Acyclovir

Molecular formula: C8H11N5O3 Molecular weight: 225.2 CAS Registry No.: 59277-89-3, 69657-51-8 (sodium salt)

SAMPLE Matrix: blood Sample preparation: 100 |xL Plasma + 10 |xL water + 100 |xL 200 mM pH 7 sodium phosphate buffer, mix well, inject a 5 |xL aliquot. HPLCVARIABLES Guard column: ixBondapak CN guard-PAK Column: 300 X 3.9 ixBondapak C18 Mobile phase: MeCN: 50 mM ammonium acetate 2:98 Flow rate: 1 Injection volume: 5 Detector: UV 254 CHROMATOGRAM Retention time: 18 Internal standard: acyclovir OTHER SUBSTANCES Extracted: ceftibuten Noninterfering: acetaminophen, amoxicillin, ampicillin, aspirin, aztreonam, caffeine, cefamandole, cefotiam, cefsulodin, ceftazidime, ceftriaxone, cefuroxime, cephaloridine, cephalothin, chlorpheniramine, gentamicin, moxolactam, nafcillin, piperacillin, pseudoephedrine, theophylline, ticarcillin, vancomycin KEYWORDS plasma; acyclovir is IS; column-switching REFERENCE
Lim, J.M.; Kim, H.; Marco, A.; Mojaverian, P.; Lin, C-C. Liquid chromatographic determination of ceftibuten, a new oral cephalosporin, in human plasma and urine. J.Pharm.Biomed.AnaL, 1994, 12, 699-703

SAMPLE Matrix: blood Sample preparation: Condition a 1 mL 100 mg Sep-Pak Vac trifunctional C18 SPE cartridge with 1 mL MeOH and 1 mL buffer. 0.5 mL Plasma + 0.5 mL buffer, vortex, add to SPE cartridge, wash with 0.5 mL buffer, elute with 300 |xL MeOH: 5 mM sodium octanesulfonate 20:80 adjusted to pH 8.50 with 4 M NaOH, inject a 130 JJLL aliquot of the eluate. (Procedure was automated (ASPEC system). Buffer was 5 mM sodium octanesulfonate adjusted to pH 2.85 with concentrated orthophosphoric acid.) HPLCVARIABLES Guard column: 20 X 2 30-40 juim Perisorb RP-18 (change each day) Column: 150 X 3.9 4 |xm Nova Pak C18 Mobile phase: MeOH: 10 mM Na2HPO4 + 10 mM sodium octanesulfonate 7:93 with the final apparent pH adjusted to 2.80 with concentrated orthophosphoric acid Column temperature: 40 Flow rate: 1

Injection volume: 130 Detector: UV 250

CHROMATOGRAM

Retention time: 4 Limit of quantitation: 10 ng/mL

KEYWORDS

plasma; SPE
REFERENCE
Swart, K.J.; Hundt, H.K.L.; Groenewald, A.M. Automated high-performance liquid chromatographic method for the determination of acyclovir in plasma. J.Chromatogr.A, 1994, 663, 65-69

SAMPLE

Matrix: blood Sample preparation: Heat-inactivate serum at 56 for 1 h. 500 jxL Serum + 50 |xL 100 |jig/mL guanosine, filter (Centrisart IM1.5000 cut-off), dilute ultrafiltrate 1:30 with mobile phase buffer, inject an aliquot.
HPLCVARIABLES

Column: 125 X 4 5^m RP8e (Merck) Mobile phase: MeOH: 100 mM pH 3.0 phosphate buffer containing 50 mM 1-octanesulfonic acid 5:95 Flow rate: 1 Detector: UV 254
CHROMATOGRAM

Retention time: 6.36 Internal standard: guanosine (7.35) Limit of detection: 50 ng/mL OTHER SUBSTANCES Noninterfering: gangiclovir, zidovudine
KEYWORDS

serum; ultrafiltrate
REFERENCE
Nebinger, P.; Koel, M. Determination of acyclovir by ultrafiltration and high-performance liquid chro matography. J.Chromatogr., 1993, 619, 342-344

SAMPLE

Matrix: blood Sample preparation: 1 mL Plasma + 300 pX. 3 M perchloric acid, vortex 15 s, centrifuge at 2000 g for 2 min, inject a 20 \xL aliquot of the supernatant.
HPLCVARIABLES

Column: 80 X 4 3 yun Nucleosil 120 3C18 Mobile phase: Gradient. A was 20 mM perchloric acid. B was MeCN: 20 mM perchloric acid 45:55. A:B 100:0 for 3 min, 0:100 for 3 min (step gradient). Column temperature: 30 Flow rate: 1.5 Injection volume: 20 Detector: F ex 260 em 375

CHROMATOGRAM Retention time: 2.3 Limit of detection: 6-10 ng/mL KEYWORDS plasma REFERENCE
Mascher, H.; Kikuta, C; Metz, R.; Vergin, H. New, high-sensitivity high-performance liquid chromatographic method for the determination of acyclovir in human plasma, using fluorometric detection. J.Chromatogr., 1992, 583, 122-127

SAMPLE Matrix: blood Sample preparation: 200 jxL Blood + 600 |mL 16% trichloroacetic acid, centrifuge, inject an aliquot of the supernatant. HPLCVARIABLES Guard column: C18 Guard-Pak (Waters) Column: 100 X 8 Nova-Pak C18 (in a Z module) Mobile phase: Gradient. A was 50 mM sodium hydrogen phosphate. B was MeOH: water 80:20 containing 5 mM NaH2PO4. A:B 99:1 for 1.5 min, to 5:95 over 18.5 min, re-equilibrate at initial conditions for 10 min. Flow rate: 1.6 Detector: UV 254 CHROMATOGRAM Retention time: 11.6 Limit of detection: 250 ng/mL OTHER SUBSTANCES Extracted: famciclovir (as penciclovir, the active metabolite) KEYWORDS mouse; pharmacokinetics REFERENCE
Boyd, 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, 358-363

SAMPLE Matrix: blood Sample preparation: Inject an aliquot directly onto the column. HPLCVARIABLES Guard column: 20 X 4.6 16 |xm Spheron Micro 300 Column: 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 sodium sulfate Column temperature: 45 Flow rate: 1 Injection volume: 10 Detector: F ex 285 em 370

CHROMATOGRAM Retention time: 2 Limit of detection: 100 ng/mL KEYWORDS plasma; effluent cooled to 2 before entering detector; GPC; dog; pharmacokinetics; direct injection REFERENCE
Salamoun, J.; Sprta, V.; Sladek, T.; Smrz, M. Determination of acyclovir in plasma by column liquid chromatography with fluorescence detection. J.Chromatogr., 1987, 420, 197-202

SAMPLE Matrix: blood, dialysate, urine Sample preparation: 100 |xL Plasma, urine, or dialysate + 100 |xL 200 mM pH 7 phosphate buffer, inject a 5 (JLL aliquot. HPLCVARIABLES Column: ixBondapak C18 Mobile phase: MeCN: 50 mM ammonium acetate 2:98 Flow rate: 1 Injection volume: 5 Detector: UV 254 CHROMATOGRAM Internal standard: acyclovir OTHER SUBSTANCES Extracted: ceftibuten KEYWORDS plasma; acyclovir is IS REFERENCE
Kelloway, J.S.; Awni, W.M.; Lin, C.C.; Lim, J.; Affrime, M.B.; Keane, W.F.; Matzke, G.R.; Halstenson, CE. Pharmacokinetics of ceftibuten-czs and its trans metabolite in healthy volunteers and in patients with chronic renal insufficiency. Antimicrob.Agents Chemother., 1991, 35, 22672274

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

KEYWORDS

plasma; pharmacokinetics
REFERENCE
Vergin, H.; Kikuta, C; Mascher, H.; Metz, R. Pharmacokinetics and bioavailability of different formulations of aciclovir. Arzneimittelforschung, 1995, 45, 508515

SAMPLE

Matrix: blood, urine Sample preparation: Plasma. 200 |xL Plasma + 200 |xL 200 mM pH 7.0 sodium phosphate, vortex, allow to sit for 15 min, add 800 JJLL MeCN, vortex for 20 s, centrifuge at 2500 g at 25 for 2 min. Remove the supernatant and add it to 1.6 mL dichloromethane, vortex for 20 s, centrifuge at 2500 g at 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, let stand for 5 min, inject an aliquot.
HPLCVARIABLES

Column: 300 X 3.9 ^Bondapak C18 Mobile phase: MeCN: 50 mM ammonium acetate 2:98 Column temperature: 30 Flow rate: 1 Injection volume: 20 Detector: UV 262 (plasma); UV 254 (urine)
CHROMATOGRAM

Internal standard: acyclovir OTHER SUBSTANCES Extracted: ceftibuten

KEYWORDS

plasma; acyclovir is IS
REFERENCE
Kearns, G.L.; Reed, M.D.; Jacobs, R.F.; Ardite, M.; Yogev, R.D.; Blumer, J.L. Single-dose pharmacokinetics of ceftibuten (SCH 39720) in infants and children. Antimicrob.Agents Chemother., 1991, 35, 2078-2084

SAMPLE Matrix: perfusate

HPLCVARIABLES

Column: Pecosphere 5C-C18 Mobile phase: MeCN: 0.1% acetic acid 2:98 Flow rate: 1 Detector: UV 254
REFERENCE
Volpato, N.M.; Santi, P.; Colombo, P. Iontophoresis enhances the transport of acyclovir through nude mouse skin by electrorepulsion and electroosmosis. Pharm.Res., 1995, 12, 1623-1627

ANNOTATED BIBLIOGRAPHY

Shao, Z.; Park, G.-B.; Krishnomoorthy, R.; Mitra, A.K. The physicochemical properties, plasma enzymatic hydrolysis, and nasal absorption of acyclovir and its 2'-ester prodrugs. Pharm.Res., 1994, 11, 237-242 [nasal perfusate]

Shao, Z.; Mitra, A.K. Bile salt-fatty acid mixed micelles as nasal absorption promoters. III. Effects on nasal 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 and plasma by micellar liquid chromatography with fluorimetric determination. 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 method for acyclovir determination in plasma. J.Liq.Chromatogr., 1990, 13, 981-989 [plasma; acetaminophen is IS] Cronqvist, J.; Nilsson-Ehle, I. Determination of acyclovir in human serum by high-performance liquid chromatography. J.Liq.Chromatogr., 1988, 11, 2593-2601 [serum; non-interfering acetaminophen, allopurinol, baclofen, carbacholine, cefuroxime, chlorpropamide, cilastatin, cloxacillin, diazepam, dicumarol, digoxin, flucloxacillm, furosemide, fusidic acid, fusidic, glipizide, heparin, hydrochlorothiazide, imipenem, insulin, isoniazid, ketoprofen, metronidazole, naproxen, perphenazine, phenytoin, prednisolone, propranolol, pyrazinamide, pyridoxine, ranitidine, rifampicin, rifampin, spironolactone, streptomycin, sulfamethoxazole, trimethoprim, warfarin] Bouquet, S.; Regnier, B.; Quehen, S.; Brisson, A.M.; Courtois, P.; Fourtillan, J.B. Rapid determination of acyclovir in plasma by reversed phase high-performance liquid chromatography. J.Liq. Chromatogr., 1985, 8, 1663-1675 [plasma; acetaminophen is IS; LOD 250 ng/mL] Smith, R.L.; Walker, D.D. High-performance liquid chromatographic determination of acyclovir in serum. 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, using ion-pair extraction and high-performance liquid chromatography. IV. Application to solid pharmaceutical dosage forms. J.Liq.Chromatogr, 1983, 6, 2521-2541 [capsules; tablets; also benzocaine, caffeine, carbinoxamine, fenfluramine, flupentixol, lidocaine, melitracen, mepivacaine, phenylephrine, 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, 629-632 Marini, D.; Pollino, G.; Balestrieri, F. Liquid chromatographic determination of acyclovir. Boll. Chim.Farm., 1991, 130, 101-104