Click to edit Master title style EVALUATION AND INNOVATION OF OUR - - PowerPoint PPT Presentation

▶
click to edit master title style
SMART_READER_LITE
LIVE PREVIEW

Click to edit Master title style EVALUATION AND INNOVATION OF OUR - - PowerPoint PPT Presentation

Mar 22, 2023 127 likes •553 views

Click to edit Master title style EVALUATION AND INNOVATION OF OUR PROFICIENCY TESTING PROGRAM. Henk Herbold Leen van Ginkel Saskia Sterk October 2008 Content of the presentation. 1. Evaluation of the results, proficiency test Gestagens in

slide-1
SLIDE 1

Henk Herbold Leen van Ginkel Saskia Sterk October 2008

EVALUATION AND INNOVATION OF OUR PROFICIENCY TESTING PROGRAM.

Click to edit Master title style

slide-2
SLIDE 2

ARO: Food and Residues ARO: Food and Residues

2

Content of the presentation.

  • 1. Evaluation of the results, proficiency test ‘Gestagens in porcine

kidney fat’

  • 2. Short evaluation of the results, proficiency test ‘Thyreostatics in

samples of porcine urine’

  • 3. Proficiency testing program 2008 - 2009: ‘future plans’
slide-3
SLIDE 3

ARO: Food and Residues ARO: Food and Residues

3

Overview of the proficiency test “Gestagens in porcine kidney fat” October 2008

slide-4
SLIDE 4

ARO: Food and Residues ARO: Food and Residues

4

‘’Molecular structure of natural progesterone and MPA”.

General information.

slide-5
SLIDE 5

ARO: Food and Residues ARO: Food and Residues

5

Animal experiment, preparation of incurred sample material.

Porcine Kidney fat

slide-6
SLIDE 6

ARO: Food and Residues ARO: Food and Residues

6

Comparing of the analytical possibilities.

Kidney fat Liquid/Liquid SPE Defatting HPLC Fractionation Alkaline Hydrolysis Derivatisation

GCMS LCMS

(ppb) LC-MSMS GC-MS average 4.00 2.69 STD 0.8 0.9 CV% 21 34 N 17 6

slide-7
SLIDE 7

ARO: Food and Residues ARO: Food and Residues

7

  • Batch A. High concentration (approximately 5 ppb) of MPA.
  • Batch B. Low concentration (approximately 1 ppb) of MPA.
  • Batch C. Blank samples.

Samples of porcine kidney fat, prepared for this study.

slide-8
SLIDE 8

ARO: Food and Residues ARO: Food and Residues

8

Homogeneity study.

Single Mix Single Mix Batch A Batch A Batch B Batch B 1 5.6 4.5 1.8 1.1 2 6.9 4.4 1.4 1.1 3 5.1 4.8 1.4 1.1 4 6.2 4.5 1.4 1.1 5 5.5 4.2 1.6 1.0 6 6.6 4.7 1.4 0.8 Average 6.0 4.5 1.5 1.0 STD 0.7 0.2 0.2 0.1 CV% 11.6 4.7 11.2 11.7 F-test 6.060 0.906 Critical value of f(p=0.05) is 6.256 (N-1)

slide-9
SLIDE 9

ARO: Food and Residues ARO: Food and Residues

9

Stability study

Batch A

1 2 3 4 5 6 7 8 9 10 1 2 4 6 8 Weeks Concentrations (ppb) 4 °C 20 °C 37 °C

Batch B

0,2 0,4 0,6 0,8 1 1,2 1,4 1,6 1,8 1 2 4 6 8 Weeks

Concentrations (ppb)

4 °C 20 °C 37 °C

slide-10
SLIDE 10

ARO: Food and Residues ARO: Food and Residues

10

Preparation of test materials, conclusions

  • The use of the LCMS method is preferred, because in case of GCMS, an

alkaline hydrolysis and derivatisation step is necessary.

  • Comparison of the CV values of the samples single and mixed by F-test,

shows no significant differences. Therefore the homogeneity is acceptable.

  • The stability is acceptable (CV<20%), with no significant differences

between storing conditions, during the period of testing.

slide-11
SLIDE 11

ARO: Food and Residues ARO: Food and Residues

11

Total number of participants

Participants (Total 30) NRL Remarks Analysed 28 21 Number of participants that submitted results. Confirmed 25 Number of participants that submitted confirmatory results. No results 2 Number of participants that didn’t send us any results.

slide-12
SLIDE 12

ARO: Food and Residues ARO: Food and Residues

12

Quantification Labs

  • Liquid/Liquid extraction

all

  • Solid Phase Extraction

all

  • LC-MS or LCMS-MS

16

  • GC-MS, GC-HRMS

8

  • Elisa-kit

4

Analytical steps in the procedures used by the participants.

slide-13
SLIDE 13

ARO: Food and Residues ARO: Food and Residues

13

Confirmation Labs

  • Liquid/Liquid extraction

all

  • Solid Phase Extraction

all

  • LC-MS or LCMS-MS

18

  • GC-MS, GC-HRMS

7

Analytical steps in the procedures used by the participants.

slide-14
SLIDE 14

ARO: Food and Residues ARO: Food and Residues

14

Results MPA in Batch A.

ppb Std CV% ppb CRL concentration: 4.57 Mean all laboratories (n=27) 3.59 1.13 32 Median 3.88 Mean all laboratories day 1 (n=27) 3.57 1.15 32 Max. 6.00 Mean all laboratories day 2 (n=24) 3.70 1.11 30 Min. 1.07

Sample Batch A

0,00 1,00 2,00 3,00 4,00 5,00 6,00 7,00 51 52 53 54 55 56 58 59 61 62 64 65 66 67 68 69 70 71 72 73 75 76 77 78 80 82 84 CRL

  • lab. code

conc.

Av1 Av2 CRL value Trimmean 2*horwitz 2*horwitz

slide-15
SLIDE 15

ARO: Food and Residues ARO: Food and Residues

15

Results MPA in Batch B.

ppb Std CV% ppb CRL concentration: 1.00 Mean all laboratories (n=28) 0.97 0.23 24 Median 1.00 Mean all laboratories day 1 (n=28) 1.01 0.28 28 Max. 2.00 Mean all laboratories day 2 (n=24) 0.94 0.23 24 Min. 0.39

Sample Batch B

0,00 0,20 0,40 0,60 0,80 1,00 1,20 1,40 1,60 1,80 2,00 51 52 53 54 55 56 58 59 61 62 64 65 66 67 68 69 70 71 72 73 75 76 77 78 80 82 84 CRL

  • lab. code

conc.

Av1 Av2 CRL value Trimmean 2*horwitz 2*horwitz

slide-16
SLIDE 16

ARO: Food and Residues ARO: Food and Residues

16

Confirmation Gestagens in porcine kidney fat.

10 20 30 40 50 60 70 80 90 Batch A Batch B Batches Number of Labs performing a positiv result, expressed as a percent of all the participants. Confirmed Not confirmed

slide-17
SLIDE 17

ARO: Food and Residues ARO: Food and Residues

17

Summary of the total number of results.

Sample code Min. Max. Mean Median s.d. CV% Batch A 1.07 6.00 3.59 3.88 1.13 32 Batch B 0.39 2.00 0.97 1.00 0.23 24

Sample code Number of participants Analysed No results Outlier 0 ≤ ⎜ Z ⎜≤ 1 1 ≤ ⎜ Z ⎜≤ 2 ⎜ Z ⎜> 2 Batch A 28 27 1 20 7 Batch B 28 28 26 2 Batch C 28 27 1

slide-18
SLIDE 18

ARO: Food and Residues ARO: Food and Residues

18

Evaluation of results from individual samples in the proficiency test (A). Batch A.

  • The overall average value (mean of means) obtained during the

proficiency test was 3.59 ppb (S.D. 1.13), with no observed difference between day one and day 2. Twenty four participants had mean values above 2.0 (range 2.05 – 5.50) and 3 participants had mean values below 2.0 (range 1.16 – 1.78).

  • Twenty-four laboratories were able to confirm residues MPA according to

the EU criteria, (Commission Decision 2002/657/EC). Four laboratories did not send us results about the confirmation of MPA.

slide-19
SLIDE 19

ARO: Food and Residues ARO: Food and Residues

19

Evaluation of results from individual samples in the proficiency test (B). Batch B.

  • The overall average value (mean of means) obtained during the

proficiency test was 0.97 ppb (S.D. 0.23), with no observed difference between day one and day 2. In view of the mean of means, there is a good agreement with the expected value.

  • Fourteen participants had mean values above 1.0 (range 1.00 – 1.38)

and 14 participants had mean values below 1.0 (range 0.60 – 0.99).

  • Twenty-two laboratories were able to confirm MPA according to the EU

criteria, (Commission Decision 2002/657/EC). Four laboratories did not send us results about the confirmation of MAP.

slide-20
SLIDE 20

ARO: Food and Residues ARO: Food and Residues

20

Evaluation of results from individual samples in the proficiency test (C).

Batch C

  • This sample was focused on a concentration of <0.5 ppb MPA.

There were no false-positive results.

  • In view of the mean of means, there is a good agreement with the

expected value.

slide-21
SLIDE 21

ARO: Food and Residues ARO: Food and Residues

21

Scores individual labs.

Graphical representation

2 4 6 8 10 12 14 16 5 7 6 8 9 scores Frequentie

slide-22
SLIDE 22

ARO: Food and Residues ARO: Food and Residues

22

Conclusion.

  • In total 30 laboratories registered as participant for the test and

28 laboratories reported their results. A limited number of participants (24 labs) was able to quantify and confirm MPA in porcine kidney fat.

  • 21 laboratories reported excellent results (score 8 - 9) and 6

laboratories reported good results (score 6 -7).

  • Only one laboratory report unsatisfactory results (scores 5).
  • Most of the laboratories demonstrated to be able to confirm and

to quantify MPA, only four laboratories did not perform confirmatory analyses.

slide-23
SLIDE 23

ARO: Food and Residues ARO: Food and Residues

23

Compared results obtained by LCMS versus GCMS. Batch A.

(ppb) LC-MSMS GC-MS average 4.00 2.69 STD 0.8 0.9 CV% 21 34 N 17 6

  • 1. Most of the participants used for detection LCMS or LCMS/MS.
  • 2. Only six laboratories sent us results obtained by GCMS/MS.
  • 3. The results obtained by GCMS were lower then the expected value.
slide-24
SLIDE 24

ARO: Food and Residues ARO: Food and Residues

24

Overall: Excellent result!

slide-25
SLIDE 25

ARO: Food and Residues ARO: Food and Residues

25

Overview of the proficiency test “Thyreostatics in porcine samples of urine” November 2008

slide-26
SLIDE 26

ARO: Food and Residues ARO: Food and Residues

26

Preparation of sample materials: Excretion pattern

Animal 3454 500 1000 1500 2000 2500 3000 3500 2005M17702005M17712005M17722005M17732005M17752005M17762005M1777 Sample nr. Concentration (ppb 1000 2000 3000 4000 5000 6000 Tapazol-IBBR Methylthiouracil-IBBR

slide-27
SLIDE 27

ARO: Food and Residues ARO: Food and Residues

27

Materials distributed

  • Batch A. High concentration (50 ± 10 ppb) of MTU and Tapazol.
  • Batch B. Low concentration (10 ± 5 ppb) of MTU and Tapazol.
  • Batch C. Blank samples.
slide-28
SLIDE 28

ARO: Food and Residues ARO: Food and Residues

28

Stability of Methylthiouracil in lyophilised urine.

2008m0496 Methylthiouracil (ng) Temperatuur t (weken) 37 0C 20 0C 4 0C

  • 70 0C

8 53,14 56,41 49,78 48,03 7 67,19 54,70 54,93 44,33 6 64,55 62,96 49,60 51,34 5 62,04 58,56 41,50 51,24 4 54,32 52,88 55,49 44,93 3 62,41 58,13 45,18 51,81 2 66,47 46,82 49,70 45,95 1 57,03 50,79 43,96 48,48 Totaal Gemiddeld 60,89 55,16 48,77 48,26 53,27 Standaarddeviatie 5,07 4,70 4,67 2,80 6,90 CV (%) 8,3 8,5 9,6 5,8 13,0

Methylthiouracil

0,00 20,00 40,00 60,00 80,00 2 4 6 8 10 t (weken) M (n g ) 37 0C 20 0C 4 0C

  • 70 0C
slide-29
SLIDE 29

ARO: Food and Residues ARO: Food and Residues

29

Summarises results of sample Batch A

Concentration of MTU in ppb.

ppb Std CV% Trimmean Horwitz ppb

CRL concentration: 44.5 78.15 18.35 Mean all laboratories (n=28) 84.0 28.8 34 Median 73.5 Mean all laboratories day 1 (n=28) 83.0 28.4 34 Max. 212.0 Mean all laboratories day 2 (n=24) 83.9 29.8 36 Min. 38.5

Sample Batch A

50 100 150 200 250 50 51 52 53 55 58 59 60 62 63 64 66 67 68 69 71 72 73 74 75 76 77 78 79 80 83 CRL

  • lab. code

conc.

Av1 Av2 CRL value Trimmean 2*horwitz 2*horwitz Copy results

slide-30
SLIDE 30

ARO: Food and Residues ARO: Food and Residues

30

Summarises results of sample Batch B

Concentration of MTU in ppb.

ppb Std CV% Trimmean Horwitz ppb

CRL concentration: 13.1 28.28 5.84 Mean all laboratories (n=28) 21.3 8.54 40 Median 20.0 Mean all laboratories day 1 (n=28) 21.7 8.93 41 Max. 46.3 Mean all laboratories day 2 (n=24) 21.1 8.81 42 Min. 8.0

Sample Batch B

5 10 15 20 25 30 35 40 45 50 51 52 53 55 58 59 60 62 63 64 66 67 68 69 71 72 73 74 75 76 77 78 79 80 83 CRL

  • lab. code

conc.

Av1 Av2 CRL value Trimmean 2*horwitz 2*horwitz Copy results

slide-31
SLIDE 31

ARO: Food and Residues ARO: Food and Residues

31

Stability of Tapazol in lyophilised urine.

2008m0496 Tapazol (ng) Temperatuur t (weken) 37 0C 20 0C 4 0C

  • 70 0C

8 13,17 16,99 25,37 20,31 7 17,18 14,75 27,80 20,63 6 12,72 18,59 23,31 22,73 5 12,50 17,18 20,55 21,24 4 12,12 16,38 27,64 22,83 3 12,19 19,46 22,08 19,92 2 13,78 16,29 24,68 19,21 1 14,02 20,65 24,87 22,43 Totaal Gemiddeld 13,46 17,54 24,54 21,16 19,17 Standaarddeviatie 1,55 1,79 2,36 1,29 4,57 CV (%) 11,5 10,2 9,6 6,1 23,8

Tapazol

0,00 5,00 10,00 15,00 20,00 25,00 30,00 2 4 6 8 10 t (weken) M (ng) 37 0C 20 0C 4 0C

  • 70 0C
slide-32
SLIDE 32

ARO: Food and Residues ARO: Food and Residues

32

Summarises results of sample Batch A

Concentration of Tapazol in ppb.

ppb Std CV% Trimmean Horwitz ppb

CRL concentration: 57.73 36.95 9.71 Mean all laboratories (n=28) 47.31 59.3 125 Median 31.5 Mean all laboratories day 1 (n=28) 47.93 59.7 124 Max. 301.0 Mean all laboratories day 2 (n=24) 44.65 59.3 133 Min. 11.1

Sample Batch A

50 100 150 200 250 50 51 52 53 55 58 59 60 62 63 64 66 67 68 69 71 72 73 74 75 76 77 78 79 80 83 CRL

  • lab. code

conc.

Av1 Av2 CRL value Trimmean 2*horwitz 2*horwitz Copy results

slide-33
SLIDE 33

ARO: Food and Residues ARO: Food and Residues

33

Summarises results of sample Batch B

Concentration of Tapazol in ppb.

ppb Std CV% Trimmean Horwitz ppb

CRL concentration: 17.55 12.35 3.83 Mean all laboratories (n=28) 15.75 13.4 85 Median 11.4 Mean all laboratories day 1 (n=28) 14.54 12.3 84 Max. 72.5 Mean all laboratories day 2 (n=24) 16.26 14.8 91 Min. 3.0 Sample Batch B

10 20 30 40 50 60 70 50 51 52 53 55 58 59 60 62 63 64 66 67 68 69 71 72 73 74 75 76 77 78 79 80 83 CRL

  • lab. code

conc.

Av1 Av2 CRL value Trimmean 2*horwitz 2*horwitz Copy results

slide-34
SLIDE 34

ARO: Food and Residues ARO: Food and Residues

34

Evaluation report of the proficiency test “Thyreostatics in porcine samples of urine”

  • Conclusions and recommendations.
  • Evaluation of the methods used by the participants.
  • Evaluation of the results.
slide-35
SLIDE 35

ARO: Food and Residues ARO: Food and Residues

35

Future plans (1).

  • Proficiency test, Methylprednisone and metabolites in

samples of bovine urine

  • Research study, metabolites of 1-Testosterone in

samples of bovine urine

slide-36
SLIDE 36

ARO: Food and Residues ARO: Food and Residues

36

Research study versus proficiency test.

Research study.

  • 1. We invite interested NRLs to analyse the samples, to agree a

common understanding of a specific analytical question.

  • 2. Continue the development of generic analytical methods

suitable for metabolites of growth promoters. Proficiency test.

  • 1. The purpose is to evaluate current analytical methodology of

the NRLs, for the quantification of low levels steroids and their metabolites.

slide-37
SLIDE 37

ARO: Food and Residues ARO: Food and Residues

37

Future plans (2).

Treatment of a heifer by three substances, (animal experiments)

  • Testosteron-cypionate

analyses of natural hormones in plasma and hair

  • Chloortestosterone-acetate

analyses of CLAD and metabolites

  • Zeranol

fusarium metabolites, research study

slide-38
SLIDE 38

ARO: Food and Residues ARO: Food and Residues

38

slide-39
SLIDE 39

ARO: Food and Residues ARO: Food and Residues

39

http://www.rivm.nl/residues

slide-40
SLIDE 40

ARO: Food and Residues ARO: Food and Residues

40

Thank you for your attention. Thank you for your attention.