Soft tissue from cultured farm fresh oysters (Crassostrea madrasensis) was subjected to two standard enzymatic peptide extraction procedures using pepsin and papain. The crude extracts obtained were partially purified by column chromatography and were freeze-dried. The hydrolysates obtained were compared with respect to their degree of hydrolysis (DH), antioxidant potential (AP) and total phenolic content (TPC). The hydrolysate showing better antioxidant property was further subjected to purification by high performance liquid chromatography and characterized by LC-MS/MS. Papain-digested oyster protein (OPHpap) hydrolysate showed higher DH, AP and TPC. OPHpap was further subjected to ultrafiltration and fractionated into 3 sizes namely, above 10, 3-10 and 1-3 kDa according to the molecular size. Antioxidant capacity of\3 kDa fraction OPHpap-3 evaluated by DPPH free radical scavenging assay, metal chelating activity, linoleic acid autoxidation assay showed maximum effectiveness. Of the seven fractions collected by purification of OPH-pap-3 on semi-preparative RP-HPLC, fraction 7 that showed the highest antioxidant activity was further characterized by LC-ESI-MS/MS and its sequence determined. An antioxidant peptide molecule with thirteen amino acids was identified in oyster protein hydrolysate obtained by papain digestion that may find application as a nutraceutical or may be utilized in food industry for prevention of rancidity in foods.
This research describes a new technological process for sesame oil extraction. The process deals with the effect of ohmic heating on enhancement of oil recovery and quality of cleaned and graded sesame seed. The effect of ohmic heating parameters namely electric field strength (EFS), end point temperature (EPT) and holding time (HT) on oil extraction process were investigated. Three levels of electric field strength (600, 750 and 900 V/m), end point temperature (65, 75 and 85 °C) and holding time (5, 10 and 15 min.) were taken as independent variables using full factorial design. Percentage oil recovered from sesame seed through mechanical extracted oil by application of ohmic heating varies from 39.98 to 43.15 %. The maximum oil recovery 43.15 % was obtained when the sample was heated and maintained at 85 °C using EFS of 900 V/m for a holding time of 10 min as against 34.14 % in control sample. The free fatty acid (FFA) of the extracted oil was within the acceptable limit (1.52 to 2.26 % oleic acid) of 0.5 to 3 % as prescribed respectively by Prevention of Food Adulteration (PFA) and Bureau of Indian Standards (BIS). The peroxide value of extracted oil was also found within the acceptable limit (0.78 to 1.01 meq/kg). The optimum value for maximum oil recovery, minimum residual oil content, free fatty acid (FFA) and peroxide value were 41.24 %, 8.61 %, 1.74 % oleic acid and 0.86 meq/kg, respectively at 722.52 V/m EFS at EPT 65 °C for 5 min. holding time which was obtained by response surface methodology.
Despite the potential of LC with tandem MS (MS/MS) in improving sensitivity and selectivity, analytical methods are scarce for the determination of protein-bound and phosphorylated forms of B vitamins in food. This prompted us to develop a method for LC-MS/MS determination of naturally occurring nicotinamide, nicotinic acid, thiamine, pyridoxine, riboflavin, pantothenic acid, biotin, folic acid, and cyanocobalamin in fish. Baseline separation of the vitamins was achieved in a hydrophilic interaction LC condition. An ultrasonication-assisted enzymatic extraction protocol for sample preparation was optimized and validated. The time required for extraction was significantly reduced (to 4 h), while maintaining good extraction efficiency. Acetonitrile content (80%, v/v) in the prepared sample was found to be optimum for excellent peak shape and sensitivity. The dynamic linear range of the vitamins ranged from 2.5 to 500 ng/g, and the regression coefficient values were greater than 0.99. LOQ values ranged from 0.4 to 50 ng/g for the different vitamins. The spike recovery values at 50 and 100 ng/g ranged from 87.5 to 97.5%. The intra- and interday precision values were satisfactory. Accuracy of the developed method was determined by analysis of a Certified Reference Material. The method could also be used for unambiguous determination of the natural content of the target vitamins in fish.
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