Precipitation Of Milk Proteins Research Articles

Application of chemometrics for detection and modeling of adulteration of fresh cow milk with reconstituted skim milk powder using voltammetric fingerpriting on a graphite/ SiO2 hybrid electrode

In the present work, an analytical approach for the voltammetric detection and prediction of adulteration of fresh cow milk with reconstituted skim milk powder is developed. After precipitation of milk proteins upon addition of ethanol and centrifugation, the supernatant liquid of the samples was analyzed by cyclic voltammetry on a novel graphite/SiO2 hybrid working electrode (GSiHE) using LiClO4 as electrolyte. Under these conditions, fresh milk samples gave broadened peaks/plateaus in both forward and backward potential scanning, attributed mainly to oxidases. Such peaks were not evident in the case of reconstituted skim milk powder samples due to inactivation of enzymes and breakdown of certain antioxidants caused by heat and pressure-treatments. The differences between fresh and reconstituted skim milk powder samples in their voltammetric profile were exploited for the detection of fresh milk adulteration by submitting voltammetric data to chemometrics. As datapoints, the differences between forward and backward current values, recorded at the same potentials, were determined and submitted to multivariate analysis. Principal Component Analysis (PCA) provided a clear differentiation between fresh milk and reconstituted skim milk powder samples. Soft independent modeling of class analogy (SIMCA) was employed to model the class of fresh milks, using samples from 12 commercially available fresh milk brands. Prediction of fresh milk adulteration with reconstituted skim milk powders was achieved by means of Partial Least Squares (PLS) regression analysis. Detection limit of the technique was found to be below 6% (v/v) and the linearity of model in terms of observed/predicted values was confirmed up to 100% (v/v). Validation and applicability of both SIMCA and PLS models were confirmed using a suitable test set, consisting of commercial fresh milk and skim milk powder samples as well as synthetic adulterated fresh milk samples.

In matrix formation of deep eutectic solvent used in liquid phase extraction coupled with solidification of organic droplets dispersive liquid-liquid microextraction; application in determination of some pesticides in milk samples

In this work, a liquid-phase extraction procedure and dispersive liquid–liquid microextraction method based on deep eutectic solvents were combined and used for the simultaneous extraction of different classes of pesticides; including carbaryl, hexythiazox, pretilachlor, iprodione, famoxadone, sethoxydim and fenazaquin from milk samples. In the first step, a deep eutectic solvent was synthesized in milk sample and simultaneously, was used for extraction of the analytes along with precipitation of milk proteins. To assist the formation of the deep eutectic solvent and increasing the mass transfer rate of the analytes, ultrasonic irradiations was used. In the second step, the collected organic phase from pervious step was mixed (as dispersive solvent) with a water-immiscible deep eutectic solvent (ChCl: decanoic acid) and injected into deionized water. The cloudy solution was placed into an ice bath and the extraction solvent was solidified on the top of the solution. After removing the solid phase by a spatula, it was melted at room temperature and 1 μL of the extraction solvent was injected into the separation system. Under the optimum extraction conditions, low limits of detection and quantification within the ranges of 0.90–3.9 and 3.1–13 ng mL−1 were achieved, respectively. Precision of the method expressed as relative standard deviation was in the ranges of 3.8–5.3 and 4.8–6.9 for intra– and inter–day (n = 5) precision, respectively, at a concentration of 50 ng mL−1 of each analyte. Extraction recoveries and enrichment factors were between 64 and 89% and 320 and 445, respectively. Lastly, several milk samples were successfully analyzed using the proposed method.

Surveillance study of novobiocin and phenylbutazone residues in raw bovine milk using liquid chromatography-tandem mass spectrometry

A simple method permitting the simultaneous determination of trace residues of novobiocin and phenylbutazone in raw milk samples using liquid chromatography-tandem mass spectrometry was developed. Raw milk samples were mixed with acetonitrile to facilitate the concurrent precipitation of milk proteins and extraction of both veterinary drugs. Without additional clean-up or concentration of the resulting extract, the analytes could be quantified at concentrations as low as 0.0025 and 0.001 µg ml−1 for phenylbutazone and novobiocin, respectively. The analysis of a series of fortified raw milk samples at analyte concentrations ranging from 0.005 to 0.1 µg ml−1 and from 0.01 to 0.2 µg ml−1 for phenylbutazone and novobiocin, respectively, yielded average recoveries ranging from 89.2% to 104.3% with standard deviations below 7%. The analytical method was applied to the analysis of raw milk samples collected from transport trucks upon delivery at dairy-processing plants throughout Alberta, Canada. Novobiocin was detected in 13 of 1072 samples tested at concentrations ranging from 0.001 to 0.007 µg ml−1. Phenylbutazone was not detected in any of the samples tested.

Study of robustness based on n-way models in the spectrofluorimetric determination of tetracyclines in milk when quenching exists

An important step in the validation of an analytical procedure is the study of its robustness. In the case of spectrofluorimetric determinations, quenching introduces specific problems which are approached in this paper for the particular case of tetracyclines determination in milk. Quenching can be detected with excitation emission matrices (EEM) signals and a three-way Parallel Factor (PARAFAC) decomposition and modelled by means of a four-way PARAFAC decomposition which reproduces the physical model of this effect. The robustness of the method is evaluated by including changes in seven experimental variables: trichloroacetic acid (TCA) volume solution used in the precipitation of milk proteins, revolutions per minute, time and temperature in the centrifugation step, pH and emission-excitation slit width in the fluorimetric analyte determination and the analyst. The robustness analysis is carried out by means of a Plackett-Burman experimental design as it is suggested by European Decision 2002/657/EC (European Decision (EC) No. 2002/657/EC of 12 August 2002, implementing Council Directive 96/23/EC, concerning the performance of analytical methods and the interpretation of results, Off. J.L 221, 17/8/2002, 8). The analyte concentration will be taken as response in the Plackett-Burman experimental design instead of the signal as it is habitual in these cases. Therefore, a three-way Partial Least Squares (3-PLS) calibration models with EEM signal is needed. When an analogous study is carried out for tetracycline (TC) in the absence of chlorotetracycline (CTC) as interferent, univariate calibration is employed, being able to conclude that in the robustness analysis, different factors are significantly active when quenching exists.

Determination of iodide in dairy products and table salt by ion chromatography with electrochemical detection

Precipitation of milk proteins and most of the fat is effected by addition of methanol. The remaining organics are removed by means of a C 18 solid-phase extraction cartridge before ion chromatographic separation and electrochemical detection of iodide. Table salt is simply diluted, filtered and directly analysed by high-performance liquid chromatography

Improved microbiological assay procedures for dihydrostreptomycin residues in milk and dairy products.

Because dihydrostreptomycin can remain as a slowly removed antibiotic residue in dairy animals and because of the need for more sensitive procedures with which to provide information concerning antibiotic residues in food products, procedures were developed in more sensitive assays of dihydrostreptomycin in milk and some representative dairy products. The cleanup procedures that aided these improvements were (i) precipitation of milk proteins by acidification and (ii) centrifugation to extract cheeses and to remove physical barriers to diffusion in the cylinder plate assay. In addition, a thin, single seeded assay layer was used to maximize diffusion. Levels of dihydrostreptomycin as low as 0.06 microgram/ml in milk and 0.2 to 0.4 microgram/g in cheeses were measurable; these levels were some fourfold more sensitive than those presently recommended by the Food and Drug Administration.

693. A rejection test for raw milk

The determination of apparent lactic acid has been used as a basis for a rejection test for raw milk. The method consists of precipitation of milk proteins with barium chloride, sodium hydroxide and zinc sulphate, and the addition of ferric chloride to the filtrate to produce the yellow colour of ferric lactate. Lactic acid is not solely responsible for the production of the yellow colour, which, nevertheless, gives a good relationship with keeping quality of milk (measured as hours to the C.O.B. end-point at 22° C.) and the values have been expressed as ‘apparent lactic acid’.The relationship of winter and summer milks to keeping quality has been studied, and a value of 0·03% apparent lactic acid in milk is equivalent to an average keeping quality of 5¾ and 8½ hr. for winter and summer milks, respectively. A value of 0·03% is recommended as the earliest value of apparent lactic acid at which milk could be rejected.The apparent lactic acid in colostrum and late-lactation milk and in milk from cows suffering from mastitis has been determined, and only in late-lactation milk were the values found to be significantly higher than usual in fresh raw milk, and an inverse relationship between yield and apparent lactic acid is suggested.Permanent glass matching disks have been prepared for use in a Lovibond comparator. This permits the intensity of the yellow colour produced with 1% ferric chloride to be determined and the apparent lactic acid in milk estimated.Grateful acknowledgement is made to the management and staff of the Dairy Department of the Reading Co-operative Society and the Farmer's Clean Milk Dairy, Reading, and local milk producers for supplying samples for experiments; to the N.M.T.S. staff in Reading for help in finding suitable farmers, and to the Dairy Husbandry Department of the N.I.R.D. for information about and samples of abnormal and late-lactation milk. Our particular thanks are due Miss Marie Gruber for technical assistance, to Dr N. J. Berridge for the suggestion and help on the work on pH change as an indication of keeping quality (given in the appendix), and to Dr A. T. R. Mattick for the advice given in this work.