Determination of Antibiotics in Meat Using Bacillus stearothermophilus Spores

1981 ◽  
Vol 44 (3) ◽  
pp. 194-200 ◽  
Author(s):  
M. BIELECKA ◽  
J. D. BALDOCK ◽  
A. W. KOTULA
Keyword(s):  
Sample Size ◽  
Bacillus Stearothermophilus ◽  
Test Organism ◽  
Optimal Concentration ◽  
Antibiotic Residues ◽  
Plate Method ◽  
Assay Medium ◽  
Organism Growth ◽  
Ph Variations

Ten parameters affecting sensitivity, accuracy and simplicity of the diffusion plate method for determining antibiotic residues in meat were evaluated with spores of Bacillus stearothermophilus as the test organism. Eight antibiotics were studied and included penicillin, bacitracin, tetracycline, chlortetracycline, oxytetracycline, streptomycin, erythromycin and neomycin. Sensitivity of the method was most influenced by concentration of inoculum, quantity of assay medium on the plate and sample size. The optimal concentration of inoculum was established as 2 × 105 spores/ml of medium, quantity of the assay medium on plate/100 mm dia., as 6 ml and quantity of sample poured on disc/12.7 mm dia., as 100 μl. The pH of the assay medium was also important to both antibiotic potency and test organism growth. The activity of streptomycin and erythromycin was the most sensitive to pH variations.

Modified Assay Medium for the Turbidimetric Assay of Chlortetracycline in Feeds

1976 ◽  
Vol 59 (3) ◽  
pp. 536-539
Author(s):  
Hussein S Ragheb
Keyword(s):  
Growth Rate ◽  
Excellent Agreement ◽  
Test Organism ◽  
Plate Method ◽  
Assay Sensitivity ◽  
Assay Medium ◽  
Turbidimetric Method ◽  
Turbidimetric Assay ◽  
Commercial Feeds ◽  
Lower Recovery

Abstract In previous experiments, the turbidimetric method for determining chlortetracycline-HCI (CTC-HCl) in feeds showed lower recovery than the AOAC plate method. Although the addition of vitamins to the turbidimetric medium improved results, values by the turbidimetric method remained about 10% lower than by the plate method. A modified (1.7 × the weight recommended by the manufacturer) turbidimetric assay medium decreased assay sensitivity but did not significantly change the slope of S. aureus response to CTC-HCl. There was no evidence that vitamin fortification of the modified medium had any significant effect on the growth rate of test organism. Examination of about 100 samples of commercial feeds containing CTC-HCl showed excellent agreement in results between the turbidimetric and plate methods.

Determination of Residual Cephalexin in Chick Tissues by Bacillus stearothermophilus Paper Disc Assay

1988 ◽  
Vol 71 (2) ◽  
pp. 337-340
Author(s):  
Junya Okada ◽  
Ikuji Higuchi ◽  
Sadao Kondo ◽  
Bun-Ichi Saito
Keyword(s):  
Detection Limit ◽  
Trichloroacetic Acid ◽  
Bacillus Stearothermophilus ◽  
Test Organism ◽  
Acid Extract ◽  
Average Recovery ◽  
Ad Libitum ◽  
Paper Disc ◽  
Disc Assay

Abstract A paper disc method is described for determination of residual cephalexin (CEX) in chick tissues. A trichloroacetic acid extract of plasma and tissues is chromatographed on a macroreticular resin (Diaion HP-20) column to remove endogenous antibacterial substances interfering with the assay. The eluate is evaporated to dryness and the residue, dissolved in methanol-water (1 + 2), is subjected to a paper disc assay using Bacillus stearothermophilus var. calidolactis C953 NIZO as a test organism. The detection limit was 0.0375 ppm in tissue; the average recovery of CEX ranged from 72.4% in skin to 90.4% in plasma. Water containing 200 or 500 mg/L of CEX was given ad libitum to 2-week-old chicks for 10 days; the highest levels of CEX were found in the kidney, and the lowest were found in muscle at 0 h of withdrawal. CEX disappeared from most tissues at 24 h after withdrawal except from skin of chicks given 500 mg/ L. However, the drug was not detected in the skin at 48 h after withdrawal.

Microbiological Determination of Thiram

1973 ◽  
Vol 56 (6) ◽  
pp. 1517-1518
Author(s):  
Andre Rappe ◽  
Guy Mauquoy ◽  
Serge Baur
Keyword(s):  
Bacillus Licheniformis ◽  
Agar Plate ◽  
Test Organism ◽  
Test Strain ◽  
Food Preparation ◽  
Plate Method ◽  
Saccharomyces Carlsbergensis ◽  
Dietetic Food

Abstract A rapid and simple agar plate method has been developed for the microbiological determination of thiram, using Saccharomyces carlsbergensis ATCC 9080 as the test organism. With this organism, as little as 25 ng thiram/ well or 200 ng/disk can be detected, well below the levels that can be detected with either Bacillus licheniformis or Flavobacterium as the test strain. The assay has been applied to a dietetic food preparation.

scholarly journals Microbiological Assay for Lomefloxacin in Coated Tablets

2006 ◽  
Vol 89 (4) ◽  
pp. 1077-1079 ◽  
Author(s):  
Greici Cristiani Gomes ◽  
Hérida Regina Nunes Salgado
Keyword(s):  
Bacillus Subtilis ◽  
Standard Deviation ◽  
Relative Standard Deviation ◽  
Method Validation ◽  
Test Organism ◽  
Microbiological Assay ◽  
Plate Method ◽  
Relative Standard ◽  
Bacillus Subtilis Atcc

Abstract The validation of a microbiological assay, applying cylinder plate method for determination of the activity of lomefloxacin in coated tablets is described. Using a strain of Bacillus subtilis ATCC 9372 as the test organism, lomefloxacin was measured in concentrations ranging from 2.0 to 8.0 μg/mL. The method validation showed that it is linear (r = 0.9999), precise (relative standard deviation = 1.15%), and accurate (it measured the added quantities). The excipients did not interfere in the determination. It was concluded that the microbiological assay is satisfactory for quantitation of lomefloxacin in tablets.

Collaborative Study of Microbiological Assay of Oxytetracycline in Feeds

1968 ◽  
Vol 51 (3) ◽  
pp. 548-552
Author(s):  
D C Billman ◽  
H Clark
Keyword(s):  
Collaborative Study ◽  
Test Organism ◽  
Plate Method ◽  
Average Recovery ◽  
Extraction Flask ◽  
Average Coefficient ◽  
Similar Materials ◽  
Aoac Method ◽  
Assay Precision

Abstract A collaborative study was made of a modification of the official AOAC method for the determination of oxytetracycline in feeds. This cylinder plate method uses Bacillus cereus var. mycoides, ATCC No. 11778, as the test organism. The modification consists of using a 20 g instead of a 10 g sample, of using an extraction flask instead of a mortar and pestle or a blender, and of filtering the assay solution. The average recovery of oxytetracycline by 12 collaborators was 92.4% with an average coefficient of variation of 13.5%. Assay precision, determined by computing standard deviations on the differences between pairs of similar materials, was estimated to be 7.4% at 10 g/ton and 5.9% at 25 g/ton. The method is recommended for adoption as official, first action.

scholarly journals Microbiological Assay for Cefoxitin Sodium in Dosage Form

2007 ◽  
Vol 90 (2) ◽  
pp. 452-455 ◽  
Author(s):  
Hrida Regina Nunes Salgado ◽  
Greici Cristiani Gomes Tozo
Keyword(s):  
Staphylococcus Epidermidis ◽  
Dosage Form ◽  
Test Organism ◽  
Microbiological Assay ◽  
Plate Method

Abstract A microbiological assay applying the cylinder-plate method is described for determination of the activity of cefoxitin sodium in injectables. Using a strain of Staphylococcus epidermidis ATCC 12226 as the test organism, cefoxitin sodium was measured in concentrations ranging from 50.0 to 200.0 g/mL. The validation showed that the method was linear (r = 0.9998), precise (RSD = 0.81%), and accurate. It was concluded that the microbiological assay is satisfactory for quantitation of cefoxitin sodium in injectables.

scholarly journals A Bridging Study Between Liquid Chromatography and Microbial Inhibition Assay Methods for Determining Amoxicillin Residues in Catfish Muscle

1998 ◽  
Vol 81 (1) ◽  
pp. 33-39 ◽  
Author(s):  
Catharina Y W Ang ◽  
Wenhong Luo ◽  
Connie R KiessLing ◽  
Karen Mckim ◽  
Rebecca Lochmann ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Bacillus Stearothermophilus ◽  
Test Organism ◽  
Precolumn Derivatization ◽  
Inhibition Assay ◽  
Microbial Inhibition ◽  
Assay Methods ◽  
Liquid Chromatographic ◽  
Bridging Study

Abstract A bridging study was conducted to establish the correlation between a liquid chromatographic (LC) method and a microbial inhibition (Ml) method for analysis of amoxicillin residues in catfish muscle. The LC procedure involved precolumn derivatization with formaldehyde followed by LC separation with fluorescence detection. The Ml procedure used Bacillus stearothermophilus as the test organism and was validated in this study before the bridging investigation. The 2 methods were compared for determination of both fortified and incurred samples. No significant differences were found between the methods when all data were included in statistical computations. The linear correlation of LC means versus Ml means had a slope of 0.972 and a negligible intercept (1.0 ng/g), with a correlation coefficient of 0.9962. LC was more specific and showed better sensitivity than Ml for amoxicillin residues at ≤10 ng/g. For practical purposes, values obtained by the 2 methods can be considered equivalent.

scholarly journals Comparison of Micellar Electrokinetic Chromatography, Liquid Chromatography, and Microbiologic Assay for Analysis of Cephalexin in Oral Suspensions

2003 ◽  
Vol 86 (4) ◽  
pp. 707-713 ◽  
Author(s):  
Martin Steppe ◽  
María S Aurora Prado ◽  
Marina F M Tavares ◽  
Teresinha J A Pinto ◽  
Erika R M Kedor-Hackmann ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Micellar Electrokinetic Chromatography ◽  
Test Organism ◽  
Electrokinetic Chromatography ◽  
European Pharmacopoeia ◽  
Plate Method ◽  
Commercial Sample ◽  
Relative Standard ◽  
Routine Quality Control

Abstract Two well-accepted methodologies, based on a microbiologic assay (MA) and liquid chromatography (LC), and a novel methodology using micellar electrokinetic chromatography (MEKC), were compared for the determination of cephalexin in commercially available and simulated samples of oral suspensions. The MA, described in the BrazilianPharmacopeia, was performed with a strain of Staphylococcus aureus ATCC 6538 as the test organism, following the cylinder-plate method. The LC analysis followed the European Pharmacopoeia, 3rd Ed., and was used with minor modifications. The MEKC analysis was based on a previous work of the group. Estimates of the repeatability relative standard deviations of the MA, LC, and MEKC methods in the analysis of a commercial sample were 0.34, 0.42, and 0.37%, respectively. The recovery obtained with LC was 99.90 ± 1.11%; for MEKC, it was 100.09 ± 0.56%. Although the 3 methodologies were statistically equivalent for the determination of cephalexin in oral suspensions, MA gave suitable repeatability despite being nonspecific and time-consuming. MEKC provided faster analysis and higher column efficiency, whereas LC presented superior sensitivity. The results indicated that MEKC can be used as an alternative method to MA and LC in routine quality control laboratories.

scholarly journals Comparison of detection sensitivity of five microbial inhibition tests for the screening of aminoglycoside residues in fortified milk

2012 ◽  
Vol 30 (No. 4) ◽  
pp. 314-320 ◽  
Author(s):  
Z. Sýkorová Goffová ◽  
I. Kožárová ◽  
D. Máté ◽  
S. Marcinčák ◽  
Z. Gondová ◽  
...  
Keyword(s):  
Bacillus Stearothermophilus ◽  
Test Strain ◽  
Detection Sensitivity ◽  
Antibiotic Residues ◽  
Milk Samples ◽  
Microbial Inhibition ◽  
Fortified Milk ◽  
Maximum Residue Limits ◽  
Commission Regulation

The assessment of detection sensitivity of five microbial inhibition tests (MITs), STAR (screening test for antibiotic residues) with the test strain Bacillus subtilis BGA, Delvotest<sup>&reg; </sup>S&nbsp; P-NT, Total Antibiotics, Kalidos TB, and Kalidos MP with the test strain Bacillus stearothermophilus var. calidolactis to five aminoglycosides (AMGs), gentamicin, neomycin, streptomycin, kanamycin, and spectinomycin in fortified milk samples were studied. The sensitivity of MITs to AMGs was evaluated on the basis of experimental determination of detection limits (LODs) of MITs for AMGs. The LODs of these tests were compared with the maximum residue limits (MRLs) established for milk by the Commission Regulation (EU) No. 37/2010. LODs of STAR for AMGs in fortified milk samples were at the levels of MRL for neomycin (1.50 &micro;g/g), gentamicin (0.10&nbsp;&micro;g/g), streptomycin (0.20 &micro;g/g) and kanamycin (0.15 &micro;g/g). Spectinomycin (0.20 &micro;g/g) was not detected at the level of MRL. The LODs determined by Delvotest<sup>&reg; </sup>SP-NT, Total Antibiotics and Kalidos MP were comparable, but only gentamicin and neomycin were reliably detected at the levels of MRL. Kalidos TB was more sensitive to AMGs than Delvotest<sup>&reg; </sup>SP-NT, Total Antibiotics and Kalidos MP. Gentamicin, neomycin and streptomycin were detected at the levels of MRL. &nbsp;

Identification and Determination of Four β-Lactam Antibiotics in Milk

1982 ◽  
Vol 45 (5) ◽  
pp. 450-451 ◽  
Author(s):  
DORIS V. HERBST
Keyword(s):  
Bacillus Stearothermophilus ◽  
Test Organism ◽  
Penicillin G ◽  
Diffusion Method ◽  
Phase Method ◽  
Preliminary Screening ◽  
Diffusion Technique ◽  
Three Phase ◽  
Layer Chromatography

A coordinated three-phase method was developed to determine residues of penicillin G, ampicillin, cephapirin and cloxacillin, the four β-lactam antibiotics most frequently used in mastitis preparations, in milk. An agar well diffusion technique in bioassay trays with Bacillus stearothermophilus as the test organism was used for preliminary screening. Positive samples were subjected to thin-layer chromatography followed by bioautography, and the residues were identified. An agar well diffusion method with standard levels of the specific β-lactam antibiotics was used for quantitation.

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