2003 61 8, 1287 1293 ACTA CHIMICA SINICA Vol 61, 2003 No 8, 1287 1293 a Ξ, a, b a Ξ ( a 400715) ( b 400715), (TR) ( KANA) (NEO) ( GEN) (TOB) (RRS), RRS RRS, 400 535 nm, 400 nm, 01013 610 g ml - 1 RRS, (3 ) 1219 1716 ng ml - 1, KANA > NEO > TOB > GEN,, RRS RRS, Resonance Rayleigh Scattering Spectra of Interaction of Aminoglycoside Antibiotics with Trypan Red and Their Analytical Applications HU, Xiao2Li a LIU, Shao2Pu Ξ a, b LUO, Hong2Qun a ( a School of Chemistry and Chemical Engineering, Southwest China Normal University, Chongqing 400715) ( b Institute of Environmental Chemistry, Southwest China Normal University, Chongqing 400715) Abstract In weak acid medium, some aminoglycoside antibiotics, such as kanamycin ( KANA), gentamicin ( GEN), tobramycin (TOB) and neomycin (NEO), or acid bisazo dye trypan red (TR) alone can only produce very weak resonance Rayleigh scattering ( RRS) signals However, when one of the antibiotics and TR react with each other to form the ion association complexes, the RRS intensity can be enhanced greatly and a new RRS spectrum and a significant enhancement of the RRS intensity in the wavelength range from 400 to 535 nm can be observed The maximum scattering peak is at 400 nm and there is a linear relationship between the RRS intensity and the antibiotics concentration in the range of 01013 610 g ml - 1 An RRS method for the determination of aminoglycoside antibiotics at trace amounts of levels has been developed The detection limits (3 ) of the four antibiotics, whose order of sensitivity from high to low ranks as KANA > NEO > TOB > GEN, are 1219 1716 ng ml - 1 This method has good selectivity and has been successfully applied to the quick determination of antibiotics not only for injections and ear drips, but also for clinic serum samples as well In addition, the reaction mechanism by using quantum chemistry method and the influencing factors of the RRS spectra and the enhancement reasons of RRS have been discussed Keywords resonance Rayleigh scattering, aminoglycoside antibiotics, trypan red, determination, ( Polyhydroxy antibiotics), [1] Ξ E2mail : liusp @swnu edu cn Received November 7, 2002 ; revised February 12, 2003 ; accepted March 25, 2003 (Nos 2985019, 20175018)
1288 Vol 61, 2003 ( Kanamycin) ( Gentamicin) ( Tobramycin) (Neomycin) (Streptomycin), 210 10-3 mol L - 1, 210 10-4,, mol L - 1 BR :0104 mol L - 1 H 3 PO 4,, H 3 BO 3 HAc 012 mol L - 1 NaOH,,, ph, ph,, (MA) [2,3] ( HPLC) [4,5] 1 2 (UV2vis) [6,7] ( IA) [8,9] MA 10 ml 110 ml, ph BR 110 ml, 110 ml, ;HPLC,,, ex = ;UV, ; IA, em, RRS,,, RRS I RRS I 0, I = I - I 0 20 90, [10] [11] RRS 2 1 [12 16] [17 20], RRS, (,trypan red,,, TR) RRS,,, RRS,, RRS,, 400 nm, 470 535 nm RRS, RRS, RRS KANA > NEO > TOB > GEN 2 KANA TR2 RRS RRS, KANA RRS, RRS KANA, RRS,, 2 2, 2 2 1, 3 BR ph,ph RRS,, RRS RRS ( TR, Switzerland, Fluka ) : 2 1-1 1 1 RF2540 (, ), :, 5, 5 nm ; UV28500 (, ) ;phs23c (, ) ( Kanamycin Sulphate, KANA ; Sigma ) (Neomycin Sulphate, NEO ; Sigma ) ( Gentamicin Sulphate, GEN ; Sigma ) (Tobramycin Sulphate, TOB ; ) : 40010 g ml - 1 ; 2010 g ml - 1, 1010 g ml - 1 1 TR2 RRS Figure 1 RRS spectra of TR2aminoglyside antibiotics 1, TR ; 2, KANA ; 3, TR2GEN ; 4, TR2TOB ; 5, TR2NEO ; 6, TR2KANA Antibiotics concentration : 316 g ml - 1 ; TR : 210 10-5 mol L - 1 ; ph 312
No 8 : 1289 2 TR2KANA RRS Figure 2 RRS spectra of TR2KANA 1, TR ; 2, KANA ; 3, TR2KANA (212 g ml - 1 ) ; 4, TR2KANA (312 g ml - 1 ) ; 5, TR2KANA (412 g ml - 1 ) ; TR : 210 10-5 mol L - 1 ; ph 312 4 Figure 4 Effect of ionic strength on I RRS Antibiotics concentration : 410 g ml - 1 ; TR : 210 10-5 mol L - 1 3 Figure 3 Effect of acidity on I RRS Antibiotics concentration : 410 g ml - 1 ; TR : 210 10-5 mol L - 1 2 4 1 RRS, KANA, TR NEO, TOB, GEN ph 210 410, 210 5 - KANA 2 +, TR 610, 210 610, 210 610, KANA (ph 2), SO 3 H,,, ; ph (ph 6), H + ( CT), AM1 TR, 5 - KANA I RRS 2 +, TR 5 - - 510-3192, KANA 2 2 2 NaCl, I RRS ( 4),, I RRS, KANA, 200 nm, Na +, Cl - ; TR 505 nm,,, 2 3 ph 312,, 5 Na +,, TR, RRS TR, n ( KANA) n ( TR) = 1 1 TR, AM1 TR KANA 5 TR 5-5, SO 3 -, 3 KANA,, KANA 2 +, 3, 6 TR 5 - KANA 2 + 1 1 7 2 4 RRS RRS 210 0192,, max 570 nm, = 65 nm, CT, E
1290 Vol 61, 2003 5 TR 5 - Figure 5 TR 5 - charge distribution at ground state 6 KANA Figure 6 KANA charge distribution at ground state 7 TR 5 - KANA 2 + Figure 7 Diagram sketch of ion association complex of TR 5-2 KANA 2 + 8 RRS Figure 8 RRS and absorption spectra,,, 400 470 nm, RRS RRS - 400 535 nm 396 570 nm -, RRS, 400 nm, RRS ( 8) RRS,, 535 nm RRS, RRS, RRS,,, 470 nm RRS, 2 4 2 RRS ( (1), - ), RF2540, -,
No 8 : 1291 (2), 3 Na +, Mg 2 +, Ca 2 +, Sr 2 +, Mn 2 + NH 4 +, RRS, Rayleigh [21] TritonX2100 C, I RRS = KcMI 0 I RRS, I 0, c, K, M, 3 1, 48616 137414, RRS, (3),,, 4, 3 2, RRS 15 1100 ml, 210 mol L - 1 2 5 0125 ml,,3000 r min - 1 5 min,, [27] 0150 ml, I RRS, I RRS, TR 0150 ml ph 312 BR 0150 ml,, 1, 019968 019991, 5100 ml, 5, 5, KANA, 1219 ng ml - 1 5, (4 12 g KANA, NEO, TOB, GEN ml - 1 ) [28], 113 % 215 % ;, 2, Sn 2 +, Pb 2 + Cu 2 + 3, 9910 % 10318 % 2 6,, 1, Table 1 Correlation coefficient, linear ranges and detection limits for standard curves / nm ( r) ( c, g ml - 1 ) / ( g ml - 1 ) (3, ng ml - 1 ) TR2KANA 400 I = 415 + 1319 c 019991 01013 610 1219 TR2NEO 400 I = 112 + 1218 c 019987 01014 610 1411 TR2TOB 400 I = 218 + 1111 c 019968 01016 610 1612 TR2GEN 400 I = 117 + 1012 c 019972 01018 610 1716 2 Table 2 Comparison of the sensitivities of RRS with other methods for the determination of aminoglycoside antibiotics / ( g ml - 1 ) (3, ng ml - 1 ) 421 nm 8 128 200 22 Cu 2 + 640 nm 200 2000 20000 23 HPLC2IPD 268 nm 015 10 20 24 ph 1010 1115 011 1715 115 25 1 15 100 26 011 515 50 8 400 nm 01013 610 1219
1292 Vol 61, 2003 / ( mol L - 1 ) 3 Table 3 Effects of coexistent substances / % / ( mol L - 1 ) / % Na + 2000 115 NO - 3 1300-118 Zn 2 + 385 018 SiO 2-3 110-218 Ba 2 + 50 116 HAsO 2-4 10 116 UO 2 + 2 25 216 SO 2-4 1500 213 NH 4 + 100 310 EDTA 570 315 C 2 O 2 4-90 113 SLS 45 415 Cu 2 + 115 511 TritonX2100 1000-612 PO 3 4-650 - 514 Mal 500 217 Sn 2 + 212 216 Lac 500 714 Cr 3 + 40 611 SDS 40 513 Co 2 + 30 217 800 413 Mg 2 + 400-113 125 417 Ca 2 + 1000-316 560 319 Pb 2 + 1192 313 a 10 515 Be 2 + a 85 419 337-212 Sr 2 + 4000-016 a 123 614 Mn 2 + 1000-111 L2 500 415 Al 3 + 16-215 410-711 Fe 3 + 36 613 2 316-318 Cl - 2000 313 C 3300 514 H 2 PO 4-2000 518 560-211 a g ml - 1 4 Table 4 Results for the determination of commercial aminoglycoside antibiotics / (mg ml - 1 ) / (mg ml - 1 ), n = 5 / %, n = 5 RSD/ %, n = 5 215 2146 9814 116 250 248 9912 212 40 3910 9715 211 40 4018 10210 118 5 Table 5 Results for the determination of serum samples / ( g ml - 1 ), n = 5 / / ( g ml - 1 ) ( g ml - 1 ), n = 5 / % n = 5 RSD/ % n = 5 1 0 610 612 10313 215 2 0 810 813 10318 113 3 0 1010 919 9910 212 References 1 Zhao, M Chin J Antibiotics 1999, 24, 319 (in Chinese) (,, 1999, 24, 319 ) 2 Barnes, W G In The Aminoglycoside Antibiotic : A Guide to Therapy, 1st ed, Ed : Barnes, W G, CRC Press Inc, Boca Raton Florida, 1984, p 201 3 The Pharmacopoeia Committee of the People s Republic of China, The Pharmacopoeia of the People s Republic of China, 2nd Version [ S ], Chemical Industry Press, Beijing, 2000, p 312 ( in Chinese) (,,
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