Health risk assessment of phthalic acid esters contamination in market-available yellow rice wine in Huzhou City / 预防医学

Objective@#To investigate the contamination of phthalic acid esters (PAEs) and assess the health risk of PAEs contamination in market-available yellow rice wine in Huzhou City, Zhejiang Province, so as to provide the safety safeguard for consuming yellow rice wine.@*Methods@#Yellow rice wine samples were collected from markets in Huzhou City from 2021 to 2022, and 16 PAEs were determined in yellow rice wine using magnetic solid-phase extraction coupled with gas chromatography-tandem mass spectrometry. The carcinogenic and non-carcinogenic risks of PAEs were evaluated using the health risk models proposed by United States Environmental Protection Agency.@*Results@#A total of 75 yellow rice wine samples were collected, and 44 samples were detected with PAEs contamination (58.67%). Dimethyl phthalate (DMP), di-isobutyl phthalate (DIBP) and di-butyl phthalate (DBP) were detected, and there were 17 samples (22.67%) detected with DBP overdose (DMP and DIBP had no limit standard). DMP, DBP and DIBP, which were not classified as Class 2B and higher carcinogens by WHO's International Agency for Research on Cancer, had no definitive carcinogenic risks. Under mean PAEs, the five types of yellow rice wine all had no carcinogenic risks. Under 75% percentile of PAEs concentrations, the DBP in beverage wine with plastic packaging had a carcinogenic risk score of 1.207 5, with a gross carcinogenic risk score of 1.207 5. Under the maximum PAEs concentration, the ross carcinogenic risk scores of cooking wine with plastic packaging, beverage wine with plastic packaging, beverage wine with glass bottle packaging, and beverage wine with jar packaging were 2.751 0, 2.782 0, 1.298 2 and 2.944 0, presenting non-carcinogenic risks.@*Conclusion@#There is PAEs contamination in market-available yellow rice wine in Huzhou City, and no carcinogenic risk is evaluated. Non-carcinogenic health risk requires to be given a high priority.

Analysis on Quality Transfer Law of Rhei Radix et Rhizoma Steamed with Rice-wine During Processing Based on Correlation Between External Appearance Color and Internal Component / 中国实验方剂学杂志

Objective:The correlation between the appearance color of cooked rhubarb samples and the components characterized by high performance liquid chromatography （HPLC） fingerprint was studied to reveal the quality transfer law in the steaming process of processed products with rice-wine. Method:The visual analyzer was used to analyze the change of the appearance color of cooked rhubarb sample powder at different processing time， the common components and their relative peak areas of processed products with rice-wine were identified by HPLC fingerprint， as well as multivariate statistics and Pearson correlation analysis were used to cluster， discriminate and analyze the appearance color and the component variables in HPLC fingerprint. Result:During the processing of cooked rhubarb， the <italic>a</italic>^* （red-green value） of sample powder had no obvious change， but the <italic>L</italic>^* （lightness value）， <italic>b</italic>^*<italic> </italic>（yellow-blue value） and <italic>E</italic>^*<italic>ab </italic>（total chromaticity value） showed a decreasing trend， and the appearance color changed from bright to dark， from yellow to brown. A total of 46 chromatographic peaks in the fingerprint were identified at 254 nm and 280 nm， and 18 of them were identified by comparison with reference standards. The change trend of <italic>L</italic>^*，<italic> b</italic>^*<italic> </italic>and <italic>E</italic>^*<italic>ab </italic>were positively correlated with the contents of tannins （catechin， epicatechin and ethyl gallate）， stilbene glycoside （<italic>trans</italic>-3，5，4′-trihydroxystyryl-4′-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucoside）， phenylbutanone glycoside of 4′-hydroxyphenyl-2-butanone-4′-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-［2ʺ-<italic>O</italic>-gallic-6ʺ-<italic>O</italic>-（4ʺ-hydroxy）-cinnamoyl）-glucoside， conjugated anthraquinones （aloe emodin-8-<italic>O</italic>-glucoside， rhein-8-<italic>O</italic>-glucoside， emodin-8-<italic>O</italic>-glucoside） and <italic>ω</italic>-hydroxyemodin （<italic>P</italic><0.05， <italic>P</italic><0.01）， and negatively correlated with the contents of free anthraquinones （emodin， aloe emodin and physcion）. Compared with 254 nm， the similarities of chromatographic peaks at 280 nm was more obvious， and the number of detected common peaks was more， which could reflect more subtle differences in chemical composition. Conclusion:Tannins， stilbene glycosides and phenylbutanone glycosides are strongly correlated with <italic>L</italic>^*， while anthraquinones are strongly correlated with <italic>b</italic>^*， the decrease of <italic>E</italic>^*<italic>ab</italic> is mainly related to the increase of free anthraquinone content and the decrease of catechins， <italic>ω</italic>-hydroxyemodin， stilbene glycosides， conjugated anthraquinone and phenylbutanone glycosides. The change of appearance color index of process samples can reflect the change trend of the contents of medicinal components in cooked rhubarb to some extent.

Dynamic changes of total acid and bacterial communities during the traditional fermentation of Hong Qu glutinous rice wine

Background: Hong Qu glutinous rice wine (HQGRW) is brewed under non-aseptic fermentation conditions, so it usually has a relatively high total acid content. The aim of this study was to investigate the dynamics of the bacterial communities and total acid during the fermentation of HQGRW and elucidate the correlation between total acid and bacterial communities. Results: The results showed that the period of rapid acid increase during fermentation occurred at the early stage of fermentation. There was a negative response between total acid increase and the rate of increase in alcohol during the early fermentation stage. Bacterial community analysis using high-throughput sequencing technology was found that the dominant bacterial communities changed during the traditional fermentation of HQGRW. Both principal component analysis (PCA) and hierarchical clustering analysis revealed that there was a great difference between the bacterial communities of Hong Qu starter and those identified during the fermentation process. Furthermore, the key bacteria likely to be associated with total acid were identified by Spearman's correlation analysis. Lactobacillus, unclassified Lactobacillaceae, and Pediococcus were found, which can make significant contributions to the total acid development (| r| N 0.6 with FDR adjusted P b 0.05), establishing that these bacteria can associate closely with the total acid of rice wine. Conclusions: This was the first study to investigate the correlation between bacterial communities and total acid during the fermentation of HQGRW. These findings may be helpful in the development of a set of fermentation techniques for controlling total acid.

Classification of Rice-wine Processed Coptidis Rhizoma Decoction Piece Based on Subjective and Objective Combination Weighting Method and Quality Constant Method / 中国药房

OBJECTIVE：To provide refere nce f or the quality control and evaluation of rice-wine processed Coptidis Rhizoma decoction piece. METHODS ：Taking 17 batches of rice-wine processed Coptidis Rhizoma decoction piece from different manufacturers as samples ，HPLC method was adopted to determine the contents of 4 kinds of alkaloids as epiberberine ，coptisine， palmatine and berberine. The compound weights of epiberberine ，berberine，palmatine and berberine were calculated by the subjective and objective combination weighting method （AHP combined with variation coefficient ）. Then the quality evaluation method was used to evaluate the quality of decoction pieces combined with the appearance of decoction pieces and the contents of 4 alkaloids. The percent mass constant was calculated and the grade of rice-wine processed Coptidis Rhizoma decoction piece was classified. RESULTS ：According to the results of content determination of 4 kinds of alkaloids ，among 17 batches of samples ，a total of 13 batches of samples met the requirements of 2015 edition of Chinese Pharmacopoeia （part Ⅳ）. Mass constants of 13 batches of qualified samples were 10.03-26.96，and the percent mass constants were 37.20％-100％. If the percent mass constant ≥ 80％ of the sample was listed as the first-class product ，the sample with the percent mass constant between 50％-＜80％ was classified as the second-class product ，and the rest was listed as the third-class product ，therefore the quality constant of first-class product was ≥21.57，that of second-class product was 13.48-＜21.57，and that of third-class product was ＜13.48. According to the grading standard ，3 batches of 13 batches of qualified samples are classified as first-class products ，6 batches are classified as second-class products ，4 batches are classified as third-class products. CONCLUSIONS ：The established subjective and objective combination weighting method and quality constant method can more scientifically and reasonably classify rice-wine processed Coptidis Rhizoma decoction piece.

Changes in Cold and Hot Syndrome and Gastrointestinal Bacterial Community Structure in Mice by Intervention with Food of Different Nature / 中国结合医学杂志

OBJECTIVE@#To reveal the effect of foods with different natures on cold or hot syndrome and gastrointestinal bacterial community structure in mice.@*METHODS@#Forty-five 6-week-old male ICR Kunming mice of clean grade were divided into 5 groups, 9 per group, including the control (CK), hot nature herbs (HM), Hong Qu glutinous rice wine (RW), tea rice wine (TW), and cold nature herbs (CM) groups. Distilled water or corresponding herbs were administered to mice (0.01 mL/g body weight) in the 5 groups by gastric infusion respectively, once daily for 28 d. Appearance, behavior, and serum biochemical indicators, including 5-hydroxytryptamine (5-HT), thyroid stimulating hormone (TSH), noradrenaline (NE), cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), the hot nature index, as well as the gastrointestinal bacterial community structure were analyzed in all groups after treatment.@*RESULTS@#After supplementation for 28 d, CM and TW mice showed different degrees of cold syndrome, and HM and RW mice showed different degrees of hot syndrome. Compared with the HM and RW mice, the TSH, NE, cAMP levels and hot nature indices in the CM and TW mice were significantly decreased and 5-HT and cGMP levels were significantly increased (P<0.05). There was no obvious change in appearance or behavior in CK mice. Results of clustering analysis showed that the gastrointestinal bacterial community structures were highly similar in TW and CM mice as well as in RW and HM mice, and that they were from the same branch, respectively, when the distance was 0.02. The key microbes associated with cold syndrome were Lachnospiraceae uncultured, Lactococcus, etc., and the key microbes associated with hot syndrome were S24-7 norank, Ruminococcaceae uncultured, etc. CONCLUSION: The interventions with different nature foods could change cold or hot syndrome in mice, leading to changes in gastrointestinal bacterial community structure.

Improving applicability of urease from Bacillus amyloliquefaciens JP-21 by site-directed mutagenesis / 生物工程学报

Ethyl carbamate (EC) is a carcinogen detected in fermented foods and alcohol beverages. Excessive intake of EC is possibly harmful to health. Enzymatic degradation is one of the most effective approaches for reducing EC in fermented foods. Urease catalyzes the hydrolysis of both EC and urea. This confers urease a good application prospect in reducing EC and its precursor urea in fermented foods. Currently, degradation of EC in alcohol beverages by urease is inefficient due to its low urethanase activity and poor affinity to EC. Urease from Bacillus amyloliquefaciens JP-21 was successfully expressed in Escherichia coli at the level of 3 292 U/L urease and 227.3 U/L urethanase. Two key residues M326 and M374 were characterized that might block the binding of enzyme to EC, through simulating docking the structure of catalytic subunit UreC of urease with EC. Three mutants (M374A, M374T and M326V) of urease with improved urethanase activity were obtained by performing point saturated mutagenesis approach. Using EC as the substrate, Km values of M374A, M374T and M326V were detected to be 101.8 mmol/L, 129.5 mmol/L and 121.7 mmol/L, respectively, which were decreased by 37.47%-50.82% compared with that of the wild type urease. These mutants can degrade more than 97% of urea in rice wine and mutant M374T shows the highest degradation of EC in rice wine. EC content in rice wine was reduced from 525 μg/L to 393 μg/L by using M374T, and the EC degradation rate of it is 0.97 folds higher than that of the wild type urease. The results are of great significance for engineering the catalytic properties of urease and improving its industrial properties, and lays a good foundation for developing strategies to reducing microbial metabolic ammonia (amine) hazards in fermented foods.

Advances in microbial enzymatic elimination of ethyl carbamate in Chinese rice wine / 生物工程学报

Ethyl carbamate (EC), a carcinogenic and teratogenic chemical that is widely distributed in various alcoholic beverages, has attracted much attention. Microbial enzymatic degradation of EC in rice wine is always efficient and attractive. In this review, we summarize the research progress and problems of microbial enzymatic elimination of EC in rice wine from three aspects: the mechanisms of EC formation in rice wine, the research progress of acid urease, and the research progress of urethanase. Then, we propose the corresponding strategies to solve the problems: screening new urethanase with satisfied enzyme properties, food-grade expression and directed evolution of the bifunctional Fe³⁺-dependent acid urease and acid urease used in combination with urethanase to eliminate both urea and EC in rice wine.

Effect of rice wine on degradation of 3,29-dibenzoyl karounitriol in Gualou-Xiebai in human intestinal flora / 中国药学杂志

OBJECTIVE: To study the effects of rice wine on the metabolism of 3, 29-dibenzoyl karounitriol as the main active component in Gualou-Xiebai in human intestinal flora, exploring the scientific connotation of Gualou-XiebaiBaijiu Decoction from the angle of metabolism. METHODS: The metabolic rate of 3, 29-dibenzoyl karounitriol in Gualou-Xiebai rice wine solution and aqueous solution at different time points were compared,respectively,by intestinal flora experiments. RESULTS: Compared to Gualou-Xiebaiaqueous solution group, the metabolic rate of 3, 29-dibenzoyl karounitriol in Gualou-Xiebai rice wine solution group decreased significantly (P<0.05). CONCLUSION: Rice wine as cites drug can inhibit the metabolism of 3, 29-dibenzoyl karounitriol in Gualou-Xiebai.

Determination of four coumarin constituents before and after Angelicae Dahuricae Radix stewed with yellow rice wine and research on its mutual transformation mechanism / 中国中药杂志

To determine the contents of oxypeucedanin, oxypeucedanin hydrate, byakangelicol and byak-angelicin both before and after Angelicae Dahuricae Radix was stewed with yellow rice wine by high-performance liquid chromatography, and study the mutual transformation mechanisms of oxypeucedanin into oxypeucedanin hydrate, as well as byakangelicol into byak-angelicin. The research results indicated that the contents of oxypeucedanin and byakangelicol were decreased, but the contents of oxypeucedanin hydrate and byak-angelicin were increased after Angelicae Dahuricae Radix was processed with yellow rice wine. The contents' changes of these chemical compounds were due to the ring opening reaction of epoxy compounds, such as oxypeucedanin and byakangelicol under the weak acidity and heating conditions of yellow rice wine. This research could provide a scientific basis for the processing mechanism of Angelicae Dahuricae Radix with yellow rice wine stewing.

Simultaneous Determination of 5-Hydroxymethylfurfural and Nine Kinds of Phenolic Compounds in Rice Wine Using High Performance Liquid Chromatography / 分析化学

An analytical method of reversed phase high performance liquid chromatography (RP-HPLC) was developed for simultaneous determination of 5-hydroxymethylfurfural and nine phenolic compounds (including (+)-catechin, (-)-epicatechin, chlorogenic acid, rutin, caffeic acid, protocatechuic acid, syringic acid, ferulic acid and p-coumaric acid) in 20 min.Ten components were detected and separated successfully by Diamonsil C_(18) column (150 mm x4.6 mm, 5 μm) at wavelength of 280 nm and column temperature of 42 ℃, with acetonitrile and 3% acetic acid solution as the mobile phase in gradient elution.The resultant correlation coefficients of the ten compounds were between 0.9911 and 0.9995 with detection limit from 0.2 to 0.5 mg/L, the RSD less than 2.4% and the average recoveries from 89.4% to 98.3%.These experimental results demonstrate that 5-hydroxymethylfurfural and the nine phenolic compounds in different rice wine samples can be determined with the new method for practical uses.

Screening of Functional Rhizopus stolonifer for Alcohol Fermentation and Production of High Quality Korean Traditional Rice Wine

Different strains of mold were screened for the production of high quality Korean traditional rice wine with anti-hypertension and good acceptability. We isolated 867 nuruk mold strains and selected 24 for further study based on measurement of amylase activity. Among them, mold No. 17 showed high ethanol production upon fermentation with Saccharomyces cerevisiae as well as anti-hypertensive properties. The No. 17 strain was therefore selected as the functional mold and later identified as Rhizopus stolonifer based on molecular biological characteristics. Optimal fermentation conditions for the brewing of anti-hypertensive traditional rice wine comprised the addition of R. stolonifer No. 17 koji at a concentration of 35 sp/g and a fermentation period of 10 days at 25degrees C using S. cerevisiae.