8Q5SAC) 8Q5SAC UV2Vis 8500 ( ) ; PHS23C ) ;721 ( ) :1 4. ;8Q5SAC : molπl ;Britton2Robinson Q5SAC BSA Britton2Robinson,

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1 (FENXI HUAXUE) 8 Chinese Journal of Analytical Chemistry ( )21,82 23,62 3 (, ) 22( )21,82 23,62 (BSA), 8Q5SAC BSA 298K, LΠmol 8Q5SAC, ph = 3. 34, mgπl,,22( )21,82 23,62,,, 1, ( )21,82 23,62 ( 4 8Q5SAC), 8Q5SAC UV2Vis 8500 ( ) ;721 ( ) ; PHS23C ( ) ( BSA, 67000, ) 1 4 ;8Q5SAC molπl ;Britton2Robinson ml 8Q5SAC BSA Britton2Robinson,, 15 min,, 670 nm 572 nm Q5SAC BSA 1 ph 3. 34,8Q5SAC 572 nm BSA,, 670 nm Q5SAC BSA, 15 min 3 h 3. 3, ph = ,, ml, 2. 0 ml Britton2 Robinson ; (QX02030)

2 8 22( )21,82 23, Q5SAC BSA Fig. 1 Absorption spectra of 22( 82hydroxyquinoline252 sulfoacid272azo) 21, 82dihydroxy23, 62naphthalene ( 8Q5SAC) 2 bovine serum albumin (BSA) complex ph 3. 34, C 8Q = molπl C BSA ( 10-7 molπl) a. 0 ; b ; c ; d ; e ph 8Q5SAC BSA Fig. 2 Effect of ph on the interaction of 8Q5SAC2BSA complex C 8Q = molπl, C BSA ( molπl) 3. 4, 8Q5SAC BSA Britton2Robinson Scatchard 5, 6 Scatchard,, nπc F = KN - Kn, n, C F,, n, nπc F, n nπc F,, n nπc F, (specific binding) (non2specific binding) Scatchard, ( ) ;,, ( ) 8Q5SAC BSA (1) 8Q5SAC ( F ) BSA , 8Q5SAC BSA,pH Britton2Robinson, 8Q5SAC,, 670 nm 572 nm, 8Q5SAC, 670 nm A = C (molπl), r = , F = 2850 (L mol - 1 cm - 1 ) ; 572 nm A = C (molπl), r = , F = (L mol - 1 cm - 1 ) 8Q5SAC BSA,pH Q5SAC, 572 nm,, A = C (molπl) 8Q5SAC (2) 5 BSA (ph 3. 34),, 3. 0 ml molπl BSA 15 cm, 8Q5SAC Britton2Robinson, 20 ml, ( ) 48 h, 572 nm, 8Q5SAC C F 8Q5SAC C B BSA n,, 3 3 1, 8Q5SAC,8Q5SAC BSA Scatchard, 35 ; 8Q5SAC, nπc F, n

3 nπc F, Plasvento, K d = nπc F, 8Q5SAC BSA BSA,pH 5. 60, 3 2 nπc F, ph,8q5sac BSA, 8Q5SAC, 3 ph 4 ph Fig. 3 The binding plots of 8Q5SAC2BSA system by using equilibrium dialysis method at different ph 1. ph ; 2. ph (detection wavelength) = 572 nm Fig. 4 The binding plots of 8Q5SAC2BSA system by using spectrophotometric method at different ph 1. ph ; 2. ph (detection wavelength) = 670 nm (1) 298K, 8Q5SAC, BSA, nm, A = C L - k - 1 AΠC P (1) n = KC L - kn Cp (2) A ; C L C P ;= B - F, B F, K, n, (2) KC L n, N (1) A = AΠC P (molπl), r = y = 5070 (L mol - 1 cm - 1 ) (2) BSA (ph 3. 34), BSA 10, 8Q5SAC,, 670 nm, 8Q5SAC C B = ( B C T - A)Π, C T = C B + C F, n nπ C F, 4 4 1, 8Q5SAC,8Q5SAC BSA Scatchard, 37 ; 8Q5SAC, nπc F, nπc F n, Plasvento, BSA (ph 5. 60) 8Q5SAC BSA, 4 (2) nπc F, ph,8q5sac, Q5SAC BSA 8, 8Q5SAC, BSA, 670 nm A, A AΠC P, 5A = C L - k - 1 AΠC P,298. 0K, LΠmol ;311. 5K, LΠmol,, H, ln k 2 Πk 1 = HΠR (1ΠT 1-1ΠT 2 ), G = H - T S, H S 298K H - 756J, S 32977J BSA

4 8 22( )21,82 23, H < 0, S > 0, 8Q5SAC BSA 5 A AΠC p 6 NaCl A AΠC P Fig. 5 Determination of the binding constant at different temperature by A AΠC p correlogram Fig. 6 The correlogram of A AΠC P adding the NaCl as adding or not K; K ph 3. 34, C 8Q = molπl 8Q5SAC BSA 1. NaCl (without NaCl) ; 2. NaCl (the mass percent of NaCl) 0. 5 % ph 3. 34, C L = molπl, NaCl NaCl 0. 5 % A = C L - k - 1 AΠC P NaCl 0. 5 %, 6 (2), A = AΠC P (molπl) r = k = LΠmol, N = kc L = 20 A = C L - k - 1 AΠC P NaCl, 6 (1) A = AΠC P (molπl) r = k = (LΠmol), N = kc L = 37,NaCl 0. 5 % N k, NaCl 8Q5SAC BSA Cl -, Q5SAC molπl,bsa 20 mgπl, 5 %, 1 1 Table 1 Tolerance of foreign substances Foreign substances (mgπl) Tolerable concentration Foreign substances (mgπl) Tolerable concentration Lysozyme 3. 3 L2 L2Phenylalanine L2 L2Tyrosine L2 L2Tryptophan Fe 3 +, ( SO ) 3. 7 Ca 2 +, (Cl - ) Cu 2 +, (SO ) Zn 2 +, (Cl - ) Mg 2 +, (Cl - ) , 572 nm,,8q5sac BSA mgπl, A = C(molΠL), r = , 3 S b ΠS, mgπl 2

5 Table 2 Analytical results of synthetic samples Composition of samples (mgπl) Found (mgπl, n = 5) Recovery ( %, n = 5) BSA 32. 0, Ca , Mg L2 L2Tryptophan L2 L2Tyrosine RSD ( %, n = 5) References 1 Congdon R W, Muth G W. Anal. Biochem., 1993, You W W, Hangland R P. Anal. Biochem., 1997, Chi Yanhua( ), Zhuang Jia( ), Dong Faqin( ), Gen Wei ( ), Li Kean( ), Tong Shenyang ( ). Acta Chimica Sinica ( ), 2000, 58 (1) Yu Ying ( ), Huang Jianfeng ( ), Hong Chaohui ( ). Physical testing & Chemical Analysis, Part B Chemical Analysis ( 2 ), 1996, 32 (1) Scatchard G, Scheiberg I I, Armstrong S H. J. Am. Chem. Soc., 1950, Taira Z, Terada H. Biochem. Pharmacol., 1985, Plsavento M, Profumo A. Talanta, 1991, 38 (10) Shang Zhenhua( ), Zhou Dongmei( ), Guo Wei ( ), Yu Yinian( ), Zhou Liangmo ( ), Pan Mingchen( ), Zhou Kang( ). Journal of Instrumental Analysis ( ), 1998, 17 (2) Wei Yongju( ), Tong Shenyang( ), Li Kean( ). Acta Chimica Sinica ( ), 1995, 53 (1) Li Na ( ), Li Kean( ), Tong Shenyang( ). Chinese J. of Anal. Chem. ( ), 1996, 24 (1) Zhu Keng( ), Tong Shenyang( ). Acta Chimica Sinica ( ), 1997, Wei Yongju( ), Li Kean( ), Tong Shenyang( ). Chemical Journal of Chinese Universities ( ), 1999, 17 (4) Interaction Studies of 22( 82Hydroxyquinoline252sulfoacid272azo) 21,82dihydroxy 23,62naphthalene disulfoacid with Bovine Serum Albumin Zhou Qiuyun, Yu Ying 3 ( Department of Chemistry, South China Normal University, Guangzhou ) Abstract The binding of 22(82hydroxyquinoline252sulfoacid272azo)21, 82dihydroxy23, 62naphthalene disulfoacid (8Q5SAC) to bovine serum albumin (BSA) in acid solution was studied by using spectrophotometric method and equilibrium dialysis method, It is considered that the noncovalent binding forces(mainly the electrostatic force) are the binding force between 8Q5SAC and BSA. The reaction mode was also discussed, the maximum binding number was determined to be ; The binding constant was LΠmol at 298k. The absorption character of 8Q5SAC2BSA complex at various condition (such as acidity,ionic strength etc. ) was also studied. At ph 3. 34, the linear range of standard curve was mgπl. Keywords Equilibrium dialysis method, spectrophotometric method, 22(82hydroxyquinoline252sulfoacid272azo)21, 82dihydroxy23, 62naphthalene disulfoacid, bovine serum albumin, binding constant, maximum binding number (Received 8 September 2002 ; accetped 25 March 2003)