Pharmacokinetics and the Elimination Regularity of Difloxacin and its Metabolite Sarafloxacin in Eriocheir sinensis

Chinese Journal of Zoology 2009,44 (2) :12 20 3 ( 201306) :,, ( Eriocheir sinensis) 20 mgπkg, 2,, V d 31170 LΠkg 21122 LΠkg 11045 LΠkg 11051 LΠkg 01203 LΠkg ;, t 1Π2 961316 h 881228 h 1371524 h 671021 h 1241679 h ; CL s 01783 LΠh kg 01040 LΠh kg 01013 LΠh kg 01011 LΠh kg 01008 LΠh kg, 4 4,,(100 gπkg), 30 gπkg,24 d, : ; ; ; :S941 :A :025023263 (2009) 02212209 Pharmacokinetics and the Elimination Regularity of Difloxacin and its Metabolite Sarafloxacin in Eriocheir sinensis LI Hai2Di YANG Xian2Le 3 HU Kun WANG Xiang2Ling ( Aquatic Pathogen Collection Center of Ministry of Agriculture, Shanghai Ocean University, Shanghai 201306, China) Abstract :The pharmacokinetics of difloxacin (DIF) and its metabolite sarafloxacin ( SAR) in hemolymph, muscle, hepatopancreas, spermary and ovary of Chinese Mitten2handed Crab ( Eriocheir sinensis) were studied by RP2HPLC, after administration by single oral garage of 20 mgπkg. The concentrations versus time in hemolymph, muscle, hepatopancreas, spermary and ovary of E. sinensis were well described by a two2department open model. The results of present studies showed that DIF was rapidly absorbed and extensively distributed in E. sinensis. The apparent volume of distribution (V d ) of DIF of hemolymph, muscle, hepatopancreas, spermary and ovary was 31170 LΠkg, (No. 2006BAD03B04) ; 3, E2mail : xlyang @shfu. edu. cn ;,, ; : ; E2mail : heidi - 610 @yahoo. cn :2008209208, : 2008211210

2 : 13 21122 LΠkg,11045 LΠkg,11051 LΠkg and 01203 LΠkg respectively. DIF eliminated slowly. The elimination half2time (t 1Π2 ) was 961316 h,881228 h,1371524 h,671021 h and 1241679 h, respectively, and the total body clearance (CL s ) was 01783 LΠh kg, 0104 LΠh kg, 01013 LΠh kg, 01011 LΠh kg and 01008 LΠh kg. The metabolite SAR concentration2time profiles of the hemolymph, muscle, hepatopancreas, spermary and ovary presented similar trend. The multipeak phenomenon were observed, but the peak time of the metabolite SAR in hemolymph, muscle, hepatopancreas, spermary and ovary was the time that the eliminating rate of DIF was very low. Considering that the eliminating rate of DIF and metabolite SAR were very low. Withdrawal time was more 24 d when the maximum residue of DIF is 100 gπkg and of SAR is 30 gπkg after single oral administration of DIF in E. sinensis. Key words : Eriocheir sinensis ; Difloxacin ; Sarafloxacin ; Pharmacokinetics ( difloxacin,dif),, 1984 Abbott [1 ],, [2 ],,,, [3 5 ] [3 ] [4 8 ] [9 ],(25 015),( Eriocheir sinensis),, 1 111, 150,(9515 1013) g(2510 015),24 h, 112 ( 9919 %) (sarafloxacin,sar) ( 9916 %),Sigma ; (9913 %,:20061129), HPLC ;, 113 Agilent21100, DAD 114 : ZORBAX SB2C 18 (150 mm 416 mm,5 m) ; 01030 molπl ( ph 311) = 5 95 (vπv) ; 276 nm ; 115 mlπmin ;40 ; 10 l,91365 min, 101617 min 115 11511 10 mgπml,20 mgπkg 15 20 min, 11512 01083 h 015 h 1 h 3 h 6 h 9 h 12 h 24 h 48 h 72 h 96 h 144 h 192 h 240 h 360 h 480 h 528 h 576 h 600 h 8,015 ml ACD 115 ml, - 70 116 11611, 1 ml ( = 250 1 1) 5 ml, 2

14 Chinese Journal of Zoology 44 min,4 8 000 rπmin 10 min,,45, 1 ml, 0145 m,4 HPLC 11612 1100 g,, 1 molπl NaCl 012 molπl PBS (ph 714) 1 ml,5 ml, 10 min,4 8 000 rπmin 10 min, ; 5 ml,,, 45 0145 m,4 HPLC 117 Excel, DAS 211 2 211,,,, 91365 min 101617 min( 1), 1 ml, 5 ml 212 116 1 Fig. 1 HPLC chromatogram of DIF and SAR in the tissues of in E. sinensis A. (5 gπml) ;B. 3 h ;C. 6 h ;D. 015 h ; E. 3 h ;F. 9 h a, b A. HPLC chromatogram from the standard sample of DIF and SAR ; B. Chromatogram from the hemolymph of E. sinensis at 3 h ; C. Chromatogram from the muscle of E. sinensis at 6 h ; D. Chromatogram from the hepatopancreas of E. sinensis at 015 h ; E. Chromatogram from the spermary of E. sinensis at 3 h ; F. Chromatogram from the ovary of E. sinensis at 9 h. Peak a means DIF, peak b means SAR.

2 : 15, ( 94125 3177) % (75153 2130) %; 417 %, 518 %, 213, 2 3 01083 01500 h,,015 h (C max = 261255 gπml),,6 h (C = 5198 gπml),,480 h (20 d), 01032 gπml ( 2),, 015 h (C max = 171004 gπml),, 6 h (C = 8104 gπml),,480 h (20 d), 01023 gπml,528 h (22 d), ( 3),, 1 h (C max = 261187 gπml),,3 h 6 h, 12 h (C max = 211234 gπ ml), 2 96 h (4 d), 96 h(4 d),576 h(24 d),01021 gπ ml,600 h (25 d), ( 3),,24 h (C max = 24196 gπml),,600 h (25 d), 01032 gπml 3,,, 9 h ( C max = 761354 gπml),96 h (4 d) (C max = 51199 gπml),,,240 h (10 d),,600 h (25 d), 01052 gπml ( 3) 2 (25 ) Fig. 2 The hemolymph concentration of DIF in E. sinensis following a single oral gavage of 20 mgπkg 214 DAS, 1,,,

16 Chinese Journal of Zoology 44 3 (25 ) Fig. 3 The muscle, hepatopancreas, spermary, and ovary concentration of DIF in E. sinensis following a single oral gavage of 20 mgπkg Parameter 1 Table 1 Pharmacokinetic parameters of DIF in tissues of E. sinensis Hemolymph Muscle Tissue Hepatopancreas Spermary C max ( gπml) 261255 171004 261187 241960 761354 T max (h) 015 015 110 2410 910 t 1Π2 (h) 01263 01123 211329 671016 31853 t 1Π2 (h) 961316 881228 1371524 671021 1241679 t 1Π2 ka (h) 01029 01031 01026 01730 31095 V d (LΠkg) 31170 21122 11045 11051 01203 CL s (LΠh kg) 01783 01040 01013 01011 01008 AUC 0-600 ( gπml h) 5001907 5051866 1 5501485 1 8581779 2 5971570 T lag (h) 01074 01080 01000 01000 01000 Ovary 215, 4 5, 6 h,,12 48 h,,96 h,,,72 h (3 d),, 96 h (4 d), ; 48 h,144 h (6 d) 4, 01860 gπ ml 11570 gπml 81589 gπml 191284 gπml

2 : 17 121213 gπml,, 360 h (15 d), 360 h (15 d), 576 h (24 d),,576 600 h (24 25 d), 01017 01027 gπml ( 5) 4 (25 ) Fig. 4 The hemolymph concentration of SAR in E. sinensis following a single oral gavage of 20 mgπkg 5 (25 ) Fig. 5 The muscle, hepatopancreas, spermary, and ovary concentration of SAR in E. sinensis following a single oral gavage of 20 mgπkg 3 311,, [3 9 ], ( Sus domestica) [4 ] ; ( Gallus gallus) [6 ] ; 20 ( Carassius auratus) [9 ], [6 ] ;10 [9 ],,,,

18 Chinese Journal of Zoology 44 [10 ], (9 h 96 h),, C max T max t 1Π2ka 20 20 mgπkg [9 ], t 1Π2ka 0154 h, C max 10128 gπml, T max 2121 h10 mgπkg [6 ], t 1Π2ka 1146 h, C max 110 gπml, T max 4134 h5 mgπkg [4 ], t 1Π2ka 0154 h, C max 2129 gπml, T max 1141 h 20 mgπkg, t 1Π2ka 01029 h, C max 261255 gπml,t max 015 hc max, (T max ) ( Capra hircus),, AUC V d 20 mgπkg [9 ], 20,AUC 355119 gπml h,v d 4102 LΠkg ; 10,AUC 1 361179 gπml h,v d 2102 LΠkg 5 mgπkg [7 ],,AUC 24123 gπml h,v d 1168 LΠ kg ;,AUC 23162 gπml h ;,AUC 19140 gπml h [6 ] 10 mgπkg,auc 29116 gπml h,v d 3110 LΠ kg ;,AUC 22162 gπml h ;, AUC 15182 gπml h [4 ] 5 mgπkg,auc 27124 gπml h, V d 4191 LΠkg ;, AUC 24198 gπml h ;, AUC 26159 gπml h, 20 mgπkg, AUC 5001907 gπml h 5051866 gπml h 1 5501485 gπml h 1 8581779 gπml h 2 597157 gπml h, (AUC) V d 3117 LΠkg t 1Π2 CL s [4 ] 5 mgπkg t 1Π2 17114 h 25179 h 16167 h,cl s 012 LΠh kg [6 ] 10 mgπkg, t 1Π26111 h, CL s 0137 LΠh kg [7 ] 5 mgπkg t 1Π2 11143 h 13189 h 12102 h, CL s 0121 LΠh kg [9 ] 20 mgπkg, 10 20, CL s 0101 LΠh kg 01056 LΠh kg,20 t 1Π248193 h,, t 1Π2 961316 h 881228 h 1371524 h 671021 h 1241679 h,, ; CL s 01783 LΠh kg 0104 LΠh kg 01013 LΠh kg 01011 LΠh kg 01008 LΠh kg,,,,, [11 ],,,,

2 : 19, 312,, [3 ] ( Canis f amiliaris) [12 ],, C max 01033 gπml,, 5, C max 01860 gπml 11570 gπml 81589 gπml 191284 gπml 121213 gπml,, 3128 % 9123 % 32179 % 77126 % 15199 %,,, [13 15 ], 4 4,,,, ( Oryctolagus cuniculus) [16,17 ] (unhomogeneous),,,, 313, (MRL), 800 gπkg, 300 gπkg, 600 gπkg, 100 gπkg ; 30 gπkg, : 240 h (10 d), 576 h (24 d),576 h (24 d),,,(100 gπ kg), 24 d [ 1 ],..,2004,9(3) :12 14. [ 2 ].. :,2005, 211 212. [ 3 ] Granneman G R, Snyder K M, Shu V S. Difloxacin metabolism and pharmacokinetics in humans after single oral doses. Antimicrobial Agents and Chemotherapeutics, 1986, 30 (3) :689 693. [ 4 ],,..,2003,36 (7) : 846 850. [ 5 ] Garcia M A, Solans C, Aramayona J J, et al. Simultaneous determination of difloxacin and its primary metabolite sarafloxacin in rabbit plasma. Chromatographia,2000,51 :487 490. [ 6 ],,..,2004,34 (6) :20 24. [ 7 ] Marin P, Fernandez2Varon E, Escudero E, et al. Pharmacokinetics after intravenous, intramuscular and subcutaneous administration of difloxacin in sheep. in Veterinary Science,2007, 83 :234 238. Research [ 8 ] Chu D T W, Grannerman G. Difloxacin metabolian and pharmacokinetics in some animals after single oral doses. Vet Pharmacol Thera,2001,24(suppl 2) :151 154. [ 9 ] Ding F K, Cao J Y, Ma L B, et al. Pharmacokinetics and Commission Regulation of EEC, No. 2377Π90. Modified in the Reg. 2908Π00 Brussels,2000. Commission Regulation ( EC) No 508Π1999. Official Journal of the European Communities,1999,03-09. J

20 Chinese Journal of Zoology 44 tissue residues of difloxacin in crucian carp ( Carassius auratus) after oral administration. Aquaculture, 2006,256 : 121 128. [10 ],..,2007,28(1) :1 3. [11 ],,,.,2004,8 ( ) :25 29. [12 ] Heinen E. Comparative pharmacokinetics of enrofloxacin and difloxacin as well as their main metabolites in dogs. Compendium on Continuing Education for the Practicing Veterinarian, 1999, 21(suppl) :12 18 [13 ],,..,2004,28 ( ) :35 41. [14 ] Tang J, Yang XL, Zheng Z, et al. Pharmacokinetics and the active metabolte of enrofloxacin in Chinese mitten2handed crab ( Eriocheir sinensis). Aquaclture,2006,260 :69 76. [15 ],,.., 2007,14(4) :622 629. [16 ],,..,1992,13 (2) :180 182. [17 ],,..,2001,17(2) :225 229. 2007 4 6,(118 48 00 118 53 04 E, 32 01 57 32 16 15 N),() 2 ( Garrulax monileger) 20 m,15 min,,, 2007 4 13 ( G. pectoralis),5,,,, ;, 2007 10 11, 10, 2 3 2007 4 27 1 ( G. berthemyi) 3 2007 11 5 12,,,, (Passeriformes) (Timaliidae) (,2006),,3, 2007 2008, ( 210037 ; 230001)