Lidia Dorantes-Alvarez

Purification and partial biochemical characterization of polyphenol oxidase from mamey (Pouteria sapota)

While a long shelf life for fruit products is highly desired, enzymatic browning is the main cause of quality loss in fruits and is therefore a main problem for the food industry. In this study polyphenol oxidase (PPO), the main enzyme responsible for browning was isolated from mamey fruit (Pouteria sapota) and characterized biochemically. Two isoenzymes (PPO 1 and PPO 2) were obtained upon ammonium sulfate precipitation and hydrophobic and ion exchange chromatography; PPO 1 was purified up to 6.6-fold with 0.28% yield, while PPO 2 could not be characterized as enzyme activity was completely lost after 24 h of storage. PPO 1 molecular weight was estimated to be 16.1 and 18 kDa by gel filtration and SDS-PAGE, respectively, indicating that the native state of the PPO 1 is a monomer. The optimum pH for PPO 1 activity was 7. The PPO 1 was determined to be maximum thermally stable up to 35°C. Kinetic constants for PPO 1 were K(m)=44 mM and K(m)=1.3 mM using catechol and pyrogallol as substrate, respectively. The best substrates for PPO 1 were pyrogallol, 4-methylcatechol and catechol, while ascorbic acid and sodium metabisulfite were the most effective inhibitors.

Effect of Thermal Treatment on the Antioxidant Activity and Content of Carotenoids and Phenolic Compounds of Cactus Pear Cladodes (Opuntia ficus-indica)

Cactus pears (Opuntia ficus-indica) are draught resistant plants originated in Mexico. Their flattened stem segments, called cladodes, have moisture, protein and fibre contents of 92, 1-2 and 4-6% respectively, and a pectin content in the range of 0.8-3.3% depending on the species. They also contain certain concentration of carotenoids which are of special interest because of their antioxidant activity. This work is aimed to identify and quantify the main carotenoids present in the stems and to evaluate the effect of thermal treatments on the antioxidant activity and concentration of carotenoids and phenolic compounds. The carotenoids -cryptoxanthin, -carotene and lutein were identified in the cladodes, the latter having the highest concentration. Thermal treatments increased the extractability of these pigments and the antioxidant activity was related to the carotenoids concentration. Total phenolic content decreased after the thermal treatments; however this result had little effect on the antioxidant activity. Mucilage present in the stems decreased the extractability of the carotenoids.

Antioxidant, Antinociceptive, and Anti-Inflammatory Effects of Carotenoids Extracted from Dried Pepper (Capsicum annuum L.)

Carotenoids extracted from dried peppers were evaluated for their antioxidant, analgesic, and anti-inflammatory activities. Peppers had a substantial carotenoid content: guajillo 3406 ± 4 μg/g, pasilla 2933 ± 1 μg/g, and ancho 1437 ± 6 μg/g of sample in dry weight basis. A complex mixture of carotenoids was discovered in each pepper extract. The TLC analysis revealed the presence of chlorophylls in the pigment extract from pasilla and ancho peppers. Guajillo pepper carotenoid extracts exhibited good antioxidant activity and had the best scavenging capacity for the DPPH+ cation (24.2%). They also exhibited significant peripheral analgesic activity at 5, 20, and 80 mg/kg and induced central analgesia at 80 mg/kg. The results suggest that the carotenoids in dried guajillo peppers have significant analgesic and anti-inflammatory benefits and could be useful for pain and inflammation relief.

Evaluation of Phenylpropanoids in Ten Capsicum annuum L. Varieties and Their Inhibitory Effects on Listeria monocytogenes Murray, Webb and Swann Scott A

Extracts of pepper from ten different varieties of Capsicum annuumL. were prepared and their phenylpropanoids content was analysed by HPLC. There were seven different compounds identified at varying concentrations depending on pepper variety (L-phenylalanine, t-cinnamic acid, o-coumaric acid, mcoumaric acid, ferulic acid, caffeic acid and capsaicin) which are intermediates of the capsaicinoids pathway. The inhibitory action of the pepper extracts was evaluated against Listeria monocytogenes Murray, Webb and Swann Scott A by the agar diffusion test. Results showed that three of the 10 extracts had an antibacterial effect. Guajillo San Luis pepper presented the highest inhibitory effect, and contained the major concentration of cinnamic and caffeic acids. In order to analyse the individual contribution of every phenylpropanoid to the bacteriostatic effect, different concentrations were tested on the growth and survival of L. monocytogenesSwann Scott A.There were no significant differences (p> 0.05) in the t-cinnamic, o-coumaric, m-coumaric, ferulic acids and capsaicin among the samples. However, a significant difference was found between these compounds and the bactericidal action of caffeic acid. The results confirmed that L. monocytogenesSwann Scott A is inhibited by some pepper extracts, and also that some specific phenylpropanoids had a bacteriostatic effect.

FT-MIR and Raman spectroscopy coupled to multivariate analysis for the detection of clenbuterol in murine model

A fast and simple screening method for the determination of clenbuterol at the ppb level in a murine model was demonstrated by Mid Infrared (MIR) and Raman spectroscopy in conjunction with multivariate analysis. In order to build the calibration models to quantify clenbuterol in rat meat, mixtures of rat meat and clenbuterol were prepared in a range of 5-10,000 ppb. Partial Least Square (PLS) analysis was used to build the calibration model. The results shown that Mid Infrared and Raman spectroscopy were efficient, but Mid Infrared (R(2) = 0.966 and SEC = 0.27) were superior to Raman (R(2) = 0.914 and SEC = 1.167). The SIMCA model developed showed 100% classification rate of rat meat samples with or without clenbuterol. The results were confirmed with contaminated meat samples from animals treated with clenbuterol. Chemometric models represent an attractive option for meat quality screening without sample pretreatments which can identify veterinary medicinal products at the ppb level.

Biochemical and Molecular Analysis of Some Commercial Samples of Chilli Peppers from Mexico

The genus Capsicum provides antioxidant compounds, such as phenolics and carotenoids, into the diet. In Mexico, there is a wide diversity of species and varieties of chilli peppers, a fruit which has local cultural and gastronomic importance. In the present study, the relationship of the carotenoid and phenolic profiles with the RAPD fingerprint of three different commercial cultivars of chilli peppers of seven regions of Mexico was investigated. Through RAPD, the species of chilli were differentiated by means of different primers (OPE-18, MFG-17, MFG-18, C51, and C52). The genetic distance found with OPE 18 was in the order of 2.6. The observed differences were maintained when the chromatographic profile of carotenoids, and the molecular markers were analyzed, which suggest a close relationship between carotenoids and the genetic profile. While the chromatographic profile of phenols and the molecular markers were unable to differentiate between genotypes of chilli peppers. In addition, by using infrared spectroscopy and statistical PCA, differences explained by geographic origin were found. Thus, this method could be an alternative for identification of chilli species with respect to their geographic origin.

A traditional intermediate moisture meat: Beef cecina

Cecina is an intermediate moisture meat produced and consumed to a large extent in Mexico. Four samples of cecina coming from different States of this country, were tested for water activity, colour, texture, fat, protein, moisture and chloride content. Sensory and microbiological analyses were also performed. Different fabrication methods for producing cecina were identified, involving large variations in the formulation of the product. There was a significant difference (P < 0·05) among samples regarding fat and chloride content, colour and texture. Differences in colour and saltiness were recorded through sensory analysis. Microbiological analysis showed higher counts than those recommended in the Mexican Official Standard for chopped and raw meat, due to poor sanitary conditions during production and marketing.

Halotolerance and Survival Kinetics of Lactic Acid Bacteria Isolated from Jalapeño Pepper (Capsicum annuum L.) Fermentation

The microbiota associated with spontaneous fermentation of vegetables in a saline substrate may represent an important group of bacteria in the food industry. In this work, the lactic acid bacteria (LAB) Weissella cibaria, Lactobacillus plantarum, Lactobacillus paraplantarum, and Leuconostoc citreum were identified by partial 16S rRNA gene sequence analysis. In addition, entophytic bacteria such as Pantoea eucalypti, Pantoea anthophila, Enterobacter cowanii, and Enterobacter asburiae were detected, but they were irrelevant for the fermentation process and were inhibited after 12 h of fermentation when the pH decreased from 6.5 to 4.9. Moreover, 2 species of yeast were isolated and identified as Hanseniaspora pseudoguilliermondii and Kodamaea ohmeri by their partial 26S rRNA gene sequence. The growth of LAB was evaluated at different sodium chloride contents. L. citreum was the most halotolerant species followed by L. plantarum and W. cibaria with a concentration index to obtain a 50% population reduction (IC(50)) of 7.2%, 6.6%, and 5.2%, respectively. Furthermore, the growth of LAB and Escherichia coli O157:H7 was evaluated in the presence of the main phenylpropanoids from chilli peppers such as p-coumaric and ferulic acid. It was determined that LAB can grow in both acids at 4 mM, unlike E. coli O157:H7, whose growth is inhibited in the presence of these acids.

Determination of Trichinella spiralis in pig muscles using Mid-Fourier Transform Infrared Spectroscopy (MID-FTIR) with Attenuated Total Reflectance (ATR) and Soft Independent Modeling of Class Analogy (SIMCA).

The aim of this work was to study the feasibility of detection of Trichinella spiralis in swine meat using Middle Infrared Spectroscopy Fourier Transform with Attenuated Total Reflectance (ATR) and Soft Independent Modeling of Class Analogy (MID-FTIR-ATR-SIMCA). Five male Pigs were orally infected at different larvae concentrations (13,000, 6500, 3500, 1625, 812 larvae/pig) and after 24 weeks the animals were euthanized. Five types of muscles were studied (leg, loin, rib, masseter, and diaphragm). Results showed that MID-FTIR-ATR-SIMCA was useful to determine the presence of T. spiralis in the samples, as the interclass distance between infected and non infected muscles varied from 13.5 to 36.8. This technique was also useful to discriminate among pig muscles, where masseter showed the largest interclass distance, while rib presented the smallest one. In all cases the recognition and rejection rates were 100%, which means that the methodology is capable of accurately separating T. spiralis infected from non infected swine meat.

Effects of Storage Temperature and Water Activity on the Degradation of Carotenoids Contained in Microencapsulated Chili Extract

The aim of this work was to evaluate the effects of storage temperature and water activity on degradation of carotenoids contained in microencapsulates of non-aqueous extracts from chili (NAEC). Total carotenoids content and adsorption isotherms of microencapsulated NAEC in a 1:1 weight ratio with gum Arabic-maltodextrin DE 20 (GA 50%–MD50%) were determined at 25, 35, and 40°C. The isotherms were fitted using the Guggenheim-Anderson-de-Boer model and their enthalpies and entropies, both differential and integral, were estimated by the Clausius-Clapeyron method. The minimum integral entropy was considered as the point of maximum stability at which water less readily participates in degradation reactions. Zones of minimum integral entropy were found between 7.56–8.30, 6.10–6.95, and 5.15–6.04 kg H2O/100 kg dried solids, corresponding to water activity (aw) of 0.210–0.239, 0.238–0.277, and 0.262–0.313 at 25, 35, and 40°C, respectively. Total carotenoids content (CT) degraded over time, but degradation of carotenoids was lower in microcapsules stored at 25°C than those stored at 35 or 40°C. The morphology of microcapsules was altered at aw > 0.6, including swelling of the polysaccharide matrix was presented, and possible subsequent dissolution of the wall material, which indicates a high rate of carotenoid degradation. When microencapsulated NAEC were storage between 0.2–0.6 of water activity, the highest glass transition temperatures were achieved. In this range, the wall materials of the microcapsules suffer less microstructural modifications, associated with the minimum level of degradation of carotenoids. Sometimes, in this water activity range, the zones of minimal entropy were observed.