, Azure Blue VX, Isosulfan Blue and Alphazurine A. Unification of nomenclature would reduce the risk of misidentification of dyes of Patent Blue group.

Currently, consumers are very anxious about many chemical substances contained in foods. To maintain food safety, the Ministry of Health, Labour and Welfare of Japan establishes standards and specifications on toxic chemical substances in foods, establishes analytical methods for surveillance, and investigates the daily dietary intake of food contaminants every year. This paper describes what sorts of standards and specifications for toxic chemical substances in foods have been established and what kinds of research on daily dietary intake have been performed. As the subjects for description, pesticide residues, toxic metals, dioxins, acrylamide, food additives, genetically modified food products, so-called health foods, and food allergens are included.

The mean concentrations and daily intake of four antifungal agents were estimated based on the results of an analysis of 7,005 samples of food obtained in official inspections by Japanese local governments in fiscal year 1998. The mean concentration of diphenyl was 0.0004% of the allowable limit, and those of imazalil, o-phenylphenol, and thiabendazole were 14.0%, 3.5%, and 5.7%, respectively. The daily intakes of these antifungal agents per person, estimated from their concentrations and the daily consumption of the foods, were 0.000326, 1.89, 11.5, and 23.3 micrograms, respectively, and assuming a body weight of 50 kg, the amounts of these antifungal agents consumed were 0.000013%, 0.15%, 0.12%, and 0.47% of the acceptable daily intake, respectively. These values are similar to the values obtained on the basis of the results of the official inspection in fiscal years 1994 and 1996, except that the amount of diphenyl is much lower (1/100).

Recommended Dietary Allowances (RDAs) are allocated to each food article as published in the Denmark Budget Methods in the Codex Alimentalius of the WHO/FAO Joint Committee when standards of pollutants in food are needed. When daily intakes of Food Additives and Contaminants need to be calculated, the Theoretical Maximum Daily Intake. (TMDI) and the Estimated Maximum Daily Intake (EMDI) have been generally used. TMDI and EMDI are calculated using the formulae shown below: [formula: see text] i: food article (i = 1, ....., n) A: standard value for food additives and contaminants X: mean weight of food article consumed daily l: rate of residue after cooking Exposure assessment should be more exact in order to meet social health needs and to help avoid unnecessarily strict regulations. The U. S. Food and Drug Administration (FDA) is continually improving its estimates of the dietary intakes of pesticides and essential minerals, and comparing these intakes with established safe or recommended dietary intake levels. Dietary survey methods have also improved in parallel, with examples being the USDA's Household Food Consumption Survey (1955 and 1965) and National Health and Nutrition Examination Survey II (1976-1980). In Japan, TMDI and EMDI have received more attention as methods of estimating the daily intakes of food additives and contaminants than has the Total Diet Study, even though the former are not as exact as the latter. The Japanese National Nutrition Survey is one of the most respected nutrition surveys in the world, because it has continued nationwide yearly since 1946. Nevertheless, it is very unfortunate that no one utilizes the Household Food Consumption Survey data for the estimation of intakes of food additives and contaminants, because that is not the primary purpose of the Japanese National Nutrition Survey. Practically, there are neither foods which have an uniform of food additives and contaminants nor individuals who consume uniform amounts of each food item. In this report the authors propose a revised estimation method for the daily intake of food contaminants and additives, based on food consumption data of 159 female volunteers, without using the National Nutrition Survey data. The results obtained are as follows: 1) This method succeeded in making clear the intakes of food additives and contaminants. Mean, maximum and minimum values and distribution curves for the target population were obtained. 2) The suggested name for this method is "Estimated Ecological Daily Intake (EEDI)", which is processed in terms of the food consumption structure for calculation, and methodologically estimated by food ecology.