Microsoft Word 徐静-二校

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1 7 4 Vol. 7 No Journal of Food Safety and Quality Apr., : * 徐静, 卲筠乔, 赵禹, 张琳, 汪霖, 郑江, 曹际娟 (, ) 摘要 : 目的, 方法, 12,,,, ( ) 结果 a AS d AS M TP n IT, 结论,, 关键词 : ; ; Sampling for genetically modified organism content analysis in agricultural products-part 2: process from analytical sample to test portions XU Jing, SHAO Yun-Qiao, ZHAO Yu, ZHANG Lin, WANG Lin, ZHENG Jiang, CAO Ji-Juan * (Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian , China) ABSTRACT: Objective To study the effects of the process from analytical sample to test portions on the accuracy of results during sampling of GMO content analysis in agricultural products. Methods Taking non-gmo maize as matrix, 12 strains of GM maize were added to produce standard analytical samples. According to the systematic sampling method, GMO content in these samples were tested in random order and the relative standard deviation (RSD) was calculated as the measure to total RSD (single analysis). Results Factors affecting RSD included GMO content a AS, particle size d AS, test portion mass M TP and the number of increments composing the test portion n IT. The main ways to reduce total RSD were to reduce the particle size, increase the mass of test portion and the number of increments. Conclusion According to the experimental results, the recommended value including the particle size, mass of test portion and the number of duplicate analysis are given for the process from analytical sample to test portions. KEY WORDS: genetically modified organisms; sampling; agricultural products 基金项目 : (2014ZX ) Fund: Supported by the Genetically Modified Organisms Breeding Major Projects and Genetically Modified Organisms Sample Preparation and Accurate Detection Technology Projects(2014ZX ) * 通讯作者 :,,, cjj0909@163.com *Corresponding author: CAO Ji-Juan, Ph.D., Professor, Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian , China. cjj0909@163.com

2 4, : - 2 : 引言 30, (genetically modified organisms, GMOs) [1], ;,, ;,,,,, [2-7] ;, [8] [9-12] ISO [13,14] [15] GMO,,,, 3 : (process from lot to laboratory sample), (from laboratory sample to analytical sample), (from analytical sample to test portion) 3, 2 材料与方法 2.1 实验材料 T25 MON88017 MON87640 American Oil Chemists Society TC1507 NK603 MON863 MON810 GA21 BT176 BT11 ES3272 MIR604 Sigma 2.2 仪器设备 DNA, DNA extraction kit of genetically modified organism (GMO) detection Ver2.0 (No. D9093) TaKaRa PCR, ABI7500, Applied Biosystems 2.3 标准分析样品的制备, ,,, MON810 Bt11 Bt176 T25 GA21 MON863 NK603 TC1507 ES3272 MON88017 MIR604 MON ( ) : 10 1 ~10 6 ( 10%~0.0001%), 10 1 MON810 Bt11 Bt176 T25 4 V 1 MON810 Table 1 表 1 标准样品的制备 The preparation of standard samples, g GMO(MON810), g M1:, 60, g , % GMO(MON810), g M2:, 100, g , % GMO(MON810), g M3:, 200,g ,%

3 试料抽取方法及结果, 5 mg 5, 25 mg 1 10, 5 mg 10, 10 mg 5, 25 mg 2, 50 mg 1 10, 5 mg 20, 10 mg 10, 20 mg 5, 50 mg 2, 100 mg mg 20, 20 mg 10, 40 mg 5, 100 mg 2, 200 mg mg, 25 mg 20, 50 mg 40, 100 mg 50, 200 mg 50 DNA DNA,,, ( ), DNA, DNA PCR 1,, MON810 2~4 3 结果与讨论 3.1 影响 RSD 的参数 2 3 4, RSD a AS (analytical sample, AS) d AS M TP (test portion, TP) n IT ( increase composition of test portion, IT) n IT, 3 RSD a AS, d AS M TP. 3.2 分析样品粒度的优化 d AS ( ) RSD 2, 1, RSD (10 1 ~10 3 ), RSD, ; (10 4 ~10 6 ), Table 2 表 2 各种试料抽取方法的相对标准偏差 -M1 RSD results of different sample extraction methods-m1 M1: 60, 25 mg % 19.4% 20.9% 65.8% 416.8% 931.6% % 28.1% 29.5% 104.0% 422.7% 931.6% M1: 60, 50 mg % 14.8% 12.2% 51.1% 159.9% 785.8% % 18.6% 17.6% 62.7% 393.5% 931.6% % 21.5% 20.9% 76.6% 314.0% overflow % 19.7% 18.6% 77.4% 262.3% 852.7% M1: 60, 100 mg % 11.9% 9.6% 35.0% 149.4% 598.2% % 13.4% 15.6% 46.2% 244.2% 784.3% % 12.1% 14.4% 47.5% 164.8% 794.9% % 13.5% 13.2% 44.6% 194.6% 854.1% % 15.6% 15.1% 50.2% 317.7% 931.6% M1: 60, 200 mg % 8.2% 8.1% 31.1% 127.0% 574.4% % 9.1% 7.8% 26.7% 125.2% 637.0% % 10.2% 10.4% 33.8% 137.7% 575.6% 2 8.8% 9.8% 10.0% 36.4% 151.9% 779.3% % 10.0% 9.2% 37.0% 144.1% 794.9% : overflow-

4 4, : - 2 : 1507 Table 3 表 3 各种试料抽取方法的相对标准偏差 -M2 RSD results of different sample extraction methods-m2 M2: 100, 25 mg % 19.7% 20.7% 27.3% 129.2% 784.3% % 27.4% 27.4% 39.8% 185.0% 570.1% M2: 100, 50 mg % 14.6% 13.6% 21.0% 85.2% 413.7% % 17.9% 16.1% 24.2% 122.9% 476.5% % 19.9% 19.4% 25.0% 101.3% 726.5% % 19.1% 20.4% 30.0% 139.9% 716.0% M2: 100, 100 mg % 8.7% 11.1% 16.9% 56.5% 206.3% % 12.9% 12.7% 17.6% 78.4% 243.5% % 13.0% 13.0% 18.7% 75.5% 477.3% % 13.5% 13.8% 18.7% 90.7% 407.4% % 13.6% 13.9% 16.5% 87.2% 398.5% % 9.1% 9.6% 12.8% 49.8% 174.1% % 9.8% 9.4% 14.7% 59.2% 345.7% 5 9.5% 7.8% 10.7% 13.6% 59.9% 258.1% 2 9.5% 8.6% 11.3% 14.9% 69.3% 245.1% 1 9.3% 9.5% 10.1% 13.5% 80.2% 381.4% Table 2 表 4 各种试料抽取方法的相对标准偏差 -M3 RSD results of different sample extraction methods-m3 M3: 200, 25 mg % 18.5% 21.4% 21.6% 41.1% 204.6% % 27.8% 31.2% 29.8% 54.3% 444.8% M3: 200, 50 mg % 14.8% 15.1% 14.5% 30.8% 153.1% % 17.7% 17.2% 16.8% 29.7% 190.4% % 17.9% 17.9% 17.6% 42.3% 279.8% % 20.0% 16.4% 23.3% 35.1% 373.1%

5 续表 4 M3: 200, 100 mg % 9.1% 10.8% 10.0% 18.7% 120.5% % 13.3% 13.1% 13.3% 27.3% 119.1% % 13.1% 12.8% 15.0% 23.7% 177.8% % 12.1% 13.2% 13.9% 23.4% 132.0% % 17.5% 14.0% 13.3% 26.2% 112.6% M3: 200, 200 mg % 8.6% 9.4% 9.6% 16.7% 93.9% % 8.8% 9.2% 9.4% 18.1% 103.2% 5 9.4% 9.6% 8.7% 10.8% 16.3% 119.4% 2 8.9% 8.7% 10.3% 10.3% 18.6% 101.0% 1 9.3% 9.5% 10.3% 10.6% 18.8% 106.6% 1 RSD Fig. 1 Influence of partical size on RSD

6 4, : - 2 : 1509, 100,,, (10 1 ~10 3 ) RSD, 100 mg, (10 4 ), (10 5 ~10 6 ) RSD 4~ , 10 4, 3.3 最小试料量 (25~200 mg),, RSD 100 mg, RSD 100 mg 200 mg RSD mg 100 mg, RSD 2.3, 200 mg, 构成试料的份样数 100, n IT RSD 3, n IT RSD (10 5 ~10 6 ), n IT =5, 3.5 试料抽取方差的计算机模拟 M1 M2 M3, mg,,, 4 Fig. 2 2 RSD Influences of the test portion mass on RSD

7 Fig. 2 2 RSD Influences of the test portion mass on RSD 3 RSD Fig. 3 Influences of the number of increments on RSD Fig. 4 4 RSD RSD results in the process from homogeneous analytical sample to test portion

8 4, : - 2 : 1511 Fig. 4 4 RSD RSD results in the process from homogeneous analytical sample to test portion 4 结论和建议 RSD(TP), RSD(TP) RSD(TAE),, 100, mg, (>0.01%) RSD(TAE) 10% ; (<0.01%), 200, mg, RSD(TAE) 10% 100, RSD(TAE), 1, 5,,, 参考文献 [1] ISAAA Brief [EB/OL] [2],,. [J]., 2013, 12: Sheng Y, Xu WT, Luo YB. Commercialization of genetically modified organisms [J]. J Agric Biotech, 2013, 12: [3],,. [J]., 2009, 5: Liu PL, Li N, Zhou YL. Safety administration system for transgenic organism in America and its enlightenment for China [J]. J Agric Sci

1512 7 Technol, 2009, 5: 49 53. [4] James C. 2014 / [J]., 2015, 1: 1 14. James C. The 2014 global biotech/gm crops commercialization development situation [J]. J Chin Biotechnol. 2015, 1: 1 14. [5] Barbau-Piednoir E, Stragier P, Roosens N, et al.

Uninformed and disinformed society and the GMO market [J]. Trends Biotechnol, 2015, 1: 1 3. [7],,. [J]., 2013, 11: 1482 1485. Han YM, Zhai GQ, Xu JF.

9 Technol, 2009, 5: [4] James C / [J]., 2015, 1: James C. The 2014 global biotech/gm crops commercialization development situation [J]. J Chin Biotechnol. 2015, 1: [5] Barbau-Piednoir E, Stragier P, Roosens N, et al. Inter-laboratory testing of GMO detection by combinatory SYBR green PCR screening(co SYPS) [J]. Food Anal Methods, 2014, 8: [6] Twardowski T, Malyska A. Uninformed and disinformed society and the GMO market [J]. Trends Biotechnol, 2015, 1: 1 3. [7],,. [J]., 2013, 11: Han YM, Zhai GQ, Xu JF. The evolution of the eu regulation system of transgenic organisms and the enlightenment to our country [J]. J Zhejiang Agric Sci, 2013, 11: [8] Emslie KR, Whaites L, Griffiths KR, et al. Sampling plan and test protocol for the semiquantitative detection of genetically modified canola(brassica napus)seed in bulk canola seed [J]. J Agric Food Chem, 2007, 11: [9] Kay S, Paoletti C. Sampling Strategies for GMO detection and/or quantification, EC directorate general JRC [Z] [10] ISO 542:1990 Oilseeds-Sampling [S]. [11] ISO 13690:1999 Cereals, pulses and milled products-sampling of static batches [S]. [12] Kay S, Eede GVD. The limit of GMO detection [J]. Nat Biotechnol, 2001(19): [13] ISO 24333:2009 Cereals and cereal products-sampling [S]. [14] ISO DTR29263 Cereals and cereal products-sampling studies [S]. [15] GB/T [S]. GB/T Detection of genetically modified organisms and derived products-methods for sampling preparation [S]. 作者简介 ( 责任编辑 : 金延秋 ) 徐静, 博士, 高级工程师主要研究方向为食品安全色谱检测的研究 jingxu99@163.com 曹际娟, 研究员, 主要研究方向为食品安全检测与研究 cjj0909@163.com

Microsoft Word 曹际娟

Microsoft Word 曹际娟 7 4 Vol. 7 No. 4 016 4 Journal of Food Safety and Quality Apr., 016-1 : 曹际娟 *, 张琳, 赵禹, 徐静, 卲筠乔, 郑江 (, 116001) 摘要 : (genetically modified organism, GO), GO, GO ; GO, GO 4,,,,, 关键词 : ; ; Sampling for genetically