Application of experimental design as a statistical approach to recover bioactive peptides from different food sources.

Bioactive peptides (BAPs) derived from samples of animals and plants have been widely recommended and consumed for their beneficial properties to human health and to control several diseases. This work presents the applications of experimental designs (DoE) used to perform factor screening and/or optimization focused on finding the ideal hydrolysis condition to obtain BAPs with specific biological activities. The collection and discussion of articles revealed that Box Behnken Desing and Central Composite Design were the most used. The main parameters evaluated were pH, time, temperature and enzyme/substrate ratio. Among vegetable protein sources, soy was the most used in the generation of BAPs, and among animal proteins, milk and shrimp stood out as the most explored sources. The degree of hydrolysis and antioxidant activity were the most investigated responses in obtaining BAPs. This review brings new information that helps researchers apply these DoE to obtain high-quality BAPs with the desired biological activities.

Structural characterization of the human DjC20/HscB cochaperone in solution.

J-domain proteins (JDPs) form a very large molecular chaperone family involved in proteostasis processes, such as protein folding, trafficking through membranes and degradation/disaggregation. JDPs are Hsp70 co-chaperones capable of stimulating ATPase activity as well as selecting and presenting client proteins to Hsp70. In mitochondria, human DjC20/HscB (a type III JDP that possesses only the conserved J-domain in some region of the protein) is involved in [FeS] protein biogenesis and assists human mitochondrial Hsp70 (HSPA9). Human DjC20 possesses a zinc-finger domain in its N-terminus, which closely contacts the J-domain and appears to be essential for its function. Here, we investigated the hDjC20 structure in solution as well as the importance of Zn+2 for its stability. The recombinant hDjC20 was pure, folded and capable of stimulating HSPA9 ATPase activity. It behaved as a slightly elongated monomer, as attested by small-angle X-ray scattering and SEC-MALS. The presence of Zn2+ in the hDjC20 samples was verified, a stoichiometry of 1:1 was observed, and its removal by high concentrations of EDTA and DTPA was unfeasible. However, thermal and chemical denaturation in the presence of EDTA led to a reduction in protein stability, suggesting a synergistic action between the chelating agent and denaturators that facilitate protein unfolding depending on metal removal. These data suggest that the affinity of Zn+2 for the protein is very high, evidencing its importance for the hDjC20 structure.

Solubility of Cd, Cr, Cu, Ni, and Pb and Its Correlation with Total Polyphenols and Soluble Melanoidins in Hot Infusions of Green and Roasted Mate.

The total and soluble contents of three potentially toxic elements (PTEs) (Cd, Cr, and Pb) and two essential elements (Cu and Ni) in commercial green and roasted mate (Ilex paraguariensis St. Hil.) and their hot infusions were accessed by inductively coupled plasma mass spectrometry. Total polyphenolic (TP) contents in the infusions were accessed by Folin-Ciocalteu method, and the presence of soluble melanoidins (SM) was evaluated by measuring absorbances at 420 nm. The soluble element contents in hot infusions of green mate were present in the following increasing order: Pb < Cd < Cr < Cu, and Ni. For hot infusions of roasted mate, the increasing order was: Cu < Pb < Cd < Cr < Ni. Except for Cr, hot infusions of roasted mate supply lower contents of the soluble PTEs than hot infusions of green mate. Cadmium was the only of evaluated trace element found in hot infusions of green and roasted mate as well as in some of their mate samples above the maximum allowed contents established by Brazilian legislation. Despite this, polyphenol and melanoidin contents in green and roasted mate infusions may reduce Cd bioavailability. The soluble trace elements that correlated directly with total polyphenols in hot infusions of green or roasted mate are indirectly correlated with the presence of soluble melanoidins. Moreover, this is the first study to evaluate the correlations of soluble forms of Cd, Cr, Cu, Ni, and Pb with the presence of SM and TP contents in hot infusions of green and roasted mate.

Multi-elemental determination in meat samples using multi-isotope calibration strategy by ICP-MS.

The multi-isotope calibration (MICal) is a fast and accurate calibration strategy for multi-elemental determination that uses only two solutions, (1) 50% (v v-1) sample plus 50% (v v-1) analytical blank solution, and (2) 50% (v v-1) sample plus 50% (v v-1) standard solution (containing the analytes). The calibration curves were built as a function of the intensity signals of solution 1 (y-axis) versus solution 2 (x-axis), employing the various isotopes of each monitored mass/charge. The concentration of the analytes was calculated from the slope and the concentrations of the standards added to solution 2. The validation of the method was evaluated using standard reference materials of bovine liver and bovine muscle. The recoveries varied from 87 to 113% for MICal, which were similar to the conventional standard additions calibration procedure (83 to 120%).

Multielemental Determination of As, Bi, Ge, Sb, and Sn in Agricultural Samples Using Hydride Generation Coupled to Microwave-Induced Plasma Optical Emission Spectrometry.

A microwave-induced plasma optical emission spectrometer with N2-based plasma was combined with a multimode sample introduction system (MSIS) for hydride generation (HG) and multielemental determination of As, Bi, Ge, Sb, and Sn in samples of forage, bovine liver, powdered milk, agricultural gypsum, rice, and mineral fertilizer, using a single condition of prereduction and reduction. The accuracy of the developed analytical method was evaluated using certified reference materials of water and mineral fertilizer, and recoveries ranged from 95 to 106%. Addition and recovery experiments were carried out, and the recoveries varied from 85 to 117% for all samples evaluated. The limits of detection for As, Bi, Ge, Sb, and Sn were 0.46, 0.09, 0.19, 0.46, and 5.2 µg/L, respectively, for liquid samples, and 0.18, 0.04, 0.08, 0.19, and 2.1 mg/kg, respectively, for solid samples. The method proposed offers a simple, fast, multielemental, and robust alternative for successful determination of all five analytes in agricultural samples with low operational cost without compromising analytical performance.

Determination of As and in mineral waters by fast sequential continuous flow hydride generation atomic absorption spectrometry.

In this study, a novel analytical method which consists of a combination of Fast Sequential Flame Atomic Absorption Spectrometry (FS-FAAS) and Continuous Flow Hydride Generation Atomic Absorption Spectrometry (CF-HGAAS) is proposed. The method developed was employed for the sequential determination of As and Sb at sub-µg L-1 levels in bottled mineral waters. A strong enhancement in the analytical throughput was obtained when compared with the traditional mono-element CF-HGAAS with a quartz tube atomizer (QTA). Variables which would affect the method performance such as Ar flow rate, HCl and NaBH4 concentrations as well as delay and integration time were optimized. A flame atomic absorption spectrometer working in fast sequential mode was used in all experiments. After just 20 s of read delay, As and Sb were sequentially determined in 6 s (3 s each element). A 26-2 fractional factorial design was employed for studies of potential interferents with transition metals that could be present in mineral water samples. Limits of detection obtained for As and Sb were 0.15 and 0.14 µg L-1, respectively. The accuracy of the proposed method was checked by the use of 2 certified reference materials: Trace elements in water (NIST 1643e) and Trace metals in drinking water (HPS TMDW). Good agreement between certified and found concentrations was observed. Finally, As and Sb were determined in commercial bottled mineral water samples. Adequate sensitivity, high throughput and minimization of reagents and sample consumption are the attractive features of this new method.

A compact miniaturized continuous flow system for the determination of urea content in milk.

A multicommutation-based flow system with photometric detection was developed, employing an analytical microsystem constructed with low temperature co-fired ceramics (LTCC) technology, a solid-phase reactor containing particles of Canavalia ensiformis DC (urease source) immobilized with glutaraldehyde, and a mini-photometer coupled directly to the microsystem which monolithically integrates a continuous flow cell. The determination of urea in milk was based on the hydrolysis of urea in the solid-phase reactor and the ammonium ions produced were monitored using the Berthelot reaction. The analytical curve was linear in the urea concentration range from 1.0 x 10(-4) to 5.0 x 10(-3) mol L(-1) with a limit of detection of 8.0 x 10(-6) mol L(-1). The relative standard deviation (RSD) for a 2.0 x 10(-3) mol L(-1) urea solution was lower than 0.4% (n = 10) and the sample throughput was 13 h(-1). To check the reproducibility of the flow system, calibration curves were obtained with freshly prepared solutions on different days and the RSD obtained was 4.7% (n = 6). Accuracy was assessed by comparing the results of the proposed method with those from the official procedure and the data are in close agreement, at a 95% confidence level.

Tungsten permanent chemical modifier for fast estimation of Se contents in soil by graphite furnace atomic absorption spectrometry.

A tungsten carbide coating on the integrated platform of a transversely heated graphite atomizer was used as a modifier for the direct determination of Se in soil extracts by graphite furnace atomic absorption spectrometry. Diethylenetriaminepentaacetic acid (0.0050 mol L(-1)) plus ammonium hydrogencarbonate (1.0 mol L(-1)) extracted predominantly available inorganic selenate from soil. The formation of a large amount of carbonaceous residue inside the atomizer was avoided with a first pyrolysis step at 600 degrees C assisted by air during 30 s. For 20 microL of soil extracts delivered to the atomizer and calibration by matrix matching, an analytical curve (10.0-100 microgram of L(-1)) with good linear correlation (r = 0.999) between integrated absorbance and analyte concentration was established. The characteristic mass was approximately 63 pg of Se, and the lifetime of the tube was approximately 750 firings. The limit of detection was 1.6 microgram L(-1), and the relative standard deviations (n = 12) were typically <4% for a soil extract containing 50 microgram of L(-1). The accuracy of the determination of Se was checked for soil samples by means of addition/recovery tests. Recovery data of Se added to four enriched soil samples varied from 80 to 90% and indicated an accurate method.