Kinetic study of a commercial lipase for hydrolysis of semi-refined oil of anchovy (Engraulis ringens) [Estudio cinético de una lipasa comercial para la hidrólisis de aceite semirrefinado de anchoa (Engraulis ringens)]

Lipases due to their ecological nature and catalytic versatility, are ideal for their application in the fish oil hydrolysis industry due to their selective property, which allows the preservation of polyunsaturated fatty acids (PUFAs) in the lipid structure. The objective of this research was to determine the activity and kinetic parameters of a commercial AY AMANO "30SD" lipase, as well as the temperature and time values ​​to achieve an optimal degree of hydrolysis in semi-refined anchovy oil. The experiments were carried out in a jacketed minireactor with a working volume of 400 mL (oil-water-enzyme) with temperature control and pH 7.00, enzyme concentration 350 U/mL and stirring 160 rpm. A 3x3 factorial design and the response surface methodology were used. The results obtained from the study of the enzyme were: activity = 37 384.55 ± 395.07 U/g and kinetic parameters: Km = 7.98 g/L and Vmax. = 0.038887 g/Lxmin. Correspondingly, the following optimal parameters were obtained: Degree of hydrolysis 4.01%, temperature 46.86 °C and hydrolysis time 90 minutes, with a confidence level of 95% (p <0.05). Conclusions: The study allowed us to kinetically characterize the commercial lipase and determine the optimum degree of hydrolysis of the semi-refined anchovy oil.

Microencapsulation of Anchovy Fish Oil (Engraulis encrasicolus) with Fish Protein (Equulites klunzingeri) Isolate: Nutritional Assessment

Fish protein isolates extracted from underutilised fish species were used for coating material of anchovy oils and their nutritive value was investigated in this study. For this purpose, Klunzinger’s ponyfish (Equulites klunzingeri) proteins were extracted by using pH shifting process. Micro particles were prepared with anchovy oil (Engraulis encrasicolus) as core material (10%), and as wall materials a ratio of 5% and 10% fish protein isolate (FPI) was used. Maltodextrin (DE: 18:20) was added to both groups in a ratio of 10%. The emulsions were fed immediately into a Buchi Mini Spray Dryer (B290, Switzerland). The inlet temperatures, feed rate and aspiration rate were maintained at 160 oC, 15 mL/min and 35m3/h, respectively. The lipid, protein and moisture contents of anchovy oil microcapsules containing 5% FPI and 10% FPI were found as 43.76-43.09%, 4.34- 9.82% and 3.95-3.92%, respectively. The main amino acids in microcapsule samples were lysine, glutamic acid, and leucine which constituted in the range of 349-578 mg/100 g sample for microcapsules containing 5% FPI, and 805-1547 mg/100 g sample for microcapsules containing 10% FPI. In addition to that, essential and non-essential amino acids (E/NE) ratio for microencapsulated fish oil with 5% FPI and 10% FPI were determined 0.92 and 0.95, respectively. As a result of this study, it can be concluded that the addition of fish protein isolate enhanced the nutritive value of microencapsulated fish oil

n-3 LC-PUFA Enrichment Protocol for Red Earthworm, Eisenia fetida: A Cheap and Sustainable Method

This study assessed the potential of omega-3 long chain polyunsaturated fatty acids (n-3 LC-PUFA) enrichment in red earthworm (Eisenia fetida, REW), via the addition of fish oil (FO, anchovy oil) to the culture-compost. FO addition to compost was tested at graded inclusion doses (0, 2.5, 5 and 10 g/100 g compost) and for different timeperiods, up to 96h. Dose, time, as well as dose and time interactions of FO compost enrichment had significant effects on all fatty acids of earthworm bodies. The enrichment with 10 g FO/100 g compost sharply increased DHA levels of the worms (14.01 mol%) within just 24 h, compared to the control group (0.92 mol%). Similar increases (ranging between 10.99 and 15.55 mol%) occurred only after 48 and 96 h in lower FO enrichment levels (2.5 and 5.0 g/100 g compost, respectively). Therefore, it was concluded that, to obtain maximum n-3 LC-PUFA enrichment efficiency in REW, an enrichment period of 24 h is ideal and recommended for the 10 g FO/ 100 g compost, whereas longer enrichment periods (48-96 h) are more suitable for lower levels of FO inclusions (2.5 or 5 g/100 g compost).

Determination of Inorganic Arsenic in Fish Oil and Fish Oil Capsules by LC-ICP-MS

Background As inorganic arsenic is a highly toxic compound, its concentration in foods should be determined. Objective Develop an analytical method for determining inorganic arsenic in fish oil and fish oil capsules. Method Inorganic arsenic was extracted from fish oil by heating at 80°C in 1.6% tetramethylammonium hydroxide-ethanol. The concentration of inorganic arsenic in fish oil was determined by liquid chromatography (LC) inductively coupled plasma (ICP) MS using an octadecylsilane (ODS) column with a mobile phase containing an ion-pair reagent. Results The LOD (0.005, 0.004 mg/kg), LOQ (0.016, 0.011 mg/kg), repeatability (4.2, 3.5%), intermediate precision (5.4, 3.5%), and trueness (recoveries 94–109% based on spiked samples) of the proposed method were satisfactory for inorganic arsenic in fish oil and fish oil capsules. A low level of inorganic arsenic was detected only in anchovy oil among all fish oil samples that were used in this study. Inorganic arsenic levels were below the quantitation limit in all fish oil capsules. Conclusions Inorganic arsenic was determined after extraction from fish oil by heating at 80°C in 1.6% tetramethylammonium hydroxide–ethanol. The level of inorganic arsenic in all fish oil samples examined in this study was lower than 0.1 mg/kg of the maximum level defined in the Codex. Highlights Arsenic speciation in fish oil and fish oil capsules was analyzed by LC-ICP-MS using an ODS column with a mobile phase containing an ion-pair reagent. A low level of inorganic arsenic was detected only in anchovy oil. No inorganic arsenic was detected in fish oil capsules.

Enrichment of Omega-3 from Anchovy (Stolephorus sp.) Fish Oil by Enzymatic Hydrolysis

Anchovy (Stolephorus sp.) is an economically important fish in Indonesia. Anchovy contains Omega-3 that important to maintain the health of the heart and brain. This study aimed to enrich the Omega 3 content of anchovy oil from the North Sea of West Java. The extraction of anchovy oil was carried out by the soxhlet method. Enrichment of omega 3 from anchovy fish oil is carried out by hydrolysis with a commercial lipase enzyme at concentration 500, 1000, 1500 and 2000 unit/600g fish oil, for 5, 10, 15 and 20 hour. Before hydrolysis, fish oil was added with solvent water, ethanol, toluene, and n-hexane. Omega 3 content of fish oil products were analyzed by using Gas Chromatography (GC) with FID detector with retention time 14.068 min and 15.506 min for α-Linolenat (ALA) and eicosapentaenoic (EPA), respectively. The results showed the highest omega 3 content (ALA 0.54% and EPA 1.103% ) was produced by addition n-hexane with a ratio 1:6 with the concentration of lipase was 1000 units for 20 hours.

Application of Plackett–Burman Design in Screening of Natural Antioxidants Suitable for Anchovy Oil

Considering the safety of synthetic antioxidants, more and more natural antioxidants have been developed and utilized in foods. This study aimed to screen out a natural antioxidant combination from many antioxidants, which could significantly affect the oxidation stability of anchovy oil, while Plackett–Burman design (PBD) methodology was employed in this screening. According to the statistical results of this design, sesamol, dihydromyricetin, teapolyphenol, and rosemary acid were four significant parameters on the oxidation stability of anchovy oil. Moreover, dihydromyricetin presented the best antioxidant effect among nine kinds of selected antioxidants when they were used alone in anchovy oil. Meanwhile, a combination including sesamol (0.02%), teapolyphenol (0.02%). and rosemary acid (0.02%) was adopted, and its antioxidant ability was similar to that of tert-butylhydroquinone (TBHQ). Additionally, phytic acid as a synergist was used and combined with sesamol, and the antioxidant ability of this combination was better than that of TBHQ. This study presented a reference for the industrial applications of natural antioxidants and synergists in anchovy oil.

Investigating the Mechanism for the Enhanced Oxidation Stability of Microencapsulated Omega-3 Concentrates

Enzymatically concentrated anchovy oil (concentrate) is known to be much less stable than unconcentrated anchovy oil. However, we previously showed that concentrate surprisingly forms more stable microcapsules, when produced by complex coacervation, than does unconcentrated anchovy oil. Here we investigate the mechanism of this unexpected stability. We also investigate whether or not incorporation of concentrate can be used as an additive to improve the stability of unconcentrated anchovy oil microcapsules. Results showed that microcap stability increased as the amount of added concentrate increased. Decreased emulsion droplet size, lower positively charged zeta potential, and higher surface hydrophobicity were observed in the oil/water (O/W) emulsion, with the incorporation of concentrate in the oil phase, compared with the unconcentrated anchovy oil O/W emulsion. Both the decreased zeta potential and the increased hydrophobicity of concentrate in the mixed oil phase may improve droplet agglomeration, leading to enhanced oxidative stability of the concentrate-containing microcapsules. Decreased repulsive forces between droplets result in a more compact structure, thicker outer shell, and smoother surface, resulting in enhanced oxidation stability of the concentrate-containing microcapsules.