Olga N. Campas-Baypoli

HPLC method validation for measurement of sulforaphane level in broccoli by-products

A simple and specific analytical method was developed and tested for the determination of sulforaphane in broccoli by-products. The method includes the optimization of the conversion of glucoraphanin to sulforaphane, followed by purification of extracts using solid-phase extraction and high-performance liquid chromatography (HPLC) analysis. The response surface methodology was used to find optimum conditions for the preparation and purification procedure. Chromatographic conditions for reversed-phase HPLC with UV photodiode array detection were as follows: column, Exil ODS C(18), 25 x 0.46 cm, 5 microm; column temperature, 36 degrees C; mobile phase, a 30 : 70 (v/v) mixture of acetonitrile:water; flow rate, 0.6 mL/min. The detection wavelength was UV 202 nm. Under these conditions, excellent linearity was obtained (r(2) = 1), and the overall recovery was 97.5 and 98.1% for fresh florets and lyophilized florets, respectively. The precision results showed that the relative standard deviation of the repeatability for florets fresh and lyophilized was 3.0 and 4.0%, respectively. Sulforaphane contents were determined in the edible portion of fresh broccoli, and broccoli crop remains.

Biochemical composition and physicochemical properties of broccoli flours

The objective of this research was to study the biochemical composition and physicochemical properties of three different flours prepared from broccoli crop remains. Florets, leaves and stalks of broccoli were dried at 60°C, and the flours obtained were analysed for proximate composition, amino acid profile, fatty acid composition, and physicochemical properties. The florets flour showed the highest protein content (22.41 g/100 g dry weight); ash was higher in leaves flour (14.67 g/100 g dry weight), and the lipid content was similar in the flours of leaves and stalks. The stalks flour had high crude fibre content and low protein content. All flours presented a high water absorption index. Tyrosine, aspartic acid, glutamic acid, proline and valine were found in larger concentration. The most abundant fatty acids in the lipids were linolenic acid (C18:3n3), palmitic acid (C16:0) and linoleic acid (C18:2n6). Broccoli flours prepared in this study are good source of nutrients and could be utilized as dietary supplements.

Microencapsulation of sulforaphane from broccoli seed extracts by gelatin/gum arabic and gelatin/pectin complexes

Sulforaphane is a phytochemical that has received attention in recent years due to its chemopreventive properties. However, the uses and applications of this compound are very limited, because is an unstable molecule that is degraded mainly by changes in temperature and pH. In this research, the use of food grade polymers for microencapsulation of sulforaphane was studied by a complex coacervation method using the interaction of oppositely charged polymers as gelatin/gum arabic and gelatin/pectin. The polymers used were previously characterized in moisture content, ash and nitrogen. The encapsulation yield was over 80%. The gelatin/pectin complex had highest encapsulation efficiency with 17.91%. The presence of sulforaphane in the complexes was confirmed by FTIR and UV/visible spectroscopy. The materials used in this work could be a new and attractive option for the protection of sulforaphane.

Antimicrobial Activity of Chitosan Membranes Against Staphylococcus Aureus of Clinical Origin

Healthy human skin has beneficial microflora and many pathogens causing infections. Staphylococcus aureus is the most prevalent and can have multiresistance to antibiotics. Chitosan is a polysaccharide composed of glucosamine and N-acetyl-D-glucosamine, which is biodegradable and has antimicrobial activity. As part of a national scientific research project for the development and application of biomaterials, we decided to study the effect of different membranes based on chitosan against strains of S. aureus isolated from infected ulcers. The study found that seven of nine strains of S. aureus are sensitive to rifampin and the least eight of nine strains were multiresistant to more than ten antibiotics. All chitosan-based membranes confirm its antimicrobial effect on direct contact with an increase in its diameter. The contact area of the membranes is increased according to the concentration of chitosan. The highest average area increase was the chitosan membranes with honey and glycerin, 88.32%. Chitosan membranes have shown their effectiveness against S. aureus strains of clinical origin. Thus, these materials can be applied for the treatment of chronic ulcers without toxic hazards and resistance caused by antibiotics.

CUANTIFICACIÓN DE RIBOFLAVINA (VITAMINA B2) EN PRODUCTOS LÁCTEOS POR HPLC

La riboflavina (B2) es una vitamina hidrosoluble del complejo B, cuyo aporte nutritional es imprescindible para el buen funcionamiento del organismo. La leche es una de las principales fuentes de riboflavina en la dieta humana, sin embargo, en la mayoria de los alimentos se degrada durante los procesos termicos convencionales y en el almacenamiento. En el presente trabajo se determino el contenido de riboflavina en productos lacteos comerciales, leches para consumo directo y yogurt, por cromatografia liquida de alta resolucion (HPLC) con deteccion por fluorescencia. La preparacion de la muestra incluye una hidrolisis acida (HC10. IN), seguida por una digestion enzimatica, y por ultimo, la precipitacion de proteinas (TCA100 %). En el analisis cromatografico se utilizo un sistema isocratico (acetato de amonio 5 mM-metanol, 72:28 v/v). El contenido de riboflavina de las muestras de yogurt estuvo en el rango de 0,289 - 3,078 µ/g BS, mientras que en la leche ultrapasteurizada y pasteurizada de 0,61-13,64 µg/gBS y 11,73-15,41 µg/ g BS, respectivamente. Las leches para consumo directo y el yogurt comercial son fuentes de riboflavina, y su consumo regular puede ayudar a satisfacer los requerimientos diarios.

Antioxidant and chelating capacity of Maillard reaction products in amino acid-sugar model systems: applications for food processing

BACKGROUND Maillard reaction products (MRP) have gained increasing interest owing to their both positive and negative effects on human health. Aqueous amino acid-sugar model systems were studied in order to evaluate the antioxidant and chelating activity of MRP under conditions similar to those of food processing. Amino acids (cysteine, glycine, isoleucine and lysine) combined with different sugars (fructose or glucose) were heated to 100 and 130 °C for 30, 60 and 90 min. Antioxidant capacity was evaluated via ABTS and DPPH free radical scavenging assays, in addition to Fe2+ and Cu2+ ion chelating capacity. RESULTS In the ABTS assay, the cysteine-fructose model system presented the highest antioxidant activity at 7.05 µmol mL-1 (130 °C, 60 min), expressed in Trolox equivalents. In the DPPH assay, the cysteine-glucose system presented the highest antioxidant activity at 3.79 µmol mL-1 (100 °C, 90 min). The maximum rate of chelation of Fe2+ and Cu2+ was 96.31 and 59.44% respectively in the lysine-fructose and cysteine-glucose systems (100 °C, 30 min). CONCLUSION The model systems presented antioxidant and chelating activity under the analyzed temperatures and heating times, which are similar to the processing conditions of some foods.

Quantitative HPLC Analysis of Riboflavin and Aromatic Amino Acids in Three Forms of Shrimp Hydrolysates

Abstract This paper presents a high performance liquid chromatography (HPLC) method for the determination of riboflavin and free aromatic amino acids. To determine riboflavin, the method includes an acid hydrolysis (HCl 0.1 N) followed by an enzymatic digestion and a protein precipitation with trichloroacetic acid. An analytical column, Chrom SEP SS C18 5 µm (4.6 mm × 150 mm) was used with an isocratic flow using a mobile phase consisting of 5 mM ammonium acetate-methanol (72:28, vv−1), at a flow rate of 1.0 mL min−1 to 28°C. The method showed adequate linearity, repeatability, reproducibility, accuracy, and limits of detection. The method was used to quantify the cited analytes in three different forms of hydrolysate obtained from fermented shrimp remnants.

PCR Assay for Detection of Staphylococcus aureus in Fresh Lettuce (Lactuca sativa)

The growth in food demand and production growth of vegetables have led to the development of intensive production systems with the aim of having regular access to enough high‐quality food. The aim is to determine the incidence of Staphylococcus aureus in fresh lettuce by PCR in order to enhance the efficiency for detection and identification process. The Baird‐Parker method was used for isolating pathogens from 54 lettuce samples. Genomic DNA extraction was performed according the Mericon DNA Bacteria Plus Kit. The detection by PCR was performed using the pair of primers: coa gene (5′‐ ATAGAGCTGATGGTACAGG‐3′ and 5′‐GCTTCCGATTGTTCGATGC‐3′). The phyloge‐ netic tree was constructed by comparing conserved sequences from the adjacent 16S gene, using the F2C 5′‐AGAGTTTGATCATGGCTC‐3′ and C 5′‐ACGGGCGGTGTGTAC‐3′ primers. To test the antimicrobial effect, we used the disk diffusion method (Kirby‐Bauer) using Mueller‐Hinton agar and five antibiotics with different concentrations. The incidence of S. aureus was 1.7%. All the isolates were situated in the ATCC 11632 clade in accordance with other reported sequences belonging to this pathogen in the NCBI database. All the isolates seemed to be resistant to penicillin (10U). The molecular techniques used in this study are suitable for the identification of S. aureus isolated from lettuce, increasing our capability of detecting this pathogen by improving the process and increasing the efficiency contributing to the safety of this vegetable.

Astaxanthin and Its Esters in Pigmented Oil from Fermented Shrimp By-Products

ABSTRACT This study focuses on the isolation and quantification of astaxanthin and its esters in the oils extracted from shrimp waste. The content of astaxanthin was determined by high performance liquid chromatography and ranged from 1.5 to 2.1 mg g−1. The astaxanthin and its esters were isolated by thin layer chromatography in three fractions: free astaxanthin, monoester, and diester of astaxanthin, with percentages of 44, 32, and 26%, respectively. Oleic acid was the major fatty acid in the monoester fraction, while palmitic, oleic, and stearic acids were present predominantly in the diester fraction. Free radical-scavenging activity of the oils and the three fractions was evaluated. All oils showed an effect on the decrease in the absorbtion of 1,1-diphenyl-2-picrylhydrazyl solutions, while among the fractions no significant difference occurred. Pigmented oil from shrimp waste is an organic source of pigment and antioxidant astaxanthin, the esters, and fatty acids, such as eicosapentaenoic and docosahexaenoic.