Stability assessment of anthocyanins from black soybean, grape, and purple sweet potato under in vitro gastrointestinal digestion.

Anthocyanins are glycosylated derivatives of anthocyanidins, whose hydroxyl groups are occasionally acylated with organic acids. The effect of anthocyanin composition on their stability under in vitro gastrointestinal digestion was investigated. Black soybean had all glycosylated anthocyanins with monosaccharide, grape contained glycosylated anthocyanins with disaccharide (23%) and acylated anthocyanins bound with two sugars (77%), and purple sweet potato had all acylated anthocyanins bound with three sugars. The order of total anthocyanins content was purple sweet potato, grape, and black soybean. Gastric digestion did not significantly decrease anthocyanins content in three samples, while intestinal digestion resulted in the significant decrease of total anthocyanins content in black soybean (40%), grape (45%), and purple sweet potato (25%). This indicates that the degree of glycosylation and acylation of anthocyanins affects their stability under the gastrointestinal conditions. Phenolic acids derived from anthocyanin degradation increased total phenolic content as well as ABTS radical scavenging activity. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01071-6.

Cellular uptake of anthocyanins extracted from black soybean, grape, and purple sweet potato using INT-407 cells.

This study combined in vitro digestion and INT-407 cells to evaluate the bioaccessibility of anthocyanins in the small intestinal epithelial cells. Black soybean, grape, and purple sweet potato were chosen as they have a different anthocyanin composition. After the aqueous extract was digested under in vitro gastric and intestinal conditions, the digested mixture was incubated in the media of INT-407 for 2 h at 37 °C. Low proportion (< 0.3%) of anthocyanins in black soybean and grape passed through cell membranes. Cyanidin-3-O-glucoside and pelargonidin-3-O-glucoside in black soybean and cyanidin-3-O-(6-O-p-coumaroyl)-5-O-diglucoside and delphinidin-3-O-(6-O-p-coumaroyl)-5-O-diglucoside in grape were found inside the cell. However, acylated anthocyanins containing three sugar moieties in purple sweet potato were not detected inside the cell. p-Coumaric acid was detected in the cells incubated with grape, but not in the media. These indicate that chemical structure of anthocyanins affected their cellular uptake and antioxidant activity in INT-407 cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-00976-y.

Stability of anthocyanins in bokbunja (Rubus occidentalis L.) under in vitro gastrointestinal digestion.

The stability of anthocyanins in bokbunja (Rubus occidentalis L.) extract was investigated using an in vitro simulated gastrointestinal digestion. Ethanolic extract from bokbunja was digested with pepsin/HCl for 2 h at 37 °C, followed by pancreatin/bile salts for 2 h at 37 °C. Four anthocyanins including cyanidin-3-glucoside, cyanidin-3-sambubioside, cyanidin-3-xylosylrutinoside and cyanidin-rutinoside were identified in the bokbunja extract. The total anthocyanin content of bokbunja was 3.76 mg cyanidin-3-glucoside equivalents/g fresh weight (FW). Gastric digestion had no significant effect on anthocyanins. However, intestinal digestion substantially decreased anthocyanins up to 1.70 mg/g FW, corresponding to 45% of that in the bokbunja extract. This indicates that about half of anthocyanins can reach an intestinal tract. In addition, a new compound comprised of cyanidin-3-glucosdie, catechin and acetaldehyde in a separated study was observed after in vitro gastrointestinal digestion. This shows that anthocyanins could be transformed into other compounds with different biochemical properties.

Application of response surface methodology to acidified water extraction of black soybeans for improving anthocyanin content, total phenols content and antioxidant activity.

Acidified water extraction of total anthocyanin content, total phenol content and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activity from black soybeans (Glycine max [L.] Merr. Cheongja4ho) was optimized using response surface methodology (RSM). A five-level, three-factor central composite design was employed to optimize the conditions for the maximum yields of these three components using19 different experiments. The HCl concentration (0.3-0.5%), solid-liquid ratio (1/30-1/50 g/mL) and extraction temperature (30-50 °C) significantly affected three components. The optimized conditions were HCl concentration of 0.359%, solid-liquid ratio of 1/54.2 g/mL and extraction temperature of 56.8 °C for total anthocyanin content (136.68 mg/100 g), total phenol content (1197.09 mg/100 g) and ABTS+ scavenging activity (211.65 mg/100 g). These experimental values fit well with the predicted values. Among three anthocyanins found in the black soybean extracts, cyanidin-3-O-glucoside was the major one (65-73% of the total), followed by petunidin-3-O-glucoside (17-23%) and delphinidin-3-O-glucoside (10-12%).

Validation of analytical methods for ethyl carbamate in nine food matrices.

Ethyl carbamate (EC) is formed through a reaction between ethanol and either cyanate or N-carbamyl compounds. This study aimed to develop and validate analytical methods for EC determination in nine food matrices: apple juice, soju, milk, corn oil, rice porridge, peanut butter, beef, flat fish, and sea mustard. Differences from AOAC procedure were use of d5-EC as internal standard, dilution of alcohol content, removal of lipophilic compounds by hexane, and removal of hydrophilic polysaccharides by gel-forming. Standard curves had a good linearity (R(2)>0.997) in all matrices tested. LOD and LOQ were in the range of 0.69-6.08µg/kg and 2.10-18.43µg/kg, respectively. Recovery rates ranged from 80.75 to 121.82% for intra-day and ranged from 78.84 to 116.98% for inter-day. The relative standard deviations (RSDs) were below 14%. These results indicate that the established methods can be applied to determine EC in a variety of foods.

Determination of Ethyl Carbamate in Alcoholic Beverages and Fermented Foods Sold in Korea.

Ethyl carbamate (EC) classified as a probable human carcinogen (Group 2A) is naturally formed in alcoholic beverages and fermented foods during fermentation process and/or during storage. The objective of this study was to analyze EC in 34 food items including 14 alcoholic beverages and 20 fermented foods sold in Korea. Each food was collected from 18 supermarkets in 9 metropolitan cities in Korea, and then made into composite. According to food composition and alcohol content, samples were divided into four matrices such as apple juice, milk, Soju (liquor containing about 20% alcohol), and rice porridge. The maximum EC value of 151.06 µg/kg was found in Maesilju (liquor made from Maesil and Soju). Whisky and Bokbunjaju (Korean black raspberry wine) contained 9.90 µg/kg and 6.30 µg/kg, respectively. EC was not detected in other alcoholic beverages. Of 20 fermented foods, Japanese-style soy sauce had highest level of 15.59 µg/kg and traditional one contained 4.18 µg/kg. Soybean paste had 1.18 µg/kg, however, EC was not found in other fermented foods.