322 (Chin J New Drugs Clin Rem)2012 6 31 [ ]1007-7669(2012)06-0322-06 1 1 2 2 1 3 (1. 210009; 2. 213200; 3. 210009) [ ] ; ; ; ; ; ; [ ] LC-MS / MS 184 LC-MS / MS Ultimate XB-Phenyl (100 mm 3.0 mm 3 μm) -0.01 mol L -1 ( 0.1% ) (75 25 V / V) SRM : m / z 748 m / z 590 m / z 749 m / z 591 10.128 ~ 8 102.4 μg L -1 (r = 0.998 7); 10.128 μg L -1 ; RSD 10%; 85% ~ 115% t max 2 h 2 h ρ max 22. 29 μg g -1 21.80 μg g -1 AUC 1-192 528.0 μg h g -1 536.5 μg h g -1 t 1 /2 25.5 h 24.1 h t max 8 h 8 h ρ max 53.10 μg g -1 51.62 μg g -1 AUC 1-192 1 584.9 μg h g -1 1 379.4 μg h g -1 t 1 /2 28.8 h 23.7 h [ ] R969.1; R988.1 [ ] A Determination and pharmacokinetics of azithromycin in rabbit conjunctiva DI Bin 1 SUN Xiao-yang 1 MAO Bai-yang 2 ZHAO Xiao-hong 2 SU Meng-xiang 1 WANG Xiaobin 3 ( 1. Department of Pharmaceutical Analysis China Pharmaceutical University Nanjing JIANGSU 210009 China; 2. Changzhou Yabang Pharmacy Research Institute Co.Ltd Changzhou JIANGSU 213200 China; 3. Laboratory Animal Center Southeast University Nanjing JIANGSU 210009 China) [ KEY WORDS] azithromycin; conjunctiva; chromatography high pressure liquid; tandem mass spectrometry; rabbits; pharmacokinetics; eye drops [ ABSTRACT] AIM To establish a LC-MS / MS method for the determination of azithromycin in rabbit conjunctiva and to study the pharmacokinetics of azithromycin eye drops after single and multiple dose administration. METHODS Following a deproteinization procedure the azithromycin concentrations in conjunctiva were determined by HPLC-MS / MS and the pharmacokinetics were evaluated in 184 New Zealand albino [ ] 2011-11-29 [ ] 2012-04-16 [ ] (1973 ) Phn: 86-25-8327-1269 E-mail: ddw888@vip.sina.com [ ]
(Chin J New Drugs Clin Rem)2012 6 31 323 rabbits after single and multiple dose of azithromycin eye drops respectively. Clarithromycin was taken as internal standard and the samples were eluted utilizing a mobile phase containing of 0.01 mol L -1 ammonium (containing 0.1% acetic acid) -acetate (25 75 V / V) and an Ultimate XB-Phenyl column (100 mm 3.0 mm 3 μm). ESI and SRM were used with positive ion scans and the mass transition pairs of m / z 748 m / z 590 and m / z 749 m / z 591 were used to detect internal standard and azithromycin. RESULTS The method demonstrated that good linearity ranged from 10.128 to 8 102.4 μg L -1 with r = 0.998 7; the lower limit of quantification for azithromycin in conjunctiva tissues was 10.128 μg L -1 ; the intra-and inter-batch precision (RSD) values were below 10% and the accuracy ranged from 85% to 115%. The main pharmacokinetics of standard formula-tion and test formulation after single administration were as follows: t max 2 h versus 2 h; ρ max 22.29 μg g -1 versus 21.80 μg g -1 ; AUC 1-192 528.0 μg h g -1 versus 536.5 μg h g -1 t 1 /2 25.5 h versus 24.1 h respectively. The parameters after multiple administration were as follows: t max 8 h versus 8 h; ρ max 53.10 μg g -1 versus 51.62 μg g -1 ; AUC 1-192 1 584.9 μg h g -1 versus 1 379.4 μg h g -1 t 1 /2 28.8 h versus 23.7 h respectively. There was no significant difference of the pharmacokinetics characteristics between two formulations. CONCLUSION The method is convenient and sensitive. There is a gentle accumulation of azithromycin after multiple administrations. (azithromycin) (PALL ); TGL-16G ( ); CM-1000 [1-4] ( ) 2 ~ 4 d [5 6] ( : 130593-200901); ( : 130558-200501); ( [7 8] : 20101207 : 5 ml: 50 mg; - ( AZASITE 1% Inspire Pharmaceuticals Inc : IN10E12B); [9-11] (LC-MS/MS) [10] ( ROE ); ( ); ( ) 184 2.0 ~ 2.5 kg : SCXK ( ) 2010-0002 [ Finnigan TSQ - - 4 (8 ); (R) (ESI) (T); 25 μl Xcalibur 1.4 ; BS 21 S (Sartorius ); XHF-1 : SYXK ( ) 2010-0004] 1 80 40 40 10 ( ); PL5242 2 104
324 (Chin J New Drugs Clin Rem)2012 6 31 52 52 13 (3 (1 h 10 ) 4 (8 ); (R) (T); d 1 d 2 2 12 h 25 μl -1 ; d 3 ~ 7 1 25 μl -1 d 5 d 6 d 7 4 d 7 1 2 4 8 12 24 48 72 120 192 h (10 1 4 ) 1 (3%) 1 ml ; 10 μl 2 ml -20 50 μl Ultimate XB-Phenyl (100 mm 3.0 mm 3 μm Welch ); : 30 ; : -0.01 mol L -1 ( 0.1% ) (75 25 V / V); : 0.4 ml min -1 : (ESI); : (SRM); : ; : [M+H] + m / z 749 m / z 591 [ M +H ] + m / z 748 m/z 590; : 4 500 ev; : : 10.128 ~ 8 102.4 μg L -1 310.3 kpa; : 13.8 kpa; : 10.128 μg L -1 34.5 kpa; : 350 ; : 50 μl 2 ml 10 ev; : 28 ev; : 0.20 Pa 32 409.6 μg L -1 ) 13.73 mg ( 12.66 mg) 25 ml 6 481.92 μg L -1 ) 3 5 506.4 mg L -1 20 μl 50.64 101.28 253.2 ; 506.4 2 532 10 128 32 409.6 40 512 μg L -1 50 μl 1 ml 12.93 mg ( 12.61 mg) 50 ml 1.5 ml 3 252.2 mg L -1 30 s 3 2 522 μg L -1 2 5 μl : = R i / R s 100 % R i 50 μl 2 ml 10 μl R s ( ) 20 μl 30 s 1 ml 3 min -20 10 min 16 880 g 5 min 5 μl 192 h) DAS2.1.1 3.2 min 2.5 min 10.128 20.256 50.64 101.28 506.4 2 025.6 6 481.92 8 102.4 μg L -1 5 μl a (As / Ai) ( 5 ) ρ (μg L -1 ) : ρ = 470.79a - 0.370 7 r = 0.998 7 (n = 8) : w = 1 y -2 ; (101.28 2 532 10 μl (20.256 506.4 10 μl 20 μl 3 (n = 5) (89.1 ± 3.3) % (97.4 ± 2.7) % (95.3 ± 1.1) %; (86.7 ± 3.2) %
(Chin J New Drugs Clin Rem)2012 6 31 325 1 ( ) HPLC ( : [M+H] + m / z 749 m / z 591 [M+H] + m / z 748 m / z 590) A: B: (506.4 μg L -1 ) C: ( 1 ) D: 1 h (1 000.7 μg L -1 ) 85% 50 μl 3 min -20 10 min 2 ml 16 880 g 5 min ( 101.28 2 532 32 409.6 μg L -1 ) 10 μl 30 s 3 20 : 2 ml 10 μl 20 μl 5 1 ml 3 min 3 20 μl 5 : 5 μl 2 1 3 (101.5 ± 7.4) % : 5 (99.7 ± 1.3) % (99.4 ± 2.2) %;
326 (Chin J New Drugs Clin Rem)2012 6 31 t 3 (101.2 ± 1.4) % 10 1 (P < 3 ( 0.05) 10 0.75 ml) ( 2 (P < 0.05) 101.28 2 532 32 409.6 μg L -1 ) 150 μl 4 30 s 20.256 506.4 6 481.92 μg L -1 3 50 μl 2 ml 10 2 20 μl 0 h ; 2 5 h 2 3 2 20 d 2 2 25 μl 40 d (n = 40 x ± s) 20 μl 5 μl 0 h 2 : 5 h 3 20 d - 2 104 (52 ) (52 ) - 3 3 25 μl (n = 52 x ± s) 80 (40 ) (40 )
(Chin J New Drugs Clin Rem)2012 6 31 327 ρ max 2 AUC 1-192 2 ~ 3 R 2.36 2.57 [ ] HPLC-UV ; [1] [J]. LC-MS / MS 200019(3): 175-177. [10] [2] C 18 2003 22 (6): 328-330. C 18 [3] 45 2004 23(9): 619-621. [4] [J]. 2004 10(1): 51-52. 3.2 min [5] FOULDS G SHEPARD RM JOHNSON RB. The pharmacokinetics of azithromycin in human serum and tissues[j]. J Anti- 2.5 min microb Chemother 1990 25 (Suppl A): 73-82. [6] SHEPARD RM FALKNER FC. Pharmacokinetics of azithromycin in rats and dogs[j]. J Antimicrob Chemother 1990 25(Suppl A): [M+H] + 1 49-60. [7] AMAR T CAILLAUD T ELENA PP. Ocular pharmacokinetic study following single and multiple azithromycin administrations 1 in pigmented rabbits[j]. Curr Eye Res 2008 33(2): 149-158. SRM [8] SI EC CHEUNG PS BOWMAN L et al. Ocular pharmacokinetics of AzaSite Xtra -2% azithromycin formulated in a DuraSite delivery system[j]. Curr Eye Res 2009 34(6): 485-491. [9] AKPEK EK VITTITOW J VERHOEVEN RS et al. Ocular surface distribution and pharmacokinetics of a novel ophthalmic [12] 1% azithromycin formulation[j]. J Ocul Pharmacol Ther 2009 MIC 0.5 mg L -1 25(5): 433-439. MIC 90 4 mg L -1[7] [10] SHEN Y YIN C SU M et al. Rapid sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS / MS) 1% 25 μl ( method for the quantification of topically applied azithromycin in ) rabbit conjunctiva tissues[j]. J Pharm Biomed Anal 2010 52 MIC 90 (4 mg L -1 (1): 99-104. ) 40 h (d1 ~ [11] GAIL T TERRENCE PO BREIN. Conjunctival tissue pharmacokinetic properties of topical azithromycin 1% and moxifloxacin 2 2 d 3 ~ 7 1 ) 1% 25 μl ( ) 0.5% ophthalmic solutions: a single-sose randomized open- active-controlled trial in healthy adult volunteers[j]. Clin ρ max label Ther 2008 30(11): 2005-2014. [12] MARK BA WARREN H ARON MS et al. Clinical cure of d 5 d 6 d 7 MIC 90 bacterial conjunctivitis with azithromycin 1%: vehicle-controlled (4 mg L -1 ) double-masked clinical trial[j]. Am J Ophthalmol 2008 145(6): 959-965. 4.. 106 [J].. [J]..