HPLC Analysis of Acyclovir

7/28/2019 HPLC Analysis of Acyclovir

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SAMPLEMatrix: bloodSample preparat ion: 100 |xL Plasma + 10 |xL water + 100 |xL 200 mM pH 7 sodiumphosphate buffer, mix well, inject a 5 |xL aliquot.HPLCVARIABLESGuard column: ixBondapak CN guard-PAKColumn: 300 X 3.9 ixBondapak C18Mobile phase: MeCN : 50 mM am monium acetate 2:98Flow rate: 1Inject ion volume: 5Detector: UV 254CHROMATOGRAMRetent ion t ime: 18Internal s tandard: acyclovirOTHER SUBSTANCESExtracted: ceftibutenNoninterfering: acetaminophen, amoxicillin, ampicillin, aspirin, aztreonam, caffeine, ce-famandole, cefotiam, cefsulodin, ceftazidime, ceftriaxone, cefuroxime, cephaloridine, ce-phalothin, chlorpheniramine, gentamicin, moxolactam, nafcillin, piperacillin, pseudoe-phe drine, theoph ylline, ticarcillin, vancomycinKEYWORDSplasma; acyclovir is IS; column-switchingREFERENCELim, J.M.; Kim, H.; Marco, A.; Mojaverian, P.; Lin, C-C. Liquid chromatographic determination of cef-tibuten, a new oral cephalosporin, in human plasma and urine. J.Pharm.Biomed.AnaL, 1994, 12,699-703SAMPLEMatrix: bloodSample preparat ion: Condition a 1 mL 100 mg Sep-Pak Vac trifunctional C18 SPE car-tridge with 1 mL M eOH and 1 mL buffer. 0.5 mL P lasm a + 0.5 mL buffer, vortex, addto SPE cartridge, wash with 0.5 m L buffer, elute with 300 |xL MeO H: 5 mM sodium

octanesulfonate 20:80 adjusted to pH 8.50 with 4 M NaOH, inject a 130 JJLL aliquot ofthe eluate. (Procedure was automated (ASPEC system). Buffer was 5 mM sodium octa-nesulfonate adjusted to pH 2.85 with concentrated orthophosphoric acid.)HPLCVARIABLESGuard column: 20 X 2 30-40 juim Perisorb RP-18 (change each day)Column: 150 X 3.9 4 |xm Nova Pak C18Mobile phase: MeOH : 10 mM Na 2HPO 4 + 10 mM sodium octanesulfonate 7:93 with thefinal apparent pH adjusted to 2.80 with concentrated orthophosphoric acidColumn temp erature: 40Flow rate: 1

AcyclovirMolecular formula: C8H11N5O3Molecular weight: 225.2CAS Reg istry No.: 59277-89-3, 69657-51-8 (sodium salt)


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Injection volume: 130Detector: UV 250CHROMATOGRAMRetention time: 4Limit of quantitation: 10 ng/mLKEYWORDSplasma; SPEREFERENCESwart, K.J.; Hundt, H.K.L.; Groenewald, A.M. Automated high-performance liquid chromatographicmethod for the determination of acyclovir in plasma. J.Chromatogr.A, 1994, 663, 6 5 - 6 9SAMPLEMatrix: bloodSample preparation: Heat-inactivate serum at 56 for 1 h. 500 jxL Serum + 50 |xL 100|jig/mL guanosine, filter (Centrisart IM 1.5000 cut-off), dilute ultrafiltra te 1:30 with mobilephase buffer, inject an aliquot.HPLCVARIABLESColumn: 125 X 4 5^m RP8e (Merck)Mobile phase: MeOH: 100 mM pH 3.0 phosphate buffer containing 50 mM 1-octanesulfonicacid 5:95Flow rate: 1Detector: UV 254CHROMATOGRAMRetention time: 6.36Internal standard: guanosine (7.35)Limit of detection: 50 ng/mLOTHER SUBSTANCESNoninterfering: gangiclovir, zidovudineKEYWORDSserum; ultrafiltrateREFERENCENebinger, P.; Koel, M. Determination of acyclovir by ultrafiltration and high-performance liquid chromatography. J.Chromatogr., 1993, 619, 342-344SAMPLEMatrix: bloodSample preparation: 1 mL Plasma + 300 pX. 3 M perchloric acid, vortex 15 s, centrifugeat 2000 g for 2 min, inject a 20 \xL aliquot of the supernatant.HPLCVARIABLESColumn: 80 X 4 3 yun Nucleosil 120 3C18Mobile phase: Gradient. A was 20 mM perchloric acid. B was MeCN: 20 mM perchloricacid 45:55. A:B 100:0 for 3 min, 0:100 for 3 min (step gradient).Column temperature: 30Flow rate: 1.5Injection volume: 20Detector: F ex 260 em 375


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CHROMATOGRAMRetent ion t ime: 2 .3Limit of detection: 6-10 ng/mLKEYWORDSplasmaREFERENCEMascher, H.; Kikuta, C; Metz, R.; Vergin, H. New, high-sensitivity high-performance liquid chromato-graphic method for the determination of acyclovir in human plasma, using fluorometric detection.J.Chromatogr., 1992, 583, 122-127SAMPLEMatrix: bloodSample preparat ion: 200 jxL Blood + 600 |mL 16% trich loroace tic acid, c entrifuge, injectan aliquot of the supernatant.HPLCVARIABLESGuard co lum n: C18 Guard-Pak (Waters)Column: 100 X 8 Nova-Pak C18 (in a Z module)Mobi le phase : Gradient. A was 50 mM sodium hydrogen phosph ate. B was M eOH: water80:20 containing 5 mM NaH 2PO 4. A:B 99:1 for 1.5 min, to 5:95 over 18.5 min, re-equil-ibrate at initial conditions for 10 min.Fl ow rate : 1.6Detector: UV 254CHROMATOGRAMR ete ntio n t ime : 11.6Limit of detect ion: 250 ng/mLOTHER SUBSTANCESExtracted: famciclovir (as penciclovir, the active metabolite)KEYWORDSmouse; pharmacokineticsREFERENCEBoyd, M.R.; Bacon, T.H.; Sutton, D. Antiherpesvirus activity of 9-(4-hydroxy-3-hydroxymethylbut-l-yl)guanine (BRL 39123) in animals. Antimicrob.Agents Chemother., 1988, 32, 3 5 8 - 3 6 3SAMPLEMatrix: bloodSample preparat ion: Inject an aliquot directly onto the column.HPLCVARIABLESGuard column: 20 X 4.6 16 |xm Spheron Micro 300Column: 150 X 3.2 12.5 |jim Spheron Micro 300 (glass column) (Lachema)Mobile phase: pH 1.8 Buffer containing 100 mM phosphoric acid and 100 mM sodiumsulfateColumn tem peratur e: 45Flow rate: 1Inject ion vo lum e: 10D etector : F ex 285 em 370


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CHROMATOGRAMRetention time: 2Limit of detection: 100 ng/mLKEYWORDSplasma; effluent cooled to 2 before entering detector; GPC; dog; pharmacokinetics; directinjectionREFERENCESalamoun, J.; Sprta, V.; Sladek, T.; Smrz, M. Determination of acyclovir in plasma by column liquidchromatography with fluorescence detection. J.Chromatogr., 1987,420, 197-202SAMPLEMatrix: blood, dialysate, urineSample preparation: 100 |xL Plasma, urine, or dialysate + 100 |xL 200 mM pH 7 phos-

phate buffer, inject a 5 (JLL aliquot.HPLCVARIABLESColumn: ixBondapak C18Mobile phase: MeCN: 50 mM ammonium acetate 2:98Flow rate: 1Injection volume: 5Detector: UV 254CHROMATOGRAMInternal standard: acyclovirOTHER SUBSTANCESExtracted: ceftibutenKEYWORDSplasma; acyclovir is ISREFERENCEKelloway, J.S.; Awni, W.M.; Lin, C.C.; Lim, J.; Affrime, M.B.; Keane, W.F.; Matzke, G.R.; Halstenson,C E . Pharmacokinetics of ceftibuten-czs and its trans metabolite in healthy volunteers and in patientswith chronic renal insufficiency. Antimicrob.Agents Chemother., 1991, 35, 22672274

SAMPLEMatrix: blood, urineSample preparation: 1 mL Plasma or urine + 300 jxL 3 M perchloric acid, vortex for 15s, centrifuge at 2000 g for 2 min, inject a 20 |xL aliquot of the supernatant.HPLCVARIABLESColumn: 80 X 4 3 |xm Nucleosil 3C-18Mobile phase: Gradient. MeCN: 20 mM perchloric acid 0:100 for 3 min, 45:55 for 3 min(step gradient).Column temperature: 30Flow rate: 1.5Injection volume: 20Detector: F ex 260 em 375CHROMATOGRAMLimit of detection: 6-10 ng/mL (plasma); 25 ng/mL (urine)


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KEYWORDSplasma; pharmacokineticsREFERENCEVergin, H.; Kik uta, C ; Mascher, H.; M etz, R. Ph arm aco kine tics and bioavailability of different formu-lations of aciclovir. Arzneimittelforschung, 1995, 45 , 508515SAMPLEMatrix: blood, urineSample preparation: Plasma. 200 |xL Plasma + 200 |xL 200 mM pH 7.0 sodium phos-phate, vortex, allow to sit for 15 min, add 800 JJLL MeCN, vortex for 20 s, centrifuge at2500 g at 25 for 2 min. Remove the supernatan t and add it to 1.6 mL dichloromethane,vortex for 20 s, centrifuge at 2500 g a t 25 for 1 min, inject an aliquot of the organic layer.Urine. Dilute urine samples 10-20-fold with water, treat with Nonidet P-40 detergent, letstand for 5 min, inject an aliquot.HPLCVARIABLESColumn: 300 X 3.9 ^Bondapak C18M obile p ha se : MeCN: 50 mM ammonium acetate 2:98Column temperature: 30Flow rate: 1Injection volume: 20D ete ctor : UV 262 (plasma); UV 254 (urine)C H R O M A T O G R A MInternal standard: acyclovirOTHER SUBSTANCESExtracted: ceftibutenKEYWORDSplasma; acyclovir is ISREFERENCEKearns, G.L.; Reed, M.D.; Jacobs, R.F.; Ardite, M.; Yogev, R.D.; Blumer, J.L. Single-dose pharmacoki-netics of ceftibuten (SCH 39720) in infants and children. Antimicrob.Agents Chemother., 1991, 35,

2078-2084SAMPLEMatrix: perfusateHPLCVARIABLESColumn: Pecosphere 5C-C18Mobile phase: MeCN: 0.1% acetic acid 2:98Flow rate: 1Detec tor: UV 254REFERENCEVolpato, N.M.; Santi, P.; Colombo, P. Iontophoresis enhances the transport of acyclovir through nudemouse skin by electrorepulsion and electroosmosis. Pharm.Res., 1995, 12, 1623-1627 ANNOTATED BIBLIOGRAPHYShao, Z.; Park, G.-B.; Krishnomoorthy, R.; Mitra, A.K. The physicochemical properties, plasma enzy-matic hydrolysis, and nasal absorption of acyclovir and its 2'-ester prodrugs. Pharm.Res., 1994, 11 ,

237-242 [nasal perfusate]


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Shao, Z.; Mitra, A.K. Bile salt-fatty acid mixed micelles as nasal absorption promoters. III. Effects onnasal transport and enzymatic degradation of acyclovir prodrugs. Pharm.Res., 1994, 11, 243-240[nasal perfusate]Macka, M.; Borak, J.; Semenkova, L.; Popl, M.; Mikes, V. Determination of acyclovir in blood serum andplasm a by micellar liquid chromatography with fluorimetric d etermination. J.Liq.Chromatogr., 1993,16, 2359-2386 [serum; plasma; fluorescence detection; LOD 80 ng/mL]Molokhia, A.M.; Niazy, E.M.; El-Hoofy, S.A.; El-Dardari, M.E. Improved liquid chromatographic methodfor acyclovir determination in plasma. J.Liq.Chromatogr., 1990, 13, 981-989 [plasma; acetamino-phen is IS]Cronqvist, J.; Nilsson-Ehle, I. Determination of acyclovir in human serum by high-performance liquidchromatography. J.Liq.Chromatogr., 1988, 11, 2593-2601 [serum; non-interfering acetaminophen,allopurinol, baclofen, carbacholine, cefuroxime, chlorpropamide, cilastatin, cloxacillin, diazepam, di-cumarol, digoxin, flucloxacillm, furosemide, fusidic acid, fusidic, glipizide, heparin, hydrochlorothia-zide, imipenem, insulin, isoniazid, ketoprofen, metronidazole, naproxen, perphenazine, phenytoin,prednisolone, propranolol, pyraz inam ide, pyridoxine, ran itidin e, rifampicin, rifamp in, spironolactone,streptomycin, sulfamethoxazole, trimethoprim, warfarin]Bouquet, S.; Regnier, B.; Quehen, S.; Brisson, A.M.; Courtois, P.; Fourtillan, J.B. Rapid determinationof acyclovir in plasma by reversed phase high-performance liquid chromatography. J.Liq.Chromatogr., 1985, 8, 166 3-16 75 [plasma; acetaminophen is IS; LOD 250 ng/mL]Smith, R.L.; Walker, D.D. High-performance liquid chromatographic determination of acyclovir in se-rum. J.Chromatogr, 1985, 343, 203-207 [serum; LOQ 1.2 fxg/mL]Hoogewijs, G.; Massart, D.L. Development of a standardized analysis strategy for basic drugs, usingion-pair extraction and high-performance liquid chromatography. IV. Application to solid pha rm a-ceutical dosage forms. J.Liq.Chromatogr, 1983, 6, 2521-2541 [capsules; tablets; also benzocaine,caffeine, carbinoxamine, fenfluramine, flupentixol, lidocaine, melitracen, mepivacaine, phenyle-phrine, piperocaine, procaine, tetracaine; normal phase]Nilsson-Ehle, I. High-performance liquid chromatography for analyses of antibiotics in biological fluids.J.Liq.Chromatogr, 1983, 6, Supp. 2, 251-293 [review]Zhang, C; Dong, S.N. [Determination of acyclovir in human plasma by RP-HPLC]. Yao.Hsueh.Hsueh.Pao., 1993, 28, 629-632Marini, D.; Pollino, G.; Balestrieri, F. Liquid chromatographic determination of acyclovir. Boll.Chim.Farm., 1991, 130, 101-104

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HPLC Analysis of Acyclovir