Bitter Substances Research Articles

Metabolomics Analysis Reveals Bitter Taste Formation in off-Season Crab Hepatopancreas Marketed in June of the Lunar Calendar.

Crabs marketed off-season in lunar June often have a brown hepatopancreas (BH) that tastes bitter, unlike the nonbitter orange hepatopancreas (OH). We conducted nontargeted and widely targeted metabolomics analyses on both raw and cooked orange and brown hepatopancreas of Eriocheir sinensis. A total of 115 bitter metabolites were identified, including oxylipins, amino acids, small peptides, nucleotides, vitamins, and coenzymes. Targeted quantitative analysis revealed that seven oxylipins were major contributors to the bitterness of BH. The primary metabolic pathways affecting hepatopancreas bitterness involved the production of unsaturated fatty acids and α-linolenic acid metabolism. Furthermore, specific odorants were produced from OH and BH groups during heat treatment, namely ethyl caproate and methional, respectively. Moreover, differential metabolites were found to act as odor precursors, resulting in a brown and bitter hepatopancreas with poor odor quality. This study elucidates the generation of bitter substances in the hepatopancreas of Eriocheir sinensis through metabolic pathways and thermal reaction pathways.

Combining non-targeted and targeted metabolomics to study key bitter and astringent substances of Liupao tea

Liupao tea is a post-fermented dark tea with bitterness and astringency as key sensory traits, though its chemical composition is not fully understood. Six Liupao tea samples with significant differences in bitterness and astringency were analyzed using non-targeted metabolomics and sensory evaluation. Thirty finished and five semi-finished Liupao tea samples were analyzed using UHPLC-MS-PRM for targeted quantification of bitter and astringent compounds. The results show that 477 non-volatile compounds were detected, including 18 potential bitter compounds and 22 potential astringent compounds. Six key bitter compounds (epigallocatechin gallate, catechin gallate, caffeine, quinic acid, neochlorogenic acid, and caffeic acid) and 11 key astringent compounds (e.g., epigallocatechin gallate, gallic acid, chlorogenic acid, ellagic acid) were identified. After fermentation, flavonoid glycosides and flavanols were reduced by 62.41 % to 97.46 %, while phenolic acids showed varied trends. Different rates of change in key compounds during fermentation resulted in variations in bitterness and astringency. This study offers insights for improving Liupao tea quality.

Integrated Metabolome and Transcriptome Analysis Reveals the Key Bitter Substances of Pleioblastus amarus Shoots at Different Developmental Stages

Pleioblastus amarus (bitter bamboo) shoots are a traditional forest vegetable in China, renowned for its nutritional composition and associated health benefits. This study aimed to investigate the dynamic changes in nutritional quality and bitterness-related metabolites, as well as corresponding gene expression in P. amarus shoots at three distinct growth stages, specifically at heights of 10 cm (BT1), 20 cm (BT2), and 40 cm (BT3). The results showed that the content of soluble sugars decreased while the content of total phenols and flavonoids, which contribute to bitterness, increased during the growth of P. amarus shoots. In addition, comparative analysis of BT2 vs. BT1, BT3 vs. BT1, and BT3 vs. BT2 revealed 14, 43, and 36, respectively, differentially accumulated metabolites (DAMs). Notably, BT2 exhibited upregulation of flavonoids, such as apigenin, while bitter amino acids, including phenylalanine and arginine, showed a marked increase in BT3. Moreover, differential gene expression analysis revealed 3161, 13268, and 3742 differentially expressed genes (DEGs) in the BT2 vs. BT1, BT3 vs. BT1, and BT3 vs. BT2 comparisons, respectively. An integrated analysis of metabolomic and transcriptomic data indicated that the metabolites and genes associated with the biosynthesis pathways of phenylalanine, tyrosine, tryptophan, arginine, and the branched-chain amino acids valine, leucine, and isoleucine were significantly enriched during the growth of P. amarus shoots. The study indicates that early-stage (≤20 cm) P. amarus shoots are mainly bitter due to flavonoids like apigenin, while the bitterness in later-stage (around 40 cm) shoots is largely due to bitter amino acids such as phenylalanine, arginine, valine, and isoleucine. This research highlights key compounds and genes related to bitterness, providing a basis for future research on enhancing the nutritional value and flavor of P. amarus shoots.

Ambient temperature-dependent variations in bitter compounds and microbial dynamics in traditional huangjiu fermentation

Huangjiu, a traditional Chinese fermented wine, often develops excessive bitterness under semi-open fermentation conditions. This study examines the impact of ambient temperature on bitterness and microbial composition during Huangjiu fermentation. Using high−performance liquid chromatography (HPLC), gas chromatography−olfactometry (GC−O), and metagenomic sequencing, we identified seven bitter amino acids and eleven bitter volatile substances. Our results indicate that lower ambient temperatures (8.21 ± 0.69 °C to 9.88 ± 0.83 °C) suppress the accumulation of bitter substances, with the total concentration of bitter amino acids reduced by up to 3.94% compared to higher temperatures (16.36 ± 0.97 °C to 4.75 ± 0.74 °C). Partial least squares (PLS) regression established correlations between electronic tongue bitterness values and the presence of bitter compounds, revealing that higher temperatures promote the formation of specific compounds, such as benzaldehyde, furfural, ethyl nonanoate, and phenylalanine, thereby enhancing bitterness perception. Metabolic pathway analysis highlighted the enzymatic processes and intermediates involved in the biosynthesis of these bitter compounds. Correlation analysis showed that Saccharomyces and Lactobacillus are closely linked to the synthesis of bitter amino acids, while Aspergillus and Rhizopus contribute to the formation of furfural and ethyl octanoate. This study offers a theoretical foundation for reducing flavor variations between Huangjiu batches and provides practical guidance for optimizing the traditional fermentation process.

Effects of single fermentation of Lactobacillus sakei and compound fermentation with Staphylococcus carnosus on the metabolomics of beef sausages

To illustrate the mechnism of the better flavor and color in combined fermented sausages than single fermentation with L. sakei, the growth behavior, pH, and metabolomics of L. sakei in single fermentation and in combination with S. carnosus at 0, 3, 6, 12, 24, and 48 h were studied, and the sensory evaluation of fermented beef sausage was conducted. Through KEGG topology analysis found that L. sakei is related to caffeine metabolism and citrate cycle, L. sakei and S. carnosus are related to metabolism of purine metabolism, caffeine metabolism, and alanine, aspartate and glutamate metabolism. Compared with L. sakei fermentation alone, the compound fermentation with S. carnosus increased the content of asparagine. The content of the bitter substance tyrosine decreased during the compound fermentation. As starter cultures for the L. sakei applied to provide a basis for the fermented beef sausage.

Potentiometric Electronic Tongue for the Evaluation of Multiple-Unit Pellet Sprinkle Formulations of Rosuvastatin Calcium.

Sprinkle formulations represent an interesting genre of medicinal products. A frequent problem, however, is the need to mask the unpleasant taste of these drug substances. In the present work, we propose the use of a novel sensor array based on solid-state ion-selective electrodes to evaluate the taste-masking efficiency of rosuvastatin (ROS) sprinkle formulations. Eight Multiple Unit Pellet Systems (MUPSs) were analyzed at two different doses (API_50) and (API_10), as well as pure Active Pharmaceutical Ingredient (API) as a bitter standard. Calcium phosphate-based starter pellets were coated with the mixture containing rosuvastatin. Some of them were additionally coated with hydroxypropyl methylcellulose, which was intended to separate the bitter substance and prevent it from coming into contact with the taste buds. The sensor array consisted of 16 prepared sensors with a polymer membrane that had a different selectivity towards rosuvastatin calcium. The main analytical parameters (sensitivity, selectivity, response time, pH dependence of potential, drift of potential, lifetime) of the constructed ion-selective electrodes sensitive for rosuvastatin were determined. The signals from the sensors array recorded during the experiments were processed using Principal Component Analysis (PCA). The results obtained, i.e., the chemical images of the pharmaceutical samples, indicated that the electronic tongue composed of the developed solid-state electrodes provided respective attributes as sensor signals, enabling both of various kinds of ROS pellets to be distinguished and their similarity to ROS bitterness standards to be tested.

Modelling and nondestructive quality prediction of Guanxi Honey Red Pomelo (<i>Citrus grandis</i> (L.) Osbeck) based on shape characteristics and bitterness

Currently, it is difficult to distinguish the inner quality of pachycarpous fruits using nondestructive instruments, and trace nutrients and flavor compounds are rarely considered in fruit sorting. In this study, the external features and internal quality indices of an extensively farmed citrus fruit, the Guanxi honey red pomelo, were measured. The correlation between the inner and external quality was analyzed using Pearson correlation, canonical correlation, and grey correlation analyses. This research innovatively introduced bitter substances and revealed that the height-diameter ratio could serve as a predictor for ultra-size fruits, which partially reflects inner quality indices. Several grey formulas were developed to express these correlations and were fitted to a matrix, which demonstrated accuracies at all level 1 (C < 0.35, P > 0.95). Based on the matrix, fruit quality analysis of white and red pomelo showed high prediction accuracy in TSS and bitter substance indices (R2 = 0.6 – 0.8) and indicated that mature and symmetrical pomelo (height-diameter ratio of 0.9 – 1.1) exhibit a similar quality pattern. This study achieved rapid quality assessment using easy measurable shape characteristics and explored the application of characteristic bitter substances in the quality evaluation model, providing theoretical references and potential application guidance for pachycarpous fruit sorting, as well as introducing a new idea to incorporating characteristic quality into modeling index.

Investigating the effects of thermal processing on bitter substances in atemoya (Annona cherimola × Annona squamosa) through sensory-guided separation

Atemoya (Annona cherimola × Annona squamosa) is a specialty crop in Taiwan. Thermal treatment induces bitterness, complicating seasonal production adjustments and surplus reduction. In this research, sensory-guided separation, metabolomics, and orthogonal partial least squares discrimination analysis (OPLS-DA) are used for identifying the bitterness in atemoya which originates from catechins, epicatechin trimers, and proanthocyanidins. Different thermal treatments (65 °C, 75 °C, and 85 °C) revealed that the glucose and fructose contents in atemoya significantly decreased, while total phenols, flavonoids, and tannins significantly increased. The concentration of 5-hydroxymethylfurfural (5-HMF) increased from 23.16 ng/g in untreated samples to 400.71 ng/g (AP-65), 1208.59 ng/g (AP-75), and 2838.51 ng/g (AP-85). However, these levels are below the 5-HMF bitterness threshold of 3780 ng/g. Combining mass spectrometry analysis with sensory evaluation, OPLS-DA revealed that atemoya treated at 65 °C, 75 °C, and 85 °C exhibited significant bitterness, with the main bitter components being proanthocyanidin dimers and trimers.

Insights into “Yin Rhyme”: Analysis of nonvolatile components in Tieguanyin oolong tea during the manufacturing process

Tieguanyin (TGY) is renowned for its distinctive “Yin Rhyme” flavor. To elucidate the underlying formation mechanism, we conducted sensory evaluations, electronic tongue analysis, and widely-targeted metabolomics. Our sensory evaluations and electronic tongue results indicated that TGY exhibits a thick and mellow taste profile, contributing to the “Yin Rhyme” flavor. Metabolomics analysis of tea products revealed that TGY shows significantly higher concentrations of umami substances (L-glutamate, L-theanine) and bitter substances (valine, catechins) compared to Jinguanyin (JGY). Additionally, metabolomic analysis during different oolong tea processing stages revealed significant differences in 21 substances, including L-glutamate, L-theanine, valine, and catechins, between fresh leaves of both varieties. These substances exhibited distinct fluctuation patterns during processing, indicating that the cultivar plays a crucial role in developing the “Yin Rhyme” flavor, which was enhanced throughout processing. This study provides a theoretical foundation for understanding the formation of the unique “Yin Rhyme” flavor of TGY.

Decoding the bitter taste of Idesia polycarpa var. vestita Diels fruit: Bitterness contribution and mechanisms

To comprehensively explore the contribution and mechanisms of identified low-threshold bitter substances in Idesia polycarpa var. vestita Diels (I. vestita) fruit, we performed quantification and elucidated their interactions with main bitter taste receptors through molecular docking. The established method for quantifying bitter compounds in I. vestita fruit was validated, yielding satisfactory parameters for linearity, stability, and accuracy. Idescarpin (17.71–101.05 mg/g) and idesin (7.88–77.14 mg/g) were the predominant bitter compounds in terms of content. Taste activity values (TAVs) exceeded 10 for the bitter substances, affirming their pivotal role as major contributors to overall bitterness of I. vestita fruit. Notably, idescarpin with the highest TAV, played a crucial role in generating the bitterness of I. vestita fruit. Hydrogen bonds and hydrophobic interactions were the main driving forces. This study holds potential implications for industrial development of I. vestita fruit by providing novel insights into the mechanism underlying its bitterness formation.

The quality characteristics and microbial communities of three components in traditional split-fermented red sour soup.

Red sour soup is a Guizhou specialty condiment made by the natural fermentation of tomatoes and chili. In this study, three components (tomato acid, chili acid, and tomato and chili mixed acid) of split-fermented red sour soup were explored to compare the quality characteristics and microbial communities in the middle and late fermentation stages. The titratable acidity of mixed acids was lower than that of tomato acid and chili acid in the middle stage, but it was significantly increased in the late stage. The cell viability of lactic acid bacteria was mostly higher than that of yeasts during the whole fermentation. Also significantly increased in the late stage of fermentation were sensory scores and the signal intensity of sour substances. However, the signal intensity of both bitter and salty substances decreased, and the total amount of free amino acids was reduced. In addition, the antioxidant capacity of the samples and the dominant microorganisms were different between the two fermentation stages, Lactobacillus and Kazachstania were the key common genus of the different components of split-fermented red sour soup. It is anticipated that this study would provide us an insight into the quality characteristics and microbial communities of split-fermented red sour soup.

Advancing investigations into the effects of sweet and bitter tastes on agreeableness and hostility

In the studies to date that have examined the effect of taste on social affect, sweet taste has been reported to increase agreeableness (Meier et al., 2012), and bitter taste has been reported to increase hostility (Sagioglou & Greitemeyer, 2014). However, no study to date has examined changes to both agreeableness and hostility after tasting sweet and bitter substances, nor how the order of self-reporting these feelings, or individual differences in taste sensitivity, may influence the results. Our pre-registered study addressed these questions in 99 female undergraduates. Participants were randomly assigned to taste one of six sweet, bitter and neutral tastants, including those used in prior research on this topic, and then immediately completed the agreeableness and hostility scales used in past research, the order of which was counterbalanced across participants. Participants then evaluated all six tastants on various hedonic scales, and a 6-n-propylthiouracil (PROP) test of taste sensitivity was administered. Statistical analyses did not reveal any association between sweet, or any tastant and agreeableness scores. Hostility ratings were increased after participants tasted bitter, but only when this scale was administered before the agreeableness scale. Sensitivity to PROP did not moderate any of the findings. Based on prior work on self-affirmation and affect (e.g., Lindsay & Creswell, 2014), our results suggest that priming a prosocial self-schema (e.g., agreeableness) can eliminate the hostility induced by bitter taste. However, other explanations are possible and future work should investigate factors including measures of personality states vs. traits, and the timing between tasting and affect assessments.

Research on bitter taste and efficacy of Ginkgo biloba extract based on UPLC-Q-TOF-MS~E integrated with molecular docking

Based on high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS~E) and molecular docking technique, bitter compounds of Ginkgo biloba extract(GBE) were characterized, and their relationship with bitter efficacy was investigated. Firstly, UPLC-Q-TOF-MS~E was used for qualitative analysis of GBE components, and 60 chemical components were identified. These chemical components were molecular-docked with bitter receptors, and 26 bitter substances were selected, mainly flavonoids. Secondly, sensory and electronic tongue bitterness evaluation techniques were used to verify that total flavones of GBE were the main bitter substances, which was consistent with the molecular docking results. Finally, network pharmacology was used to predict and analyze bitter substances. The relationship between the target of bitter substance and bitter effect was explored. The key targets of bitter substances are CYP2B6, ALOX15, and PTGS2, etc., and bitter substances may exert a bitter efficacy by ac-ting on related disease targets, indicating that bitter substances of GBE are the material basis of the bitter effect. In summary, the study indicated that the molecular docking technique had a guiding effect on the screening of bitter substances in traditianal Chinese medicine(TCM), and bitter substances of GBE had a bitter efficacy. It provides ideas and references for the study of the "taste-efficacy relationship" of TCM in the future.

Assessment of quality differences between wild and cultivated fruits of Rosa roxburghii Tratt

Cultivated and wild Rosa roxburghii Tratt were investigated to determine the intrinsic causes of the quality differences. The cultivated fruit, Guinong No. 5 (GNNF) was larger, heavier, and had less color difference. Its juice contained higher levels of soluble solids and tannins. The wild variety (WRRT) had higher concentrations of condensed tannins, polyphenols and flavonoids. Artificial sensory and electronic taste tests indicated GNNF had a lower astringent and bitter taste compared to WRRT. A total of 80 flavour substances were identified by gas chromatography‒mass spectrometry (GC-MS). The relative contents of esters, organic acids, ketones, alcohols, aldehydes and aromatic hydrocarbons were higher in GNNF than in WRRT. Caffeylalcohol, catechin, quinidine, pantothenic acid and five bitter amino acids were high in GNNF. The high contents of bitter substances such as limonin, daidzein, and rutaevin were responsible for the quality differences between WRRT and GNNF.

Structure revision of tricholomenyn B, an antimitotic geranylcyclohexenone rediscovered as a bitter and antibacterial substance, from a basidiomycete Tricholoma japonicum (Shiro-shimeji).

Tricholomenyn B, an antimitotic geranylcyclohexenone originally discovered from a basidiomycete Tricholoma acerbum, was isolated as a bitter and antibacterial constituent from fruiting bodies of T. japonicum. Careful comparison of NMR, MS, and other physicochemical properties of the isolated substance with the literature values revised a previously proposed macrolide structure 1 to a macrodiolide 2. Compound 2 was perceived bitter at a minimum dose of 37.5 μg, showed weak antimicrobial activity against Kocuria rhizophila and Staphylococcus aureus, and was marginally cytotoxic (IC50 2.6 µM) against P388 murine leukemia cells.

Assessment of Bitterness in Non-Charged Pharmaceuticals with a Taste Sensor: A Study on Substances with Xanthine Scaffold and Allopurinol.

Taste sensors with an allostery approach have been studied to detect non-charged bitter substances, such as xanthine derivatives, used in foods (e.g., caffeine) or pharmaceuticals (e.g., etofylline). In this study, the authors modified a taste sensor with 3-bromo-2,6-dihydroxybenzoic acid and used it in conjunction with sensory tests to assess the bitterness of non-charged pharmaceuticals with xanthine scaffolds (i.e., acefylline and doxofylline), as well as allopurinol, an analogue of hypoxanthine. The results show that the sensor was able to differentiate between different levels of sample bitterness. For instance, when assessing a 30 mM sample solution, the sensor response to acefylline was 34.24 mV, which corresponded to the highest level of bitterness (τ = 3.50), while the response to allopurinol was lowest at 2.72 mV, corresponding to relatively weaker bitterness (τ = 0.50). Additionally, this study extended the application of the sensor to detect pentoxifylline, an active pharmaceutical ingredient in pediatric medicines. These results underscore the taste sensor's value as an additional tool for early-stage assessment and prediction of bitterness in non-charged pharmaceuticals.

Bitter taste TAS2R14 activation by intracellular tastants and cholesterol.

Bitter taste receptors, particularly TAS2R14, play central roles in discerning a wide array of bitter substances, ranging from dietary components to pharmaceutical agents1,2. TAS2R14 is also widely expressed in extragustatory tissues, suggesting its extra roles in diverse physiological processes and potential therapeutic applications3. Here we present cryogenicelectron microscopy structures of TAS2R14 in complex with aristolochic acid, flufenamic acid and compound 28.1, coupling with different G-protein subtypes. Uniquely, a cholesterol molecule is observed occupying what is typically an orthosteric site in class A G-protein-coupled receptors. The three potent agonists bind, individually, to the intracellular pockets, suggesting a distinct activation mechanism for this receptor. Comprehensive structural analysis, combined with mutagenesis and molecular dynamic simulation studies, elucidate the broad-spectrum ligand recognition and activation of the receptor by means of intricate multiple ligand-binding sites. Our study also uncovers the specific coupling modes of TAS2R14 with gustducin and Gi1 proteins. These findings should be instrumental in advancing knowledge of bitter taste perception and its broader implications in sensory biology and drug discovery.

Membrane-bound chemoreception of bitter bile acids and peptides is mediated by the same subset of bitter taste receptors

The vertebrate sense of taste allows rapid assessment of the nutritional quality and potential presence of harmful substances prior to ingestion. Among the five basic taste qualities, salty, sour, sweet, umami, and bitter, bitterness is associated with the presence of putative toxic substances and elicits rejection behaviors in a wide range of animals including humans. However, not all bitter substances are harmful, some are thought to be health-beneficial and nutritious. Among those compound classes that elicit a bitter taste although being non-toxic and partly even essential for humans are bitter peptides and l-amino acids. Using functional heterologous expression assays, we observed that the 5 dominant human bitter taste receptors responsive to bitter peptides and amino acids are activated by bile acids, which are notorious for their extreme bitterness. We further demonstrate that the cross-reactivity of bitter taste receptors for these two different compound classes is evolutionary conserved and can be traced back to the amphibian lineage. Moreover, we show that the cross-detection by some receptors relies on “structural mimicry” between the very bitter peptide l-Trp-Trp-Trp and bile acids, whereas other receptors exhibit a phylogenetic conservation of this trait. As some bile acid-sensitive bitter taste receptor genes fulfill dual-roles in gustatory and non-gustatory systems, we suggest that the phylogenetic conservation of the rather surprising cross-detection of the two substance classes could rely on a gene-sharing-like mechanism in which the non-gustatory function accounts for the bitter taste response to amino acids and peptides.

A New Culture Medium Rich in Phenols Used for Screening Bitter Degrading Strains of Lactic Acid Bacteria to Employ in Table Olive Production.

The olive oil industry recently introduced a novel multi-phase decanter with the "Leopard DMF" series, which gives a by-product called pâté, made up of pulp and olive wastewater with a high content of phenolic substances and without pits. This study aims to create a new culture medium, the Olive Juice Broth (OJB), from DMF pâté, and apply it to select bacteria strains able to survive and degrade the bitter substances normally present in the olive fruit. Thirty-five different bacterial strains of Lactiplantibacillus plantarum from the CREA-IT.PE Collection of Microorganisms were tested. Seven strains characterized by ≥50% growth in OJB (B31, B137, B28, B39, B124, B130, and B51) showed a degradation of the total phenolic content of OJB ≥ 30%. From this set, L. plantarum B51 strain was selected as a starter for table olive production vs. spontaneous fermentation. The selected inoculant effectively reduced the debittering time compared to spontaneous fermentation. Hydroxytyrosol, derived from oleuropein and verbascoside degradation, and tyrosol, derived from ligstroside degradation, were produced faster than during spontaneous fermentation. The OJB medium is confirmed to be useful in selecting bacterial strains resistant to the complex phenolic environment of the olive fruit.

Discovery of bitter masking compounds from Allspice (Pimenta dioica) using sensory guided isolation

Bitter substances in functional foods and beverages can act as nutraceuticals, offering potential health benefits. However, their unpleasant sensory impact reduces the consumption of these foods. Consequently, the discovery of bitter masking compounds is crucial for enhancing the intake of bioactive compounds in functional foods and beverages. Bitter taste is mediated by TAS2Rs, a sub-family of G-protein-coupled receptors. TAS2R14 is especially pivotal in the perception of bitterness, as it is one of the most broadly tuned bitter receptors. In this study, allspice was extracted and purified to yield five single compounds based on sensory guided fractionation. The structures of each compound were determined based on nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HR-MS). In a sensory evaluation, compound 1 exhibited bitter masking activity against quinine. Molecular docking analysis revealed that compound 1 could act as an antagonist of the TAS2R14 bitter receptor.