Optimization of ultrasonic pretreatment and analysis of chlorogenic acid in potato leaves.

Chlorogenic acid (CA) is an effective ingredient that can strengthen immunity during following the COVID-19 era. The current cost of CA is high owing to its complex purification process and low yield (approximately 2%). In this study, a one-step path orthogonal experiment was designed based on the results from Gauss calculation, which consisted of acidity, coordination, and hydrolysis in molecules. The optimized extraction conditions were 60 â„ƒ, 60 min, 1:20 liquid ratio, and 40% ethanol in a nitrogen atmosphere controlled using a device of our own design, which led to CA yields of up to 6.35% from potato leaves. The purified CA was analyzed using high-performance liquid chromatography, thin-layer chromatography, ultraviolet-visible spectroscopy, and molecular fluorescence. This accurate and reproducible method can not only be used to obtain high yields of CA but can also be used for the quality control of active plant products and their isomers.

Chemical composition and sensory profiling of coffees treated with asparaginase to decrease acrylamide formation during roasting.

Acrylamide is an amide formed in the Maillard reaction, with asparagine as the primary amino acid precursor. The intake of large amounts of acrylamide has induced genotoxic and carcinogenic effects in hormone-sensitive tissues of animals. The enzime asparaginase is one of the most effective methods for lowering the formation of acrylamide in foods such as potatoes. However, the reported sensory outcomes for coffee have been unsatisfactory so far. This study aimed to produce coffees with reduced levels of acrylamide by treating them with asparaginase while retaining their original sensory and bioactive profiles. Three raw samples of Coffea arabica, including two specialty coffees, and one of Coffea canephora were treated with 1000, 2000, and 3000 ASNU of the enzyme. Asparagine and bioactive compounds (chlorogenic acids-CGA, caffeine, and trigonelline) were quantified in raw and roasted beans by HPLC and LC-MS, while the determination of acrylamide and volatile organic compounds was performed in roasted beans by CG-MS. Soluble solids, titratable acidity, and pH were also determined. Professional cupping by Q-graders and consumer sensory tests were also conducted. Results were analyzed by ANOVA-Fisher, MFA, PCA and Cluster analyses, with significance levels set at p ≤ 0.05. Steam treatment alone decreased acrylamide content by 18.4%, on average, and 6.1% in medium roasted arabica and canefora coffees. Average reductions of 32.5-56.0% in acrylamide formation were observed in medium roasted arabica beans when 1000-3000 ASNU were applied. In the canefora sample, 59.4-60.7% reductions were observed. However, steam treatment primarily caused 17.1-26.7% reduction of total CGA and lactones in medium roasted arabica samples and 13.9-22.0% in canefora sample, while changes in trigonelline, caffeine, and other evaluated chemical parameters, including the volatile profiles were minimal. Increasing enzyme loads slightly elevated acidity. The only sensory changes observed by Q-graders and or consumers in treated samples were a modest increase in acidity when 3000 ASNU was used in the sample with lower acidity, loss of mild off-notes in control samples, and increased perception of sensory descriptors. The former was selected given the similarity in chemical outcomes among beans treated with 2000 and 3000 ASNU loads.

Evaluation of Environmental Factor Effects on the Polyphenol and Flavonoid Content in the Leaves of Chrysanthemum indicum L. and Its Habitat Suitability Prediction Mapping.

The leaves of Chrysanthemum indicum L. are known to have various bioactive compounds; however, industrial use is extremely limited. To overcome this situation by producing high-quality leaves with high bioactive content, this study examined the environmental factors affecting the phytochemical content and antioxidant activity using C. indicum leaves collected from 22 sites in Kochi Prefecture, Japan. Total phenolic and flavonoid content in the dry leaves ranged between 15.0 and 64.1 (mg gallic acid g-1) and 2.3 and 11.4 (mg quercetin g-1), while the antioxidant activity (EC50) of the 50% ethanol extracts ranged between 28.0 and 123.2 (µg mL-1) in 1,1-Diphenyl-2-picrylhydrazyl radical scavenging assay. Among the identified compounds, chlorogenic acid and 1,5-dicaffeoylquinic acid were the main constituents in C. indicum leaves. The antioxidant activity demonstrated a positive correlation with 1,5-dicaffeoylquinic acid (R2 = 0.62) and 3,5-dicaffeoylquinic acid (R2 = 0.77). The content of chlorogenic acid and dicaffeoylquinic acid isomers varied significantly according to the effects of exchangeable magnesium, cation exchange capacity, annual temperature, and precipitation, based on analysis of variance. The habitat suitability map using the geographical information system and the MaxEnt model predicted very high and high regions, comprising 3.2% and 10.1% of the total area, respectively. These findings could be used in future cultivation to produce high-quality leaves of C. indicum.

Cytotoxic and Antitumoral Effects of Methanolic Extracts of Avocado Fruit Mesocarp in Colorectal Cancer Cell Line HT29.

Colorectal cancer is a widespread neoplasia with high ratios of chemoresistance. Phytochemicals in plant-based extracts could be useful to treat colorectal cancer, and/or reduce chemoresistance. Methanolic extract of avocado mesocarp (MEAM) has demonstrated antitumoral properties, depending on the fruit ripening stage (RS). The aim of this study was to analyze the effects of methanolic extracts of "Hass" avocado fruit at different RS on cytotoxicity, antioxidative, anti-inflammatory, anti-invasive, cell cycle, and epithelial-mesenchymal transition inhibition in colorectal adenocarcinoma cell line HT29. The MEAM showed an increasing concentration of total phenolic compounds as the RS progressed, which was correlated with antioxidant capacity measured by the Ferric Reducing Antioxidant Power assay but not with the 2.2-diphenyl-1-picrylhydrazyl assay. The specific phenolic compounds of MEAM were determined by high-performance liquid chromatography, and it was found that concentrations of epicatechin decreased while concentrations of chlorogenic acid increased as the RS progressed. The HT29 cell line was treated with MEAM for 48 h, and all MEAM had a cytotoxic effect, reported by MTT assay, nevertheless, the strongest effect was associated with the presence of chlorogenic acid. MEAM induced apoptosis and cell cycle arrest in phase G0/G1, reported by flow cytometry. Moreover, MEAM inhibited cell migration evidenced by the wound healing assay. On the other hand, MEAM significantly reduced expression of mRNA of tumor necrosis factor-alpha and cyclooxygenase 2. These effects comprise important inhibition of some hallmarks of cancer. This, in turn, may provide interesting guidelines for developing antitumoral intervention agents.

Influence of maturation and roasting on the quality and chemical composition of new conilon coffee cultivar by chemometrics.

Coffee is one of the most consumed beverages worldwide. Espírito Santo is the largest Brazilian producer of conilon coffee, and invested in the creation of new cultivars, such as "Conquista ES8152", launched in 2019. Therefore, the present study aimed to evaluate the effects of maturation and roasting on the chemical and sensorial composition of the new conilon coffee cultivar "Conquista ES8152". The coffee was harvested containing 3 different percentages of ripe fruits: 60%, 80%, and 100%, and roasted at 3 different degrees of roasting: light, medium, and dark, to evaluate the moisture and ash content, yield of soluble extract, volatile compound profile, chlorogenic acid and caffeine content, and sensory profile. "Conquista ES8152" coffee has a moisture content between 1.38 and 2.62%; ash between 4.34 and 4.72%; and yield between 30.7 and 35.8%. Sensory scores ranged between 75 and 80 and the majority of volatile compounds belong to the pyrazine, phenol, furan, and pyrrole groups. The content of total chlorogenic acids was drastically reduced by roasting, with values between 2.40 and 9.33%, with 3-caffeoylquinic acid being the majority. Caffeine was not influenced by either maturation or roasting, with values between 2.16 and 2.41%. The volatile compounds furfural, 5-methylfurfural, and 2-ethyl-5-methylpyrazine were positively correlated with the evaluated sensory attributes and 5-methylfurfural was the only one significantly correlated with all attributes. Ethylpyrazine, furfuryl acetate, 1-furfurylpyrrole, 4-ethyl-2-methoxyphenol, and difurfuryl ether were negatively correlated. The stripping did not affect the quality and composition of this new cultivar, however, the roasting caused changes in both the chemical and sensorial profiles, appropriately indicated by the principal component analysis.

Quantitative analysis of the quality constituents of Lonicera japonica Thunberg based on Raman spectroscopy.

Quantitative analysis of the quality constituents of Lonicera japonica (Jinyinhua [JYH]) using a feasible method provides important information on its evaluation and applications. Limitations of sample pretreatment, experimental site, and analysis time should be considered when identifying new methods. In response to these considerations, Raman spectroscopy combined with deep learning was used to establish a quantitative analysis model to determine the quality of JYH. Chlorogenic acid and total flavonoids were identified as analysis targets via network pharmacology. High performance liquid chromatograph and ultraviolet spectroscopy were used to construct standard curves for quantitative analysis. Raman spectra of JYH extracts (1200) were collected. Subsequently, models were built using partial least squares regression, Support Vector Machine, Back Propagation Neural Network, and One-dimensional Convolutional Neural Network (1D-CNN). Among these, the 1D-CNN model showed superior prediction capability and had higher accuracy (R2 = 0.971), and lower root mean square error, indicating its suitability for rapid quantitative analysis.

Rapid screening and evaluation of natural antioxidants from leaf, stem, and root of Artemisia argyi by online liquid microextraction combined with HPLC-based antioxidant assay system coupled with calibration quantitative analysis.

To reveal the utilization value of leaf, stem, and root of Artemisia argyi, a rapid online liquid microextraction combined with a high-performance liquid chromatography coupled with 2,2-nitrogen-di (3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt antioxidant assay system was established for analysis of antioxidants in the leaf, stem, and root of A. argyi, and a calibration quantitative method of antioxidant activity with equivalent chlorogenic acid was proposed. Thirty-three positive peaks were identified; among them, 12 compounds were found that possess good antioxidant activity including eleven organic acids (components 2-4, 8, 11-14, 17, 19, and 21) and one flavonoids (component 22). The proposed calibration quantitative method avoided the influence of content of compound and compared the extent of radical scavenging capacity of five antioxidant compounds, which were ranked as follow: 3,5-dicaffeoylquinic acid > 3,4-dicaffeoylquinic acid ≈ 4,5-dicaffeoylquinic acid > 1,4-dicaffeoylquinic acid > chlorogenic acid. In conclusion, this study provided composition and biological potential for the future development of the leaf, stem, and root of A. argyi. It is believed that the online liquid microextraction combined with high-performance liquid chromatography based antioxidant assay system can be widely used for the rapid screening of natural antioxidant components in the different parts of natural products.

Using underutilized residues of coffee to obtain valuable dietary and antioxidant bioactive compounds.

BACKGROUND: Coffee cultivation and agroindustry generate residues that are rich in several metabolites. These compounds, such as phenolic compounds and alkaloids, are known for their antioxidant activity and are usually consumed as nutraceuticals. The purpose of this study was to evaluate the occurrence of chemical and antioxidant components of low-pruned coffee stems under different fertilizer regimes. Extractives and lignin composition, histochemical, chromatographic, and antioxidant analyses were performed. RESULTS: Multiple compounds were found to accumulate in the stems of coffee trees. Furthermore, the presence of phenolic compounds such as chlorogenic acid, vanillin, resveratrol, and the alkaloids caffeine and trigonelline varied depending on the type of fertilization. In all samples examined, optimal performance was observed at the highest tested concentration (500 µg mL-1 ). All samples analyzed presented a great performance at the highest concentration tested (500 µg mL-1 ), with the dose 70% and the dose 100%, which is the recommended for the culture, showing the highest values for most of the concentrations and the best half-maximal inhibitory concentration (IC50 ) when compared with the other samples tested. CONCLUSION: As shown in the results, the reuse of stem residues as antioxidant material, with the potential to be profitable, and has the added benefit of providing a sustainable destination for material that until now has been underutilized. © 2023 Society of Chemical Industry.

Src Tyrosine Kinase Inhibitory and Antioxidant Activity of Black Chokeberry and Bilberry Fruit Extracts Rich in Chlorogenic Acid.

Edible berries such as the fruits of black chokeberry (Aronia melanocarpa (Michx.) Elliott) and bilberry (Vaccinium myrtillus L.) are considered to be rich in phenolic compounds, which are nowadays attracting great interest due to their promising health benefits. The main objective of our study was to investigate, for the first time, their inhibitory properties on Src tyrosine kinase activity, as this enzyme plays an important role in multiple cellular processes and is activated in both cancer and inflammatory cells. In hydroethanolic fruit extracts, 5.0-5.9% of total polyphenols were determined spectrophotometrically, including high amounts of hydroxycinnamic acid derivatives. HPLC analysis revealed that the black chokeberry and bilberry extracts contained 2.05 mg/g and 2.54 mg/g of chlorogenic acid, respectively. Using a time-resolved fluorescence resonance energy transfer (TR-FRET) assay, the extracts studied were found to have comparable inhibitory effects on Src tyrosine kinase, with IC50 values of 366 µg/mL and 369 µg/mL, respectively. The results also indicated that chlorogenic acid contributes significantly to the observed effect. In addition, both fruit extracts exhibited antioxidant activity by scavenging DPPH and NO radicals with SC50 values of 153-352 µg/mL. Our study suggested that black chokeberry and bilberry fruits may be beneficial in cancer and other inflammation-related diseases.

Light spectra manipulation stimulates growth, specialized metabolites and nutritional quality in Anethum graveolens.

Light-Emitting Diodes (LED) play a major role in manipulating light spectra that helps in regulating the growth and specialized metabolite synthesis relevant to the plant defence system. In this study, we assessed photosynthetic performance, phytonutrients, and anatomical variations of an aromatic herb Anethum graveolens (also known as dill), grown under various combinations of LED lights viz. red (100R:0B), red:blue (50R:50B); blue (0R:100B) and warm white (WW, served as control). Exposure to 0R:100B LED lights led to the tallest stem height, whereas, the number of leaves were highest under 50R:50B LED lights. The photosynthetic performance was observed to be highest under 50R:50B LED lights. HPLC analysis revealed chlorogenic acid and rosmarinic acid as the major phenolic compounds accumulated under different spectral irradiations. The highest chlorogenic acid content was observed in 50R:50B LED treated dill plants, while 100R:0B light showed the highest accumulation of rosmarinic acid. Dill plants grown under 50R:50B light displayed a relatively higher content of volatile compounds including, myristicin (phenylpropene), psi-limonene, and α-phellandrene (monoterpenoids). Expression analyses of candidate genes of phenylpropanoid and monoterpenoid biosynthetic pathways showed good correlations with the enhanced phenolic compounds and monoterpenes detected under appropriate light treatments. Further, the stem anatomy revealed higher vascularization under the influence of 0R:100B LED lights, whereas, intense histochemical localization of specialized metabolites could be correlated with enhanced accumulation of phenolic compounds and terpenoids observed in this study. Taken together, these studies suggest that proper combinations of blue and red spectra of light could play important role to augment the growth and phytochemical characteristics of dill, thus improving its value addition in the food industry.

Chlorogenic acid as an indispensible partner of caffeic acid in coffee via selective regulation of prooxidative actions of caffeic acid.

Chlorogenic acid (CGA) and caffeic acid (CA) are two major phenolic acids in coffee. Though the International Agency for Research on Cancer has classified CA as a Group2B carcinogen, coffee consumption seems generally safe within the usual levels of intake and is more likely to benefit health than to harm it. We thus speculated that CGA may effectively suppress the carcinogenic potential of CA. In a molar ratio achievable in vivo, this study shows that CGA can inhibit (i) copper reduction caused by CA, (ii) CA oxidation caused by copper, (iii) the formation of hydroxyl radicals by CA and copper, and (iv) DNA damage induced by CA, quercetin or (-)-epigallocatechin-3-gallate in the presence of copper. CA tends to undergo autoxidation to produce hydrogen peroxide and quinone, which further reacts with proteins to form quinoproteins. This autoxidation at a tolerable level normally induces beneficial adaptive responses. This study shows that CGA is less efficient than CA in producing hydrogen peroxide and quinoprotein; however, together they synergistically produce hydrogen peroxide and quinoprotein in vitro at a molar ratio achievable in vivo. In conclusion, CGA can selectively regulate the prooxidant activities of CA depending on whether copper is involved or not. CGA could be viewed as an indispensable partner of CA in coffee, given its dual role in suppressing the carcinogenic potential of CA and boosting CA autoxidation which is beneficial for disease prevention.

LC-MS methods combination for identification and quantification of trans-sinapoylquinic acid regioisomers in green coffee.

The present study aims to both identify and quantify trans-sinapoylquinic acid (SiQA) regioisomers in green coffee by combined UHPLC-ESI-QqTOF-MS/MS and UHPLC-ESI-QqQ-MS/MS methods. Among the various mono-acyl chlorogenic acids found in green coffee, SiQA regioisomers are the least studied despite having been indicated as unique phytochemical markers of Coffea canephora (known as Robusta). The lack of commercially available authentic standards has been bypassed by resorting to the advantages offered by high-resolution LC-MS as far as the identification is concerned. SiQA regioisomers have been identified in several samples of Robusta and Coffea arabica (known as Arabica) commercial lots from different geographical origin and, for the first time, in different samples of coffee wild species (Coffea liberica and Coffea pseudozanguebariae). Quantification (total SiQA ranging from 3 to 5 mg/100 g) let to reconsider these chlorogenic acids as unique phytochemical markers of Robusta being present in the same quantity and distribution in C. liberica as well. Gardeniae Fructus samples (fruits of Gardenia jasminoides) have additionally been characterized as this matrix is recognized as one of the few naturally occurring SiQA sources. The SiQA regioisomer content (total SiQA about 80 mg/100 mg) fully supports the proposal to use this matrix as a surrogate standard for further studies.

Unbiased and biased chemometric analysis of LC-MS data from human urine following coffee intake.

We carried out a human volunteer study with 14 participants, eight of whom were asked to consume one cup of coffee at four different time points. Urine samples were collected at eight time points and analyzed by HPLC-MS analysis. The LC-MS data were subjected to unsupervised multivariate statistical analysis (principal component analysis) followed by supervised multivariate analysis (linear discriminant analysis). In an unbiased approach, in the absence of data preselection and filtering, the most important features explaining differences between coffee consumers and the control group observed showed variations in endogenous human hormonal steroid metabolites as well as xanthine derivatives. Only after a biased data treatment data revealed differences between the sample groups based on literature reported chlorogenic acid metabolites resulting directly from coffee intake. Such analysis could confirm the presence of 21 previously reported chlorogenic acid plasma metabolites as urinary metabolites. The application of tandem MS molecular networking revealed the presence of five bioavailable chlorogenic acid derivatives in urine previously not reported, including both quinic acid lactone and dimethoxy caffeoyl esters. Selected cinnamic acids were quantified in urine.

Peculiarities of the Variation of Biologically Active Compounds in Fruit of Vaccinium oxycoccos L. Growing in the Cepkeliai State Strict Nature Reserve.

This study was carried out to analyze the accumulation patterns of anthocyanins, proanthocyanidins, flavonols, chlorogenic acid, and triterpene compounds in fruit samples of Vaccinium oxycoccos L. berries growing in the Cepkeliai State Strict Nature Reserve in Lithuania. Studies were carried out on the phytochemical composition of cranberry fruit samples during the period of 2020-2022. Anthocyanins, flavonols, chlorogenic acid and triterpene compounds were identified and quantified using UPLC-DAD methods, and proanthocyanins were determined using spectrophotometric methods. The content of identified compounds varied, as reflected in the total amounts of anthocyanins (710.3 ± 40 µg/g to 6993.8 ± 119 µg/g), proanthocyanidins (378.4 ± 10 µg EE/g to 3557. 3 ± 75 µg EE/g), flavonols (479.6 ± 9 µg/g to 7291.2 ± 226 µg/g), chlorogenic acid (68.0 ± 1 µg/g to 3858.2 ± 119 µg/g), and triterpenoids (3780.8 ± 98 µg/g to 7226.9 ± 224 µg/g). Cranberry fruit samples harvested from open oligotrophic wetland habitats contained higher levels of anthocyanins, anthocyanidins, flavonol glycosides, and proanthocyanidins. The highest levels of triterpene compounds were found in the cranberry fruits harvested in the spring of the following year after the snowmelt. The use of principal component analysis showed that cranberry plant material harvested in October and November had higher levels of bioactive compounds.

Evaluation of hackberry (Celtis australis L.) fruits as sources of bioactive compounds.

Hackberry (Celtis australis L.) is native to the Mediterranean region and is distributed in Europe, Turkey, North Africa, and Iran. To the best of our knowledge, no study has been conducted on C. australis L. in the Arasbaran region, Iran. In the present study, total phenol (TP), flavonoid (TF), antioxidant capacity based on DPPH and FRAP assays and phenolic compounds and sugars profiles were investigated. According to the results, the range of antioxidant capacity based on DPPH and FRAP assays was 14.12-88.24% and 44.35-117.87 mg Fe2+/100 g, respectively. Also, the range of gallic acid, caffeic acid, chlorogenic acid, rutin, p-coumaric acid, rosmaric acid, cinnamic acid, and apigenin content was 2.59-26.32, 2.03-9.32, 0.94-11.35, 1.80-4.857, 2.32-9.52, 4.74-51.38, 0.18-2.10 and 0.27-1.37 mg/g, respectively. The results of factor analysis showed that the C12, C14, C15, C20, C8, C16, C3, and C20 genotypes are positively characterized by the first principal component (PCA1) that have a higher caffeic acid, chlorogenic acid, rutin, p-coumaric acid, rosmaric acid, quercetin, cinnamic acid, and apigenin phenolic compounds. Based on cluster analysis, the twenty genotypes were located in 2 main clusters. In general, the obtained results can be useful for breeding programs and the introduction of cultivars in Celtis australis L.

International society of sports nutrition position stand: coffee and sports performance.

Based on review and critical analysis of the literature regarding the contents and physiological effects of coffee related to physical and cognitive performance conducted by experts in the field and selected members of the International Society of Sports Nutrition (ISSN), the following conclusions represent the official Position of the Society:(1) Coffee is a complex matrix of hundreds of compounds. These are consumed with broad variability based upon serving size, bean type (e.g. common Arabica vs. Robusta), and brew method (water temperature, roasting method, grind size, time, and equipment).(2) Coffee's constituents, including but not limited to caffeine, have neuromuscular, antioxidant, endocrine, cognitive, and metabolic (e.g. glucose disposal and vasodilation) effects that impact exercise performance and recovery.(3) Coffee's physiologic effects are influenced by dose, timing, habituation to a small degree (to coffee or caffeine), nutrigenetics, and potentially by gut microbiota differences, sex, and training status.(4) Coffee and/or its components improve performance across a temporal range of activities from reaction time, through brief power exercises, and into the aerobic time frame in most but not all studies. These broad and varied effects have been demonstrated in men (mostly) and in women, with effects that can differ from caffeine ingestion, per se. More research is needed.(5) Optimal dosing and timing are approximately two to four cups (approximately 473-946 ml or 16-32 oz.) of typical hot-brewed or reconstituted instant coffee (depending on individual sensitivity and body size), providing a caffeine equivalent of 3-6 mg/kg (among other components such as chlorogenic acids at approximately 100-400 mg per cup) 60 min prior to exercise.(6) Coffee has a history of controversy regarding side effects but is generally considered safe and beneficial for healthy, exercising individuals in the dose range above.(7) Coffee can serve as a vehicle for other dietary supplements, and it can interact with nutrients in other foods.(8) A dearth of literature exists examining coffee-specific ergogenic and recovery effects, as well as variability in the operational definition of "coffee," making conclusions more challenging than when examining caffeine in its many other forms of delivery (capsules, energy drinks, "pre-workout" powders, gum, etc.).

[Nature-effect transformation mechanism of mulberry leaves and silkworm droppings based on chemical composition analysis].

Starting with the relationship between mulberry leaves and silkworm droppings as food and metabolites, this study systematically compared the chemical components, screened out differential components, and quantitatively analyzed the main differential components based on ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) and UPLC-Q-TRAP-MS combined with principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Moreover, the in vitro enzymatic transformation of the representative differential components was studied. The results showed that(1) 95 components were identified from mulberry leaves and silkworm droppings, among which 27 components only exist in mulberry leaves and 8 components in silkworm droppings. The main differential components were flavonoid glycosides and chlorogenic acids.(2) Nineteen components with significant difference were quantitatively analyzed, and the components with significant differences and high content were neochlorogenic acid, chlorogenic acid, and rutin.(3) The crude protease in the mid-gut of silkworm significantly metabolized neochlorogenic acid and chlorogenic acid, which may be an important reason for the efficacy change in mulberry leaves and silkworm droppings. This study lays a scientific foundation for the development, utilization, and quality control of mulberry leaves and silkworm droppings. It provides references for clarifying the possible material basis and mechanism of the pungent-cool and dispersing nature of mulberry leaves transforming into the pungent-warm and dampness-resolving nature of silkworm droppings, and offers a new idea for the study of nature-effect transformation mechanism of traditional Chinese medicine.

[Effects of different drying methods on quality of male flowers of Eucommia ulmoides based on color and chemical composition].

To study the effects of different drying methods on the quality of male flowers of Eucommia ulmoides(MFOEU), we treated fresh MFOEU samples with drying in the shade(DS), vacuum freeze drying(VFD), high-or low-temperature hot air drying(HTHAD, LTHAD), microwave drying(MD), and vacuum drying(VD), respectively. The color, total flavonoid content, total polysaccharide content, and main active components such as geniposide, geniposidic acid, rutin, chlorogenic acid, galuteolin, pinoresinol diglucoside, and aucubin in MFOEU were taken as the evaluation indicators. The quality of MFOEU was comprehensively evaluated by entropy weight method combined with color index method, partial least squares discriminant analysis and content clustering heat map. The experimental results showed that VFD and DS basically kept the original color of MFOEU. The MFOEU treated with MD had higher content of total polysaccharides, phenylpropanoids, lignans, and iridoids. The MFOEU treated with LTHAD had higher content of total flavonoids and that treated with VD had lower content of active components. According to the results of comprehensive evaluation, the quality of MFOEU dried with different methods followed the order of MD>HTHAD>VFD>LTHAD>DS>VD. Considering the color of MFOEU, the suitable drying methods were DS and VFD. Considering the color, active components, and economic benefits of MFOEU, MD was the suitable drying method. The results of this study are of a reference value for the determination of suitable methods for MFOEU processing in the producing areas.

Generation and structure elucidation of a red colorant formed by oxidative coupling of chlorogenic acid and tryptophan.

In view of the poor acceptance of synthetic food colorants by consumers, there is intense interest in novel natural compounds, preferably from plant-derived sources. We oxidized chlorogenic acid using NaIO4 and reacted the resultant quinone with tryptophan (Trp) to obtain a red-colored product. The colorant was precipitated, freeze-dried, purified by size exclusion chromatography, and subsequently characterized using UHPLC-MS, high-resolution mass spectrometry, and NMR spectroscopy. Additional mass spectrometric studies were performed on the reaction product generated with Trp educts labeled with 15N and 13C. The data obtained from these studies allowed the identification of a complex compound consisting of two Trp and one caffeic acid moieties, and the proposition of a tentative pathway of its formation. Thus, the present investigation expands our knowledge about the formation of red colorants based on the reaction of plant phenols and amino acids.

Full-length transcriptome sequencing and transgenic tobacco revealed the key genes in the chlorogenic acid synthesis pathway of Sambucus chinensis L.

Chlorogenic acid is a key chemical in antioxidation and antisepsis. Sambucus chinensis L. is an herbaceous plant rich in chlorogenic acid and a potential genetic resource for breeding high-chlorogenic acid plants. However, there are few studies on the synthesis pathway of chlorogenic acid in S. chinensis. Our study found chlorogenic acid accumulation in S. chinensis to be organ-specific, higher in leaves and buds but lower in roots, stems and fruits. A total number of 546,844 CCS (circular consensus sequence), including 402,767 full-length nonchimeric (FLNC) and 39 annotated sequences related to the synthesis of chlorogenic acid, was obtained by single-molecule real-time sequencing technology (SMRT). qRT-PCR showed that a number of key genes involved in chlorogenic acid synthesis were differentially expressed in various tissues of S. chinensis. Transgenic tobacco revealed that ectopic expression of the HCT homologous gene HCT-45178 increased the content of chlorogenic acid. Our results should be the first report of full-length transcriptome data of S. chinensis, which help to understand the basis of chlorogenic acid synthesis and provide a novel strategy for breeding tobacco cultivars with higher levels of chlorogenic acid.