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Dilution Analysis (dilution + analysis)
Kinds of Dilution Analysis Selected AbstractsEvaluation of Bitterness in Enzymatic Hydrolysates of Soy Protein Isolate by Taste Dilution AnalysisJOURNAL OF FOOD SCIENCE, Issue 1 2008W.H. Seo ABSTRACT:, Although enzymatic hydrolysates of soy protein isolate (SPI) have physiological functionality, partially hydrolyzed SPI exhibits bitter taste depending on proteases and degree of hydrolysis (DH). To determine proteolysis conditions for SPI, it is important to evaluate bitterness during enzymatic hydrolysis. Taste dilution analysis (TDA) has been developed for the screening technique of taste-active compounds in foods. The objectives of the present study were to evaluate bitterness of enzyme-hydrolyzed SPI by TDA and to compare bitterness of SPI hydrolysates with respect to kinds of proteases and DH. SPI was hydrolyzed at 50 °C and pH 6.8 to 7.1 to obtain various DH with commercial proteases (flavourzyme, alcalase, neutrase, protamex, papain, and bromelain) at E/S ratios of 0.5%, 1%, and 2%. The DH of enzymatic hydrolysates was measured by trinitrobenzenesulfonic acid method. The bitterness of enzymatic hydrolysates was evaluated by TDA, which is based on threshold detection in serially diluted samples. Taste dilution (TD) factor was defined as the dilution at which a taste difference between the diluted sample and 2 blanks could be detected. As DH increased, the bitterness increased for all proteases evaluated. Alcalase showed the highest TD factor at the same DH, followed by neutrase. Flavourzyme showed the lowest TD factor at the entire DH ranges. At the DH of 10%, TD factor of hydrolysate by flavourzyme was 0 whereas those by protamex and alcalase were 4 and 16, respectively. These results suggest that TDA could be applied for the alternative of bitterness evaluation to the hedonic scale sensory evaluation. [source] A longitudinal analysis of cytotoxic T lymphocyte precursor frequencies to the hepatitis B virus in chronically infected patientsJOURNAL OF VIRAL HEPATITIS, Issue 1 2001G. K. Sing Individuals with acute hepatitis B virus (HBV) infection characteristically mount a strong, multispecific cytotoxic T lymphocyte (CTL) response that is effective in eradicating virus. In contrast, this response in chronic carriers is usually weak or undetectable. Since it is generally acknowledged that HBV pathogenesis is immune-mediated, the occurrence of episodes of active liver disease in many carriers suggests that these individuals can mount active CTL responses to HBV. To see whether the detection of circulating CTLs is related to these flare episodes, we have determined the CTL precursor (CTLp) frequencies to HLA-A2-restricted viral peptides in seven patients over a 12,24-month period of their disease. Limiting dilution analyses (LDA) were performed longitudinally to five epitopes comprising the viral capsid (HBc), envelope (HBs) and polymerase (pol) proteins. Assays were performed against a mixture of peptides, or against each individual peptide, to measure overall CTL activity and the multispecificity of the responses, respectively. Since two of the patients were treated with recombinant human interleukin-12 (rHuIL-12) at the time, with one individual achieving complete disease remission a year later after being treated with interferon-,, we were also able to examine the effects of these cytokines on HBV cytotoxicity. Our results indicate that weak but detectable CTL responses do occur in chronic carriers which are generally associated with disease flares, although CTLps were also seen occasionally during minimal disease activity. The range of specificities varied between individuals and within each individual during the course of the disease. Finally, we also provide evidence that CTL reactivity is stimulated following treatment with certain cytokines, but is dependent on the time of administration. [source] Character impact odorants from wild mushroom (Lactarius hatsudake) used in Japanese traditional foodFLAVOUR AND FRAGRANCE JOURNAL, Issue 4 2010Mitsuo Miyazawa Abstract The components of the volatile oil from wild mushroom (Lactarius hatsudake), used in Japanese traditional food, were analysed and quantified for the first time by capillary GC and GC,MS. Seventy-six components were separated from the oil and of these 71 components were identified. The main components of the oil were oxidized sesquiterpenes [cis -isolongifolanone (624.9,,g/100,g), , -cedrene epoxide (578.7,,g/100,g), humulene epoxide III (453.9,,g/100,g), clovane (425.4,,g/100,g)], aliphatic acids [linoleic acid (585.9,,g/100,g) and palmitoleic acid (333.3,,g/100,g)]. Odour evaluation of the volatile oil from L. hatsudake was also carried out using GC,MS/olfactometry (GC,MS/O) and aroma extract dilution analysis (AEDA), from which it was found that hexanal, 4-dehydroviridiflorol, myliol and phenylacetaldehyde seem to contribute to the green, spicy and sweet odour of L. hatsudake. Copyright © 2010 John Wiley & Sons, Ltd. [source] Characterization of the aroma-active compounds in five sweet cherry cultivars grown in Yantai (China)FLAVOUR AND FRAGRANCE JOURNAL, Issue 4 2010Shu Yang Sun Abstract This study was conducted to determine the aroma-active compounds of five sweet cherry cultivars grown in Yantai region, China, viz. ,Lapins', ,Rainier', ,Stella', ,Hongdeng' and ,Zhifuhong'. The samples were extracted by headspace solid phase microextraction (HS,SPME) and analysed by gas chromatography,mass spectrometry (GC,MS) on DB-wax and DB-5 columns. A total of 52 volatiles were identified. Among these, hexanal, (E)-2-hexenal, 1-hexanol, (E)-2-hexen-1-ol, benzaldehyde and benzyl alcohol were the main volatile compounds in the five cherries. Furthermore, the aroma compounds of five cherry samples were evaluated using a combination of HS,SPME and GC,olfactometry (GC,O) dilution analysis, and a total of 40 aroma-active compounds were identified. The results suggested that hexanal, (E)-2-hexenal, (Z)-3-hexenal, nonanal, benzaldehyde and geranylacetone (FD , 16), responsible for the green, orange, almond and floral characters of the cherries, were the potentially important common odorants in these cherry cultivars. Benzyl alcohol and linalool were significant aroma compounds in most cherries, with the exception of ,Stella' and ,Rainier'. In addition, (E,Z)-2,6-nonadienal (cucumber-like odour) could be important to ,Hongdeng' and ,Zhifuhong', and (E,E)-2,4-nonadienal (fatty odour) probably made great contributions to the aromas in ,Lapins' and ,Stella'. From the present result, it was concluded that the aroma profiles were similar in the five cherry cultivars, but significant variation was found in the contributions of these compounds to each cherry. Copyright © 2010 John Wiley & Sons, Ltd. [source] Evaluation of odour-active compounds in roasted shrimp (Sergia lucens Hansen) by aroma extract dilution analysisFLAVOUR AND FRAGRANCE JOURNAL, Issue 6 2005Susumu Ishizaki Abstract In an investigation of roasted shrimp odour adsorbed by Tenax TA during the roasting of sun-dried sakuraebi (Sergia lucens Hansen), 31 odour-active compounds were detected by application of an aroma extract dilution analysis and subsequently identified by GC,MS analysis. Among these odorants, the following six contributed to roasted shrimp aroma, with high flavour dilution (FD) factors: methanethiol (sulphury); 1-pyrroline (shrimp meat-like); N -(2,-methylbutyl)pyrrolidine (roasted seafood-like); N -(3,-methylbutyl)pyrrolidine (roasted seafood-like); methyl isopropyl disulphide (roasted shrimp meat-like); 3-methylpyridine (fishy, green). Copyright © 2005 John Wiley & Sons, Ltd. [source] Aroma Components of American Country HamJOURNAL OF FOOD SCIENCE, Issue 1 2008H. Song ABSTRACT:, The aroma-active compounds of American country ham were investigated by using direct solvent extraction-solvent assisted flavor evaporation (DSE-SAFE), dynamic headspace dilution analysis (DHDA), gas chromatography-olfactometry (GCO), aroma extract dilution analysis (AEDA), and gas chromatography-mass spectrometry (GC-MS). The results indicated the involvement of numerous volatile constituents in the aroma of country ham. For DHDA, 38 compounds were identified as major odorants, among them, 1-octen-3-one, 2-acetyl-1-pyrroline, 1-nonen-3-one, decanal, and (E)-2-nonenal were the most predominant, having FD-factors , 125 in all 3 hams examined, followed by 3-methylbutanal, 1-hexen-3-one, octanal, acetic acid, phenylacetaldehyde, and FuraneolŌ. For the DSE-SAFE method, the neutral/basic fraction was dominated by 1-octen-3-one, methional, guaiacol, (E)-4,5-epoxy-(E)-decenal, p-cresol as well as 3-methylbutanal, hexanal, 2-acetyl-1-pyrroline, phenylacetaldehyde, and ,-nonalactone. The acidic fraction contained mainly short-chain volatile acids (3-methylbutanoic acid, butanoic acid, hexanoic acid, and acetic acid) and Maillard reaction products (for example, 4-hydroxy-2,5-dimethyl-3(2H)-furanone). The above compounds identified were derived from lipid oxidation, amino acid degradation, and Maillard/Strecker and associated reactions. Both methods revealed the same nature of the aroma components of American country ham. [source] Evaluation of Bitterness in Enzymatic Hydrolysates of Soy Protein Isolate by Taste Dilution AnalysisJOURNAL OF FOOD SCIENCE, Issue 1 2008W.H. Seo ABSTRACT:, Although enzymatic hydrolysates of soy protein isolate (SPI) have physiological functionality, partially hydrolyzed SPI exhibits bitter taste depending on proteases and degree of hydrolysis (DH). To determine proteolysis conditions for SPI, it is important to evaluate bitterness during enzymatic hydrolysis. Taste dilution analysis (TDA) has been developed for the screening technique of taste-active compounds in foods. The objectives of the present study were to evaluate bitterness of enzyme-hydrolyzed SPI by TDA and to compare bitterness of SPI hydrolysates with respect to kinds of proteases and DH. SPI was hydrolyzed at 50 °C and pH 6.8 to 7.1 to obtain various DH with commercial proteases (flavourzyme, alcalase, neutrase, protamex, papain, and bromelain) at E/S ratios of 0.5%, 1%, and 2%. The DH of enzymatic hydrolysates was measured by trinitrobenzenesulfonic acid method. The bitterness of enzymatic hydrolysates was evaluated by TDA, which is based on threshold detection in serially diluted samples. Taste dilution (TD) factor was defined as the dilution at which a taste difference between the diluted sample and 2 blanks could be detected. As DH increased, the bitterness increased for all proteases evaluated. Alcalase showed the highest TD factor at the same DH, followed by neutrase. Flavourzyme showed the lowest TD factor at the entire DH ranges. At the DH of 10%, TD factor of hydrolysate by flavourzyme was 0 whereas those by protamex and alcalase were 4 and 16, respectively. These results suggest that TDA could be applied for the alternative of bitterness evaluation to the hedonic scale sensory evaluation. [source] Characterization of Aroma-Active Compounds in Microwave Blanched PeanutsJOURNAL OF FOOD SCIENCE, Issue 9 2006A.V. Schirack ABSTRACT:, Microwave blanching of peanuts has been explored as an alternative to conventional oven methods based on its speed of operation, energy savings, and efficiency of process control. Although processing times can be greatly reduced, the occurrence of stale/floral and ashy off-flavors has been reported at high process temperatures. This study examined the chemical compounds responsible for this off-flavor using solvent extraction/solvent assisted flavor evaporation (SAFE), gas chromatography-olfactometry (GC/O), gas chromatography-mass spectrometry (GC/MS), and aroma extract dilution analysis (AEDA). Select compounds were quantified based on AEDA results using SAFE and GC/MS. Quantification, threshold testing, and analysis of model systems revealed increased formation of guaiacol and phenylacetaldehyde in the off-flavored peanuts, which resulted in the burnt and stale/floral flavors noted by a trained panel. [source] Analysis of aldehydes via headspace SPME with on-fiber derivatization to their O -(2,3,4,5,6-pentafluorobenzyl)oxime derivatives and comprehensive 2D-GC-MSJOURNAL OF SEPARATION SCIENCE, JSS, Issue 19 2008Hans-Georg Schmarr Abstract A method for the analysis of the homologous series of alkanals, (E)-2-alkenals, and (E,E)-2,4-alkadienals is described utilizing a headspace solid-phase microextraction (HS-SPME) step and on-fiber derivatization with O -(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) hydrochloride. Oxime derivatives formed on the fiber are desorbed in the gas chromatographic injector and analyzed by comprehensive 2-D GC coupled to quadrupole MS (GC×GC-qMS). Selecting specific fragment ions within the electron impact mass spectra of the oxime derivatives provides a suitable method for the target analysis of these aldehyde classes, which furthermore benefits from the increased separation efficiency by GC×GC. The analysis of higher molecular weight aldehydes is described in wine and grape seed oil as examples. Quantification of the aldehydes utilizes a stable isotope dilution analysis (SIDA) assay with octan-d16 -al as isotopomeric internal standard. Besides the selectivity and sensitivity of aldehyde analysis using PFBHA derivatives, critical aspects on background level contamination and repeatability of the sample preparation method are discussed. Optimization of GC×GC-qMS parameters allowed a considerable saving of the cryogenic medium, involving additional (unmodulated) conditioning runs, rendering the method more amenable to routine analysis. [source] Stable isotope dilution analysis of the Fusarium mycotoxins deoxynivalenol and 3-acetyldeoxynivalenolMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 3 2006Michael Bretz Abstract Trichothecenes are secondary metabolites produced by several fungi of the Fusarium genus during their growth period. They inhibit protein biosynthesis in eukaryotic cells resulting in numerous toxic effects such as diarrhea, vomiting, and gastro-intestinal inflammation. Considering its occurrence in food and feedstuff, deoxynivalenol (DON) is one of the most important trichothecenes. We report the synthesis of stable isotope labeled 15- d1 -deoxynivalenol (15- d1 -DON) from its natural precursor 3-acetyldeoxynivalenol (3-AcDON) as starting material. Furthermore, a method for the analysis of DON and 3-AcDON using HPLC-MS/MS with stable isotope labeled 15- d1 -DON and 3- d3 -AcDON as internal standards has been developed. In total, 18 cereal product samples were analyzed with contamination levels ranging from 10,301 ,g/kg for DON and 5,14 ,g/kg for 3-AcDON. This is the first report of an isotope dilution MS method for the analysis of type B-trichothecenes. [source] A new standard for the assessment of disease progression in murine cutaneous leishmaniasisPARASITE IMMUNOLOGY, Issue 5 2000Lynden J.roberts Infection of mice with Leishmania major has been used both as a model for the cutaneous disease in humans and as a model for the more general control and function of helper T cells in immunity. In both cases, disease patterns and disease progression have been assessed by two complementary methods, lesion size and parasite burden in the draining lymph nodes. We propose a much improved method for the graphical representation of lesion development which conveys more information with better accuracy. We also describe a polymerase chain reaction method for determining parasite burden, which is faster and allows the analysis of larger numbers of experimental animals than the current limiting dilution analysis. Moreover, these methods are equally applicable to other infectious diseases, an obvious one being schistosomiasis. [source] Pretreatment With Portal Venous Ultraviolet B Irradiated Donor Alloantigen Promotes Donor-Specific Tolerance to Rat Nerve Allografts,THE LARYNGOSCOPE, Issue 3 2001Eric M. Genden MD Abstract Objective To determine if a single intraportal inoculation of ultraviolet B-irradiated (UVB) donor splenocytes can prevent nerve allograft rejection and confer donor-specific immunotolerance to rat nerve allograft segments. Methods Age-matched, class I and class II major histocompatibility complex (MHC) mismatched Buffalo (RT1b) rats were transplanted with a syngeneic nerve isograft, a Lewis (RT1l) nerve allograft, or a Brown-Norway (RT1n) rat nerve allograft segment. Control Buffalo rats in group I received a 3.0-cm Lewis (RT11) sciatic-posterior tibial interposition nerve allograft without pretreatment;group II Buffalo rats received a syngeneic Buffalo nerve isograft without pretreatment. Group III Buffalo recipients were inoculated with 2.5 × 107 UVB-irradiated Lewis donor splenocyte cells by portal venous administration 7 days before transplantation with a 3.0-cm sciatic-posterior tibial nerve allograft from a Lewis (RT11) or a third party Brown-Norway rat (RT1n) donor (group IV). Nerve graft regeneration was assessed with walking track analysis, nerve conduction studies, retrograde neural tracing, nerve graft histology, and morphometry. Recipient immune tolerance was assessed through in vitro immunological assessment. Results Pretreatment with UVB-irradiated donor splenocytes 7 days before transplantation prevented nerve allograft rejection. Pretreated animals receiving a nerve allograft recovered limb function, and demonstrated morphological, histological, and electrophysiologic parameters of nerve regeneration similar to that measured in rats receiving a nerve isograft. In vitro immunological assessment by mixed lymphocyte culture (MLC), cytotoxic T lymphocyte (CTL) assay, limiting dilution analysis (LDA) of helper (pTH) and cytotoxic (pCTL) precursor frequencies, and IL-2 production demonstrated a marked donor-specific suppression in allografted animals pretreated with intraportal UVB-irradiated donor splenocytes. These assessments correlated with indefinite acceptance of donor nerve allografts. Conclusions A single pretreatment with a single intraportal dose of UVB-modified donor antigen specifically induces tolerance to peripheral nerve allografts in rats. [source] Identification and analysis of 2-chloro-6-methylphenol, 2,6-dichlorophenol and indole: causes of taints and off-flavours in winesAUSTRALIAN JOURNAL OF GRAPE AND WINE RESEARCH, Issue 1 2010D.L. CAPONE Abstract Background and Aims:, The aim of this study was to determine the cause of taints and off-flavours in a number of commercial wines and to develop methods for quantitative analysis of the compounds responsible. Methods and Results:, Three compounds, 2-chloro-6-methylphenol (i.e. 6-chloro- ortho -cresol), 2,6-dichlorophenol and indole, were identified by gas chromatography (GC)/mass spectrometry (MS)/olfactometry as causes of taints or off-flavours in the wines. New methods for quantitative analysis of these three compounds, as well as some other chloro- and bromophenols were developed. The methods employed GC/MS and stable isotope dilution analysis, and confirmed the presence of 2-chloro-6-methylphenol, 2,6-dichlorophenol and indole in some wines at concentrations well above their odour and taste detection thresholds. Conclusions:, 2-Chloro-6-methylphenol, 2,6-dichlorophenol and indole were confirmed as causes of taints and off-flavours in commercial wines. Significance of the Study:, The development of new analytical methods will make it possible to determine whether chlorophenol (and perhaps also bromophenol) taint in wine is more widespread than previously recognised. Similarly, the simple, rapid and accurate method for quantifying indole in wine will allow large numbers of wine and ferment samples to be studied to determine those factors that can result in unacceptably high concentrations in commercial wines. [source] Stable isotope dilution analysis of N-acetylaspartic acid in urine by liquid chromatography electrospray ionization tandem mass spectrometryBIOMEDICAL CHROMATOGRAPHY, Issue 9 2007Osama Y. Al-Dirbashi Abstract N -acetylaspartic acid (NAA) is a specific urinary marker for Canavan disease, an autosomal recessive leukodystrophy. We developed a ,dilute and shoot' stable isotope dilution liquid chromatography tandem mass spectrometry (LC-MS/MS) method for determination of NAA in urine. Deuterated internal standard d3 -NAA was added to untreated urine and the mixture was injected into the LC-MS/MS system operated in the negative ion mode. Chromatography was carried out on a C8 minibore column using 50% acetonitrile solution containing 0.05% formic acid at a flow rate of 0.25 mL/min. The retention time was 1.6 min and the turnaround time was 2.2 min. NAA and d3 -NAA were analyzed in multiple reaction monitoring mode. Calibrators and quality control samples were prepared in pooled control urine. The assay was linear up to 2000 µmol/L with limit of quantification at 1 µmol/L (S/N = 12). Interassay and intraassay coefficients of variation were less than 7% and recovery at three different concentrations was 98.9,102.5%. The LC-MS/MS method for NAA as described involves no extraction and no derivatization, showed no interference and gave excellent recovery with low variability and short analytical time. The method was successfully applied for the retrospective analysis of urine from 21 Canavan disease cases. Copyright © 2007 John Wiley & Sons, Ltd. [source] | |||||||||||||