Trichloroacetic Acid Extracts Research Articles

Polar metabolomics using trichloroacetic acid extraction and porous graphitic carbon stationary phase

IntroductionAccurately identifying and quantifying polar metabolites using untargeted metabolomics has proven challenging in comparison to mid to non-polar metabolites. Hydrophilic interaction chromatography and gas chromatography–mass spectrometry are predominantly used to target polar metabolites.ObjectivesThis study aims to demonstrate a simple one-step extraction combined with liquid chromatography–mass spectrometry (LC–MS) that reliably retains polar metabolites.MethodsThe method involves a MilliQ + 10% trichloroacetic acid extraction from 6 healthy individuals serum, combined with porous graphitic carbon liquid chromatography–mass spectrometry (LC–MS). The coefficient of variation (CV) assessed retention reliability of polar metabolites with logP as low as − 9. QreSS (Quantification, Retention, and System Suitability) internal standards determined the method's consistency and recovery efficiency.ResultsThe method demonstrated reliable retention (CV < 0.30) of polar metabolites within a logP range of − 9.1 to 5.6. QreSS internal standards confirmed consistent performance (CV < 0.16) and effective recovery (70–130%) of polar to mid-polar metabolites. Quality control dilution series demonstrated that ~ 80% of annotated metabolites could be accurately quantified (Pearson’s correlation coefficient > 0.80) within their concentration range. Repeatability was demonstrated through clustering of repeated extractions from a single sample.ConclusionThis LC–MS method is better suited to covering the polar segment of the metabolome than current methods, offering a reliable and efficient approach for accurate quantification of polar metabolites in untargeted metabolomics.

A Simple and Rapid Analysis Method for Nonvolatile Amines via LC-MS/MS

In order to quickly analyze 8 types of nonvolatile amine, such as histamine, a simple analytical method was developed. A test solution was prepared only by diluting and filtering a trichloroacetic acid extract before analysis via LC-MS/MS.As a result of the additive recovery test with 11 types of food, including fresh seafood, seafood processed products, and other processed foods, all amines had an accuracy in the range of 70 to 120% with a repeatability of less than 15% RSD in 9 types of food. This confirmed the validity of the analytical method with the lower limit of quantification between 5 to 6 mg/kg.

Development of a rapid process for purification of Bowman-Birk and Kunitz inhibitors from legume seeds, and evaluation of their biophysical, insecticidal, and antimicrobial properties

Bowman-Birk inhibitor (BBI ~10 kDa) and Kunitz inhibitor (KI ~20 kDa) are serine protease/proteinase inhibitor(s) [PI(s)] ubiquitously found in several Leguminous plant species with insecticidal and therapeutic properties. Due to narrow molecular mass differences, the separation of these inhibitors from a single seed variety is tedious. The present study is aimed to develop a rapid protocol (<24 h) for purifying BBI and KI from legume seeds using mild trichloroacetic acid (TCA) extraction followed by trypsin-affinity chromatography. The mature seeds of Vigna radiata and Cajanus platycarpus are used as a model to purify BBI and KI using this protocol. The BBI and KI purified from the seeds of V. radiata are labeled as VrBBI & VrKI, and C. platycarpus are labeled as CpBBI & CpKI, respectively. These PIs are confirmed by immunodetection and MALDI-TOF studies and further characterized for their structural (CD & fluorescence spectroscopy) and functional properties (temperature & DTT stability). BBI(s) purified using the above process are effective in the management of castor semi-looper ‘Achaea janata’, while KI(s) are effective in the management of pod borer ‘Helicoverpa armigera’. Besides, both BBI(s) and KI(s) have significant potential in controlling the growth of methicillin-sensitive ‘Staphylococcus aureus’, a gram-positive pathogenic bacterium.

UCP-LF and other assay methods for schistosome circulating anodic antigen between 1978 and 2022.

Detection of circulating anodic antigen (CAA) is known for its high sensitivity in diagnosing schistosomiasis infection, even in low-prevalence settings. The Up-Converting Phosphor-Lateral Flow (UCP-LF) assay developed in 2008 presented greater sensitivity than other assay methods in use for CAA detection. Our study aims to comprehensively review all studies conducted in this area and thus generate informed conclusions on the potential for adopting the UCP-LF assay for diagnosing this important yet neglected tropical disease. Using the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) guidelines, we generated search criteria to capture all studies in English journals available in the Scopus and PubMed databases on 20 December 2022. A total of 219 articles were identified, and 84 that met the inclusion criteria were retrieved and eventually included in the study. Twelve different assay methods were identified with a noteworthy transition from enzyme-linked immunosorbent assay (ELISA) to the UCP-LF assay, a laboratory-based assay that may be applicable as a point-of-care (POC) diagnostic test for schistosomiasis. Reducing the time, cost, and dependence on specialized laboratory skills and equipment, especially relating to the trichloroacetic acid extraction step and centrifugation in the UCP-LF CAA assay may go a long way to aid its potential as a POC tool. We also propose the development of a CAA-specific aptamer (short protein/antigen-binding oligonucleotide) as a possible alternative to monoclonal antibodies in the assay. UCP-LF has great potential for POC application.

The Effects of Different Processing Methods on the Levels of Biogenic Amines in Zijuan Tea.

This study aimed to evaluate the effects of processing methods on the content of biogenic amines in Zijuan tea by using derivatization and hot trichloroacetic acid extraction with HPLC-UV. The results showed that the most abundant biogenic amine in the original leaves was butylamine, followed by ethylamine, methylamine, 1,7-diaminoheptane, histamine, tyramine, and 2-phenethylamine. However, during the process of producing green tea, white tea, and black tea, the content of ethylamine increased sharply, which directly led to their total contents of biogenic amines increasing by 184.4%, 169.3%, and 178.7% compared with that of the original leaves, respectively. Unexpectedly, the contents of methylamine, ethylamine, butylamine, and tyramine in dark tea were significantly reduced compared with those of the original leaves. Accordingly, the total content of biogenic amines in dark tea was only 161.19 μg/g, a reduction of 47.2% compared with that of the original leaves, indicating that the pile-fermentation process could significantly degrade the biogenic amines present in dark tea.

Comparative study of technologies for extraction of biologically active substances from the raw material of animal origin

Technologies of isolation and concentration of biologically active substances, developed in the middle of the 20th century, need adjustment and adaptation to modern conditions both to increase the activity of substances and for greater economic efficiency. The aim of the research is the comparison of dynamics of biologically active compounds extraction from porcines pancreas in two methods: the saline method based on 0.9% sodium chloride solution, and the acidic method based on 2.4% trichloroacetic acid solution. Also the purpose of research is to assess the possibilities for further optimization of technologies. The total protein concentration based on the biuret reaction in the samples taken during the extraction, as well as the calculation and analysis of the point degrees and rates of extraction are chosen as the controlled parameters. Local maxima of the protein yields into the extractant media at the 60th, 135th and 255th minute were recorded during saline extraction; and at the 75th and 135th minute during acid extraction. Also the proteomic profile of the extracts was studied. Wide range of compounds with molecular weight of less than 52 kDa was found in extracts based on physiological saline solution, and protein substances of whole presented range of molecular weights in trichloroacetic acid based extracts were considered. The predominance of low molecular weight protein fraction of interest was noted also in this method of extraction in comparison with the other methods of extraction. According to the UniProt database, we assume availability of probable compounds with a molecular weight of less than 30 kDa in the purified acidic extract. The presence of some proteins absent in the final saline extract was noted. The acidic erythrograms showed a weak degrading effect of both types of extracts on the membranes of rat erythrocytes, as well as the cytoprotective effect of acidic ultrafiltrates (less than 3 kDa). The obtained results prove a better efficiency of trichloroacetic acid extraction method used for obtaining a mixture of a wide range of compounds, including biologically active substances of low molecular weight.

Use of Rapid Capillary Zone Electrophoresis to Determine Amino Acids Indicators of Herring Ripening during Salting.

Currently, herring fillets are salted with acetic acid to activate muscle proteases. This causes a change in the composition of free amino acids, compared to salting of whole fish with viscera proteases. Therefore, old indicators of the ripening dynamics of salted fish based on amino acids are not current. Determination of free amino acids can be performed by many methods, but most are labor intensive and expensive. Therefore, a capillary electrophoresis method without derivatization (CZE) was used to determine the actual ripening rates of salted herring fillets. A group of hydrophobic and basic amino acids were determined in trichloroacetic acid (TCA) extracts of meat and brine to develop 16 indicators. Statistical regression analysis of the indicators (R2adj, RMSE, cluster analysis) followed by principal component analysis (PCA) correlation analysis of the indicators vs sensory evaluation parameters of texture and TPA-hardness of salted fillet meat allowed the choice of the most precise indicators. The best indicator in meat was Phe/Tyr-height, which value increased during salting. A more precise indicator of ripening was His/Tyr-height in brine, which value decreased during salting. Sensory evaluation parameters of salted herring texture correlated strongly with TPA-hardness and traditional indicators such as non-protein nitrogen and protein hydrolysis product fraction content. However, the most precise indicators were those obtained from amino acids determined by the CZE method. Results obtained in this study may be suitable for fast monitoring of the salted herring ripening process in industry.

Early diagenetic sequestration of microbial mat lipid biomarkers through covalent binding into insoluble macromolecular organic matter (IMOM) as revealed by sequential chemolysis and catalytic hydropyrolysis

Analyses of lipids in modern microbial mat ecosystems provide valuable insights into the taxonomic affinities of the dominant biological producers and consumers as well as carbon flow within the mat community. Such microbial mat biomarker assemblages also provide a window into the ancient biomarker record, facilitating paleobiological and paleoenvironmental reconstructions of analogous ecosystems on early Earth. Our focus was to investigate the covalent binding of lipid biomarkers into insoluble macromolecular organic matter (IMOM) during diagenesis in the well-studied hypersaline microbial mat ecosystems and underlying sediments at Guerrero Negro, Baja California Sur, Mexico. Following exhaustive solvent extraction, three different chemical degradation reagents of varying reactivity (acid methanolysis, trichloroacetic acid extraction, and periodate oxidation) were applied to two shallow sedimentary layers prior to fragmentation of the recalcitrant insoluble residues using catalytic hydropyrolysis (HyPy). The hydrocarbons released by HyPy included a complex variety of linear, branched and polycyclic alkane structures, including hopanes, methylhopanes and steranes. Our findings indicate that a significant fraction of the bound biomarker pool was strongly linked by covalent binding through functional groups into IMOM but that a portion of this bound pool was not recoverable by chemolysis treatment. These results provide new insights about the mode and timing of chemical binding of steroids and hopanoids, and their early diagenetic transformation products, into IMOM. The formation of sedimentary IMOM from the earliest stages of organic matter diagenesis, commencing on a timescale of only years, aids the long-term preservation of biomarker lipids in the geological record.

Sulfonamide Residues: Honey Quality in the Czech Market

In this study, we determined four sulfonamide compounds (sulfadiazine, sulfadoxine, sulfamethoxazole, and sulfathiazole) in honey marketed in the Czech Republic (Brno). The content of these compounds was monitored in 50 honeys with different botanical and geographical origin. Preanalytical treatment included acidic hydrolysis with 10% trichloroacetic acid (TCA) and double extraction with acetonitrile and dichloromethane. Chromatographic analysis was performed by ultraperformance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) using a triple quadrupole (QqQ) in positive ionization mode (ESI+). For separation, the mobile phase was a 0.05% aqueous solution of formic acid (A) and 0.05% formic acid in methanol (B) with a gradient elution of 0–3 minutes: 10% B; 3–3.6 min: 90% B; and 3.6–5 min: 10% B, flow rate 0.30 ml/min. Three positive samples (6%) were found containing sulfadiazine (232.88 and 618.87 μg/kg) and sulfamethoxazole (62.64 μg/kg). According to the country of origin, two of these honeys were from European Union (EU) countries, and one was labeled as a blend of honey from EU and non-EU countries.

Preparation of high affinity antibody for ribavirin with new haptens and residue analysis in chicken muscle, eggs and duck muscle

ABSTRACTIn this work, high affinity polyclonal antibodies for ribavirin (RBV) from new haptens were prepared and were used to analyse RBV residues in chicken muscle, eggs and duck muscle. The new haptens were synthesised with different spacers, and the best antibody was obtained with an IC50 value as low as 0.61 ng/mL in indirect competitive enzyme-linked immunosorbent assay (ELISA). The cross-reactivities with another five antiviral drugs including amantadine, rimantadine, moroxydine, zanamivir and oseltamivir were less than 0.1%, which indicated the good specificity of the antibody. An ELISA was developed based on the antibody and applied to detect RBV in multi-food matrices. The sample preparation prior to detection only needed simple dilution after trichloroacetic acid extraction. The limits of detection were 1.07, 1.18 and 1.03 μg/kg in chicken muscle, eggs and duck muscle, respectively. Recoveries ranged from 89.0% to 112.7% with coefficients of variation below 13.0%. Ten blind samples of chicken muscle were analysed simultaneously by ELISA and liquid chromatography-tandem mass spectrometry, and a good correlation between the methods was observed. The results indicated that the high affinity antibody could be applied for the simple and fast detection of RBV in multi-food matrices.

Quality changes of salmon by-products during storage: Assessment and quantification by NMR

Safe utilization of fish by-products is an important task due to increasing fish consumption. It can provide new valuable food/feed and will increase the economical profit and sustainability of the fishery industry. NMR spectroscopy is a reliable tool able to monitor qualitative and quantitative changes in by-products. In this work the trichloroacetic acid extracts of salmon backbones, heads and viscera stored at industrially relevant temperatures (4 and 10°C) were studied using NMR. Twenty-five metabolites were detected and the possibility of salmon by-products utilization as a source of anserine, phosphocreatine and taurine was discussed. Statistical data elaboration allowed determining the main processes occurring during by-products storage: formation of trimethylamine and biogenic amines, proteolysis and different types of fermentations. By-products freshness was evaluated using a multi-parameter approach: the trimethylamine and biogenic amines concentration changes were compared with Ki and H-values and safe temperatures and times for storage of salmon by-products were proposed.

Metabolite release and protein hydrolysis during the in vitro digestion of cooked sea bass fillets. A study by 1H NMR

Proton Nuclear Magnetic Resonance (1H NMR) was used to study micro-wave cooked European sea bass (Dicentrarchus labrax) fillets, along five different points of their in vitro digestion: after oral digestion, after gastric digestion and subsequent increasing of the pH to 5.0, and after 90 and 180min of duodenal digestion. During the gastric phase, the appearance of broad signals in different spectral regions, typical of amino acid protons, provided evidence for the expected release of unstructured fragments of denatured proteins, whilst smaller peptides, with sharper signals in the same spectral regions, were becoming visible only after duodenal digestion. Prior digestion, the spectral features of trichloroacetic acid extracts, from raw and cooked fish fillets, have been analysed to assess whether a significant protein hydrolysis or metabolite release (e.g., amino acids, organic acids, saccharides, and nucleotide derivatives) already occurred during cooking, by comparing the spectral profiles of these extracts. No significant differences among the molecular profile of both extracts were found. The present study demonstrates the suitability of NMR spectroscopy to follow the kinetics of fish fillets digestion under in vitro conditions simulating human digestion.

English

Genomic resources have just started to focus on the faba bean; the genes for synthesis of vicine and convicine (V-C) have not been determined, and recently developed genetic markers for these anti-nutritionals have not been used to examine these traits in very large scale in faba bean. Simple, rapid and cost-effective technologies are crucial in crop breeding programs, especially in the developing world, and in some cases, traditional methodologies are used in combination with genetic markers to assess agronomic traits and the value of gene markers. Here, two methodologies (TiCl4 assay and 274 nm absorption) are re-evaluated for their application in detection of V-C in faba beans. In comparison with TiCl4 assay, the method of 274 nm UV absorption without an HPLC analysis offers more reliable analysis for detection of V-C in immature and mature seeds of faba bean. Its application in high throughput screening by 60 min agitation of immature seeds or mature seed flour in 2% trichloroacetic acid (TCA) allows quick screening of low V-C faba beans. The level of V-C was maximum when seed moisture was 80% and V-C level was measured as 0.92% in CDC Fatima flour. Though V-C from 2% TCA extract of mature and immature seeds of CDC Fatima was detected by 274 wavelength in the TiCl4 assay reaction, a Ti-aglycone complex was not clearly detectible at 480 nm as previously suggested. Key words: Vicine, convicine, TiCl4 assay, UV assay, high throughput, seed, faba bean.

Improved proteomic analysis following trichloroacetic acid extraction of Bacillus anthracis spore proteins

Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Analysis of cellular proteins is dependent upon efficient extraction from bacterial samples, which can be challenging with increasing complexity and refractory characteristics. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrichment for certain classes of proteins. The method presented here is technically simple, does not require specialized equipment such as a mechanical disrupter, and is effective for protein extraction of the particularly challenging sample type of Bacillus anthracis Sterne spores. The ability of Trichloroacetic acid (TCA) extraction to isolate proteins from spores and enrich for spore-specific proteins was compared to the traditional mechanical disruption method of bead beating. TCA extraction improved the total average number of proteins identified within a sample as compared to bead beating (547 vs 495, respectively). Further, TCA extraction enriched for 270 spore proteins, including those typically identified by first isolating the spore coat and exosporium layers. Bead beating enriched for 156 spore proteins more typically identified from whole spore proteome analyses. The total average number of proteins identified was equal using TCA or bead beating for easily lysed samples, such as B. anthracis vegetative cells. As with all assays, supplemental methods such as implementation of an alternative preparation method may simplify sample preparation and provide additional insight to the protein biology of the organism being studied.

An efficient strategy for heterologous expression and purification of active peptide hainantoxin-IV.

Hainantoxin-IV (HNTX-IV) from the venom of the spider Selenocosmia hainana is a potent antagonist that specifically inhibits the tetrodotoxin-sensitive (TTX-S) sodium channels. The toxin peptide consists of 35 amino acids and adopts a typical inhibitory cystine knot (ICK) motif. To obtain adequate HNTX-IV peptides for further insight into the structure-activity relationships of the toxin, a novel strategy including cloning, expression and purification was developed in an E. coli expression system. For this purpose, a seamless restriction-free (RF) cloning method was employed for the construction of an expression vector to avoid introducing unwanted sequences into the target gene. Furthermore, the solubility of recombinant HNTX-IV could be promoted efficiently by the combination of a glutathione S-transferase (GST) tag and a small ubiquitin-related modifier (SUMO) tag. Finally, an affinity-chromatography-free purification strategy was developed by cut-off dialysis tubing combined with trichloroacetic acid (TCA) extraction. Further HPLC purification yielded recombinant, tag-free HNTX-IV with high yield and purity. The molecular weight of recombinant HNTX-IV (rHNTX-IV) is identical to its theoretical value according to Matrix-Assisted Laser Desorption / Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) analysis. The recombinant toxin has similar activity (IC50 value of 120 nM) on the tetrodotoxin-sensitive (TTX-S) sodium channels in adult rat dorsal root ganglion (DRG) neurons to native toxins. In the report, an efficient and cost-effective strategy for producing rHNTX-IV was developed, which paved the way for the further study of structure-activity relationships of rHNTX-IV and its pharmaceutical applications.

Seafood sold in Sweden contains BMAA: A study of free and total concentrations with UHPLC-MS/MS and dansyl chloride derivatization.

β-N-Methylamino-l-alanine (BMAA) is a potential neurotoxin associated with the aquatic environment. Validated analytical methods for the quantification of both free and total concentrations of BMAA were used in an investigation of seafood purchased from different grocery stores in Uppsala, Sweden. The analysis was performed using ultra high performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) and detection of BMAA as a dansyl derivate. The determined concentrations of free BMAA (after a simple trichloroacetic acid extraction) in mussels and scallops were up to 0.46μgg-1 wet homogenate. The total BMAA (after hydrochloric acid hydrolysis) levels were between 0.29 and 7.08μgg-1 wet mussel homogenate. The highest concentration of total BMAA was found in imported cooked and canned mussels which contained about ten times the quantity of BMAA measured in domestic cooked and frozen mussels. In this study it was also concluded that BMAA could be detected in seafood origin from four different continents. The risks associated with human exposure to BMAA through food are unknown today. However, the results of this study show that imported seafood in Sweden contain BMAA, indicating that this area needs more investigation, including a risk assessment regarding the consumption of e.g., mussels, scallops and crab.

Measurement of Evans blue dye concentration in central and peripheral tissue following administration in vitro and in vivo: dose‐ and replicate‐dependent effects on absorbance, fluorescence, and extraction by trichloroacetic acid (851.3)

The Evans blue dye (EBD) is the most commonly used inert tracer for tracking endogenous plasma protein extravasation into the brain. Even though several methods have been developed for measuring EBD concentration extravasated into central and peripheral tissues, the accuracy and precision of these methods remains poorly defined. First, we measured the absorbance and fluorescence of EBD solutions prepared in 96‐well plates using standard photo‐spectrometer. EBD absorbance increased in a dose‐dependent manner, and logarithm‐transformation revealed a sigmoidal curve with Hill coefficient of 2.086. The steepest slope of the curve lied between 5 µg/ml to 500 µg/ml, and hence this concentration range was the region at which measurements would be most precise. On the other hand, EBD fluorescence also increased in a concentration‐dependent manner, but with region of precision from 0.2 µg/ml to at least 5 µg/ml. Nevertheless, substantial photo‐bleaching of EBD fluorescence was found with increasing concentrations. Thus, our results recommends the use of fluorescent measurements to define EBD concentrations between 0.2 µg/ml to 5 µg/ml, and the use of absorbance measurements to define EBD concentrations between 5µg/ml to 500 µg/ml. Second, we determined the efficiency at which EBD can be retrieved from plasma proteins using the trichloroacetic acid (TCA) extraction technique. The degree of extraction was substantial even with 1:1 plasma‐to‐TCA ratio, and reached maximum with 1:2 ratio where increased ratios failed to further increase EBD extraction. Notably, contrary to earlier reports, we failed to completely extract EBD from plasma sample even with substantial amount of TCA. Third, we determined the sample‐to‐TCA ratio required to maximally extract EBD from central and peripheral tissue in rats receiving intravenous EBD. Taken together, the data presented here will be of great value in guiding future research using EBD as a tool to study central and peripheral vascular permeability.Grant Funding Source: Supported by the National Science Council of Taiwan

A new simple fluorimetric method to assay cytosolic ATP content: application to durum wheat seedlings to assess modulation of mitochondrial potassium channel and uncoupling protein activity under hyperosmotic stress

AbstractThe assays commonly used to determine ATP content in biological samples generally measure total cellular ATP content, but not the different subcellular pools. In this study a new simple method for measuring ATP content in a cytosol-enriched fraction (CEF) was developed, based on a rapid cytosolic ATP extraction (by an isotonic grinding medium that preserves organelle integrity) and its detection monitoring the NADPH fluorescence generated via hexokinase/glucose-6-phosphate dehydrogenase coupled reactions. Four protocols, differing for timing of NADPH generation and for either the presence or absence of some inhibitors of ATP and NADPH metabolism, were compared by determining CEF-ATP, as well as total ATP, in durum wheat (Triticum durum Desf.) etiolated seedlings. The best protocol was the one adopting both simultaneous NADPH generation and use of inhibitors during tissue homogenization. This protocol also showed higher performance than the classical trichloroacetic acid extraction. Using the new method, CEF-ATP content was assessed in control, salt- and osmotic-stressed seedlings, resulting 2.68 ± 0.04, 1.69 ± 0.12 (−40%) and 1.35 ± 0.16 (−50%) μmol/g dry weight, respectively. Finally, the effects of this stress-dependent decrease of cytosolic ATP were evaluated with respect to a possible modulation of two mitochondrial energy-dissipating systems, the uncoupling protein (PUCP) and the K+ channel (PmitoKATP), both inhibited by cytosolic ATP. Experiments carried out at different physiological ATP concentrations suggest that the decreased cytosolic ATP content occurring under hyperosmotic stress may contribute to attenuate inhibition of PmitoKATP, thus promoting its activity (up to about 90%), but not of PUCP, that appears to lose ATP sensitivity under stress condition.

Comparison in taste and extractive components of boiled dorsal muscle and broth from half-smooth golden puffer Lagocephalus spadiceus caught in Japan with those of the same fish imported

The taste and extractive components of boiled dorsal muscle and broth prepared from half-smooth golden puffer Lagocephalus spadiceus caught in Japan and from those imported from China were compared. In the sensory test, the first taste, elasticity, and saltiness of boiled muscle from Japanese (domestic) fish were higher than those of the imported fish, whereas the orthonasal fishy smell of the imported fish was higher than that of the domestic fish. The first taste, aftertaste, retronasal fishy smell, sweetness, saltiness, and umami of the broth prepared from dorsal muscle of the domestic fish were higher than those of the imported fish, whereas orthonasal fishy smell and bitterness of the imported fish were higher than those of the domestic fish. Most panelists preferred the overall taste of the domestic fish to that of the imported fish. The concentration of trimethylamine in the trichloroacetic acid extracts from boiled muscle and broth of the imported fish was higher than that of the domestic fish, suggesting that this substance contributes to the orthonasal fishy smell of the imported fish.

Changes in the amino acid composition of Bogue (Boops boops) fish during storage at different temperatures by 1H-NMR spectroscopy.

Nuclear magnetic resonance spectroscopy was employed to obtain information about the changes occurring in Bogue (Boops boops) fish during storage. For this purpose, 1H-NMR spectra were recorded at 600 MHz on trichloroacetic acid extracts of fish flesh stored over a 15 days period both at 4 °C and on ice. Such spectra allowed the identification and quantification of amino acids, together with the main organic acids and alcohols. The concentration of acidic and basic free amino acids was generally found to increase and decrease during storage, respectively. These concentration changes were slow during the first days, as a consequence of protein autolysis, and at higher rates afterward, resulting from microbial development. Two of the amino acids that showed the greatest concentration change were alanine and glycine, known to have a key role in determining the individual taste of different fish species. The concentration of serine decreased during storage, as highlighted in the literature for frozen fish samples. Differences in the amino acids concentration trends were found to be related to the different storage temperatures from day 4 onwards.