A Novel Rapid Enzyme Immunoassay (Fluorophos BetaScreen) for Detection of β-Lactam Residues in Ex-Farm Raw Milk

1998 ◽  
Vol 61 (7) ◽  
pp. 808-811 ◽  
Author(s):  
ÅSE STERNESJÖ ◽  
GÖRAN JOHNSSON
Keyword(s):  
Enzyme Immunoassay ◽  
Limit Of Detection ◽  
Raw Milk ◽  
Penicillin G ◽  
Milk Samples ◽  
Quality Testing ◽  
Response Profile ◽  
Very High ◽  
Higher Sensitivity

A novel, qualitative enzyme immunoassay based on fluorescence detection for determination of (β-lactam antibiotics in raw, commingled milk (Fluorophos BetaScreen E. U. test) was evaluated. A dose-response profile for penicillin G was constructed by analysis of spiked milk samples. The limit of detection, defined as the concentration of penicillin G that resulted in 95% of the samples being evaluated as positive, was 1.8 μg/kg. The repeatability of the assay was very high both within and between the three participating milk quality testing laboratories. In total 5,061 randomly selected tanker milk samples were analyzed with the BetaScreen test and compared with the Delvotest SP. The agreement between the two tests was 99.7%. Probably due to a higher sensitivity to penicillin G, the BetaScreen test detected almost twice as many suspect positive tanker milk samples (0.45%) as the Delvotest SP (0.26%).

scholarly journals Determination of Aflatoxin M1 in Raw Milk Using an HPLC-FL Method in Comparison with Commercial ELISA Kits—Application in Raw Milk Samples from Various Regions of Greece

2021 ◽  
Vol 8 (3) ◽  
pp. 46
Author(s):  
Martha Maggira ◽  
Maria Ioannidou ◽  
Ioannis Sakaridis ◽  
Georgios Samouris
Keyword(s):  
Limit Of Detection ◽  
Reference Method ◽  
Raw Milk ◽  
Aflatoxin M1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fluorescence Detector ◽  
Elisa Kit ◽  
Milk Samples ◽  
Of Greece

The highly toxic Aflatoxin M1 (AFM1) is most often detected in milk using an Enzyme-Linked-Immunosorbent Assay (ELISA) for screening purposes, while High-Performance Liquid Chromatography with Fluorescence Detector (HPLC-FL) is the reference method used for confirmation. The aim of the present study was the comparison between three commercially available ELISA kits and a newly developed HPLC-FL method for the determination of the AFM1 in milk samples. The developed HPLC-FL method was validated for the AFM1 and Aflatoxin M2 (AFM2), determining the accuracy, precision, linearity, decision limit, and detection capability with fairly good results. All three ELISA kits were also validated and showed equally good performance with high recovery rates. Moreover, the Limit Of Detection (LOD) and Limit Of Quantification (LOQ) values were found to be significantly lower than the Maximum Residue Limit (MRL) (50 ng kg−1). After the evaluation of all three commercial kits, the ELISA kit with the optimum performance along with the HPLC method was used for the determination of AFM1 in raw cow’s, goat’s, and sheep’s milk samples (396) obtained from producers in different regions of Greece. The evaluation of both methods showed that this ELISA kit could be considered as a faster and equally reliable alternative method to HPLC in routine analysis for the determination of AFM1 in milk.

Validation of a Rapid Lateral Flow Test for the Simultaneous Determination of β-Lactam Drugs and Flunixin in Raw Milk

2012 ◽  
Vol 75 (7) ◽  
pp. 1270-1277 ◽  
Author(s):  
DAVID DOUGLAS ◽  
KATIE BANASZEWSKI ◽  
RIMA JUSKELIS ◽  
FADWA AL-TAHER ◽  
YANG CHEN ◽  
...  
Keyword(s):  
Simultaneous Determination ◽  
Limit Of Detection ◽  
Agricultural Practices ◽  
Raw Milk ◽  
Lateral Flow ◽  
Penicillin G ◽  
Screening Programs ◽  
Flow Test ◽  
Lateral Flow Test

β-Lactam antibiotics are the most commonly used drugs on dairy farms. β-Lactam residues in milk are kept out of the human milk supply with good agricultural practices and mandatory truck screening performed by the dairy industry under Appendix N of the Pasteurized Milk Ordinance. Flunixin, a nonsteroidal and anti-inflammatory drug, appears in dairy cattle tissue residues with a frequency similar to the occurrence of penicillin G. This creates concern that flunixin residues could be in milk and would go undetected under current milk screening programs. A single test that combines mandatory β-lactam screening with voluntary flunixin screening is an economical approach for monitoring and controlling for potential flunixin or 5-hydroxyflunixin, the primary flunixin metabolite marker in milk. The objective of this study was to validate a β-lactam and flunixin rapid lateral flow test (LFT) and compare the results obtained with a liquid chromatography-triple quadrupole tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of flunixin and 5-hydroxyflunixin in raw milk with a limit of detection of <1 ppb, equivalent to 1 ng/ml. Using the LFT, three combined manufactured lots of test strips detected penicillin G at 2.0 ppb, ampicillin at 6.8 ppb, amoxicillin at 5.9 ppb, cephapirin at 13.4 ppb, ceftiofur (total metabolites) at 63 ppb, and 5-hydroxyflunixin at 1.9 ppb at least 90% of the time with 95% confidence. The LFT also detected incurred flunixin milk samples that were analyzed with the LC-MS/MS and diluted to tolerance in raw milk. The detection levels for the LFT are lower than the U.S. safe levels or tolerances and qualify the test to be used in compliance with U.S. milk screening programs.

Detection of proteolysis in raw milk stored at low temperature by an inhibition ELISA

1994 ◽  
Vol 61 (3) ◽  
pp. 395-404 ◽  
Author(s):  
Catherine Picard ◽  
Isabelle Plard ◽  
Dominique Rongdaux-Gaida ◽  
Jean-Claude Collin
Keyword(s):  
Proteolytic Activity ◽  
Limit Of Detection ◽  
Bacterial Count ◽  
Raw Milk ◽  
Enzyme Linked Immunosorbent Assay ◽  
Psychrotrophic Bacteria ◽  
Milk Samples ◽  
Inhibition Elisa ◽  
Different Strains

SummaryAn inhibition ELISA (enzyme-linked immunosorbent assay) was developed for the determination of caseinomacropeptide (CMP) in order to estimate the proteolysis of κ-casein due to the enzymes of psychrotrophic bacteria in bulk raw milk. The CMP present in milk was quantified specifically by an antibody. The limit of detection was ∽ 0·1 μg/ml and the CV was < 10%. This method was used to study the proteolytic activity of three strains of psychrotrophic Pseudomonas fluorescens in raw milk and to analyse different raw milk samples supplied by four dairy plants. The proteolytic activity for different strains of psychrotrophs and for different milk samples varied considerably, but no correlation was established between the level of microbial flora and κ-casein proteolysis. It is thus not possible to determine the extent of proteolysis from the bacterial count alone. However, by CMP determination in bulk raw milk samples after 6 d storage at 4°C, the mean κ-casein proteolysis was ∽ 4%. Among the milk samples analysed that contained < 107 cfu psychrotrophs/ml, 30% exhibited a proteolysis of κ-casein < 0·5%, i.e. < 5μg CMP/ml.

scholarly journals Simultaneous Determination of C18 Fatty Acids in Milk by GC-MS

Separations ◽  
2021 ◽  
Vol 8 (8) ◽  
pp. 118
Author(s):  
Meiqing Chen ◽  
Yangdong Zhang ◽  
Fengen Wang ◽  
Nan Zheng ◽  
Jiaqi Wang
Keyword(s):  
Fatty Acids ◽  
High Sensitivity ◽  
Raw Milk ◽  
Uht Milk ◽  
Milk Samples ◽  
Accuracy And Precision ◽  
Different Temperatures ◽  
Chromatographic Resolution ◽  
Commercial Milk

The determination of C18 fatty acids (FAs) is a key and difficult aspect in FA profiling, and a qualified method with good chromatographic separation and high sensitivity, as well as easy methylation, is required. A GC-MS method was established to simultaneously determine C18 FAs in milk. To simplify the methylation protocol for milk samples, besides a base-catalyzation methylation (50 °C for 20 min), the necessity of an additional acid-catalyzation was also studied using different temperatures (60 °C, 70 °C, 80 °C, and 90 °C) and durations (90 min and 150 min). The results showed that the chromatographic resolution was improved, although three co-eluted peaks existed. The base-catalyzation was sufficient, and an additional acid-catalyzation was not necessary. The proposed method was validated with good sensitivity, linearity, accuracy, and precision, and then applied in determining C18 FAs in 20 raw milk and 30 commercial milk samples. UHT milk presented a different profile of C18 FAs from raw milk and PAS milk samples, which indicated that excessive heating could change the profile. Overall, the proposed method is a high-throughput and competent approach for the determination of C18 FAs in milk, and which presents an improvement in chromatographic resolution and sensitivity, as well as a simplification of methylation.

Determination of Viable Bacterial Populations in Raw Milk within 20 Minutes by Using a Direct Epifluorescent Filter Technique

1997 ◽  
Vol 60 (7) ◽  
pp. 874-876 ◽  
Author(s):  
CLAUDE P. CHAMPAGNE ◽  
NANCY J. GARDNER ◽  
JULIE FONTAINE ◽  
JACQUES RICHARD
Keyword(s):  
Raw Milk ◽  
Plate Count ◽  
Bacterial Populations ◽  
Filter Technique ◽  
Milk Samples ◽  
Standard Plate ◽  
Plate Counts ◽  
Direct Epifluorescent Filter Technique ◽  
Triton X

The results from a shortened procedure for the direct epifluorescent filter technique (DEFT) determination of viable bacterial populations in raw milk were compared to standard plate counts. Shortening the prefiltration trypsin-Triton X-100 incubation period from 10 to 3 min enabled the completion of the analysis within 20 min. The short DEFT method results had a correlation coefficient (r) of 0.81 with plate counts. With respect to precision, the average difference between values of duplicate plate count analyses was 0.16 log units; that of the short DEFT was 0.14 log units. The slopes of the regressions equations were less than 1, indicating that a direct correlation is not achieved. Short DEFT values were 0.17 log units higher than those of plate counts on milk samples containing less than 10,000 CFU/ml. For milk samples containing counts over 10,000 CFU/ml, short DEFT values averaged only 0.05 log units above plate count readings. Daily preparation of the stain appears unnecessary since acridine orange solutions stored for up to 2 days at 4°C did not produce results significantly (P &gt; 0.05) different from those obtained with fresh solutions. The short DEFT method has potential for the assessment of the bacteriological quality of raw milk in tanker deliveries.

scholarly journals Determination of Four Fluoroquinolones in Milk by Liquid Chromatography

1997 ◽  
Vol 80 (5) ◽  
pp. 982-987 ◽  
Author(s):  
José E Roybal ◽  
Allen P Pfenning ◽  
Sherri B Turnipseed ◽  
Calvin C Walker ◽  
Jeffrey A Hurlbut
Keyword(s):  
Fluorescence Detection ◽  
Solid Phase ◽  
Raw Milk ◽  
Extraction Column ◽  
Isocratic Elution ◽  
Standard Curve ◽  
Milk Samples ◽  
Relative Standard ◽  
Liquid Chromatographic

Abstract A liquid chromatographic (LC) method with fluorescence detection is presented for the analysis of 4 fluoroquinolones; enrofloxacin (ENRO), ciprofloxacin (CIPRO), sarafloxacin (SARA), and difloxacin (DIFLX) in milk. The procedure consists of extraction of milk with acidified ethanol, isolation and retention on a cation exchange solid-phase extraction column, elution with basic methanol, and LC analysis with fluorescence detection. LC analysis is performed by isocratic elution using an acetonitrile-2% acetic acid (15 + 85) mobile phase and an Inertsil phenyl column with fluorescence detection at excitation and emission wavelengths of 278 and 450 nm, respectively. A target level of 10 ppb for each of the 4 fluoroquinolones has been established for this method. Average recovery from fortified raw milk samples (5-100 ppb each) based on a 5-point standard curve calculation was 70-90%, with relative standard deviations of &lt;15%.

scholarly journals Physical and microbial qualities of raw milk collected from Bangladesh Agricultural University dairy farm and the surrounding villages

1970 ◽  
Vol 6 (2) ◽  
pp. 217-221 ◽  
Author(s):  
MTG Khan ◽  
MA Zinnah ◽  
MP Siddique ◽  
MHA Rashid ◽  
MA Islam ◽  
...  
Keyword(s):  
Microbiological Quality ◽  
Dairy Farm ◽  
Raw Milk ◽  
Viable Count ◽  
Physical Parameters ◽  
Total Viable Count ◽  
Coliform Count ◽  
Milk Samples

The present study was undertaken with the aim of investigating the physical parameters (e.g. organoleptic and specific gravity of raw milk) and also to study the microbiological quality of raw milk (total viable count, Coliform count and Staphylococcal count) from different villages and Bangladesh Agricultural University (BAU) Dairy Farm of Mymensingh District of Bangladesh, during the period from July to November 2007. A total number of 100 raw milk samples were collected at morning and evening from BAU dairy farm and surrounding four villages of BAU campus. The organoleptic and bacteriological qualities of each sample were analyzed. The organoleptic examination included taste panel score to assess consumer's acceptance and the bacteriological analysis comprised enumeration of total viable count (TVC), total colifrom count (TCC) and total staphylococcal count (TSC) for the determination of sanitary quality. The organoleptic quality of the milk samples is more or less same except the Churkhai milk samples which had flat taste (in 16% milk sample). The average values of TVC/ml were log 5.920, 5.934, 6.007, 6.075 and 6.127 for BAU Dairy Farm, Boira, Shutiakhali, Churkahai and Paglabazar respectively; coliform count were log 2.501, 2.522, 2.550, 2.620 and 2.619 respectively; staphylococcal count were log 2.832, 2.812, 2.866, 2.931 and 2.988 respectively. So, it may be concluded that the raw milk samples of BAU Dairy Farm were superior to others collected from the selected villages which may be due to maintaining better hygienic condition. Key words: Raw milk, physical and microbial quality   doi: 10.3329/bjvm.v6i2.2339 Bangl. J. Vet. Med. (2008). 6 (2): 217-221

Application of a microbial sensor for determination of short-chain fatty acids in raw milk samples

1992 ◽  
Vol 195 (1) ◽  
pp. 1-2 ◽  
Author(s):  
Hiroyuki Ukeda ◽  
Gotthold Wagner ◽  
G�nther Weis ◽  
Manfred Miller ◽  
Henning Klostermeyer ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Raw Milk ◽  
Short Chain Fatty Acids ◽  
Short Chain ◽  
Milk Samples ◽  
Microbial Sensor ◽  
Chain Fatty Acids

scholarly journals Determination of carcinogenic herbicides in milk samples using green non-ionic silicone surfactant of cloud point extraction and spectrophotometry

2018 ◽  
Vol 5 (4) ◽  
pp. 171500 ◽  
Author(s):  
N. I. Mohd ◽  
N. N. M. Zain ◽  
M. Raoov ◽  
S. Mohamad
Keyword(s):  
Cloud Point ◽  
Cloud Point Extraction ◽  
Limit Of Detection ◽  
Cost Effective ◽  
Silicone Surfactant ◽  
Extraction Solvent ◽  
Milk Samples ◽  
Relative Standard ◽  
Limit Of Quantitation

A new cloud point methodology was successfully used for the extraction of carcinogenic pesticides in milk samples as a prior step to their determination by spectrophotometry. In this work, non-ionic silicone surfactant, also known as 3-(3-hydroxypropyl-heptatrimethylxyloxane), was chosen as a green extraction solvent because of its structure and properties. The effect of different parameters, such as the type of surfactant, concentration and volume of surfactant, pH, salt, temperature, incubation time and water content on the cloud point extraction of carcinogenic pesticides such as atrazine and propazine, was studied in detail and a set of optimum conditions was established. A good correlation coefficient ( R 2 ) in the range of 0.991–0.997 for all calibration curves was obtained. The limit of detection was 1.06 µg l −1 (atrazine) and 1.22 µg l −1 (propazine), and the limit of quantitation was 3.54 µg l −1 (atrazine) and 4.07 µg l −1 (propazine). Satisfactory recoveries in the range of 81–108% were determined in milk samples at 5 and 1000 µg l −1 , respectively, with low relative standard deviation, n  = 3 of 0.301–7.45% in milk matrices. The proposed method is very convenient, rapid, cost-effective and environmentally friendly for food analysis.

A Practical Evaluation of the Delvotest P Multi Plate Test in Screening Raw Milk for Antibiotics

1986 ◽  
Vol 49 (11) ◽  
pp. 868-870 ◽  
Author(s):  
LYSE LAROCQUE ◽  
G. A. NEVILLE
Keyword(s):  
Bacillus Subtilis ◽  
Staphylococcus Epidermidis ◽  
Raw Milk ◽  
The Other ◽  
Penicillin G ◽  
Antibiotic Residues ◽  
Plate Test ◽  
Milk Samples ◽  
Practical Evaluation ◽  
Bacillus Subtilis Atcc

The Delvotest P Multi plate test was evaluated by screening 100 milk samples for total antibiotic residues (penicillin G, streptomycin and neomycin). The samples were taken in conjunction with an antibiotic depletion study in milk derived from six cows treated with a multiple antibiotic, intramammary infusion product. Within the limits of sensitivity of the Delvotest, only penicillin G persisted in milk samples taken beyond 60 h, whereas in some samples, the other antibiotics appeared to be depleted as early as 48 h. More sensitive tests, however, detected neomycin (Staphylococcus epidermidis, ATCC 12228) in 50% of samples taken at 60 h and streptomycin (Bacillus subtilis, ATCC 6633) at 14.5 d after discontinuation of infusion.

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