CRL workshop 2004 summary report-final mjAl. Paryzantow 57 PL-24-100 PULAWY Poland Tel: +48 818863051/173 Fax: +48 818862595 e-mail: [email protected] Th. Dabalis Ministry of

ARODOCnr. 23693

Summary Report, Conclusions and Recommendations

“Evaluation Results Proficiency Tests

and

Technical Training”

EU CRL-NRL Workshop, 11-15 October 2004, Bilthoven, The Netherlands

EU CRL-NRL Workshop, 11-15 October 2004, Bilthoven, The Netherlands “Evaluation Results Proficiency Tests and Technical Training” Summary Report, Conclusions and Recommendations

Summary report, Conclusions and Recommendations EU-CRL Workshop; 11-15 October 2004. Page 1 of 14

Contents List of participants 2 Guest lecturers 4 Organizing CRL 4 Critical concentrations in method validation 5 Implementation of MRPLs 5 Discussion on setting new MRPLs 6 Boldenone, the end of the story? 7 The synthesis of reference compounds 7 Proficiency tests in 2004 and 2005, some results and a preview 8 Animal experiments in support of proficiency tests 8 Discussion on priorities for future CRL work programs 9 Priorities with respect to proficiency tests 10 Questionnaire on Stilbene analysis 10 Evaluation of the 2001 – 2003 ANPs for stilbenes 10 Update on CODEX / EU business and SPECLOG explained 11 Criteria compared. WADA and EU criteria in practice 11 Annex to Summary Report, Conclusions and Recommendations 12 CD-ROM contents back cover CD-ROM: CRL Workshop 2004 back cover Note: Fotograph on cover taken in front of the monument of the historical entrance of

RIV originally located in Utrecht



List of participants J.M Degroodt Ministerie van Sociale Zaken Volksgezondheid en Leefmilieu WIV - Wetenschappelijk Instituut Volksgezondheid Juliëtte Wytsmanstraat 14 B-1050 BRUSSEL België Tel: +32 26425353 Fax: +32 26425691 e-mail: [email protected] Mrs.S. Berg National Veterinary and Food Research Institute Department of Chemistry P.O.BOX 45 SF-00581 HELSINKI Finland Tel: +358 93931918 Fax: +358 93931920 e-mail: [email protected] M. Lohmus Tallinn Veterinary and Food Laboratory Väeke-Paala 3 EE-11415 TALLINN Estonia Tel: +372 6035810 Fax: +372 6035811 e-mail: [email protected] B. Sas National Food Investigation Institute P.O.Box 1740 BUDAPEST 94 H-1465 Hungary Tel: +361 4563013 Fax: +361 2151574 e-mail: [email protected] G. Ferretti Istituto Superiore di Sanità (ISS) Laboratory Vet. Med. Viale Regina Elena 299 I-00161 ROMA Italy Tel: +39 0649903078 Fax: +39 0649903079 e-mail: [email protected]

E. Bichon Laboratoire d'Etude des Résidus et Contaminants dans les Aliments (LABERCA) Route de Gachet BP 50707 F-44307 NANTES CEDEX 03 France Tel: +33 240687880 Fax: +33 240687878 e-mail: [email protected] M. Hadjigeorgiou The State General Laboratory, Veterinary Drug Residues Lab, 44 Kimonos Street, 1451 NICOSIA ACROPOLIS, Cyprus Tel: +357 22809204 Fax: +357 22316434 e-mail: [email protected] M. Pedersen National Food Agency of Denmark Morkhoj Bygade 19 DK-2860 SOBORG Denmark Tel: +45 72347429 Fax: +45 72347488 e-mail: [email protected] S.K. Mortensen National Food Agency of Denmark Morkhoj Bygade 19 DK-2860 SOBORG Denmark Tel: +45 72347471 Fax: +45 72347001 e-mail: [email protected] D. Farrugia Ministry of Agriculture and Fisheries Albertown, Marsa, MT - CMR 02 MARSA, Malta Tel: +21242694 Fax: +21238105 e-mail: [email protected]



G. Alfredsson Swedish National Food Administration Chemistry Division I P.O. Box 622 SE-75126 UPPSALA Sweden Tel: +46 18175612 Fax: +46 18105848 e-mail: [email protected] B. Wosniak National Veterinary Research Institute Dept. of Hygiene of Food of Animal Origin Al. Paryzantow 57 PL-24-100 PULAWY Poland Tel: +48 818863051/173 Fax: +48 818862595 e-mail: [email protected] Th. Dabalis Ministry of Agriculture Veterinary Laboratory of Serres Terma Omonoias, GR-62110 SERRES Greece Tel: +30 23210 35411 Fax: +30 2321035410 e-mail: [email protected] S.A. Hewitt Ministery of Agriculture and Rural Development, Veterinary Sciences Division, Stoney Road, Stormont, BELFAST BT4 3SD United Kingdom – NI Tel: +44 2890525650 Fax: +44 2890525626 e-mail: [email protected] P. Fevrier Dept. of Agriculture, Food and Forestry Central Meat Control Laboratory Abbotstown-Castleknock DUBLIN 15 Ireland Tel: +35318025800 Fax: +35318217320 e-mail: [email protected]

K. Fragalova USKVBL Medlanky Hudcova 56a CZ 6200 BRNO Czech Republic Tel: +420 541210022 Fax: +420 54121607 e-mail: [email protected] T. Kuhn AGES-VMMOE Federal Institute for Veterinary Medicine Dept. of Chemistry, Section for Growth Promoting Agents Robert Koch Gasse 17 A 2340 MÖDLING Austria Tel: + 43 2236 46640360 Fax: + 43 2236 46640368 e-mail: [email protected] R. Schuch AGES-VMMOE Federal Institute for Veterinary Medicine Dept. of Chemistry, Section for Growth Promoting Agents Robert Koch Gasse 17 A 2340 MÖDLING Austria Tel: + 43 2236 46640360 Fax: + 43 2236 46640368 e-mail: [email protected] J.M. Blanca Barrano Ministerio de Sanidad y Consumo Centro Nacional de Alimentacion Instituto de Salud Carlos III Unidad de Residuos Zoosanitarios, Carretera Majadahonda-Pozuelo Km 2 E-28220 MADRID MAJADAHONDA Spain Tel: +34 918223040 Fax: +34 915097926 e-mail: [email protected] Ch. Hjaeresen Sondervang 4 DK-4100 RINGSTED Denmark Tel: +45 57682096 Fax: +45 57682199 e-mail: [email protected]



R. Hazucha State Veterinary and Food Institute Food Residue Hormones Department Akademicka3 949 01 NITRA Slovak Republic Tel: +421376536520 e-mail: [email protected] K. Schmidt BVL Laboratory Residues of Veterinary Drugs Diedersdorfer Weg 1 D-12277 BERLIN Germany Tel: +49 18884122310 Fax: +4918884122300 e-mail: [email protected] Neringa Visciulyte National Veterinary Laboratory Department of Chemistry J. Kairiukscio 10 LT 08409 VILNIUS Lithuania Tel: +370 52780407 Fax: +370 52780471 e-mail: [email protected]

Guest lecturers A. Blass Rico European Commission DG Health and Consumers Protection Rue de La Loi 200 B-1049 Brussels BELGIUM Tel: +32 22959663 Fax: +32 22991856 e-mail: [email protected] J. Langemans Animal Sciences Group Wageningen Universiteit & Researchcentrum Runderweg 6 8219 PK Lelystad THE NETHERLANDS Tel: +31 320 293 250 Fax: +31 320 293 220 e-mail: [email protected] Organizing CRL (RIVM / ARO, Bilthoven, the Netherlands) Tel: +31 30 2742613 Fax: + 31 30 2744403 e-mail: [email protected] www.rivm.nl/crl/residues R.W. Stephany ( CRL, Director) L.A. van Ginkel ( ARO, acting Head) S.S. Sterk ( ARO, Head of Section) P. Kootstra M.A. Jonker M. Blokland H. Herbold H.J. van Rossum K. van Twillert P. Zoontjes M. Kartasasmita



The first workshop presentation was made by Rainer Stephany, Director of the CRL. Critical concentrations in method validation. During this presentation a specific problem with respect to the use of CCalpha in confirmatory analyses was addressed. The definition in Commission Decision 2002/657/EC only reflects the quantitative aspect on the analysis. The alpha error, which by definition means the probability that the tested sample is compliant, even though a non-compliant measurement has been obtained (false non-compliant decision), can only be applied after an evaluation of all the identification criteria. For group A substances, a non-compliant result is obtained the moment it is concluded that all the identifiers are present and, when applicable, have the correct characteristics (e.g. ion-ratio’s). The error-probability for this decision if not formally set but generally considered to be 1 out of 10.000 or less. The identification criteria, though not chemometrically derived, have this value as a minimum. Methods used for confirmatory analyses must be able to identify true positive samples reliably at a value equal or lower than the MRPL. The value of CCalpha as determined as described in CD 2002/657/EC has little value in this respect. For consumer protection at the end the value of CCbeta of the screening method is the most important value to be quality controlled. The full presentation is included in the RIVM/CRL workshop CD-ROM 2004. Implementation of MRPLs. Mrs Ana Blass on behalf of the European Commission presented the background and consequences of the new Commission Decision on import controls on the implementation of MRPLs. According to Council Directive 96/23/EC , the maximum residue limits established according to Regulation EC (No) 2377/90 (laying down a Community procedure for the establishment of maximum residue limits of veterinary medicinal products in foodstuffs of animal origin), are the permitted limits to be applied in food control. However, Regulation 2377/90 does not provide permitted limits for all substances and in particular not for those substances whose use is not authorized or prohibited in the European Community. To ensure a consistent approach across the Community and to avoid disruption to trade it was considered necessary to set harmonized standards for the procedures for testing for residues in imported products of animal origin under veterinary checks carried out at border inspection posts. Additionally, Commission Decision 2002/657/EC sets down the minimum required performance limits (MRPLs) for analytical methods to be used for certain substances for which no permitted residue limit has been established. In cases where permitted limits have not been established, any presence of relevant substances in a consignment was construed as presenting grounds to reject and destroy the consignment. In practice, the consignments are accepted or rejected at import on the basis of the choice of the method employed and in particular the method’s limit of detection. The opportunity should also be taken to align the existing requirements with the provisions of Regulation (EC)



178/2002 on Food Law and the Regulation (EC) 882/2004 on Official Food and Feed Controls , especially those provisions relating to the non-compliant consignments at import. According to the new Decision, where results of analytical tests are at or above the MRPLs laid down in Commission Decision 2002/657/EC the consignment considered shall be considered non-compliant with Community Legislation. Where the results of analytical tests on products are below the MRPLs laid down in Commission Decision 2002/657/EC, the products will not be prohibited from entering in the food chain. If there is any evidence indicating a pattern of misuse of particular substances or a breach of agreement of non-use of certain substances in the production of food intended for import in the Community, the Member State shall inform the Commission and the Third Country will be requested for additional information and corrective actions. For intra community controls, if the results of analytical tests on products are below the MRPLs, the sample will be considered compliant but there has been evidence of misuse and follow up actions should be taken in order to identify and prosecute the origin of the illegal use. The results should be reported to the Commission in the annual report including the follow up measures taken after non-compliant results. If the product is coming from another MS, the MS of origin shall be informed in order to allow them to take appropriate actions. The system of reporting is being developed by the European Commission. A copy of the draft Decision on import controls was distributed to the participants. The full presentation is included in the RIVM/CRL workshop CD-ROM 2004. N.B. The Decision has been published on 20.1.2005, OJ L 16/61, Commission Decision 2005/34/EC and is included on the CD-ROM. Discussion on setting new MRPLs. During the 2003 workshop a number of MRPLs were agreed upon by participants. The objective for this year workshop was a further extension of proposed MRPL values, inclusive MRPL-values for muscle tissue. With the publication of Commission Decision 2005/34/EC, MRPL values for edible (imported) matrices are of special interest.

Previously advised MRPL values Diethylstilbestrol: 1 ppb for bovine urine Thiouracil: 100 ppb for bovine urine 17beta-Boldenone: 1 ppb for bovine urine 17alpha-Nortestosterone: 2 ppb in bovine urine Methyltestosterone: 2 ppb for bovine urine 16 beta-Hydroxystanozolol: 2 ppb for bovine urine Dexamethasone: 2 ppb for bovine urine Zeranol and Taleranol: 2 ppb for bovine urine



The topics discussed were: • Extending the number of analytes for the matrix urine, and extension with ovine

and porcine species • Setting MRPL values for the matrix liver for A1, A3 and A4 compounds • Setting MRPL values for the matrix muscle, inclusive fish for A1, A3 and A4

compounds The tables with MRPL values discussed are added as an annex to this summary report and are included on the RIVM/CRL workshop CD-ROM 2004. Boldenone, the end of the story? A presentation was given by Marco Blokland on the current situation with respect to residue control for boldenone. A working group established by the EC prepared an advise with respect to the control for the illegal use of boldenone. Its recommendation is to consider conjugated forms of 17beta-boldenone (primarily glucuronides) as conclusive for the illegal use of boldenone. As a consequence, specific analytical methods are necessary to discriminate conjugated from non-conjugated forms of 17beta-boldenone. The analytical approach developed at the CRL was explained. The full presentation is included on the RIVM/CRL workshop CD-ROM 2004, inclusive SOP 478 and the EU review paper by De Brabander et al. (ARODOC nr 22941; see file 22941adc ). The full presentation is included in the RIVM/CRL workshop CD-ROM 2004. The synthesis of reference compounds. Klaas van Twillert presented his work on the synthesis of deuterated Trenbolone. This hormone was prepared starting with triendione. This compound was treated with D2O to exchange hydrogen with deuterium on the 2, 4, 6 and 16 position. Reduction of the two ketone moieties was performed with NaBD4. During this reduction two more deuterium atoms were introduced (on the 3 and 17 position). With MnO2 the hydroxyl group on the 3 position was selectively oxidized to a ketone. Because the back exchange (deuterium to hydrogen) of the enolisable positions failed under both basic and acid conditions the product turned out to be Trenbolone(d8). This was confirmed by GCMS and LCMS. It was announced that in the next future another attempt will be made to produce deuterium- labeled Trenbolone on a larger scale and that once more it will be tried to achieve the intended d3 label instead of d8. The full presentation is included in the RIVM/CRL workshop CD-ROM 2004.



Proficiency tests in 2004 and 2005, some results and a preview. Henk Herbold discussed the results of the recent proficiency tests and gave an overview of the tests planned for 2005. One proficiency test focused on Zeranol and its metabolites. The short evaluation report with the results obtained had been available for some time and the full report is under preparation. The proficiency test was very useful, especially for the matrix urine. More than 20 laboratories participated. The results for Zeranol and Taleranol were excellent at the level of 3 ppb. Most participants were able to detect and confirm the analytes at this level quantified the compounds with acceptable accuracy. At the level of 0.5 ppb, the majority of participants was not able to produce valid results. From the proficiency test it is concluded that the current MRPL value of 2 ppb, for both Zeranol and Taleranol, reflect the technical possibilities of the NRLs adequately. The results for the other Resorcylic Acid Lactones (RALs) were highly variable with only a relative small group able to produce good results. No evaluation was made for the matrix bile, with only one participant producing results. The full presentation is included on the RIVM/CRL workshop CD-ROM 2004. Animal experiments in support of proficiency tests. Presented by Jan Langemans (Animal Sciences Group, Wageningen UR, The Netherlands)) To develop and/or validate assay systems for the detection of residues in farm animals high-quality, defined samples are required. Animals kept under carefully controlled circumstances can provide such important samples. Substances for which an assay is developed and / or validated can be administered to the animals under controlled conditions and amounts. Subsequently, samples (e.g. urine, blood, feces) can be obtained at exactly defined time-points. Samples have to be taken in a way that prevents cross-contamination (e.g. urine via a catheter). To obtain reliable incurred samples for general validation and assay development, experimental animal facilities with the required facilities and personnel and experience in this type of research are essential. The full presentation is included in the RIVM/CRL workshop CD-ROM 2004.



Discussion on priorities for future CRL work programs. As on previous occasions the future priorities were discussed. Participants were requested to prioritize on the basis of ten prelisted possibilities.

1. Development / validation of confirmatory methods (TRADE protection) 2. Development / validation of screening methods (CONSUMERS protection) 3. Analytical research into new technologies / alternative approaches 4. Studies into metabolism of compounds 5. Proficiency tests 6. Documentation services 7. Assistance with implementation of QA-systems 8. General training facilities 9. Bank of Reference Standards and Materials 10. Maintaining a dedicated CRL-website

The results are summarized in the figure below.

relative priorities CRL

0%

5%

10%

15%

20%

25%

1 2 3 4 5 6 7 8 9 10

activity

rela

tive

prio

rity

Series1

Making reference standards and materials available clearly is the no. 1 priority for the NRLs. The next three priorities include metabolism studies, proficiency tests and the development of confirmatory methods. These relative priorities are in close agreement with the workprogramme for 2005. Preparing test and reference materials, in conjunction with the organization of proficiency test, are prominently included, next to the development and validation of necessary new analytical methods.



Priorities with respect to proficiency tests. In general terms participants were satisfied with the frequency with which the proficiency tests are organized, They appreciated the detailed reports. Priorities mentioned reflected the whish to include more matrices other than urine, viz. stilbenes in aqua culture, poultry liver and muscle, thyreostatics in urine or thyroid, Zeranol in muscle, gestagens in kidney fat, oestradiol in plasma. The preferred period for reporting is two months, excluding the summer period from June to August. Questionnaire on Stilbene analysis. A questionnaire on the current status of analytical methods for stilbenes, prepared by Marco Jonker, was distributed among the participants of the CRL workshop. Besides the name and coordinates of the contact person who completed the inquiry, the kind of information desired by EU CRL Bilthoven from the 25 MS is related to the following subjects: the nature of the programme for regulatory control of stilbenes, the stilbenes taken up in the control, the matrices of the materials controlled, the validation of the method used, analytical techniques used for sampling, screening and confirmation, the validation parameters of the method, the findings of the control program for stilbenes in 2004, new methods in development, other currently research subjects for stilbenes, and availability of standards and reference materials. Participants were asked to return the questionnaire before 30 November 2004. In this way the CRL is collecting information on the analytical methods applied for the analysis of stilbenes by responsible National Reference Laboratories (NRLs) in the Member States and / or the National Routine or Regional Laboratories under mandate of the NRLs. The information also is collected in preparation of a proficiency study to be organized in 2005. Evaluation of the 2001 – 2003 ANPs for stilbenes. An evaluation by Marco Jonker of the National Residue Monitoring Plans of the Member States (ANPs) in the years 2001, 2002 and 2003 showed that the stilbenes diethylstilbestrol and dienostrol were routinely done in all the 15 MS of that time for bovine and porcine animals, and sheep and goats. Hexoestrol was not tested in some MS for those animals. For poultry, diethylstilbestrol was also tested in most of the 15 MS, and hexoestrol and dienostrol in a little lower number of countries. Testing for stilbenes in other species is less complete. The potent stilbene derivative benzoestrol was not tested for in any case. These conclusions are quite similar over the years.



Update on CODEX / EU business and SPECLOG explained (R.W. Stephany) An introduction is presented about the various recent and future achievements in the Codex Alimentarius Committee for Residues of Veterinary Drugs in Food (CCRVDF, hosting country USA) on one hand and in the EU network Commission-CRL-NRL on the other hand. The still inadequately resolved qualitative problem of the validation parameter specificity ( now in other EU legislation replaced by selectivity ) was addressed. The tool SPECLOG with a potential to solve some of the problems, especially in forensic cases, was explained. Finally some illustration were given with international arbitration case for DES residues. The full presentation is included in the RIVM/CRL workshop CD-ROM 2004. Criteria compared. WADA and EU criteria in practice. (R.W. Stephany) As the Independent Observer (IO) for the World Anti-Doping Agency (WADA) in the Doping Control Laboratory during the Olympic Summer Games in Athens, Greece, the lecturer had an unique opportunity to compare WADA and EU criteria for doping and residue test methods, respectively. Although there are largely overlapping areas and similarities in both field also explicit and important differences could be observed. The most striking differences can be found in the standard setting procedures and in the test result management structure in both fields of application. The full presentation is included in the RIVM/CRL workshop CD-ROM 2004.



Annex to Summary Report, Conclusions and Recommendations. Proposed Minimum list of substances and matrices for inclusion in Annual National Plans of EU Member States. A6 compounds. ( Values in bold already were agreed upon previously ) Bovine, Ovine and Porcine species Substance Marker residue or

metabolite Matrix MRPL

A1 Stilbenes Diethylstilboestrol (DES) Diethylstilboestrol Urine 1 µg/l Dienoestrol (DE) Dienoestrol Urine 1 µg/l Hexestrol (HEX) Hexestrol Urine 1 µg/l A2 Antithyroid agents Thiouracil (TU) Thiouracil Urine

Thyroid 100 µg/l

100 µg/kg Methylthiouracil (MTU) Methylthiouracil Urine

Thyroid 100 µg/l

100 µg/kg Propylthiouracil (PTU) Propylthiouracil Urine

Thyroid 100 µg/l

100 µg/kg Tapazole (TAP) Tapazole Urine

Thyroid 100 µg/l

100 µg/kg A3 Steroids Boldenone (BOL) 17β-Boldenone Urine 1 µg/l 17β-Nortestosterone (NT) 17α-Nortestosterone Urine 2 µg/l Ethynyl estradiol (EE2) Ethynyl estradiol Urine 2 µg/l 17β-Oestradiol (E2) 17β-Oestradiol Serum 0,04 µg/l Methyltestosterone (MT) Methyltestosterone Urine 2 µg/l 17β-Trenbolone (TB) 17α-Trenbolone Urine 2 µg/l Stanozolol (STAN) 16β-Hydroxystanozolol Urine 2 µg/l Dexamethasone (DEX) Dexamethasone Urine 2 µg/l Megestrol acetate Megestrol (acetate) Kidney fat 1 µg/kg Melengestrol acetate (MGA) Melengestrol (acetate) Kidney fat 1 µg/kg Chlormadinone acetate (CMA) Chlormadinone (acetate) Kidney fat 1 µg/kg Medroxyprogesterone acetate (MPA)

Medroxyprogesterone(acetate) Kidney fat 1 µg/kg



Bovine, Ovine and Porcine species Substance Marker residue or

metabolite Matrix MRPL

A4 Resorcylic acid lactones (RAL)

Zeranol (Z) Taleranol (see Note) Urine 2 µg/l Zearalenone (F2-toxin) Zearalenone Urine 2 µg/l A6 banned compounds Chlorpromazine Chlorpromazine Kidney 5 µg/kg Note: An additional minor metabolite of Zeranol is zearalanone, not to be confused with the F2-toxin Zearalenone the most abundant RAL in case of contaminated moulded feed Proposed MRPL-values for the matrix liver Substance Marker residue or

metabolite MRPL

A1 Stilbenes Diethylstilboestrol Diethylstilboestrol 2 µg/kg Dienoestrol Dienoestrol 2 µg/kg Hexestrol Hexestrol 2 µg/kg A3 Steroids

17β-Nortestosterone 17α-Nortestosterone 2 µg/kg Ethynyl estradiol Ethynyl estradiol 2 µg/kg Methyltestosterone Methyltestosterone 2 µg/kg 17β-Trenbolone 17α-Trenbolone 2 µg/kg Stanozolol 16β-Hydroxystanozolol 2 µg/kg Dexamethasone Dexamethasone 2 µg/kg A4 Resorcylic acid lactones (RAL)

Zeranol Taleranol (see Note) 2 µg/kg Note: An additional minor metabolite of Zeranol is zearalanone not to be confused with the F2-toxin Zearalenone the most abundant RAL in case of contaminated moulded feed. .



Proposed MRPLs for the matrix muscle tissue, inclusive fish Substance MRPL A1 Stilbenes Diethylstilboestrol 0,5 µg/kg Dienoestrol 0,5 µg/kg Hexestrol 0,5 µg/kg A3 Steroids Boldenone 0,5 µg/kg 17β-Nortestosterone 0,5 µg/kg Ethynyl oestradiol 0,5 µg/kg 17β-Oestradiol 0,05 µg/kg * Methyltestosterone 0,5 µg/kg 17β-Trenbolone 0,5 µg/kg Stanozolol 0,5 µg/kg Dexamethasone 0,5 µg/kg Megestrol 0,5 µg/kg Melengestrol 0,5 µg/kg Chlormadinone 0,5 µg/kg Medroxyprogesterone 0,5 µg/kg A4 Resorcylic acid

lactones (RAL)

Zeranol 0,5 µg/kg

* Value based on recent CRL information on the distribution of 17β-oestradiol in bovine muscle tissue

Documents

CRL workshop 2004 summary report-final mjAl. Paryzantow 57 PL-24-100 PULAWY Poland Tel: +48 818863051/173 Fax: +48 818862595 e-mail: [email protected] Th. Dabalis Ministry of