Quantification of Nε-(2-Furoylmethyl)-L-lysine (furosine), Nε-(Carboxymethyl)-L-lysine (CML), Nε-(Carboxyethyl)-L-lysine (CEL) and total lysine through stable isotope dilution assay and tandem mass spectrometry

The control of Maillard reaction (MR) is a key point to ensure processed foods quality. Due to the presence of a primary amino group on its side chain, lysine is particularly prone to chemical modifications with the formation of Amadori products (AP), Nε-(Carboxymethyl)-L-lysine (CML), Nε-(Carboxyethyl)-L-lysine (CEL). A new analytical strategy was proposed which allowed to simultaneously quantify lysine, CML, CEL and the Nε-(2-Furoylmethyl)-L-lysine (furosine), the indirect marker of AP. The procedure is based on stable isotope dilution assay followed by, liquid chromatography tandem mass spectrometry. It showed high sensitivity and good reproducibility and repeatability in different foods. The limit of detection and the RSD% were lower than 5 ppb and below 8%, respectively. Results obtained with the new procedure not only improved the knowledge about the reliability of thermal treatment markers, but also defined new insights in the relationship between Maillard reaction products and their precursors

Application of Apparent Metabolizable Energy versus Nitrogen-Corrected Apparent Metabolizable Energy in Poultry Feed Formulations: A Continuing Conundrum

In the present investigation, N retention, AME, and AMEn data from six energy evaluation assays, involving four protein sources (soybean meal, full-fat soybean, rapeseed meal and maize distiller’s dried grains with solubles [DDGS]), are reported. The correction for zero N retention, reduced the AME value of soybean meal samples from different origins from 9.9 to 17.8% with increasing N retention. The magnitude of AME penalization in full-fat soybean samples, imposed by zero N correction, increased from 1.90 to 9.64% with increasing N retention. The Δ AME (AME minus AMEn) in rapeseed meal samples increased from 0.70 to 1.09 MJ/kg as N-retention increased. In maize DDGS samples, the correction for zero N retention increased the magnitude of AME penalization from 5.44 to 8.21% with increasing N retention. For all protein sources, positive correlations (p < 0.001; r = 0.831 to 0.991) were observed between the N retention and Δ AME. The present data confirms that correcting AME values to zero N retention for modern broilers penalizes the energy value of protein sources and is of higher magnitude for ingredients with higher protein quality. Feed formulation based on uncorrected AME values could benefit least cost broiler feed formulations and merits further investigation

Influence of Conditioning and Expansion Characteristics on the Apparent Metabolizable Energy and Standardized Ileal Amino Acid Digestibility of Full-Fat Soybeans for Broilers

This study investigated the influence of short-term and long-term conditioning and expansion on the nitrogen-corrected apparent metabolizable energy (AMEn) and standardized ileal digestibility (SID) of amino acids (AA) in full-fat soybeans (FFSB) for broilers. A batch of raw soybeans was used to manufacture 10 FFSB products (T0 to T9) by applying various combinations of conditioning and expansion. The AMEn and SID AA of FFSB were determined by difference and direct methods, respectively. All heat treatments increased (p p p < 0.05) SID AA was recorded for T5. The results demonstrated that the long-term conditioning of FFSB at 100 °C for 6 min prior to expansion with 18 kWh/t specific energy input enhanced metabolizable energy and SID AA. Further increases in conditioning time from 6 to 9 min or expansion of specific energy input from 18 to 28 kWh/t did not yield additional benefits to energy utilization and AA digestibility of FFSB

Bioavailability of liquid methionine hydroxy analogue-free acid relative to DL-methionine in broilers

An experiment with broiler chickens was conducted to compare the relative bioavailability of liquid methionine hydroxy analogue free acid (MHA-FA) with that of DL-methionine (DLM) during fattening to 35 days of age. Ross 308 male chicks were allotted to 9 treatments, each consisting of six replicates of 140 birds/pen. Four graded levels (0.04, 0.08, 0.16, and 0.28 %) of MHA-FA or DLM products (weight/weight comparison) were added to a maize-wheat-soyabean meal basal diet deficient in sulphur amino acids. The criteria of response were body weight, feed conversion ratio, carcass yield and breast meat yield. Significant responses to graded levels of both methionine sources were observed in all response criteria. Using a multi-exponential model describing the dose-response relationships, the bioavailability estimates of MHA-FA relative to DLM on a weight-to-weight basis were 68, 70, 54 and 59 % for body weight, feed conversion, carcass yield and breast meat yield, respectively. If MHA-FA was compared with DLM on equimolar basis its bioavailability was 77.7, 79.0, 59.3 and 64.6 for body weight, feed conversion, carcass yield and breast meat yield, respectively. The bioavailability of MHA-FA for carcass yield and breast meat yield was significantly (P &lt; 0.05) lower than that of DLM on a weight-to-weight and on equimolar basis

The quantification of free Amadori compounds and amino acids allows to model the bound Maillard reaction products formation in soybean products

The quantification of protein bound Maillard reaction products (MRPs) is still a challenge in food chemistry. Protein hydrolysis is the bottleneck step: it is time consuming and the protein degradation is not always complete. In this study, the quantitation of free amino acids and Amadori products (APs) was compared to the percentage of blocked lysine by using chemometric tools. Eighty thermally treated soybean samples were analyzed by mass spectrometry to measure the concentration of free amino acids, free APs and the protein-bound markers of the Maillard reaction (furosine, Nε-(carboxymethyl)-L-lysine, Nε-(carboxyethyl)-L-lysine, total lysine). Results demonstrated that Discriminant Analysis (DA) and Correlated Component Regression (CCR) correctly estimated the percent of blocked lysine in a validation and prediction set. These findings indicate that the measure of free markers reflects the extent of protein damage in soybean samples and it suggests the possibility to obtain rapid information on the quality of the industrial processes

Effects of gradual differences in trypsin inhibitor activity on the estimation of digestible amino acids in soybean expellers for broiler chickens

The present study investigated the effect of varying trypsin inhibitor activity (TIA) in differently processed soybean expellers on apparent prececal amino acid (AA) digestibility in male broiler chickens. Two different raw soybean batches were treated using varying processing techniques and intensities. In this way, 45 expeller extracted soybean meal (ESBM) variants were created. The processed soybean variants were then merged into a basal diet (160 g/kg crude protein (CP)) at two inclusion levels (15%, 30%) resulting in 90 different diets plus one basal diet (0.4 mg/g – 8.5 mg/g TIA). All diets contained 0.5% titanium dioxide. 5,460 day old male broilers (Ross 308) were allocated on day 14 to 546 pens (10 birds/pen) after a starter phase (CP 215 g/kg, 14 g/kg Lysine, 12.5 MJ ME/kg). The 91 experimental diets were fed ad libitum until day 22. Subsequently, birds were euthanized and digesta of the terminal ileum was collected for determination of AA digestibility. TIA depressed the prececal digestibility of every single AA significantly in a straight linear fashion (P<0.001). Sulfur-containing AA expressed the strongest suppression by TIA with cystine showing the lowest apparent prececal digestibility measured (4.94% at 23.6 mg/g TIA in raw ESBM). The present data demonstrate that TIA severely depresses digestibility of essential and non-essential AA in a straight linear fashion. On the one hand, this questions the usefulness of defined upper limits of TIA in soy products whereas on the other hand, TIA must be considered when testing raw components for their feed protein value in viv