Determination of soy in meat

A number of methods may be used for determining soy flour in meat products. Highly purified soy products are more difficult to determine because the nonprotein components used to quantify the flour are reduced. Immunoassays have been used to directly measure protein content of soy products. Immunological methods for determination of soy proteins in meat are complicated by changes in the structure of the soy proteins during processing. These changes alter the available epitopes, changing the immunoreactivity of soy proteins. The epitopes available are dictated by the details of the processing. Other workers circumvented this problem by denaturing the soy protein with urea and mercaptoethanol, and then removing these agents by dialysis; whatever the initial protein conformation, all soy samples came to the same final conformation after the denaturing agents were removed. The assay used antibody made against the "renatured protein." These steps made the assay long and laborious. Attempts to develop a rapid assay were complicated by the same protein denaturation problems. Sodium dodecylsulfate gel electrophoresis coupled with immunoblotting may be the best quantitative approach.     

Analysis of polyphenols using capillary zone electrophoresis and HPLC: detection of soy, lupin, and pea protein in meat products

The analysis of polyphenols, which are characteristic of certain legumes, enables a rapid and sensitive detection of legume proteins in meat products. Separation of specific isoflavones can be achieved by capillary zone electrophoresis (CZE) or high-performance liquid chromatography (HPLC), both coupled with a photodiode array detector (DAD). The use of CZE in the identification process is an appropriate means of rapid screening; the HPLC is less dependent on matrix effects and clearly more sensitive. Additives of soy protein isolates up to 0.1% could be detected in meat products, even in sausages heated to a high temperature or with hydrolyzed soy proteins. A solid-phase extraction procedure with polyamide cartridges has been developed to concentrate polyphenols. A similar detection of lupin protein is possible in principle. In the case of pea protein, a reliable detection was not possible depending on the coincidental appearance of polyphenols as indicating substances.     

Yeast (Saccharomyces cerevisiae) protein concentrate: preparation, chemical composition, and nutritional and functional properties

The main objective of the present study was to compare the composition and functional and nutritional properties of whole yeast cells (WY) from an ethanol distillery with those of a phosphorylated protein concentrate (PPC) prepared from the same cells. Comparisons were also made of PPC with texturized soy protein (TSP) and soy protein isolate (SPI), both acquired in the local market. Yeast (Saccharomyces cerevisiae) is a rich source of protein, soluble fiber, and some minerals. Saturated fatty acids predominated over monounsaturated and polyunsaturated in both WY and PPC. The functional properties of PPC were similar to those of SPI and TSP. Both soy products and PPC replaced 20 or 40% chuck roll protein without affecting the emulsion properties of the meat products. Amino acid scoring was high for both WY and PPC; digestibility was higher (90%) for PPC and lower (68%) for WY. The protein nutritive value of PPC did not differ from that of casein and was significantly higher than that for WY.     

Protein quality, antigenicity, and antioxidant activity of soy-based foodstuffs

Commercial soy-based foodstuffs, including beverages ( n = 15), cow's milk supplemented with soy isoflavones ( n = 1), snacks ( n = 1), and biscuits ( n = 2), were analyzed to find any link between alterations in protein quality, safety (antigenicity), functionality (antioxidant activity), and food processing. Protein content was analyzed by the Kjeldhal method and available lysine by OPA assay. Chromatographic (RP-HPLC) and electrophoretic (SDS-PAGE) protein profiles were obtained to monitor modifications in the structure of soy allergens. The antigenicity was estimated by immunoblotting against soy total antibodies. Total phenol content was measured by Folin-Ciocalteu, while peroxyl radical scavenging activity of the sample was determined by ORAC FL assay. Protein content did not differ of those declared by the producers. Lysine availability was higher in liquid soy beverages compared to that in other soy foodstuffs studied here. 7S and 11S soy allergens were detected by RP-HPLC and SDS-PAGE, respectively. Both data indicated changes in soy protein patterns due to processing of instant powdered soymilk, soy snacks, and biscuits. Immunoblotting assay showed modifications in the antigenic response of these foodstuffs based on soy, suggesting that their processing had altered the structure of soy allergens. RP-HPLC, SDS-PAGE, and immunoblotting resulted in adequate analytical approaches for detecting changes in protein structure due to processing and adulteration. Protein quality, antigenicity, and antioxidant activity of soy products can be affected as a function of the intensity of the thermal processing.     

Chemical analysis of meat products

Meat, particularly muscle tissue, is basically a 3-component system of protein, moisture, and fat. Although this seems a simple analytical system in which to monitor product composition for regulatory compliance, the simplicity quickly erodes when meat is formulated into the broad variety of products commercially available today. Alternative protein sources, as well as preservatives, binders, extenders, emulsifiers, spices, and other flavoring ingredients, add to the analytes of concern and highlight the need for analytical methods suitable to support inspection and labeling requirements to ensure product compliance. Some key issues are noted which involve protein quality analysis, rapid compositional analysis, isolated soy protein analysis, and minced fish in meat products.     

Validation of a novel estrogen receptor-based microtitration plate assay for the determination of phytoestrogens in soy-based foods.

A novel, nonisotopic microtitration plate assay based on the human estrogen receptor has been used to screen soy-based and soy-containing foods for their phytoestrogen content (measured as genistein equivalents). The validation of the assay for use with food extracts has been demonstrated by investigation of recoveries after acidic and enzymic hydrolysis, by investigation of matrix effects, and by comparison of results with HPLC analysis. Phytoestrogen levels in soy products analyzed ranged between 520 and 1872 microgram of genistein equiv/g of soy flour, 5-282 microgram/g of soy concentrates, 503-1292 microgram/g of soy-protein isolates, and 108-226 microgram/g of soy-based infant formulas. Samples of textured vegetable protein and bread containing soy and linseed gave values of 1114 and 68 microgram/g, respectively. Comparison of results for 12 samples analyzed both by the receptor assay and by HPLC showed good correlation (r(2) = 0.905). The assay, which is rapid and simple to perform, is suitable for screening phytoestrogen-containing foods in order to assess human exposure to these bioactive compounds. The assay sensitivity is 3.4 microgram/g, and 14 samples/plate can be analyzed in 4 h following hydrolysis.     

Quantification of the group B soyasaponins by high-performance liquid chromatography

High-performance liquid chromatographic methods were developed for the isolation and quantitative determination of the group B soyasaponins, including 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP)-conjugated soyasaponins alphag, betag, and betaa, and their non-DDMP counterparts, soyasaponins V, I, and II, respectively, with formononetin used as the internal standard. The limits of quantification for soy products were 0.11-4.86 micromol/g. The within-day and between-days assay coefficients of variation were <9.8 and < 14.3%, respectively. The group B soyasaponin concentrations in 46 soybean varieties ranged from 2.50 to 5.85 micromol/g. Soy ingredients (soybean flour, toasted soy hypocotyls, soy protein isolates, textured vegetable protein, soy protein concentrates, and Novasoy) and soy foods (commercial soy milk, tofu, and tempeh) contained the group B soyasaponins from 0.20 to 114.02 micromol/g. There was no apparent correlation between isoflavone and soyasaponin concentrations in the soy products examined.     

Beta-conglycinin embeds active peptides that inhibit lipid accumulation in 3T3-L1 adipocytes in vitro

Obesity is a worldwide health concern because it is a well-recognized predictor of premature mortality. The objective was to identify soybean varieties that have improved potential to inhibit fat accumulation in adipocytes by testing the effects of soy hydrolysates having a range of protein subunit compositions on lipid accumulation and adiponectin expression in 3T3-L1 adipocytes. The results showed that differences in the protein distribution of 15 soy genotypes led to different potentials for the reduction of fat accumulation. The inhibition of lipid accumulation of soy alcalase hydrolysates in 3T3-L1 adipocytes ranged from 29 to 46%. Soy hydrolysates made from genotypes with 45.3 +/- 3.3% of total protein as beta-conglycinin, on average, showed significantly higher inhibition of lipid accumulation compared to those with 24.7 +/- 1.5% of extracted total protein as beta-conglycinin. Moreover, after in vitro simulated digestion with pepsin-pancreatin of the soy alcalase hydrolysates, 86% of the original activity remained. Adiponectin expression was induced in 3T3-L1 adipocytes treated with 15 soy hydrolysates up to 2.49- and 2.63-fold for high and low molecular weight adiponectin, respectively. The inhibition of lipid accumulation calculated from a partial least squares (PLS) analysis model correlated well with experimental data (R(2) = 0.91). In conclusion, it was feasible to differentiate soy varieties on the basis of the potential of their proteins to reduce fat accumulation using a statistical model and a cell-based assay in vitro. Furthermore, beta-conglycinin embeds more peptides than glycinin subunits that inhibit lipid accumulation and induce adiponectin in 3T3-L1 adipocytes. Therefore, soy ingredients containing beta-conglycinin may be important food components for the control of lipid accumulation in adipose tissue.     

Lunasin concentration in different soybean genotypes, commercial soy protein, and isoflavone products

Lunasin is a unique and novel cancer preventive peptide originally isolated from soy. Information on lunasin concentration of soybean cultivars and commercial soy proteins would be useful in developing lunasin-enriched cultivars and soy products. We report the development of an enzyme-linked immunosorbent assay (ELISA) method to identify lunasin and quantify the variations in concentration in 144 selected, diverse soybean accessions from the U.S. Department of Agriculture Soybean Germplasm Collection, several commercially available soy protein fractions and isoflavone-enriched products. With synthetic lunasin and monoclonal antibody, ELISA shows a linear concentration range of 24-72 ng/mL, good reproducibility, a detection limit of 8 ng/mL, and a recovery of 90% on spiked soy samples. Lunasin concentrations in the tested materials range from 0.10 to 1.33 g/100 g flour. Differences that exceeded 100% have been observed among accessions of similar maturity that were grown in the same environment, indicating that genetic differences in soybeans exist for lunasin. The mean of 23 major ancestral lines of U.S. cultivars is similar to the mean of 16 modern cultivars selected to represent the current diversity of the crop, but the highest values were found within the ancestral and exotic accessions. Soy protein concentrate, isolate, and hydrolyzate contain 2.81 +/- 0.30, 3.75 +/- 0.43, and 4.43 +/- 0.59 g lunasin/100 g flour, respectively, while soy flour and soy flakes contain 1.24 +/- 0.22 g lunasin/100 g flour. Isoflavone-enriched products contain very little or no lunasin. The relative mass (M(r)) of lunasin in the samples is 5.45 +/- 0.25 kDa. The wide range of lunasin concentrations within the Glycine max species indicates that the levels of this important bioactive peptide can be genetically manipulated. Furthermore, soy isolates and hydrolyzed soy proteins contain the highest concentrations of lunasin.     

Behavior of Bradford-reactive substances is consistent with predictions for glomalin

There is considerable controversy concerning detection in soils of the protein, glomalin, which is produced by arbuscular mycorrhizal fungi. Glomalin was originally defined as a substance that cross reacts with a monoclonal antibody formed against a substance in the cell walls of an arbuscular mycorrhizal fungus. Thus, one can use an immunological approach to detect glomalin. However, it was recently shown that other proteins cross react with the antibody. The other, more common, approach involves assay of soil protein using the Bradford reaction. This approach assumes that the Bradford assay is specific to protein, and that the assayed protein is largely glomalin, either because other proteins are in low concentration, or because the extraction process eliminates the possibility of their detection. These assumptions, however, have been called into question recently. One way to test whether the Bradford assay can be useful in quantifying glomalin is to determine whether the concentrations of Bradford-reactive substances are consistent with predictions for glomalin. For example, if recently produced glomalin is more labile than older glomalin, the concentrations of the two fractions should not be highly correlated. Moreover, when a contrast is established between mycorrhizal and nonmycorrhizal vegetation, recently produced glomalin should soon occur in higher concentration in soils supporting mycorrhizal vegetation. Older glomalin should not be found in higher concentrations in the soils of mycorrhizal vegetation until some time later. We tested these predictions by employing the Bradford assay during the course of a three-year field experiment in which canola (nonmycorrhizal) and soy (mycorrhizal) were grown in separate plots in year 1, both of which were followed by maize (mycorrhizal) in years 2 and 3. The correlation between the concentrations of fraction 1 Bradford-reactive substances (also known as easily extractable glomalin and frequently assumed to be recently produced) and fraction 2 (the more difficult-to-extract fraction and frequently assumed to be older glomalin), was very poor. In year 1, the concentration of fraction 1 was significantly greater in soy plots than in canola plots. Finally, fraction 2 was only significantly higher in the former soy plots than in former canola plots in years 2 and 3. These data support the hypothesis that the Bradford assay was useful in detecting glomalin in this case.     

不同种类大豆蛋白粉对面包加工特性的影响

为探索大豆蛋白作为营养补充剂在面包中应用时，对面团物理特性和焙烤特性产生的影响，该文考察了不同种类的大豆蛋白制品，包括大豆分离蛋白、灭酶全脂粉、活性全脂粉、活性脱脂粉、灭酶脱脂粉对面团粉质特性、拉伸特性和焙烤特性的影响。结果表明，面粉的吸水率与大豆蛋白粉氮溶解指数显著相关，面团的抗拉阻力受大豆蛋白添加量的影响明显。大豆蛋白粉的加入，对面包比体积产生不利影响，下降趋势与大豆蛋白粉对面团拉伸特性的影响显著相关。大豆蛋白粉有软化面包质地的作用，活性全脂粉表现最为明显。大豆蛋白粉的加入量占面粉质量分数的3%时，对面包口感影响不明显，当加入量超过面粉质量分数的7％时，容易出现发粘和豆腥味等现象。     

Near-infrared reflectance spectroscopy enables the fast and accurate prediction of the essential amino acid contents in soy, rapeseed meal, sunflower meal, peas, fishmeal, meat meal products, and poultry meal

Near-infrared reflectance spectroscopy (NIRS) calibrations were developed to enable the accurate and fast prediction of the total contents of methionine, cystine, lysine, threonine, tryptophan, and other essential amino acids, protein, and moisture in the most important protein-rich feed ingredients. More than 1000 samples of global origin collected over four years were analyzed on amino acids following the official methods of the United States and the European Union. Detailed data and graphics are given to characterize the obtained calibration equations. NIRS was validated with independent samples for soy and meat meal products and compared to the amino acid predictions using linear crude protein regressions. With a few exceptions, validation showed that 85-98% of the amino acid variance in the samples could be explained using NIRS. NIRS predictions compared to reference results agree excellently, with relative mean deviations below 5%. Especially for meat and poultry meals, NIRS can predict amino acids much better than crude protein regressions. By enabling the amino acid analysis of many samples to be completed in a short time, NIRS can improve the accuracy of feed formulation and thus the quality and production costs of mixed feeds.     

Characterization and quantification of proteins in lecithins

Several methods for extraction and quantification of proteins from lecithins were compared. Extraction with hexane-2-propanol-water followed by amino acid analysis is the most suitable method for isolation and quantification of proteins from lecithins. The detection limit of the method is 15 mg protein/kg lecithin, and the quantification limit is 50 mg protein/kg. The relative repeatability limits for samples containing 0-500 and 500-5000 mg protein/kg sample were 12.6 and 7.5%, respectively. The protein recovery ranged between 101 and 123%. The protein content has been determined in different kinds of lecithins. The results were as follows: standard soy lecithins (between 232 and 1338 mg/kg), deoiled soy lecithin (342 mg/kg), phosphatydylcholine-enriched soy lecithins (not detectable and 163 mg/kg), sunflower lecithins (892 and 414 mg/kg), and egg lecithin (50 mg/kg). The sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein patterns of the standard soy and sunflower lecithins are very similar to those of soy flour. The protein profile of the egg lecithin shows several bands with a broad range of molecular masses. The molecular masses of the main proteins of soy lecithins and soy flour have been determined by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and ranged from 10.5 to 52.2 kDa. Most of the major proteins from soy and sunflower lecithins identified by MALDI-MS and electrospray tandem MS belong to the 11S globulin fraction, which is one of the main fractions of soy and sunflower seeds. In addition, the seed maturation protein P34 from the 7S globulin fraction of soy proteins has also been identified in soy lecithins. This protein has been reported as the most allergenic protein in soybean.     

Development of a dot blot assay for the rapid detection of central nervous system tissue on meat and contact surfaces

As a potential transmitter of bovine spongiform encephalopathy (BSE), tissue from bovine central nervous system (CNS) is not accepted in meat and meat products. Western blot analysis of the CNS marker myelin proteolipid protein (PLP) detects CNS contamination selectively and sensitively. In this study, a rapid dot blot assay using an anti-PLP antibody was developed to screen CNS contamination of meat and contact surfaces. The detection limit was 0.01% bovine brain in minced bovine muscle. When applied to a swab test, down to 0.5 mg of CNS tissue on meat or other surfaces was detectable. Other offal tissues or peripheral nerves did not interfere with the assay. The test allows a differentiation between mammalian and avian CNS but not among mammalian species. The swab test was applied immediately after slaughtering at several areas of the bovine head. CNS was not detectable at any region which may enter the food chain.     

Rheological characterization of a novel functional food: tomato juice with soy germ

The rheological properties of tomato juice containing 1.5% soy germ were compared to plain tomato juice with and without soy protein isolate. This novel product was developed to provide a delivery system of carotenoids, soy protein, and significant isoflavone content without compromising the perceived juice characteristics of tomato product. Rheological tests depicted physical gel characteristics for all three products. Dynamic tests as a function of temperature showed that the stability and the compatibility between tomato juice and soy germ were higher as compared to soy protein isolate. The hydrophobic and electrostatic interactions between pectin and protein in the tomato soy protein isolate system were weakened as the temperature was increased. In the case of tomato juice with soy germ, the viscosity did not change during heating. The addition of soy germ increased the viscosity of tomato juice reinforcing the entire system without major qualitative effects on the rheological properties of plain tomato juice.     

Improving digestibility of soy flour by reducing disulfide bonds with thioredoxin

The Kunitz trypsin inhibitor (KTI) and the Bowman-Birk inhibitor (BBI) of trypsin and chymotrypsin contain disulfide bonds. Glycinin, the major storage protein in soybeans also contains disulfide bonds. Treatment of soy white flour with a NADP-thioredoxin system (NTS) effectively reduced disulfide bonds in soy flour and increased protein digestibility by trypsin and pancreatin as measured by the pH stat method. Treatment of soy flour with NTS increased the digestibility compared to soy white flour by 29.3 and 60.6% for trypsin and pancreatin, respectively. NTS-treated soy flour had similar digestibility by trypsin to autoclaved soy flour and casein, but digestibility by pancreatin was less than autoclaved soy flour and casein. The degree of reduction by NTS was highly correlated to the degree of hydrolysis (DH) by trypsin (R(2) = 0.93) and pancreatin (R(2) = 0.99). The DH of NTS-treated soy flour by trypsin is reflective of both inactivation of trypsin inhibitors and overall protein digestibility while pancreatin hydrolysis is reflective of only overall protein digestibility.     

Optimization of growth of Lactobacillus acidophilus FTCC 0291 and evaluation of growth characteristics in soy whey medium: a response surface methodology approach

Four strains of probiotics were evaluated for their alpha-galactosidase activity. Lactobacillus acidophilus FTCC 0291 displayed the highest specific alpha-galactosidase activity and was thus selected to be optimized in soy whey medium supplemented with seven nitrogen sources. The first-order model showed that meat extract, vegetable extract, and peptone significantly (P < 0.05) influenced the growth of L. acidophilus. The second-order polynomial regression estimated that maximum growth was obtained from the combination of 7.25% (w/v) meat extract, 4.7% (w/v) vegetable extract, and 6.85% (w/v) peptone. The validation experiment showed that response surface methodology was reliable with a variation of only 1.14% from the actual experimental data. Increased utilization of oligosaccharides and reducing sugars contributed to increased growth of L. acidophilus in the soy whey medium. This was accompanied by increased production of short-chain fatty acids and a decrease in pH.     

Detection and quantification of roundup ready soy in foods by conventional and real-time polymerase chain reaction

Transgenic soybean line GTS-40-3-2, marketed under the trade name Roundup Ready (RR) soy, was developed by Monsanto (USA) to allow for the use of glyphosate, the active ingredient of the herbicide Roundup, as a weed control agent. RR soy was first approved in Canada for environmental release and for feed products in 1995 and later for food products in 1996 and is widely grown in Canada. Consumer concern issues have resulted in proposed labeling regulations in Canada for foods derived from genetically engineered crops. One requirement for labeling is the ability to detect and accurately quantify the amount of transgenic material present in foods. Two assays were evaluated. A conventional qualitative Polymerase Chain Reaction (PCR) assay to detect the presence of soy and RR soy and a real-time PCR to quantify the amount of RR soy present in samples that tested positive in the first assay. PCR controls consisted of certified RR soy reference material, single transgenic soybeans, and a processed food sample containing a known amount of RR soy. To test real-world applicability, a number of common grocery store food items that contain soy-based products were tested. For some samples, significant differences in amplification efficiencies during the quantitative PCR assays were observed compared to the controls, resulting in potentially large errors in quantification. A correction factor was used to try to compensate for these differences.     

Variations in isoflavone levels in soy foods and soy protein isolates and issues related to isoflavone databases and food labeling

The reliability of databases on the isoflavone composition of foods designed to estimate dietary intakes is contingent on the assumption that soy foods are consistent in their isoflavone content. To validate this, total and individual isoflavone compositions were determined by HPLC for two different soy protein isolates used in the commercial manufacture of soy foods over a 3-year period (n = 30/isolate) and 85 samples of 40 different brands of soy milks. Total isoflavone concentrations differed markedly between the soy protein isolates, varying by 200-300% over 3 years, whereas the protein content varied by only 3%. Total isoflavone content varied by up to 5-fold among different commercial soy milks and was not consistent between repeat purchases. Whole soybean milks had significantly higher isoflavone levels than those made from soy protein isolates (mean +/- SD, 63.6 +/- 21.9 mg/L, n = 43, vs 30.2 +/- 5.8 mg/L, n = 38, respectively, p < 0.0001), although some isolated soy protein-based milks were similar in content to "whole bean" varieties. The ratio of genistein to daidzein isoflavone forms was higher in isolated soy protein-based versus "whole bean" soy milks (2.72 +/- 0.24 vs 1.62 +/- 0.47, respectively, p < 0.0001), and the greatest variability in isoflavone content was observed among brands of whole bean soy milks. These studies illustrate large variability in the isoflavone content of isolated soy proteins used in food manufacture and in commercial soy milks and reinforce the need to accurately determine the isoflavone content of foods used in dietary intervention studies while exposing the limitations of food databases for estimating daily isoflavone intakes.     

Effect of soy milk characteristics and cooking conditions on coagulant requirements for making filled tofu

The amount of coagulant added to soy milk is a critical factor for tofu-making; particularly it affects the textural properties of tofu. Earlier research indicated that the critical point of coagulant concentration (CPCC) is a characteristic parameter of soy milk and could be used as an effective indicator of optimal coagulant concentration (OCC) for making filled tofu. The objective of this study was to investigate the possible correlations between CPCC and the characteristics of soy milk made from various soybean samples and the effect of soy milk cooking and dilution conditions on CPCC. CPCC was determined by a titration method. Calcium chloride and magnesium chloride were used as coagulants. Soy milk characteristics including solid, protein, phytate, pH, titratable acidity, mineral content, and 11S/7S protein and these characteristics as affected by heating rate, heating time, and sequence of dilution and heating were studied. The results showed that the CPCC was significantly (p < 0.05) positively correlated with phytate content (grams per gram of protein), pH, and 7S protein content but negatively correlated with protein content, 11S protein content, 11S/7S ratio, titratable acidity, and original calcium content. Within the same soybean material, more proteins required more coagulant, but higher protein concentration during cooking resulted in less coagulant required by each gram of protein during coagulation. The CPCC decreased with increasing soy milk heating time or decreasing heating rate. The sequence of heating and diluting for preparing soy milk also had an effect on CPCC.