Progetto Trieste 2011, 4Th Round, Veterinary Drug Residues

Progetto Trieste 2011, 4th round, Veterinary Drug Residues Proficiency Testing Service by Test Veritas S.r.l.

TEST MATERIAL: MI1133A-1 and MI1133A-2 (confirmatory) TETRACYCLINE and QUINOLONES IN LYOPHILIZED MILK - spiked 2011, 4th round

Test material A are two different samples: A-1 and A-2 - please test both materials!

LABORATORY: … ANALYST: ... DATE: ...

COMPOUNDS RESULTS SPECIFICATIONS DECISION CONCENTRATION DETECTION LIMIT ISO CORRECTED FOR RECOVERY CAPABILITY CC or 17025** MI1133A-1 RECOVERY* % CC cut-off yes/no (g/kg) (g/kg) (g/kg) OXYTETRACYCLINE EPIOXYTETRACYCLINE CHLORTETRACYCLINE EPICHLORTETRACYCLINE DOXYCYCLINE EPIDOXYCYCLINE TETRACYCLINE EPITETRACYCLINE ENROFLOXACIN CIPROFLOXACIN FLUMEQUINE MARBOFLOXACIN DANOFLOXACIN SARAFLOXACIN NALIDIXIC ACID OXOLINIC ACID Milk density = 1.03

Please indicate the concentration in relation to liquid milk sample (not of the powder)

A Maximum Residue Limits (MRL) of 100 g/kg for tetracycline, chlortetracycline and oxitetracyclin in milk has been definited by the European Commission (EU Regulation 37/2010). Doxycyclin is not allowed in dairy cows. A Maximum Residue Limit (MRL) in milk has been definited by the European Commission (EU Decision 37/2010) for some quinolones, as follows: 100 g/kg for enrofloxacin and ciprofloxacin (sum); 30 g/kg for danofloxacin; 50g/kg for flumequine; 75 g/kg for marbofloxacin. Oxolinic acid is not allowed in dairy cows.

TEST MATERIAL: MI1133A-1 and MI1133A-2 (confirmatory) TETRACYCLINE and QUINOLONES IN LYOPHILIZED MILK - spiked PAGINA 1 di 5 REVISIONE DATA DELLA REVISIONE DOCUMENTO AGGIORNATO IL 17/10/11 N. 0 03/01/2011 The data of your analyses will be kept for 5 years and after then will be destroyed. Progetto Trieste 2011, 4th round, Veterinary Drug Residues Proficiency Testing Service by Test Veritas S.r.l.

2011, 4th round

COMPOUNDS RESULTS SPECIFICATIONS DECISION CONCENTRATION DETECTION LIMIT ISO CORRECTED FOR RECOVERY CAPABILITY CC or 17025** MI1133A-2 RECOVERY* % CC cut-off yes/no (g/kg) (g/kg) (g/kg) OXYTETRACYCLINE EPIOXYTETRACYCLINE CHLORTETRACYCLINE EPICHLORTETRACYCLINE DOXYCYCLINE EPIDOXYCYCLINE TETRACYCLINE EPITETRACYCLINE ENROFLOXACIN CIPROFLOXACIN FLUMEQUINE MARBOFLOXACIN DANOFLOXACIN SARAFLOXACIN NALIDIXIC ACID OXOLINIC ACID Milk density = 1.03

Please indicate the concentration in relation to liquid milk sample (not of the powder)

Comments:

* Please indicate:  estimated concentration corrected with recovery if the compound is detected;  "< of…" if the compound is not detected;  "NS" if the compound is not searched (see example above reported).

**Please indicate whether the method used is in the scope of accreditation (choose “yes” if the methods is already accreditated).

PAGINA 2 di 5 REVISIONE DATA DELLA REVISIONE DOCUMENTO AGGIORNATO IL 17/10/11 N. 0 03/01/2011 The data of your analyses will be kept for 5 years and after then will be destroyed. Progetto Trieste 2011, 4th round, Veterinary Drug Residues Proficiency Testing Service by Test Veritas S.r.l.

Example: COMPOUNDS RESULTS SPECIFICATIONS CONCENTRATIO DECISION DETECTION N ISO RECOVER LIMIT CAPABILITY CORRECTED 17025** Y % CC CC FOR RECOVERY* yes/no (g/kg) (g/kg) (g/kg) Compound 1 150 90 80 95 y Compound 2 NS - - - - Compound 3 < 50 - 50 70 n

LABORATORY: … ANALYST: …

MARK YOUR CHOICE

Analytical methods used by participants Methods HPLC/MS UPLC/MS/MS HPLC/MS/MS GC/MS HPLC/DAD Other (specify)

Is the analytical method a routine one? Yes No

Source of standards for chromatographyc analysis OTC CTC DTC TC EPIMERS Sigma, Aldrich, Fluka Riedel-de Haen Dr. Ehrenstorfer Other (specify)

ENRO CIPRO FLUM MARB DAN SARA NAL. AC OXO.AC Sigma, Aldrich, Fluka Riedel-de Haen Dr. Ehrenstorfer Other (specify)

Sample preparation Amount of test sample for the extraction Sample volume (ml) 1 5 2 10 3 Other (specify)

Sample treatment Extraction dichloromethane Mc Ilvaine buffer ethyl acetate succinate buffer acetonitrile chloroform / acetone dichloromethane / trichloroacetic Other (specify) Digestion … (specify)

PAGINA 3 di 5 REVISIONE DATA DELLA REVISIONE DOCUMENTO AGGIORNATO IL 17/10/11 N. 0 03/01/2011 The data of your analyses will be kept for 5 years and after then will be destroyed. Progetto Trieste 2011, 4th round, Veterinary Drug Residues Proficiency Testing Service by Test Veritas S.r.l.

Clean up (SPE or other) SPE clean up C18 OASIS® Silica StrataTM SCX Other (specify) Other clean up n-hexane centrifugation filtration Other (specify)

HPLC methodology and conditions HPLC Gradient Isocratic Column phase C8 PFP C18 Other (specify) Column length (cm) 5 20 10 25 15 Other (specify) Column diameter (mm) 2 3.9 2.1 4.6 3 Other (specify) Particle size (m) 1.7 4 2.5 5 3 Other (specify) Cromatographyc column (type) AtlantisTM Prodigy® AcquityTM Symmetry® Beckman Synergi® GeminiTM Superspher® Hypersil® SupelcosilTM Hypercab® Ultrabase® HyPURITY X-terra® Lichrospher® YMCTM Luna® Zebron® Novapak® Zorbax® Nucleodur® Other (specify) Mobil phase acetonitrile / acetate buffer sodium acetate buffer / acetonitrile ammonium acetate / acetonitrile formic acid / acetonitrile water / acetonitrile / acetic acid formic acid / oxalic acid / acetonitrile acetate buffer Other (specify) pH of mobile phase x < 2 4 < x < 5 2 < x < 3 5 < x < 6 3 < x < 4 Other (specify) Flow rate (ml/min) 0.25 0.7 0.3 1 0.4 Other (specify) Post column reagent … (specify) TEST MATERIAL: MI1133A-1 and MI1133A-2 (confirmatory) TETRACYCLINE and QUINOLONES IN LYOPHILIZED MILK - spiked 2011, 4th round

Injection volume (l) 10 50 PAGINA20 4 di 5 REVISIONE DATA DELLA REVISIONE 100 DOCUMENTO AGGIORNATO IL 17/10/11 N. 0 03/01/2011 30 Other (specify) The data of your analyses will be kept for 5 years and after then will be destroyed. UV(nm) … (specify) Progetto Trieste 2011, 4th round, Veterinary Drug Residues Proficiency Testing Service by Test Veritas S.r.l.

GC methodology GC Capillary column length (m) … (specify) Film thickness (m) … (specify) Cromatographyc column Agilent Zorbax WCOT Other (specify) Zebron® Injection volume (l) … (specify) Injection temperature (°C) … (specify) Injection procedure Splitless Other (specify) Oven temperature (°C) … (specify)

Number of replicates (1 replicates = 1 extracion) Number of extraction 1 Other (specify) 2

Recovery (HPLC and GC) Recovery calculated by Internal standard Matrix matched calibration Spiked sample Other (specify) In-house Reference Material (RM)

Quality (HPLC and GC) Quality control implemented Control chart with in-house Duplicate analyses Reference Material (RM) Control chart with Certified Spiked sample Reference Material (CRM) Regular partecipation to Other (specify) proficency test (PT)

Reference (HPLC and GC) Reference Internal method Other (specify)

PAGINA 5 di 5 REVISIONE DATA DELLA REVISIONE DOCUMENTO AGGIORNATO IL 17/10/11 N. 0 03/01/2011 The data of your analyses will be kept for 5 years and after then will be destroyed.