Abstract
A rapid, simple, accurate, sensitive and reproducible high performance liquid chromatographic (HPLC) method for the quantitation of amoxicillin in human plasma using cefadroxil as an internal standard (IS) has been developed and validated. The procedure involves an ultrafiltration step prior to a reversed-phase liquid chromatography. The drug and the IS were eluted from Symmetry® C18 stainless steel column (5 μm, 150 × 4.6 mm I.D.) at room temperature with a mobile phase consisting of methanol: 75 mM potassium dihydrogen phosphate buffer solution (10:90,v/v) (pH adjusted to 3.0 with phosphoric acid), at a flow rate of 1.5 mL min−1. The effluent was monitored using a UV detector set at 228 nm. Each analysis required no longer than 10 min. Quantitation was achieved by measurement of the peak area ratio of the drug to the internal standard, and the limit of quantification of amoxicillin in plasma was 0.5 μg mL−1 (RSD% and DEVs% were <15%). The method showed good precision: the intraday RSD% values for amoxicillin were in the range of 1.41–6.86% whereas the values for interday were in the range of 1.13–8.16% at four different concentrations. The method is rapid (total run time <12 min) and accurate (DEVs,% <10%). The mean relative recovery was 99.67% and the mean absolute recovery was 86.68%. Stability testing shows that amoxicillin is stable in plasma for at least 4 weeks when stored at −70 °C. The method was successfully applied in a pharmacokinetic study involving healthy rabbits.
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References
Todd P, Benfield P (1990) Drugs 39:264–307
Finlay J, Miller L, Poupard J (2003) J Antimicrob Chemother 52:18–23
Sutherland R, Croydon E, Rolinson G (1972) Br Med J 3:13–16
Muth P, Metz R, Beck H, Bolten W, Vergin H (1996a) J Chromatogr A 729:259–266
Yuan Z, Russlie H, Canafax D (1995) J Chromatogr B 674:93–99
Krauwinkel W, Volkers-Kamermans N, vanZijtveld J (1993) J Chromatogr 617:334–338
Charles B, Chulavatnatol S (1993) Biomed Chromatogr 7:204–207
Mascher H, Kikuta C (1990) J Chromatogr A 506:417–421
Wibawa J, Fowkes D, Shaw P, Barrett D (2002) J Chromatogr B Analyt Technol Biomed Life Sci 774:141–148
Miyazaki K, Ohtani K, Sunada K, Arita T (1983) J Chromatogr 276:478–482
Carlqvist J, Westerlund D (1985) J Chromatogr 344:285–296
Haginaka J, Wakai J (1987) J Chromatogr 413:219–226
Mascher H, Kikuta C (1998) J Chromatogr A 812:221–226
Erdmann G, Walker K, Giebink G, Canafax D (1990) J Liq Chromatogr 13:3339–3350
Pires de Abreu L, Ortiz R, de Castro S, Pedrazzoli J (2003) J Pharm Pharm Sci 6:223–230
Hoizey G, Lamiable D, Frances C, Trenque T, Kaltenbach M, Denis J, Millart H (2002) J Pharm Biomed Anal 30:661–666
Maurer H (1998) J Chromatogr B 713:3–25
Henion J, Brewer E, Rule G (1998) Anal Chem 70:650–656
Oliveira C, Abib E, Vannuchi Y, Sucupira M, Ilha J, De Nucci G (2001) Int J Clin Pharm Ther 39:167–172
Storms M, Stewart J (2002) J Liq Chromatogr Relat Technol 25:2433–2443
Nelis H, Vandenbranden J, DeKruif A, DeLeenheer A (1992) Antimicrob Agents Chemother 36:1859–1863
Chulavatnatol S, Charles B (1993) J Chromatogr 615:91–96
Cass Q, Gomes R, Calafatti S, Pedrazolli J (2003) J Chromatogr A 987:235–241
Shah V, Midha K, Dighe S, McGilveray I, Skelly J, Yacobi A, Layloff T, Viswanathan C, Cook C, McDowall R (1992) Pharm Res 9:588–592
Muth P, Metz R, Beck H, Bolten W, Vergin H (1996b) J Chromatogr A 729:259–266
Garcia-Gonzalez J, Mendez R, Martin-Villacorta J (1998) J Chromatogr A 812:213–220
Myers C, Blumer J (1984) Antimicrob Agents Chemother 26:78–81
Garcia-Capdevila L, Lopez-Calull C, Arroyo C, Moral M, Mangues M, Bonal J (1997) J Chromatogr B Biomed Sci Appl 692:127–132
Granneman G, Sennello L (1982a) J Chromatogr 229:149–157
Granneman G, Sennello L (1982b) J Phar Sci 71:1112–1115
Breilh D, Lavallee C, Fratta A, Ducint D, Cony-Makhoul P, Saux M (1999) J Chromatogr B Biomed Sci Appl 734:121–127
Essers L (1982) Eur J Clin Microbiol 1:367–370
Papp E, Knupp C, Barbhaiya R (1992) J Chromatogr 574:93–99
Liang H, Kays M, Sowinski K (2002) J Chromatogr B Analyt Technol Biomed Life Sci 772:53–63
Overholser B, Kays M, Sowinski K (2003) J Chromatogr B Analyt Technol Biomed Life Sci 798:167–173
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Matar, K.M. Simple and Rapid LC Method for the Determination of Amoxicillin in Plasma. Chroma 64, 255–260 (2006). https://doi.org/10.1365/s10337-006-0021-9
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DOI: https://doi.org/10.1365/s10337-006-0021-9