2004 62 8, 794 798 ACTA CHIMICA SINICA Vol 62, 2004 No 8, 794 798 Ξ Ξ ( 200032),, : :H 2 O 2 2CH 3 CN2 ( ) H 2 O 2 2CH 3 CN2N2 252 ( ),,, IgG Study of Ne w Chemiluminescence Technique Recognition of Sodium Azide by External Reference Method CAO, Zhi2Juan LIU, Cai2Yun LU, Jian2Zhong Ξ ( School of Pharmacy, Fudan University, Shanghai 200032) Abstract A limitation to the widespread application of chemiluminescence (CL) technique often lies in the poor selectivity of the CL reaction A novel concept was proposed to address this selectivity problem Two CL reactions, not just one reaction as generally used before, were used for the recognition of sodium azide from other similar radical scavengers These two reactions have the same oxidant and catalyst but different enhancer By coupling two CL reactions, i1 e H 2 O 2 2acetonitrile2tetracycline ( working reaction ) and H 2 O 2 2acetonitrile2melatonin ( reference reaction), sodium azide could be clearly identified and quantitatively determined in the presence of other similar compounds It is because that sodium azide only quenches the CL signal of the working reaction, whereas other similar compounds quench the CL signal of both working reaction and reference reaction As a preliminary application, this novel technique has been employed to determine sodium azide in the reagents of the commercial insulin kit Keywords chemiluminescence, sodium azide, recognition, coupling, working reaction, reference reaction,, Cu 2 +,Co 2 + Fe (CN) 3 6 -,, [1 8] -,, Cu 2 +, Co 2 + Fe (CN) 3 6 - Luminol2H 2 O 2 : Ξ E2mail :jzlu @shmu edu cn Received September 30, 2003 ; revised November 11, 2003 ; accepted December 20, 2003
No 8 : 795, CH 3 CN2 ( ) H 2 O 2 2CH 3 CN2N2 252 ( ),, : 1a, H 2 O 2 2CH 3 CN2,, H 2 O 2 2CH 3 CN2N2 252 [9 11] 015 20 g ml - 1,, ; 0105 g ml - 1, ( 1b) : : : H 2 O 2 2CH 3 CN2 ( ) H 2 O 2 2CH 3 CN2N2 252 ( ) IgG 1 1 1, (Sigma ) 10-2 mol/ L ; N2 252 (Nakarai) 10-3 mol/ L, ( RPN2567) Amersham Biosciences IgG, Sigma BPCL ( ) 1 2 10 L (011 mmol/ L) N2 252 (1 mmol/ L) 100 L NaHCO 3 2Na 2 CO 3 (011 mol/ L, ph 10) 20 L (011 mol/ L 015 mol/ L) 20 L 50 L, 10 s 1 2 1 1 Blank ; 2 5 10-4 mol/ L ; 3 011 mol/ L ; 1 011 mol/ L ; 2, 3 2 1 blank, 5 10-4 mol/ L (a) 5 10-6 mol/ L ; ( b) 5, [12] :,CH 3 CN 10-5 mol/ L N2 252 ; 0105 mol/ L ph = 10 NaHCO 3 2 Na H 2 O 2 2, :CH 3 CN 2 CO 3 ;25 % 10 mmol/ L H 2 O 2 Figure 1 Time course of the kinetics profile of the H 2 O 2 2 H 2 O 2 2N2 252 [12] chemiluminescence reaction containing ( a ) 5 10-6 mol/ L : H 2 O 2 CH 3 CN [12 14] tetracycline and ( b) 5 10-5 mol/ L melatonin in the different N2 252 concentrations of sodium azdie ; other components are 0105 mol/ L ph = 10 NaHCO 3 2Na 2 CO 3 buffer, 25 % acetonitrile and 10 mmol/ L H 2 O 2 2 H 2 O 2
796 Vol 62, 2004, [15], CL 0 / CL = 1 + K[ SA] (4) [15,16], 1,42 2[2,2,2 ]2 [17,18] 2, 1,42 2[ 2,2,2 ]2, : 100 % DMSO : ;,, 2 2 H 2 O 2 2CH 3 CN2 ( ) : CH 3 CN + 2H 2 O 2 CH 3 CONH 2 + 1 O 2 + H 2 O 1 O 2 + SA 1 O 2 2SA ( ) 1 O 2 +, 1 O 2, SA 1 O 2 2SA - 1 O 2 2SA : K = [ 1 O 2 2SA]/ [ 1 O 2 ] [ SA] (1), 2 : ( CL 0 - CL) / CL = {[ 1 O 2 ] 0 - [ 1 O 2 ]}/ [ 1 O 2 ] (2),[ 1 O 2 ] 0 [ 1 NaHCO 3 2Na 2 CO 3 25 % 10 mmol/ L H 2 O 2 O 2 ] Figure 2 Time course of the kinetics profile of the ; CL 0 CL chemiluminescence reaction containing ( a ) 5 10-6 mol/ L tetracycline and (b) 5 10, - 5 mol/ L melatonin in the different radical scavengers : 1 No scavenger ; 2 0105 mol/ L dimethylfuran ; 3 0105 mol/ L 1, 42 [ 1 O 2 ] 0 = [ 1 O 2 ] + [ 1 O 2 2SA] (3) (a) 5 10-6 mol/ L ; (b) 5 10-5 mol/ L N2 252 ;1 ;2 0105 mol/ L ;3 0105 mol/ L 1,42 2[ 2,2,2 ]2 ;4 10 % DMSO ; 0105 mol/ L ph = 10 diazabicyclo[2,2,2 ]octane ; 4 10 % DMSO ; other components are 0105 mol/ L ph = 10 NaHCO 3 2Na 2 CO 3 buffer, 25 % acetonitrile and 10 mmol/ L H 2 O 2 (1) (3) (2) : ( CL 0 - CL) / CL = [ 1 O 2 2SA]/ [ 1 O 2 ] = K[ SA] (4) ( CL 0 / CL ) 3 ( CL 0 / CL) ( r 2 = 019958)
No 8 : 797 CL 0 / CL = 678188[ SA] + 110878 ; CL 0 CL H 2 O 2 2CH 3 CN2 ( 6 min ) 0101 mol/ L 50 mol/ L, [ 011 % ( w/ w) ] ; 200 ng/ [011 % ( w/ w) ] 10 %, IgG, 25 g/ ml IgG 1 g/ ml 25 g/ ml 97 % 105 % 3 ) 3 ( CL 0 / CL) :5 10-6 mol/ L ;0105 mol/ L ph = 10 NaHCO 3 2 Na 2 CO 3 ;25 % 10 mmol/ L H 2 O 2 Figure 3 Chemiluminscence intensity versus [ sodium azide ], other components are 5 10-6 mol/ L tetracycline, 0105 mol/ L ph = 10 NaHCO 3 2Na 2 CO 3 buffer, 25 % acetonitrile and 10 mmol/ L H 2 O 2, H 2 O 2 2CH 3 CN2N2 252 ( ) 0 0101 mol/ L,6 min, References 0101 mol/ L, 2 3 1 Lu, J 2Z ; Zhang, Z2J Acta Chim Sinica 1996, 54, 71 (in : 5 10-4 mol/ L,1000 K +, Na +, Ca 2 +, Mg 2 +, Al 3 +, Zn 2 +, F -, Cl -, NO3 -,, : H 2 O 2 2CH 3 CN2 ( ) H 2 O 2 2CH 3 CN2N2 252 ( ( ), IgG, Chinese) (,,, 1996, 54, 71 ) SO 2 4 -, PO 3 4 - ; 10 Fe 3 + Mn 2 +, 100 g/ ml 2 Zhang, Z2L ; Yang, F2Z ; Zhang, X2R Prog Biochem,10 g/ ml Biophys 2000, 27, 440 (in Chinese) 011 mmol/ L EDTA (,,,, ; 0101 mmol/ L EDTA, 2000, 27, 440 ) 3 Zhuang, H2S ; Ye, H2Z ; Wang, J Chin J Anal Lab, Cu 2 + Cd 2 + 2002, 21, 13 (in Chinese) 50 g/ ml (,,,, 2002, 21, 13 ) 15 %, 25 g/ ml 4 Liu, Y2M ; Liu, E2B ; Cheng, J 2K Chin J Anal Chem 2002, 30, 1000 (in Chinese) (,,,, 2002, 30, 1000 ) 2 4 5 Han, H 2Y ; Cui, H ; Lin, X2Q Chin J Spec Lab 2002, 19, 39 (in Chinese) (,,,, 2002, 19, 39) H 2 O 2 2CH 3 CN2 ( ) H 2 O 2 2CH 3 CN2N2 6 Lu, J 2G ; Zhang, Z2J ; Zheng, H2Z Acta Chim Sinica 252 ( ), 2002, 60, 1274 (in Chinese) ; (,,,, 2002, 60, 1274 )
798 Vol 62, 2004 7 Rao, Z2M ; Shi, J 2Z ; Zhang, X2R Acta Chim Sinica 2002, 60, 1668 (in Chinese) (,,,, 2002, 60, 1668 ) 8 Lu, J 2Z ; Qin, W ; Zhang, Z2J ; Feng, M2L ; Wang, Y2 J Anal Chim Acta 1995, 304, 369 9 Kruszyna, R ; Smith, R P ; Kruszyna, H J Forensic Sci 1998, 43, 200 10 Fujita, H ; Kuroda, H Nippon Shokukin Kagaku Gakkaishi 1999, 6, 130 11 Oshima, H ; Ueno, E ; Saito, I ; Matsumoto, H J AOAC Int 2000, 83, 1410 12 Lu, J ; Lau, C ; Morizono, M ; Ohta, K ; Kai, M Anal Chem 2001, 73, 5979 13 Almeida, E A ; Miyamoto, S ; Martinez, G R ; Medeiros, M H G ; Mascio, P D Anal Chim Acta 2003, 482, 99 14 McKeown, E ; Waters, W A Nature 1964, 4949, 1063 15 Oh, S K ; Cha, S H Anal Biochem 1994, 218, 222 16 Emteborg, M ; Ponten, E ; Irgum, K Anal Chem 1997, 69, 2109 17 Yamamoto, K ; Inoue, S ; Yamazaki, A ; Yoshinaga, T ; Kawanishi, S Chem Res Toxicol 1989, 2, 234 18 Kreitner, M ; Alth, G ; Koren, H ; Loew, S ; Ebermann, R Anal Biochem 1993, 213, 63 (A0309301 SHEN, H ; DONG, H Z )