Comparative analysis of isothiocyanates in eight cruciferous vegetables and evaluation of the hepatoprotective effects of 4-(methylsulfinyl)-3-butenyl isothiocyanate (sulforaphene) from daikon radish (Raphanus sativus L.) sprouts.

The contributions of cruciferous vegetables to human health are widely recognised, particularly at the molecular level, where their isothiocyanates play a significant role. However, compared to the well-studied isothiocyanate 4-(methylsulfinyl)butyl isothiocyanate (sulforaphane) produced from broccoli sprouts, less is known about the pharmacological effects of other isothiocyanates and the stage of vegetables preferable to obtain their benefits. We analysed the quantity and quality of isothiocyanates produced in both the sprouts and mature stages of eight cruciferous vegetables using gas chromatography-mass spectrometry (GC-MS). Additionally, we investigated the hepatoprotective effects of isothiocyanates in a mouse model of acute hepatitis induced by carbon tetrachloride (CCl4). Furthermore, we explored the detoxification enzyme-inducing activities of crude sprout extracts in normal rats. Among the eight cruciferous vegetables, daikon radish (Raphanus sativus L.) sprouts produced the highest amount of isothiocyanates, with 4-(methylsulfinyl)-3-butenyl isothiocyanate (sulforaphene) being the dominant compound. The amount of sulforaphene in daikon radish sprouts was approximately 30 times that of sulforaphane in broccoli sprouts. Sulforaphene demonstrated hepatoprotective effects similar to sulforaphane in ameliorating CCl4-induced hepatic injury in mice. A crude extract of 3-day-old daikon radish sprouts upregulated the detoxifying enzyme glutathione S-transferase (GST) in the liver, whereas the crude extract of broccoli sprouts showed limited upregulation. This study highlights that daikon radish sprouts and sulforaphene have the potential to serve as functional food materials with hepatoprotective effects. Furthermore, daikon radish sprouts may exhibit more potent hepatoprotective effects compared to broccoli sprouts.

Estimation of daily intake of food additives by Japanese young children using the market basket method in 2018.

To estimate the daily intake of food additives by young children aged 1-6 years in Japan, an intake survey was conducted in 2018 using the market basket method for food additives, including twelve types of colourants, three kinds of preservatives, three kinds of sweeteners and two kinds of food manufacturing agents. A list of the daily consumption of processed foods was prepared based on a special survey (MHLW 2011) and used for the estimation. The results of the survey showed that the food additives with the highest daily intake were phosphorus compounds (phosphoric acid and its salts; 11.2 mg/kg bw/day, expressed as phosphorus), followed by propylene glycol (0.80 mg/kg bw/day). The daily intake of other food additives ranged from 0 to 0.20 mg/kg bw/day. The estimated daily intake of each food additives by young children was compared with the acceptable daily intake (ADI) or maximum tolerable daily intake (MTDI). The highest ratio of the estimated daily intake to ADI was 3.2% for propylene glycol, whereas the ratios of the estimated daily intake to ADI for colourants, preservatives and sweeteners ranged from 0 to 1.1% (benzoic acid). The ratio of the estimated daily intake to MTDI for phosphorus compounds was 16%.

[A Screening Analysis Method for Stevia Sweetener in Processed Foods by LC-MS/MS].

A LC-MS/MS-based screening method was developed for stevia sweetener in processed foods. After extraction of stevia sweetener from processed foods by dialysis, the dialysate was diluted with water, and stevia sweetener was measured by LC-MS/MS. Recovery from 5 kinds of processed foods spiked with 10 mg/kg of stevioside （SS）, 10 mg/kg of rebaudioside A （RS）, or 100 mg/kg of α-glu-cosyltransferase-treated stevia （Gts） product was excellent, and no interfering peak was observed. Thirty-six commercial processed foods indicated as containing "stevia" were analyzed using this established method. Among them, 33 contained SS, 33 contained RS, and 11 contained Gts. Five products contained both stevia extract and Gts.

[Method of Quantitative Analysis Using Enzymatic Hydrolysis of α-Glucosyltransferase-Treated Stevia in Foods].

A quantitative analysis by HPLC of α-glucosyltransferase-treated stevia in foods was considered. This analysis is the way which hydrolyzed α-glucosyltransferase-treated stevia in the stevioside (SS) and the rebaudioside A (RS) using a glucoamylase. Recovery (%) of α-glucosyltransferase-treated stevia, spiked at 200 mg/kg in various foods, were more than 80% and the relative standard deviations were less than 5.0% as SS and RS for the rate of collection. A qualitative analysis by LC-MS/MS was performed 36 products of commercial foods containing stevia. We quantified of 11 products in which α-glucosyltransferase-treated stevia was detected. Quantitative value was at most 180 mg/kg as SS, at most 70 mg/kg as RS.

A Rapid Dialysis Method for Analysis of Stevioside and Rebaudioside A in Foods.

A rapid dialysis method for the analysis of stevioside (SS) and rebaudioside A (RS) in foods was developed. Minced samples (10 g) were packed into 30 cm net length dialysis tubing with 30% methanol to increase the dialysis efficiency. The dialysis tubing was put in a 100 mL centrifuge tube, and the total fluid volume was made up to 100 mL with 30% methanol. Dialysis was done with shaking while heating at 50℃. The dialysis times were reduced from 48-72 hr in the conventional method to 2 hr under these conditions. The dialysate was loaded on a C18 solid- phase extraction cartridge, and the cartridge was washed with 40% methanol. SS and RS were eluted from the cartridge with 80% methanol, and separated by reversed-phase HPLC. Recovery yields (%)of SS and RS, spiked at 0.02 g/kg in various foods, were 83.0-105.1% and the relative standard deviations were mostly less than 5%.