Conversion of Syringaldehyde to Sinapinic Acid through Knoevenagel-Doebner Condensation

Sinapinic acid is a widespread compound in vegetable material and is as such common in the human diet. Recently it has drawn attention because of its biological activities. Sinapinic acid can be synthesized from syringaldehyde via the Knoevenagel-Doebner condensation. However this reaction is limited by the formation of 4-vinylsyringol after a second decarboxylation. To gain more detailed information about this reaction and to improve control over the formation of sinapinic acid, the concentration time history of syringaldehyde and the reaction products of this reaction have been monitored over time at different reaction temperatures. The formation of 4-vinylsyringol was found to be inhibited by performing the reaction at temperature below 80°C. This allows the reaction to be optimized for the production of sinapinic acid, with an optimal yield of 78% after 2.5 hours at 70°C.

Glucosinolates and Sinapine in Camelina Meal

Forty seven accessions of camelina (Camelina sativa L. Crantz) were analyzed for glucosinolates (GSLs) and sinapine in defatted meal. These antinutritional compounds are undesirable in camelina meal for use in animal feeding and therefore we show their variability to identify the best varieties for future breeding programs. Total GSLs ranged from 19.6 to 40.3 mmol Kg-1 dry weight (DW) with an average of 30.3 mmol kg-1 DW. Great variability has also been observed in the levels of individual GSLs (GSL1, GSL2 and GSL3), so that the content of GSL1 and GSL3 were not correlated to each other in the accessions of camelina. Five out of six winter forms of camelina showed low content of GSLs. Sinapine ranged from 1.09 to 4.75 g Kg-1 DW with an average of 2.57 g kg-1 DW. The sinapine content was not correlated with that of GSLs. The use of camelina meal is only limited by the presence of GSLs while sinapine content can be ignored in camelina varieties.

Antinutritive Compounds in Twelve <i>Camelina sativa </i>Genotypes

Camelina sativa is an oilseed crop becoming important in North America and Europe for biodiesel production. The use of Camelina flours in animal diet may be limited by antinutritive compounds. The content of glucosinolates, phytic acid, sinapine and condensed tannins was evaluated in twelve accessions of Camelina sativa. All compounds showed significant differences among genotypes. Only the concentration of glucosinolates in the flour deserves attention, while the content of phytic acid, sinapine and condensed tannins are to acceptable levels. Camelina showed the presence of three different glucosinolates (GSL1, GSL2 and GSL3) in the flour, with differences among genotypes regarding the relative abundance of each glucosinolate. The content of glucosinolates is inversely correlated with that of sinapine. The glucosinolate content in Camelina flour has to be reduced to increase the use of this flour in animal diet, but avoiding altering the sinapine content.

Purification of a Sinapine-Glucoraphanin Salt from Broccoli Seeds

A sinapine (sinapoylcholine)-glucoraphanin salt has been isolated from broccoli seeds and characterized by NMR and mass spectrometry. This salt extraction method can be used to purify glucoraphanin free from contamination by glucoiberin.

Extraction process in Chuantieling(CTL) based on the quality by design(QbD) concept

Objective This study aimed to design and optimize the water extraction process for Chuantieling(喘贴灵,CTL)based on the concept of quality by design(QbD).Methods The single-factor experiments were used to select the best experimental points of CTL water extraction.On this basis,the transfer rate of ephedrine hydrochloride and sin-apine thiocyanate,and the yield of the extract were evaluated as the evaluation indicators.The liquid-solid ratio,extraction time,and pH value were selected as the main factors to carry out the Box-Behnken design(BBD).A mathematical model of the critical process parameters(CPPs)and critical quality attributes(CQAs)was established,the interaction between CQAs and CPPs was examined,and the CTL extraction process design space was established and optimized,which guaranteed the stability of the process.The high performance liquid chro-matography(HPLC)was used to analyze the main active compounds in the CTL extract.Results Through single-factor experiments,the best experimental parameters were found to be a liquid-solid ratio of 8∶1,extraction time of 90 min,pH value of 4,and extraction times of three.The experimental variance analysis results of the BBD showed that the P value of the re-gression model was less than 0.05,and the lack-of-fit value was greater than 0.01,indicating that the constructed model had good predictive ability.The operating space of the CPPs of the CTL water extraction process was combined with the actual production situation.In the pro-duction situation,the best extraction process was nine times of water addition,72 min of ex-traction time,and 4.5 of pH value.The HPLC results showed that the peak areas of ephedrine hydrochloride,sinapine thiocyanate,tetrahydropalmatine,methyl eugenol,cinnamaldehyde,and 6-gingerol in the CTL extract accounted for 0.94%,14.32%,0.78%,31.23%,0.34%,and 0.44%of the total peak area,respectively.Conclusion The water extraction process design space of CTL based on QbD was conducive to actual production operations,ensuring the stability of the process.

Isolation and identification of sinapine-degrading bacteria from the intestinal tract of laying hens

This study was aimed to isolate sinapine-degrading bacteria from the intestinal tract of laying hens and to identify the predominant bacteria. Thirty-week old healthy laying hens were killed, and the chyme in the digestive tract was inoculated into modified Czapek medium containing sinapin and cultivated at 37 ℃for 10 days. The optical density(OD) values of the bacterial solutions at different cultivating times were detected by a spectrophotometric method. The predominant strains were identified by 16 S rRNA gene analysis. We extracted the extracellular products of the predominant strains to determine the total protein using the Coomassie brilliant blue method, and to determine the activities of some extracellular enzymes using the agar plate diffusion method. Nine strains were isolated from the lower intestinal tract of laying hens. Among the 9 strains, 5 were from the ileum, 2 were from the ceca and 2 were from the jejunum. We could not isolate any strains from the upper intestinal tract, such as the stomach and duodenum. Eight of those 9 isolated strains were gram negative and one was gram positive. Strains YD-1 and YD-2 were better than other strains in their abilities to degrade sinapine. Strains YD-1 and YD-2 were identified as Escherichia coli and Klebsiella spp., respectively, by the 16 S rRNA sequence analysis.The total protein level of the extracellular products was 1.213 g/L for YD-1 and 1.990 g/L for YD-2. Both extracellular products of YD-1 and YD-2 had the activities of protease, amylase and urease. This study confirmed that the primary site of sinapine degradation is in the lower intestinal tract of laying hens. The sinapine-degrading strains are mainly gram negative. Strains YD-1 and YD-2 are predominant in degrading sinapine and they belong to E. coli and Klebsiella spp., respectively. Both extracellular products of YD-1 and YD-2 contain protease, amylase and urease. Strain YD-2 is better than strain YD-1 in its ability to degrade sinapine.

Serum trimethylamine-N-oxide and gut microbiome alterations are associated with cholesterol deposition in the liver of laying hens fed with rapeseed meal

Sinapine derived from cruciferous plants could be converted into trimethylamine by intestinal microbiota.Its metabolite,trimethylamine N-oxide(TMAO),is closely linked to increased risk of cardiovascular disease and fat deposition in mammals.Hens fed with rapeseed meal(RSM)suffered from fatty liver hemorrhage syndrome(FLHS).This study was conducted to investigate whether RSM-induced fatty liver is due to TMAO via altering microbiota composition and diversity.At 33 weeks of age,600 laying hens were randomly divided into 5 treatment groups,namely control and 14%RSM treatment groups(DY5,with 16.2% erucic acid [EA]and 74.66% glucosinolate[Gl]contents;MB1,with 3.50% EA and 43.23 %Gl contents;DY6,with 6.7% EA and 22.67% Gl contents;XH3,with 44.60% EA and 132.83% Gl contents)for 8 weeks.Results revealed that 3 hens died due to liver hemorrhage after ingesting 14% RSM diet.The 14% RSM decreased serum low-density lipoprotein cholesterol(LDL-C)content(P<0.01)while tended to increase serum TMAO content compared to the control group(P=0.08).The 14% RSM diet increased red oil O optical density(P<0.01),and increased total cholesterol(TC)and LDL-C content in the liver(P<0.01,and P<0.01,respectively).The 14%RSM decreased liver total bile acid(TBA)content compared to the control(P<0.01).The DY6 had a higher TBA content in the liver than the XH3(P<0.01).The 14%RSM decreased mRNA abundance of liver X receptors alpha(LXR-α,P=0.01),and increased mRNA abundance of sterol response element binding protein 2(SREBP-2,P=0.04).Results revealed that the in-feed RSM could alter richness and diversity of cecal microbiota compared to the control(P<0.05).Liver TC content and serum TMAO showed a negative relationship with Proteobacteria and Actinobacteria(P=0.04).In conclusion,14%RSM increased liver TC and induced high liver score of FLHS,which was possibly associated with the altered cecal microbiota composition,increased serum TMAO levels and LXR-αand SREBP-2 expressions.