Development and Prospect of Monitoring Technology for Pesticides and Chemical Contaminants in Edible Agricultural Products

Transcription

1 International Conference on Engineering Science and Technology 214 June 3, 214 Beijing Development and Prospect of Monitoring Technology for Pesticides and Chemical Contaminants in Edible Agricultural Products 1.Origin of Pesticide Chemical Contaminants Monitoring( ) 2. Development of Pesticide Chemical Contaminants Monitoring Techniques ( ) 3. HRMS is the developing trend of pesticide residue analytical technique Guo-Fang Pang Chinese Inspection and Quarantine Academy

2 1.Origin of Pesticide Chemical Contaminants Monitoring( ) 1.1 GEMS/Food set up by WHO-FAO-UNEP In 1976, WHO, FAO and UNEP jointly set up Global Environment Monitoring System/Food, with a view to learning about the food contamination state of each member country as well as the intake of food contaminants for the purpose of protecting human health and promoting trade development. In 21, WHO and FAO stipulated explicitly in the Food Safety Strategy Draft promulgated that monitoring chemical harmful substances serves as a key means for risk assessment, and setting up food contaminant monitoring system is the core link of the food safety management. Dating from 1976, it is now nearly 4 years, and it is also 13 years since 21. How about the current monitoring of domestic and foreign pesticide and chemical contaminant residues? 2

3 1.2 Maximum Residue Limit (MRL) for the Current Pesticide Chemical Contaminants MRL for pesticide residues is both one of the food safety standards and threshold of access for agricultural products in the international trade. With the confirmation of global food safety strategic status, there are more and pesticide residual limit standards and stricter and stricter limits (839 pesticides ) (823 pesticides ) (5 pesticides ) (387 pesticides ) 365 EU USA Japan China 欧盟 美国 日本 中国 (213) (211) (213) (214) Pesticide MRLs for EU, USA, Japan and China

4 1.3 Development of Pesticide Chemical Contaminants Monitoring USA (1) In 1962, FDA: Silent Spring was published, which awoke peoples' environmental protection consciousness, and in the three years that ensued US FDA conducted a monitor of pesticide residues in over 54 cases of food samples; hence Pesticide Program Residue Monitoring (PPRM); (2) In 1967, USDA started to implement National Residue Program (NRP); (3) In 1991, USDA started to implement Pesticide Data Program (PDP) for the purpose of satisfying the formulation of pesticide residue limit standards and the great quantity of pesticide residue data needed for risk assessment. US three main residue programs took shape 4

5 1.3 Development of Pesticide Chemical Contaminants Monitoring USA Pesticide Residue Monitoring by American PPRM (FDA) in 监测农药个数 Number of the pesticide that can be detected (Detectable) 检出农药个数 Number of the pesticides that were actually detected (Found) Trend for the Pesticides Monitored and Detected by PPRM (FDA) in With PPRM, methods based on GC are mostly adopted, including GC-ECD,GC-FPD,GC- ELCD,GC-FID,GC-NP and GC-MS, and the rest are TLC,LC-UV,LC-FL and LC-ECD. 5

6 1.4 Development of Pesticide Chemical Contaminants Monitoring EU (1) In 1971,71/1118/EEC request each member country to conduct veterinary drugs and pesticide residue inspection. (2) In , MRLs were promulgated for pesticides in 76/895/EEC (fruits and vegetables), 86/362/EEC (grains and grain processed products), 86/363/EEC (animal products), 9/642/EEC/EEC, etc. (3) In 1996, the Commonwealth Nations initiated Commonwealth Pesticide Residue Monitoring Plan. (4) In 1996, 96/23/EC were promulgated for veterinary drugs monitoring. Residue Monitoring Systems for EU and Each EU Member Country 6

7 1.4 Development of Pesticide Chemical Contaminants Monitoring The number of pesticides monitored by 15 EU countries in EU The number of pesticides monitored for the monitoring plan by 15 EU countries in 21 Detection methods mainly adopted in 28 Detection methods mainly adopted in 21 GC(ECD,NPD,FPD,ITD,MS,TOFMS,MSMS) LC(DAD,FLD,UV,MSMS) GC(ECD,NPD,FPD,FID,ITD,MS,MSMS) LC(UV,MSMS) 7

8 1.5 Development of Pesticide Chemical Contaminants Monitoring China (1) In 1999, China s Ministry of Agriculture and AQSIQ jointly formulated Monitoring Plan of Residue Matters in Animals and Animal Food by P.R. China. (2) In 24, AQSIQ initiated monitoring plan of residual matters in export plant food. (3)In 28, AQSIQ changed residual matter monitoring to harmful matters monitoring, initiating a monitoring plan on a greater scale (AQSIQ FOOD [28] No.581]. (4)In 29, Food Safety Law was promulgated in our country. Chemical contaminants determination and limit formulations were written into national standards for food safety. (5)In 21, the Ministry of Health and other 5 government departments together enacted Food safety risk monitoring and control,and put forward the national food safety risk monitoring plans. 8

9 1.5 Development of Pesticide Chemical Contaminants Monitoring China China pesticide residue monitoring Pesticide residues: test results from pesticide residues in 28: test data obtained from 4739 actual samples, with positive ratio of 1.1% and non- conformity ratio of 6.76% by Japanese standard, 1.13% by EU standard. Participating laboratories: totaling 35 (to the only exclusion of Tibetan CIQ not being listed in the Monitoring Plan) The number of target matter varieties monitored increased from 52 in 24 to 11 in 28. The main methods adopted are: 1GC(ECD,NPD,FPD,ITD,MS,MSMS); 2LC(DAD,FLD,UV,MSMS) 9

10 Paper number 2. Development of Pesticide Chemical Contaminants Monitoring Techniques ( ) 2.1 Research Scope:115 SCI journals(usa 3,England 6,Germany 3,Netherlands 3,a total of 15); 2419 papers on pesticide residue analytical techniques for edible agricultural over the past 24 years( ) A total of 419 papers for statistical analysis came from 15 journals, among which 862 from J. Chromatogr. A, 686 J. AOAC Int., 58 J. Agric Food Chem and 436 Anal. Chim. Acta totaling 2564, accounting for 62% Witnessing the Progress of World Pesticide Residue Analytical Techniques by Tracing the SCI Papers over the Past 24 Years SCI USA UK Germany UK UK Germany UK USA UK Germany UK Netherlands Netherlands USA Netherlands Retrieval unit: Information Research Center of Chinese Academy of Agricultural Science. 1

11 technical papers come from 7 countries and regions across the five continents. There are 214 papers from 32 countries in Europe, 898 from 13 countries in America, 893 from 17 countries and regions in Asia, 39 from 2 countries in Oceania, and 34 from 6 countries in Africa, which represents the academic research on the detection of pesticide chemical contaminants in foods. No. Countries and Regions Paper number 214 papers from 32 countries in Europe 1 SPAIN ITALY UK 23 4 GERMANY FRANCE BELGIUM NETHERLANDS 14 8 GREECE 98 9 SWITZERLAND 56 1 IRELAND CZECH REPUBLIC POLAND PORTUGAL SWEDEN FINLAND DENMARK AUSTRIA SLOVAKIA NORWAY 1 2 SLOVENIA 9 21 BULGARIA 6 22 CROATIA 5 23 HUNGARY 5 24 ESTONIA 4 25 RUSSIA 3 No. Countries and Regions Paper number 26 SCOTLAND 3 27 CZECHOSLOVAKIA 3 28 YUGOSLAVIA 2 29 SERBIA 2 3 LITHUANIA 1 31 ROMANIA 1 32 UKRAINE papers from 13 countries in America 33 USA CANADA BRAZIL ARGENTINA MEXICO CHILE 4 39 CUBA 2 4 COLOMBIA 1 41 NICARAGUA 1 42 TRINIDAD AND TOBAGO 1 43 VENEZUELA 1 44 JAMAICA 1 45 MARTINQIQUE papers from 17 countries and regions in Asia 46 CHINA JAPAN INDIA KOREA 37 No. Countries and Regions Paper number 5 TAIWAN IRAN THAILAND TURKEY KUWAIT 7 55 MALAYSIA 6 56 HONGKONG 4 57 JORDAN 4 58 PAKISTAN 3 59 SINGAPORE 3 6 LEBANON 2 61 SAUDIARABIA 2 62 ISRAEL 2 39 papers from 2 countries in Oceania 63 AUSTRALIA NEWZEALAND 4 34 papers from 6 countries in Africa 65 EGYPT 2 66 SOUTHAFRICA 7 67 BOTSWANA 3 68 TUNISIA 2 69 ALGERIA 1 7 TANZANIA 1 The ranking for countries with papers exceeding 1 in sequence is: Spain, USA, China, Italy, UK, Canada, Japan, Germany, France, Belgium and Holland. 11

12 2.3 Pesticide residue analytical technical papers are increasing year by year. 35 Distribution of 419 papers between : 1559 papers in the first 12 years, and 255 in the last 12 years, increasing by 64% and manifesting the common concern on pesticide residues and food safety by countries all over the world. 3 y = 12.79x

13 paper number 2. 4 Development of sample cleanup techniques: 419 papers involve 144 sample cleanup techniques Sample cleanup techniques that rank the first The firs 2 techniques take the first place in sample cleanup techniques, which boosted the development of cleanup material production all over the world. In these 144 cleanup techniques: 1 Sample cleanup techniques that rank the first 2 account for 9.7%, 2 Sample cleanup techniques that rank the first 1 account for 8.1%, 3 Sample cleanup techniques than rank the first 5 account for 63.5%, 4 SPE cleanup technique that ranks the 1 st accounts for 41.1% Sample cleanup techniques that rank the first 2 1 SPE 2 LLE 3 MSPD 4 QuEChERS 5 SFE 6 MIP 7 GPC 8 Derivatisation 9 MIP-SPE 1 SPME 11 ASE 12 DSPE 13 IAC 14 MAE 15 LC 16 SE 17 DLLME 18 SBSE 19 Solution extraction 2 SDME

14 Simplicity, rapidness and high efficiency are the general trends of development Comparison of sample cleanup techniques in the first ten years (199-21) and the last ten years (22-213) Cleanup techniques that rank the first 3 in the first 12 years were SPE, LLE and SFE. No Cleanup techniques SPE LLE SFE GPC Derivati sation MSPD LC IAC TLC SE Paper number No Cleanup techniques SPME MAE MCAC on-line microextractio n ASE dialysis DSPE affinity adsorption chromatograp chromatograp hy hic Paper number commercial kit SPME Rank No.11 Rank No Cleanup techniques thank ranked the first 3 in the last 12 years were SPE, QuEChERS and LLE. No Cleanup techniques SPE QuEChERS LLE MIP MSPD MIP-SPE GPC SPME ASE DSPE Paper number No Cleanup techniques SFE Derivati sation MAE DLLME SBSE IAC SDME Solution Exration Paper number UAE SE ASE Rank No.9 Rank No a)techniques that have always been in leading positions in the 24 years are: SPE and LLE, which rank 1 st and 2 nd. b)techniques that rose the fastest in rankings are: SPME (from Rank No.11 in the first 12 years to No.8 in the last 12 years) and ASE (from Rank No.15 in the first 12 years to No.9 in the last 12 years). c)techniques that have retained relatively stable positions: LLE, MSPD, and GPC. d)newly emerged techniques: QuEChERS, which ranked No.2 since it appeared in the last 12 years. In addition, MIP, MIP-SPE, SBSE and SDME ranked 4, 6, 15 and 18 respectively. 14

15 paper number 2.5 Analytical technical development: 176 analytical techniques are involved in these 419 papers Sample analytical techniques that rank the first In the 176 analytical techniques 1papers and times with analytical techniques than rank the first 2 account for 9.4% of total papers and times. 2The papers and times with analytical techniques that rank the first 1 account for 79.9% of total papers and times. 3The papers and times with analytical techniques that rank the first 5 account for 57.4% of total papers and times Sample analytical techniques that rank the first 2 1 LC-MS/MS 2 GC-MS 4 LC-UV 3 GC-ECD 5 LC-MS 6 LC-FLD 7 LC-DAD 8 ELISA 9 GC-MS-MS 1 GC-NPD 11 sensor 12 GC-FPD 13 LC-QTOF-MS 14 IA 15 GC-FID 16 TLC 17 CE 18 EIA 19 GC-TOFMS 2 CE-UV 21 LC-ED

16 Accuracy, rapidness and high throughput are the general trends of development Comparison of development of analytical techniques in the first 12 years (199-21) and the last 12 years (22-213) Sample analytical techniques that rank the first 3 in the first 12 years respectively are LC-UV, GC-MS and GC-ECD. No Analytical techniques LC-UV GC-MS GC-ECD LC-FLD LC-MS GC-NPD ELISA LC-DAD LC-MS/MS GC-FPD Paper number No Analytical techniques TLC GC-FID EIA IA LC-ED GC-MS-MS sensor CE CE-UV NMR Paper number LC-MS/MS Rank No.9 46 Rank No Sample analytical techniques that rank the first 3 in the last 12 years respectively are LC-MSMS, GC-MS and LC-UV No Analytical techniques LC-MS/MS GC-MS LC-UV LC-MS GC-ECD LC-DAD LC-FLD ELISA GC-MS-MS sensor Paper number No Analytical techniques LC-QTOF-MS GC-NPD IA GC-FPD CE GC-TOF/MS GC-FID CL Microbiologi cal Paper number LC-Orbitrap Rank No.1 LC-UV 264 Rank No The analytical technique that developed fastest is LC-MS/MS, which rose from Rank No.9 (46 papers) in the first 12 years to No.1 (577 papers) in the last 12 years, with its application papers and times increasing 12.5 folds; other analytical techniques that developed relatively fast include GC-MS/MS, which rose from No.16 (11 papers) to No.9 (111 papers) in the last 12 years, with its application papers and times increasing 1.1 folds. 2 LC-UV technique that ranked No.1 for the first 12 years (262) dropped to 3 rd place in the last 12 years (22). 3 For two low-resolution GC-MS and LC-MS resolution techniques, both of them were developing relatively stably in the first and last 12 years, with its growth rate being consistent with that of total papers and times. 4 The 1 analytical techniques that belonged to the top 2 in the first 12 years dropped in the last 12 years, especially such nonmass spectrometric determination techniques that are low in sensitivity and narrow in application scope as TLC, EIA were gradually getting obsolete. 16

17 2.6 Evolution of chromatographic and mass spectrometric techniques, and mass spectrometric technique surpasses the traditional chromatography. In these 419 papers, there are 179 papers that involve mass spectrometer technique, making up 43.6%, 171 papers chromatographic technique (equipped with selective detectors), accounting for 41.4%. Obviously, both these two techniques are the most widely used ones in residue analysis. In the past 24 years, these two residue analytical techniques respectively dominated the relative areas for 12 years 质谱技术 chromatographic techniques mass spectrometric techniques 色谱技术 In , there are more papers on chromatographic techniques than those on mass spectrometry, and the ratios of papers and times are 929:418. The peak for chromatographic papers appeared between , with 113 papers every year on the average, while there are fewer than 42 papers on mass spectrometer during this period. In , mass spectrometer application left chromatographic techniques far behind, while the ratios of papers and times for mass spectrometer and chromatography are 1372:772, and the peak for mass spectrometer application occurred between 29 and 211, with 142 papers and times over these two years while those for chromatography are only half of them. 17

18 paper number paper number paper number paper number 2.7 MS analytical has gained unprecedented development, and GC-LC analytical technique is developing sluggishly LC-MS LC-MS/MS TOF Increase by 42.4% LC-UV FLD DAD 486 Reduction of 6% GC-MS GC-MS/MS TOF Increase by 98.8% GC-ECD NPD FPD 38 Reduction of 33.1% On the whole, the emerging techniques are gradually taking place of the conventional ones. It is crucial for one to take the lead and seize hold of innovation. 18

19 paper number 2.8 Mass spectrometer technical papers witness an unprecedented high production There are 418 technical papers of such mass spectrometric analysis published in the first 12 years, while there are 1372 papers in the last 12, with the papers in the last 12 years being 3 times more than those in the first 12 years Increase by 228% y = x R² =

20 2.9 low-resolution mass spectrometry is thriving, and the high-resolution mass spectrometry is just on the horizon. 1 8 LC-MS/MS GC-MS HRMS 7 The technical breakthrough with ESI and APCI led the innovative development of LC-MS/MS LC-MS/MS technique was only used in 46 papers and times in the first 12 years, while 577 papers and times in the last 12 years, which was nearly 12.5 times those of the first 12 years, which could be attributed to the technical progress of ionization with ESI and APCI, making LC-MS/MS technique thrive and take a leading role. 2Since 1992, GC-MS technique has maintained a stable development for 22 years continuously. 3High-resolution mass spectrometry (HRMS) such as TOF/MS and LC-Orbitrap, etc. was not yet used in the residue analysis in the first 12 years, while in the last 12 years its applications increased markedly. HRMS is made to be the direction of future development for residue analysis on account of its unique precise mass identification capability. 2

21 3. HRMS is the developing trend of pesticide residue analytical technique 3.1 Research on HRMS for pesticide residues Our team is based on serving the foreign trade-serving the entire country-integrating with the international and has experience three stages of development from GC-LC-Low-resolution MS. Three AOAC International Collaborative Studies were organized, which was attended by 63 laboratories from 17 countries and regions across the globe, and three AOAC International technical standards were developed. 1 2 Analytical techniques for pesticides chemical contaminants have been keeping pace with the times for 4 years 4 5 In the nineties:lc/ms/ms,lc/ms emergence,gc/ms became widely used 3 In the eighties :GC/MS emergence,gc/ecd,gc/fpd,gc/npd became widely used In the seventies:lc/uv emergence,gc/ecd,gc/fpd became * widely used In the sixties:gc/ecd,uv In the first of 21 st century :GC-Q-TOFMS,LC-Q-TOFMS * * * (* ) Presently our team is meeting the new challenges of high-resolution MS that can be included in the fourth stage and strives for newer innovations, making our research level climb one storey higher. 21

22 3.2 Fundamental study on building the databases for GC-Q-TOFMS precise mass spectrometer Development of GC-Q-TOFMS precise MS databases and detection methods TOF/MS Precise mass database development Flow chart Q-TOF/MS Precise mass database development Flow chart Pesticide chemical contaminants residues monitoring technique flow chart Pesticide standards injection Sample GC-TOF/MS chromatography-mass spectra acquisition select the best fragment ion as precursor ion Pretreatment Pesticide mass spectrogram import to qualitative software Input elements information of pesticides and produce formula calculation the theory molecular weight of the fragment ions by the formula acquisition the Q-TOF/MS spectrum under 4 different CE select the spectrum at the optimal CE detect by GC-Q-TOF/MS A Search by the TOF/MS database B detect by QTOF/MS select 3-4 ions to detect Import to the Library Editor A retention time fragment ions with accurate mass acquisition Q- TOF/MS fragment ion spectrum Establish database document Input pesticide name, retention time fragment ions formula Establish Q-TOF/MS spectrum library Two or more characteristic ions satisfy the search condition, and database retrieval match score> 6 Target pesticides identify spectral difference analysis by Q-TOF/MS spectrum library Confirmatory target pesticides TOF/MS accurate mass database 22

23 3.3 Fundamental study on building the databases for LC-Q-TOFMS precise mass spectrometer Development of LC-Q-TOFMS mass spectrometer databases and detection methods 1LC-TOF/MS first-stage mass spectrometer database development flow chart for 51 pesticides Pesticide standard Detect by LC-Q-TOFMS (MS mode) Find by formula LC-Q-TOF/MS monitoring and confirmation technical method flow chart for 51 pesticide chemical contaminants sample The accurate mass The distribution of isotopic clusters The proportion of isotopic clusters Retention time Ionized forms of percursor ion (+H,+NH4,+Na) Sample preparation (extraction, cleanup, concentration) Set the weight of score for each element The score obtained by auto-match Database of TOF/MS 2LC-Q-TOF/MS Second-stage mass spectrometer database development flow chart for 51 pesticides Pesticide standard B Detect by TOF/MS (MS) A A Test sample B Detect by Q-TOF/MS (target MS/MS) B Input the precursor ion, retention time and collision energy to establish the acquisition method of Q-TOF-MS Obtain MS spectra of the compound Obtain product ions spectra of the compound Product ion spectra detected under 8 different collision energys in Q-TOF/MS mode A B Choose 4 product ion spectra based on the number of product ions Search by database of TOF/MS Search by library of Q-TOF/MS Import into the PCDL software Llibrary of Q-TOF/MS A Suspected pesticides B Identified pesticides 23

24 3.4 A tentative probe into the new high throughput analytical technique of pesticide residues using GC-Q-TOFMS and LC-Q-TOFMS for simultaneous detection of 1138 pesticides in agricultural products with one single sample preparation One single sample preparation GC-Q-TOFMS for detection of 722 compounds 271 compounds LC-Q-TOFMS for detection of 687 compounds A total of 1138 pesticides can be detected by these two techniques 1 Precise mass number has replaced the conventional method of using pesticide reference materials for comparison, which is the first breakthrough for realizing the high throughput analytical technique innovation. 2The software program developed on our own for one single sample preparation and simultaneous detection of 1138 pesticide residues using these two techniques is the second breakthrough in the technical innovation. 3Analytical technique, net technique and geographical information are merged together to constitute a strong monitoring database for pesticide residues, which may provide us with a three-dimensional digitally written report with names of agricultural products, target pesticides and places of production as well as a threedimensional visual display of the afore-mentioned information. 24

25 3.5 Comparison of low resolution mass spectrometry and high resolution mass spectrometry monitoring technical features for over 1 pesticide residues No Items High resolution (GC-Q-TOFMS/LC-Q-TOFMS) 1 Resolution(m/z).1m/z.1 m/z Low resolution (GC-MS/LC-MS/MS) 2 Pesticide data banks More than 12 pesticides about 5 pesticides 3 4 Reference materials for comparison Scanning characteristi cs Not need Unlimited targets: target, nontarget were scanned simultaneous, and with high speed 5 MS acquisition full-spectrum scanning 6 Confirmation points 7 Data processing 8 Software Above 6, with isotope percentage and isotope abundance Simple: using data bank to make a direct determination, which be processed automatically Already equipped with the software matching automatic operation need limited targets: about 1 pesticides one time, with above 6 times needed for scanning 5 pesticides, and with slow speed needing to edit acquisition method 4 Complicated: needing to edit the treatment method per each acquisition Lacking the software matching automatic operation 25

26 3.6 Establishment of big databases and general survey for pesticide Pesticide monitoring has been completed for 6978 lots of fruits and vegetables samples that folk feed on out of 91 kinds of fruits and vegetables from 18 provinces and cities. 新疆 239 山西省 318 内蒙古 26 甘肃省 239 河北省 391 贵州省 299 北京市 415 山东省 592 江西省 298 天津市 533 辽宁省 447 河南省 433 江苏省 485 安徽省 339 上海市 521 浙江省 442 黑龙江省 289 吉林省 197 Provinces and cities Samples Sampling points 1 Beijing Tianjin Shanghai Heilongjiang Haerbin Jilin Changchun Liaoning shenyang Neimentgu Huhehaote Hebei Shijiazhuang ShanXI Taiyauan Henan Zhengzhou Shandong Jinan Xinjiang Wulunmuqi Guizhou guiyang Jiangxi nanchang Anhun Hefei Jiangsu Nanjing zhejiang hangzhou Shangdong 9 cities Total From the statistical analysis of big data, many new discover can be derived, numerous forecasts can be obtained, from which food safety monitor and control is moved ahead. 26

27 3.7 Big databases may play a role in the following 8 aspects of two categories. The monitoring of over 5 lots of samples of 91 kinds of fruits and vegetables collected from the markets of 1-plus provincial capitals in China has demonstrated the technique s robust detection capability, which is an effective tool for the risk control of pesticide residues and extremely promising for future application and development. (1)Law Enforcement basis 1problem tracing 2problem liability 3product recall 4issuing warning (2)Scientific Research basis 5study on exposure assessment 6development of MRL standards 7providing scientific data for laws and regulations making and amending 8Conducting research a deeper level and international exchange Management comes only after measurement; improvement comes only after management; boosting food safety to a higher level comes only after improvement. 27

28 paper number paper number 4 Conclusion: The monitoring techniques for veterinary drug and pesticide residues over the past 24 years have gained very rapid development; LC-MS monitoring techniques for veterinary drug and pesticide residues in the future 2 years are on the rise increase of 64 % The last 12 year witnessed an increase of 64 % over the first 12 years Increased by 2.3 times 339 Increased by 2.3 times In , the increase ratio at 64% for papers over the past 24 years demonstrated a strong growth for the future. The era when mass spectrometry technique is gaining momentum has just kicked off. Owing to food safety being listed in the state strategic place by countries all over the world, this will enable analytical techniques for veterinary drugs and pesticide residues in foods to still be the hot spot of research for global food safety in the 2 years to come. 28

29 Thanks Chinese Inspection and Quarantine Academy 29