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1 ELISA (, ) :,, ELISA,AFB 1 - BSA, gΠml,1 : B + G0113 ngπml, 1150gΠkg ngπml, y = x ( R 2 = ) ; B + G 50 % 2108 ngπml ; % %, %85160 % B 1 B 2 G 1 G % 5715 % 104 %19 %, : ;,;, [3 ] [ 8,9 ] 9, : 13,13π ; 11,11π ; 9,9π 7 9,,C 30 C 18, C 30, C 30,,,, [1 ] Young A J, Britton G, eds. Carotenoids in photosynthesis [M]. London : Chapman and Hall, [2 ] Britton G, Liaaen2Jensen S, Pfander H, eds. Carotenoids (1A) Basel [ M ]. Boston and Berlin : Birkhguser, [3 ],,,. [M]. :, [4 ],,. [J ].,2002, (5) : [5 ] Riemersma R A, Wood D A, Macintyre C C, et al. Risk of angina pectoris and plasma concentrations of vitamins A, C, E and carotene[j ]. Lancer, 1991, (337) : 125. [6 ] Tsukida K, Saiki K, Takii, et al. Separation and determina2 tion of cisπtrans2 2carotenes by high2performance liquid chro2 matography[j ]. J Chromatogr,1982, (245) : [7 ] Lin C H, Chen B H. Determination of carotenoids in tomato juice by liquid chromatography [ J ]. J Chromatogr, 2003, (1012) : [8 ] Emenhiser C, Sander L C. Separation of geometrical carote2 noid isomers in biological extracts using a polymeric C 30 col2 umn in reversed2phase liquid chromatography [ J ]. J Agric Food Chem,1996, (44) : [9 ] Emenhiser C, Sander L C, Schwartz S J. Capability of a polymeric C 30 stationary phase to resolve cis2trans carotenoid isomers in reversed phase liquid chromatography[j ]. J Chro2 matogr A, 1995, (707) : [ : ] :R15 ;O :A : (2006) :(2001BA804A20) : :

2 Quantitative Analysis of Aflatoxins by EL ISA J IANG Tao, LIU Zhen, ZHENGJia, LI Min, J I Rong (National Institute for Nutrition and Food Safety, Chinese CDC, Beijing , China) Abstract : To detect the level of contamination of total aflatoxins in grains, a rapid, specific and quantitative ELISA2based method was developed using monoclonal antibodies against Aflatoxins. The coating antigen was AFB 1 2BSA and the concentration was gΠml. The working titre of antibody was 1 : For the standard Aflatoxin (B + G), the limit of detection was 0113 ngπml ; for samples, the limit of detection was 1150gΠkg ; the linear range was ngπml and the linear equation was y = x ( R 2 = ). The 50 % inhibition concentration for Aflatoxin (B + G) was 2108 ngπml. The recovery rates of spiked maize on the level of 2110gΠkg and 10140gΠkg were % and % respectively. For peanut recovery rates on the level of 2100gΠkg and 4100gΠkg were % and %, respectively. The cross reaction rates with aflatoxin B 1, B 2, G 1, G 2 were 100 %, 5715 %, 104 %, 19 % respectively, and there were no cross reactions with other my2 cotoxins. Key word : Aflatoxins ;Antibody Monoclonal ; Enzyme2Linked Immunosorbent Assay B 1 + B 2 + G 1 + G 2,, 5gΠkg [1 ],, ;;,,, ( ELISA),, ELSIA ELISA 2002, ELISA, B, CO 2 ( Queue ) () ( ) (SUNRISE) ( Mettler ) ( Hermle ) (BIO - RAD) AFB 1 - BSA (B + G) (Aflatoxin B + G) B 1 (Aflatoxin B 1 ) B 2 (Aflatoxin B 2 ) G 1 (Aflatoxin G 1 ) G 2 (Aflatoxin G 2 ) T - 2 ( T - 2 toxin) A(Ochratoxin A) ( Patulin) (Deoxynivalenol,DON) (Zearalenone,ZEN) ( Keyhole limpet he2 mocyanin,klh) (Bovine serum albu2 min, BSA) B 1 ( IgM IgG IgA IgD IgG 1 IgG 2a IgG 2b IgG 3 ), Sigma ; RPMI1640 HAT HEPES ( ) (DMF) 20 ( Tween 20) ( PEG, MW 5000),Gibco ; IgG, 30 % H 2 O 2 ELISA 0105 molπl, ph 916 ; PBS - T, 0105 % Tween molπl PBS ( ) ; 011 molπl 012 molπl,ph 510 ; 10 mg TMB 1 ml, 75 l,10 ml 10 l H 2 O 2 ; 2 molπl H 2 SO %( ) BSA 0105 molπl PBS Sp2Π0, 8 - BalBΠc BalBΠc,, ELISA 293

3 [2 ] ; SDS - PAGE [3 ] ; IgG BCA ; ELISA [2 ] :AFB 1 - BSA, ,Sp2Π0,(A - ) / ( A - ) 211 ; ELISA [4 ], B 1 B 2 G 1 G 2 (DON) A(OA) B 1 T - 2 BSA ELISA [5 ] (), 1 : , ELISA, ELISA, B 1 B 2 G 1 G 2 50 % AFB 1 - BSA, ngπml ( ) gΠml, 120 l,(b + G) ngπml, ( ), 3, 3, 1, B + G 0113 ngπml 30, ), 30 min 10 (gπkg) = CDXΠW : C (ngπml), ; D (ml) ; X ; W (g) () () ELISA HPLC ELISA AOAC HPLC, IgG 1, molπl 150 kd, IgG10 gπl B 1 B 2 G 1 G % 5715 % 104 %19 %, T - 2 A B % 1 (B + G) ( ELISA) 11312, 2, gΠkg, gΠkg 5 90 % m 5 g 100 ml, 015 g NaCl 25 ml ( B 1 ( ELISA) 3 B 2 ( ELISA)

4 4 G 1 ( ELISA) BSA ngπml,,50 %,,BSA 1 %, ELISA HPLC ELISA AOAC HPLC, 3t, P > 0105, 3 ELISA HPLC gπg ELISA G 2 ( ELISA) 212 ELISA 21211, 1 : ,AFB 1 - BSA gπml (B + G) gΠkg 2, 1 (B + G) gΠkg 2, 2 1 (B + G) % RSD :1 :2110 gπkg ; 2 : gπkg 2 (B + G) % RSD :1 :2100 gπkg ;2 : 4100 gπkg HPLC ELISA Biopharm 4 4 ELISA Biopharm ELISA ELISA Kit of Biopharm gπkg % % % AFB AFB AFG AFG , ELISA, Biopharm,, ELISA, ;Biopharm,, B 1 B 2 G 1 G 2, ELISA 0126gΠkg, 1150gΠkg, % %, %85160 % ELISA 295

5 Cry1Ie (11, ; 21, ) : Cry1Ie, Cry1Ie 72 ICR Cry1Ie (833gΠkg BW d d), ( ) ( ) Cry1Ie,, Cry1Ie : ; ; ; Effects of Cry1Ie Protein Expressed in E coli. on Biochemical Indicators and Flora of Intestinal Tract in Mice ZHANG Xiao2xia, YANG Bao2lan, YAN Wei2xing, LIU Yun2jun, WANG Gou2ying, XU Hai2bin (National Institute for Nutrition and Food Safety, Chinese CDC, Beijing , China) Abstract : Cry1Ie expressed protein is used as a model protein to examine its effects on biochemical indicators and flora of intesti2 nal tract in mice. 72 ICR mice were randomly divided into one experimental group and two control groups. The experimental group was treated with 833gΠkg BW d - 1 Cry1Ie expressed protein. The two control groups were treated with solution of bovine serum albumin and culture solution of E coli. with control plasmid without Cry1Ie, respectively. After 30 days, body weight, biochemi2 cal indicators (ASTALTTPALBBUNCRETRIGCHOL GLU) and numbers of flora of intestinal tract (enteric ba2 cilli, enterococci, Bacillus bifidus, lactobacilli) were examined. There were no significant differences between experimental group and control groups in body weight, biochemical indicators and numbers of flora of intestinal tract. In the present study, the Cry1Ie expressed protein had no toxicity to experimental animals. These results offer basic information for safety evaluation of new transgenic Cry1Ie expressed food, and establish foundation for food safety assessment of exogenous proteins derived from modern biological technique. Key word : Proteins ; Escherichia coil ; Blood Chemical Analysis ; mice, (B + G), ELISA, [1 ] GBΠT B 1 B 2 G 1 [ S]. G 2 [2 ],. [M]. 2. :, [3 ],,.J,E F, T,. [M]. 2. :, [4 ],,,. A [J ]., 2004,16 (1) [5 ],,,. A [J ].,2004,20 (5) : [ : ] :R15 ;R379 ;Q :A : (2006) : (J002C2003) : :

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