Application of instrumental methods for quality control of lactoferrin and raw materials for its production

Bovine cattle milk lactoferrin is industrially produced from cow’s milk and used in a variety of functional products, including baby food, nutritional supplements, dairy products, and veterinary drugs. The possibility of obtaining lactoferrin from lactic acid milk wheyrises a particular interest. The purpose of this study is to assess the possibility of using and adapting existing spectrophotometric methods for determining the concentration of total protein and an electrophoretic method for analyzing the protein composition of whey fractions and lactoferrin substances. A comparative study of two methods for determining the total protein in whey fractions and lactoferrin preparations biuret and Bradford method was carried out. The validation parameters of both methods were defined. An analytical range from 30 μg / ml to 1000 μg / ml characterizes the linearity of the Bradford method. The electrophoretic profile of milk, milk whey, whey fractions and substances of lactoferrinwere analyzed. The results obtained indicate a high information content of the electrophoretic method for characterizing the purification efficiency of whey fractions.

ACTIVITIES OF AMYLASE, PROTEASE and LIPASE FROM HONEY Trigona sp, Apis mellifera and Apis dorsata

Activities of amylase, protease and lipase from honey Trigona sp, Apis mellifera and Apis dorsata, determination protein concentration and the activity protease done with Bradford method, the determination of the glucose standard and activity amylase done with 3,5-dinitro salicylic acid (DNS) method and activity lipase done with acid-base titration with coconut oil substrate. The honey from Trigona sp has value of the activity amylase and lipase respectively by 0,0136 U / mg and 0,359 U/mg, whereas honey Apis mellifera has activity of protease, amylase and lipase of each 1,22 x 10-6 U/mg; 0,944 U/mg and 0,304 U/mg and then honey Apis dorsata has amylase and lipase activity of each of 0,0645 U/mg and 0,287 U/mg.

Biuret and Bradford Methods Suitability for Protein Quantification in Rapeseed Meal

Summary Being attractive for insects, non-genetically modified rapeseed is valuable for maintaining environmental biodiversity. Primarily, the rapeseed is an important industrial crop which is used for production of vegetable oil. Oil extraction from rapeseeds results in the generation of substantial amounts of rapeseed meal which is used either as a protein rich feed additive or as a source for preparation of protein containing ingredients for food industry. Both applications require frequent evaluation of protein content. Although Kjeldahl method is considered standard, it is not appropriate for routine evaluation of protein content in protein extracts. The aim of the study was to evaluate suitability of biuret and Bradford methods for protein quantification in rapeseed meal extracts. After consecutive triple extraction of proteins with water, 5% NaCl, 70% ethyl alcohol and 0.1 N NaOH, protein evaluation of each albumin, globulin, prolamin and glutelin extraction aliquot demonstrated overall lower protein content by Bradford method compared to biuret method. The most pronounced differences in protein content were observed with prolamin fraction where three fold higher protein concentrations in each extraction aliquot was observed when biuret method was applied for the evaluation. Comparative quantification of the total protein of each of the four fractions followed a similar trend of lower protein content evaluation by Bradford method. Overall results indicated biuret method as more suitable for protein quantification in rapeseed meal extracts which was confirmed by comparison with data obtained by Kjeldahl method.

Birefringence of Collagen Fibres in Rat Calcaneal Tendons Treated with Acupuncture during Three Phases of Healing

Background Birefringence is an optical anisotropy that is investigated by polarisation microscopy, and has been valuable for the study of the oriented organisation of collagen fibres in tendons. However, the application of this technology to evaluate the effect of different acupuncture points during tendon healing has not yet been described. Objectives To evaluate the concentration of non-collagenous proteins (NCP) and birefringence in rat calcaneal tendons following injury during the three different phases of healing: inflammatory (7th day), proliferative (14th day), and remodelling (21st day). Methods Tendons of 120 Wistar rats were tenotomised and left untreated (teno group, n=24), treated with manual acupuncture at ST36 (ST36 group, n=24), BL57 (BL57 group, n=24) or ST36+BL57 (SB group, n=24), or treated with electroacupuncture at ST36+BL57 (EA group, n=24). Tendon samples were collected at 7, 14 and 21 days after injury (n=8 per group). NCP concentrations were measured using the Bradford method (n=4 each) and birefringence was examined using polarisation microscopy and image analysis (n=4 each). Comparison was also made with healthy (non-tenotomised) tendons in a subgroup of rats (n=4 each). Results Manual acupuncture at ST36 and BL57 increased molecular organisation of collagen fibres on day 14 and 21 after injury. Isolated use of BL57 and ST36 also increased collagen fibre organisation when examined on day 14 and 21, respectively. No significant increase in NCP concentration was observed in any of the treated tenotomised groups. Conclusions Acupuncture, through putative anti-inflammatory and mechanotransductor effects, may have a role in strengthening tendons and increasing resistance to re-rupture.

Dairy manure protein analysis using UV-vis based on the Bradford method

Organic nitrogen in the form of protein and degraded protein fragments is one of the most important components of dairy manure.