RESUMO
Protein scarcity is the most vital cause of long-lasting diseases and even untimely deaths in some developing nations. The application of protein in food is advantageous from the point of view of non-toxicity, biocompatibility, and dietary benefits. This study aimed to determine the protein contents of the sprouts of Vigna radiates (mung beans), Lens culinaris (lentils), and Cicer arietinum (chickpeas) using the Kjeldahl and Lowry methods. The results obtained from the Kjeldahl method identified protein concentrations of 2.54, 2.63, and 2.19%, whereas the Lowry method results identified protein concentrations of 2.96%, 4.10%, and 1.6% in mung beans, lentils, and chickpeas, respectively. In both the methods, lentils were found to have the highest amount of protein followed by mung beans and chickpeas. Both the Kjeldahl and Lowry methods demonstrated good protein values and low variation in the protein amount in the analyzed samples. Furthermore, the methods had greater sensitivity and comparable experimental variability. The outcomes revealed that assays can be applied for protein analysis in legumes. In the context of a lack of suitable standard procedures for evaluating legumes' compositions, the present study is suitable for food control laboratories. In addition, the studied samples represent a significant source of protein and can be used to fulfil the daily requirements for protein intake and other food applications.
Assuntos
Cicer/química , Lens (Planta)/química , Proteínas de Plantas/análise , Plântula/química , Vigna/química , EspectrofotometriaRESUMO
Currently, a broad discussion exists in the literature regarding insect protein analysis. At its core, main difficulties and uncertainties are the inconsistent use of analysis methods and nitrogen-to-protein conversion (kP) factors. While the Kjeldahl and Dumas methods are both used in the literature, their result represents inherently different nitrogen fractions. Thus far, no correlation between them is established for insect matrices, which is a major uncertainty. Although much effort has been made towards more accurate kP factors, calculation of these was based on merely one sample while the chemical composition varies depending on rearing conditions. Using a broad variation in black soldier fly (BSF) larvae samples in the present study, a correlation between Kjeldahl and Dumas and a robust kP factor have been established. Moreover, the nitrogen distribution of BSF samples was also assessed after accurate chitin analyses. A highly significant linear correlation existed between the results of Kjeldahl and Dumas (slope, 1.009; intercept, - 0.008; R2, 0.9997). Consequently, both methods were deemed interchangeable for BSF larvae. Using amino acid data, a practical, more accurate and robust kP factor of 4.43 was obtained. Concerning the chitin content, the average of all BSF larvae samples was 5.95 ± 0.86 g N-acetylglucosamine/100 g dry matter and no correlation with the kP factor was observed. Regarding the nitrogen distribution of the samples, it was found that the contribution of nitrogenous compounds other than protein and chitin is not only high but also prone to variation (12-30% of the total nitrogen content).
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Dípteros/metabolismo , Proteínas de Insetos/metabolismo , Nitrogênio/metabolismo , Aminoácidos/química , Aminoácidos/metabolismo , Animais , Proteínas de Insetos/genética , Larva/metabolismoRESUMO
Our objectives were to determine if milk casein as a percentage of true protein (CN%TP) estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is equivalent to CN%TP estimated by Kjeldahl, and to determine the proportion of casein (CN), casein proteolysis products (CNPP), and serum protein (SP) from milk true protein (TP) that goes into the Kjeldahl noncasein nitrogen (NCN) filtrate and the proportion that stays in the NCN precipitate using SDS-PAGE. Raw milk samples were collected from 16 mid-lactation Holstein cows twice a week for 2 wk. These milks were analyzed for Kjeldahl total nitrogen, nonprotein nitrogen, and NCN content in duplicate, and by SDS-PAGE. The CN%TP determined by Kjeldahl was compared with the CN%TP estimated by SDS-PAGE calculated in 2 ways: as a percentage of only intact caseins divided by TP and as a percentage of both intact caseins and CNPP divided by TP. Three milks varying in fat, lactose, TP, CN, and SP content were formulated. These milks were analyzed in duplicate for Kjeldahl total nitrogen, nonprotein nitrogen, and NCN content, and each of the NCN filtrate and NCN precipitate were analyzed in duplicate by SDS-PAGE for relative quantity (%) of CN, CNPP, and SP. We found that the estimate of CN%TP by Kjeldahl was higher than the estimate of CN%TP by SDS-PAGE that was calculated as only intact CN divided by the total of all protein bands. However, no difference was detected in the estimate of CN%TP by Kjeldahl compared with CN%TP by SDS-PAGE when CNPP were included as CN in the calculation of SDS-PAGE results. Based on SDS-PAGE results, we found that a majority (89%) of the CNPP from the milk (approximately 10.13 out of 11.41% TP) were retained in the Kjeldahl NCN precipitate. Thus, CN%TP measured by Kjeldahl underestimates the amount of proteolytic damage that has been done to CN in milk. It is important for the dairy industry to correctly and rapidly measure the extent of proteolytic damage to milk protein to correctly value milk from a product quality and yield point of view. A rapid and quantitative measure of proteolytic damage to milk protein is needed.
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Caseínas , Leite , Animais , Bovinos , Eletroforese em Gel de Poliacrilamida/veterinária , Feminino , Leite/química , Proteínas do Leite/análise , Dodecilsulfato de SódioRESUMO
To clarify the genetic mechanism underlying grain protein content (GPC) and to improve rice grain qualities, the mapping and cloning of quantitative trait loci (QTLs) controlling the natural variation of GPC are very important. Based on genotyping-by-resequencing, a total of 14 QTLs were detected with the Huanghuazhan/Jizi1560 (HHZ/JZ1560) recombinant inbred line (RIL) population in 2016 and 2017. Seven of the fourteen QTLs were repeatedly identified across two years. Using three residual heterozygote-derived populations, a stably inherited QTL named as qGPC1-1 was validated and delimited to a ~862 kb marker interval JD1006-JD1075 on the short arm of chromosome 1. Comparing the GPC values of the RIL population determined by near infrared reflectance spectroscopy (NIRS) and Kjeldahl nitrogen determination (KND) methods, high correlation coefficients (0.966 and 0.983) were observed in 2016 and 2017. Furthermore, 12 of the 14 QTLs were identically identified with the GPC measured by the two methods. These results indicated that instead of the traditional KND method, the rapid and easy-to-operate NIRS was suitable for analyzing a massive number of samples in mapping and cloning QTLs for GPC. Using the gel-based low-density map consisted of 208 simple sequence repeat (SSR) and insert/deletion (InDel) markers, the same number of QTLs (fourteen) were identified in the same HHZ/JZ1560 RIL population, and three QTLs were repeatedly detected across two years. More stably expressed QTLs were identified based on the genome resequencing, which might be attributed to the high-density map, increasing the detection power of minor QTLs. Our results are helpful in dissecting the genetic basis of GPC and improving rice grain qualities through molecular assisted selection.
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Genoma de Planta , Técnicas de Genotipagem , Proteínas de Grãos/metabolismo , Oryza/genética , Locos de Características Quantitativas/genética , Análise de Sequência de DNA , Mapeamento Cromossômico , Cruzamentos Genéticos , Ligação Genética , Heterozigoto , Endogamia , Fenótipo , Reprodutibilidade dos TestesRESUMO
In the summer of 2005 and 2010, moss samples were collected from 72 sampling sites evenly distributed all over the territory of Republic of North Macedonia. Kjeldahl method was used to determine the nitrogen content in the samples. Descriptive statistics and distribution maps were prepared. Data obtained from these two surveys were compared, and additional comparison was done with data obtained from similar studies in the South-Eastern European countries and Finland as a clean area. The median value of N content in the samples collected in 2005 is 1.21%, varies from 0.70% to 1.54%, while the content of N in samples collected in 2010 ranges between 0.68% and 1.75% with the median value of 1.06%. High contents of N were found in the Northern and Central parts of the country mainly as a result of agricultural activities, industry and traffic.
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Poluentes Atmosféricos/análise , Poluição do Ar/análise , Monitoramento Biológico/métodos , Briófitas/química , Nitrogênio/análise , Indústrias , República da Macedônia do Norte , Análise EspacialRESUMO
Nitrogen concentrations in feeds, feces, milk, and urine samples were measured using 2 analytical methods following different drying procedures. Ten samples of corn silage, alfalfa silage, and concentrates collected from 2017 to 2018 at Krauss Dairy Research Center, The Ohio State University (Wooster), were used. A 4-d total collection digestion trial provided fecal samples from 10 cows (1 sample/cow), and another 10 cows were used to collect milk samples (1 sample/cow) and spot urine samples (1 sample/cow). Spot urine samples were acidified immediately to pH <3.0 when collected. Feed samples were oven dried (55°C) or lyophilized and analyzed using the Kjeldahl (KJ; copper sulfate as a catalyst) method and a combustion method (elemental analyzer; EA). Feces, urine, and milk samples were analyzed for N using the following methods: (1) fresh samples by KJ (referred to as wet KJ), (2) lyophilization (urine and milk for 8 h; feces for 120 h) followed by EA (LYO-EA), and (3) oven drying (milk and urine for 1 h; feces for 72 h at 55°C) followed by EA (OD-EA). Additionally, changes in N content of acidified urine at -20° over 180 d of storage were examined. Nitrogen concentrations in corn silage, alfalfa silage, and concentrates were greater for EA by 6.1, 4.8, and 8.3%, respectively, compared with KJ. Analysis of dried samples via EA compared with wet KJ resulted in lower fecal N content (27.8 vs. 29.3 g/kg of DM). Nitrogen concentration in fecal samples via KJ after lyophilization was lower by 5% compared with wet KJ but did not differ from LYO-EA, suggesting that N losses occurred during drying. Nitrogen determination with EA after drying of samples resulted in greater milk N (5.70 vs. 5.50 g/kg) and urinary N (9.16 vs. 9.06 g/kg) content compared with wet KJ. However, drying method (i.e., lyophilization vs. oven drying) did not affect N content of milk, urine, or feces. The use of EA resulted in lower percentage deviation of N content from duplicate sample assays for most samples (no difference was found for concentrate and fecal N), suggesting that EA was more precise than KJ. In conclusion, drying of feces caused N losses regardless of drying methods. For urine and milk samples, if drying is necessary (i.e., EA), oven drying at 55°C can be used rather than lyophilization. The N content was greater in feeds, milk, and urine when determined with EA versus KJ. In addition, N content in acidified and undiluted urine at -20° changed and should be analyzed within 90 d of storage. The results in the current study, however, did not account for laboratory-to-laboratory variation.
Assuntos
Ração Animal , Bovinos , Leite/química , Nitrogênio/análise , Animais , Dessecação , Dieta/veterinária , Fezes/química , Feminino , Lactação , Medicago sativa/química , Nitrogênio/urina , Silagem/análise , Zea mays/químicaRESUMO
Concentrations of essential (Cu, Mn, and Zn) and toxic (Cr, Cd, and Pb) trace metals in 30 raw cow's milk samples were quantified using flame atomic absorption spectrometry. The samples were collected from the Nara-Awudarda, Tana-Abo, and Kosoye Amba-Rass sites in North Gondar, Ethiopia, preserved in a deep freezer (-20 °C), and then digested by Kjeldahl apparatus with HNO3/H2O2 (5:2; v/v) at 300 °C for 2.5 h. The data were subject to principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA). Overall hazard quotient (HQ) and carcinogenic risk (CR) values were also estimated to assess metal-related health risks. The mean concentrations of Cr, Mn, Cu, Zn, Cd, and Pb in the milk samples ranged 0.468-0.828, 1.614-2.806, 0.840-1.532, 1.208-5.267, ND-0.330, and ND-0.186 mg/kg, respectively. The lowest values were obtained for Kosoye Amba-Rass milk samples, while the highest were found for those collected from Nara-Awudarda milk samples, probably due to high mineral enrichment and metal leaching (especially Cd and Pb) from coal deposits. PCA revealed clustering of samples with respect to their geographic origin. Validation of PLS-DA model showed 100% classification efficiency using external validation samples and detected Cd and Cu as trace metal markers. The HQ and CR values were within the safe level; however, the former is close to the alert threshold level for Nara-Awudarda milk samples. Thus, further studies on common foodstuffs, constituting a higher proportion in the local diet, are required in this area to provide a complete risk assessment.
Assuntos
Poluentes Ambientais/análise , Contaminação de Alimentos/análise , Metais Pesados/análise , Leite/química , Oligoelementos/análise , Animais , Bovinos , Etiópia , Fazendas , Feminino , Leite/normas , Medição de Risco , Espectrofotometria AtômicaRESUMO
Macronutrient contents in human milk are the common basis for estimating these nutrient requirements for both infants and lactating women. A mid-infrared human milk analyser (HMA, Miris, Sweden) was recently developed for determining macronutrient levels. The purpose of the study is to compare the accuracy and precision of HMA method with fresh milk samples in the field studies with chemical methods with frozen samples in the lab. Full breast milk was collected using electric pumps and fresh milk was analyzed in the field studies using HMA. All human milk samples were thawed and analyzed with chemical reference methods in the lab. The protein, fat and total solid levels were significantly correlated between the two methods and the correlation coefficient was 0.88, 0.93 and 0.78, respectively (p < 0.001). The mean protein content was significantly lower and the mean fat level was significantly greater when measured using HMA method (1.0 g 100 mL-1 vs 1.2 g 100 mL-1 and 3. 7 g 100 mL-1 vs 3.2 g 100 mL-1 , respectively, p < 0.001). Thus, linear recalibration could be used to improve mean estimation for both protein and fat. There was no significant correlation for lactose between the two methods (p > 0.05). There was no statistically significant difference in the mean total solid concentration (12.2 g 100 mL-1 vs 12.3 g 100 mL-1 , p > 0.05). Overall, HMA might be used to analyze macronutrients in fresh human milk with acceptable accuracy and precision after recalibrating fat and protein levels of field samples.
Assuntos
Leite Humano/química , Valor Nutritivo , Espectrofotometria Infravermelho , Calibragem , Gorduras na Dieta/análise , Feminino , Análise de Alimentos , Humanos , Lactose/análise , Proteínas do Leite/análiseRESUMO
Our goal was to determine the feasibility of combining proficiency testing, analytical method quality-assurance system, and production of reference samples for calibration of infrared milk analyzers to achieve a more efficient use of resources and reduce costs while maximizing analytical accuracy within and among milk payment-testing laboratories. To achieve this, we developed and demonstrated a multilaboratory combined proficiency testing and analytical method quality-assurance system as an approach to evaluate and improve the analytical performance of methods. A set of modified milks was developed and optimized to serve multiple purposes (i.e., proficiency testing, quality-assurance and method improvement, and to provide reference materials for calibration of secondary testing methods). Over a period of years, the approach has enabled the group of laboratories to document improved analytical performance (i.e., reduced within- and between-laboratory variation) of chemical reference methods used as the primary reference for calibration of high-speed electronic milk-testing equipment. An annual meeting of the laboratory technicians allows for review of results and discussion of each method and provides a forum for communication of experience and techniques that are of value to new analysts in the group. The monthly proficiency testing sample exchanges have the added benefit of producing all-laboratory mean reference values for a set of 14 milks that can be used for calibration, evaluation, and troubleshooting of calibration adjustment issues on infrared milk analyzers.
Assuntos
Laboratórios/normas , Leite/química , Animais , Calibragem , Controle de Qualidade , Valores de Referência , Reprodutibilidade dos TestesRESUMO
The objective of our research was to modify the current indirect casein method for bovine milk to enable it to be applied to bovine milk, bovine milk concentrates, and milks of other species that contain a protein concentration up to 9% (wt/wt). Our work used a series of bovine milk concentrates from about 3 to 9% protein with the same casein as a percentage of true protein to determine the amount of buffer required and pH of the noncasein nitrogen (NCN) filtrate to achieve consistent estimates of casein and casein as percent of true protein. As the concentration of protein in milk increased (either in bovine milk concentrates or in milks of other species), the amount of buffer needed for the NCN sample preparation method to achieve a filtrate pH of 4.6 increased. In the first part of the study using a series of bovine milk concentrates, it was demonstrated that the method gave more consistent predictions of casein as a percentage of true protein when the final NCN filtrate pH was between 4.5 and 4.6 at 38°C. When the amount of buffer added to the sample was not sufficient (i.e., the filtrate pH was too high), the filtrates were not clear. A polynomial equation was developed for prediction of the amount of acetic acid or sodium acetate buffer required to achieve pH 4.5 to 4.6 for milk protein concentrations from 3 to 9% protein using bovine milk and milk concentrates. When the equation developed using cow milk was applied to goat, sheep, and water buffalo milks, it correctly predicted the volume of reagents needed to achieve a final NCN filtrate pH of 4.6 at 38°C. We also verified as part of this work that the ability to measure NPN content of milk was not influenced by protein content of milk in the range from 3 to 9% protein. The results of this study will be used as the basis for proposed changes in the official methods for measurement of the casein content of milk to expand the scope of the method so it can be used to achieve accurate results for milk concentrates and milks of other species.
Assuntos
Caseínas/análise , Leite/química , Nitrogênio/análise , Animais , Búfalos , Bovinos , Filtração , Cabras , Concentração de Íons de Hidrogênio , Proteínas do Leite/análise , OvinosRESUMO
During the summer and autumn of 2006 moss samples were collected from 98 sampling sites evenly distributed all over the territory of Croatia. Moss sampling was repeated in 2010 when additional sampling sites were added and a total of 121 samples were collected. Kjeldahl method was used to determine the nitrogen content in the samples. Descriptive statistics and distribution maps were prepared. Data obtained from these two surveys were compared, and additional comparison was done with data obtained from similar studies in neighbouring countries and Finland as a clean area. The median value of N content in the samples collected in 2006 is 1.60% and varies from 0.79% to 3.16%. The content of N in samples collected in 2010 ranges between 0.71% and 2.93% with the median value of 1.49%. High contents of N (2.32% - 3.17%) were found in the regions of Slavonia, Podravina, Posavina and cities Zagreb and Sisak as a result of agricultural activities, industry and traffic.
Assuntos
Briófitas/química , Nitrogênio/análise , Poluentes Atmosféricos/análise , Briófitas/metabolismo , Cidades , Croácia , Monitoramento Ambiental/métodos , Nitrogênio/metabolismoRESUMO
Brassica juncea depends heavily on nitrogen (N) fertilizers for growth and accumulation of seed protein. However, it is an inefficient mobilizer of applied N which leads to accumulation of excess N in the soil, posing environmental risks. Hence, it is imperative to systematically examine spatial-temporal pattern of crop N to efficiently manage N application. The Kjeldahl method is commonly used to estimate N status of crops but it is a destructive method that entails the use of perilous and expensive chemicals. Near-infrared reflectance spectroscopy (NIRS) offers a safe, accurate, and non-destructive alternative for large-scale screening of seed metabolites. Currently, no NIRS model exists to quickly estimate N content in shoots and roots from large germplasm sets in any rapeseed-mustard crop. Developing such a model is essential to breed for enhanced nitrogen use efficiency (NUE). We used 738 shoot and 346 root samples from a B. juncea diversity set to construct the NIRS models. A diverse range of genetic variation in N content was recorded in the stem (0.21-6.61%) and root (0.15-3.04%) tissues of the crop raised on two different N levels (N0 and N100). Modified partial least squares (MPLS) method was employed to establish a regression equation linking reference N values with spectral changes. The developed models exhibited strong associations with reference values, with RSQ values of 0.884 for stem and 0.645 for roots. Furthermore, external validation confirms the reliability of the developed models. The developed models have strong predictive capabilities for rapid and reliable N estimation in various tissues of B. juncea plants.
Assuntos
Mostardeira , Nitrogênio , Raízes de Plantas , Caules de Planta , Espectroscopia de Luz Próxima ao Infravermelho , Mostardeira/química , Mostardeira/metabolismo , Nitrogênio/análise , Nitrogênio/metabolismo , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Raízes de Plantas/química , Raízes de Plantas/metabolismo , Caules de Planta/química , Caules de Planta/metabolismo , Análise dos Mínimos QuadradosRESUMO
Accurately determining the macronutrient profile of mare milk is a precursor to studying how milk composition affects foals' growth and development. This study optimized and validated an extraction and quantification method for mare milk oligosaccharides, which make up a portion of the carbohydrate fraction of mare milk. Mare milk was extracted with chloroform and methanol, and oligosaccharides were selectively isolated from the carbohydrate fraction using porous-graphitized carbon solid-phase-extraction (SPE). Good recovery rates for milk oligosaccharides (between 70 and 100%) were achieved with the optimized method. This study also compared the use of Fourier-Transform infrared (FTIR) spectroscopy versus wet chemistry quantification methods for protein, fat, and lactose. The FTIR method produced statistically equivalent protein contents to the wet chemistry method, along with substantial savings in both analyst time and consumable consumption. FTIR analysis slightly underestimated the fat content of mare milk relative to the official wet chemistry method, with the difference between the methods increasing at higher fat contents. FTIR also overestimated the lactose content of mare milk and appeared to generate "lactose" values that included the milk oligosaccharides and thus represented the total carbohydrate (lactose and milk oligosaccharides) content of mare milk.
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Understanding transient nitrogen (N) storage and transformation in the deep vadose zone is critical for controlling groundwater contamination by nitrate. The occurrence of organic and inorganic forms of carbon (C) and nitrogen and their importance in the deep vadose zone is not well characterized due to difficulty in sampling and the limited number of studies. We sampled and characterized these pools beneath 27 croplands with different vadose zone thicknesses (6-45 m). We measured nitrate and ammonium in different depths for the 27 sites to evaluate inorganic N storage. We measured total Kjeldahl nitrogen (TKN), hot-water extractable organic carbon (EOC), soil organic carbon (SOC), and δ13C for two sites to understand the potential role of organic N and C pools in N transformations. Inorganic N stocks in the vadose zone were 21.7-1043.6 g m-2 across 27 sites; the thicker vadose zone significantly stored more inorganic N (p < 0.05). We observed significant reservoirs of TKN and SOC at depths, likely representing paleosols that may provide organic C and N to subsurface microbes. The occurrence of deep C and N needs to be addressed in future research on terrestrial C and N storage potential. The increase of ammonium and EOC and δ13C value in the proximity of these horizons is consistent with N mineralization. An increase of nitrate, concurrent with the sandy soil texture and the water-filled pore space (WFPS) of 78 %, suggests that deep vadose zone nitrification may be supported in vadose zones with organic-rich layers such as paleosol. A profile showing the decrease of nitrate concentrations, concurrent with the clay soil texture and the WFPS of 91 %, also suggests denitrification may be an important process. Our study shows that microbial N transformation may be possible even in deep vadose zone with co-occurrence of C and N sources and controlled by labile C availability and soil texture.
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Milk ultrafiltration is a widely used membrane filtration process that allows the recuperation of whey proteins in a concentrate high in total solids, which can later be transformed in multiple healthy dairy products with great prospects for the food industry. Protein content is a decisive factor for the technological performance of milk concentrates and currently, the ISO standard method for its determination is Kjeldahl, which is time-consuming and requires specific instrumentation. For this reason, the use of rapid methods to quantify protein would greatly facilitate the monitoring of the milk ultrafiltration process. In this study, the bicinchoninic acid assay (BCA), the detergent compatible Bradford assay and the Dumas method were compared to Kjeldahl protein determination to select a quick and accurate methodology suitable for milk of different species and its ultrafiltration products (retentates and permeates). The protein content obtained from Bradford assay and Dumas method in origin milk and retentate samples was consistent with Kjeldahl values. In contrast, BCA protein levels were significantly different when compared to Kjeldahl and no method was proved to be suitable for protein determination in permeate samples. The use of sodium dodecyl sulfate was also examined to improve protein measurements without success. In comparison with the official method, Bradford assay quantitatively provided the best results, and it would be recommended for a quick, economic and easy determination of total protein content in milk and retentate samples.
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Peptides with different lengths or amino sequences could have specific tastes or bio-activities. So far, either the quantity or pattern differences of peptide among various of teas were unknown. Here, firstly, we developed a method for tea oligopeptide quantification and made comparison of their contents. Secondly, we applied ultra-high performance liquid chromatography coupled with quadrupole-orbitrap ultra-high resolution mass spectrometry (UHPLC-Quadrupole-Orbitrap-UHRMS) to sequence oligopeptides. As a result, the total amount of oligopeptides in white tea and dark tea were higher, followed by black tea and green tea, finally with oolong tea. It suggested that withering which undergoes with endogenous protease and post-fermented that undergoes with a participation of exotic micro-organisms were key in oligopeptide enrichment. Thirdly, a total of 902 abundant identified peptides, most of which were tripeptide, tetrapeptide, pentapeptide, and hexapeptide were screened against several existing peptide databases. There were a series of taste peptides and bio-active peptides existing.
Assuntos
Oligopeptídeos , Chá , Cromatografia Líquida de Alta Pressão , Fermentação , Espectrometria de MassasRESUMO
The number of diatoms detected in the kidneys and liver varies considerably when comparing various reports, but the reason remains unclear. We previously showed that the reuse of Kjeldahl flasks is susceptible to generating false positives; false-positive results were observed in 17 of 60 tested flasks, and a maximum of 290 diatoms were detected. Furthermore, these data suggested that the number of diatoms in kidney and liver tissue may be much lower than previously thought. Therefore, only new Kjeldahl flasks were used to perform diatom tests on 80 autopsy cases (drowning and non-drowning victims) in this study. When combined with the 20 cases in a previous study, a total of 100 cases were analyzed. Four grams of lung, 10-30 g each of kidney and liver, 10 mL of blood, and 50-100 mL of water (discovery site) were examined. A large number of diatoms (n = 400) were found in organs other than the lungs in only 1 of 56 cases of suspected drowning (excluding cases of suspected drowning in baths). Together with the previous 20 cases, this single case represented only 1 of 76 cases (1.3 %). In closed organs and blood samples, 3 diatoms were found in 2 of 56 victims (3.6 %), and 1 or 2 diatoms were found in 8 victims (14 %). However, 5 diatoms were found in 1 of 10 bathtub drowning victims and 12 non-drowning victims (4.5 %), and 1 or 2 diatoms were found in 6 victims (27 %). Moreover, 1 or 2 diatoms were found in negative control flasks in 3 of 80 victims (3.8 %). The results showed that distinguishing false-positive results is difficult when a small number of diatoms are found in the kidneys and liver. Thus, the presence of diatoms in closed organs and blood seems to be unreliable as evidence of death by drowning, at least in the water environments in our region (freshwater areas: 23-26,000 diatoms/mL, median 200 diatoms; brackish water areas: 26-1200 diatoms/mL, median 210 diatoms; seawater area: 2-1000 diatoms/mL, median 24 diatoms). These results also reconfirmed that the reuse of flasks is the most important cause of false positives. Conversely, the presence of a large number of diatoms in lung samples (1-850,000 diatoms/g lung tissue; median (right lung): 440 diatoms, (left lung): 320 diatoms) is very informative and is an effective diagnostic aid in many cases.
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Diatomáceas , Afogamento , Humanos , Autopsia , Afogamento/diagnóstico , Água , PulmãoRESUMO
Protein is one of the most abundant substances in plants and plays a major role in human health hence standardization of its analytical quantification method is essential. Various methods for protein quantification exist, such as Kjeldahl, Bradford, Lowry, bicinchoninic acid assay (BCA), Biuret, and total amino acid content methods. These methods are widely applied; however, the development of the rapid and efficient method is the need of the time hence the objective of this research was to analyze and comparing compare the modification of the Kjeldahl method for the determination of protein content in oilseed crops. The study was performed to improve the sample preparation method (processing and digestion) for protein quantification. Generally, the method initially requires homogenization of grains to a fine flour, which involves time and increases the risk of sample cross-contamination and partial loss of oil from the sample during grinding. Moreover at times, it becomes challenging to homogenize oil seeds to fine flour due to high oil content. However, in the present research, the whole grain was digested in place of grounded flour to accomplish quick protein quantification and compared it with the flour matrix of different oil seeds. To further reduce the digestion time and avoid frothing, we have used the modified digestion mixture. The developed method was statistically validated using analysis of variance (ANOVA), Pearson correlation reliability test, paired T-test, and different types of plot analysis. The validation of the sample preparation method in protein quantification demonstrated non-significant differences that the protein content from whole grain of all the five oilseed crops shows 100% non-significant results compared with the flour matrix in both the digestion mixtures. The developed novel method could be used to prepare the sample for protein analysis and reduces the overall analysis time while ensuring the accuracy of the results.
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There is an ever-expanding number of high protein dietary supplements marketed as beneficial to athletes, body builders, infant formulas, elder care, and animal feed. Consumers will pay more for products with high protein per serving data on their nutritional labels, making the accurate reporting of protein content critical to customer confidence. The Kjeldahl method (KM) is the industry standard to quantitate dairy proteins, but the result is based on nitrogen content, which is an approximation of nitrogen attributable to protein in milk. Product tampering by third-party manufacturers is an issue, due to the lack of United States Food and Drug Administration regulation of nutraceutical products, permitting formulators to add low-cost nitrogen-containing components to artificially inflate the KM approximated protein content in products. Optical spectroscopy is commonly used for quality control measurements and has been identified as having the potential to complement the KM as a more nuanced testing measure of dairy protein. Mid-infrared (MIR) spectroscopy spectra of eight protein standards provided qualitative characterization of each protein by amide I and amide II peak absorbance wavenumber. Protein doping experiments revealed that as protein amounts were increased, the amide I/II peak shape changed from the broad protein powder peaks to the narrower peaks characteristic of the individual protein. Amino acid doping experiments with lysine, glutamic acid, and glycine, determined the limit of detection by MIR spectroscopy as 25%, suggesting that MIR spectroscopy can provide product quality assurance complementary to dairy protein measurement by the KM.