Quantitation of Antibiotics by High-Pressure Liquid Chromatography: Cephalothin

Nilsson-Ehle, Ingrid; Yoshikawa, Thomas T.; Schotz, Michael C.; Guze, Lucien B.

doi:10.1128/aac.13.2.221

Cited by 23 publications

(8 citation statements)

References 16 publications

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“…Original reports of cephalosporin quantitation by HPLC typically describe the analysis of a single cephalosporin in biological fluid (2,7,9,10,13,14). Each of these methods presents a unique preparatory and chromatographic protocol for a single cephalosporin or its metabolite or both.…”

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High-pressure liquid chromatographic method for analysis of cephalosporins

Signs¹,

File²,

Tan³

1984

Antimicrob Agents Chemother

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A high-pressure liquid chromatographic method is described for the analysis of a wide range of cephalosporin congeners, using only three reagents for extraction and drug analysis. Plasma was treated with cold methanol-0.1 M sodium acetate to precipitate protein. Cephalosporins were resolved on a C-18 reverse-phase column, utilizing a mobile phase of various percentages of 0.01 M sodium acetate and acetonitrile-methanol. Compounds analyzed included cephalexin, cefamandole, cephalothin, cefotaxime, cefazolin, cephaloridine, cefoxitin, cefaclor, cephapirin, and cefoperazone. Each antibiotic demonstrated excellent linearity throughout the therapeutic range. The method of standard additions revealed recoveries of 93 to 101%, with detection limits ranging from 0.2 to 1.0 ,ug/ml for these drugs. Retention times ranged from 4 to 6 min. This method offers a rapid and simple means by which this group of cephalosporins may be reliably quantitated.The application of high-pressure liquid chromatography (HPLC) to the analysis of antibiotics introduces a powerful tool for therapeutic drug monitoring as well as clinical research (1). The advantages of short turnaround time, method reliability, method sensitivity, and drug specificity justify the use of HPLC for various groups of antibiotics (8,15).Original reports of cephalosporin quantitation by HPLC typically describe the analysis of a single cephalosporin in biological fluid (2,7,9,10,13,14). Each of these methods presents a unique preparatory and chromatographic protocol for a single cephalosporin or its metabolite or both. The analysis of a wide range of different cephalosporins would be complicated by extensive method changeover of extraction procedure, mobile phase, stationary phase, and UV detection wavelength (6, 15). Research on a large class of compounds such as the cephalosporins would be expedited by the availability of a readily adjustable method of analysis that used a limited number of reagents.With this goal in mind, we describe the development of a reverse-phase HPLC method for the analysis of cephalothin, cephaloridine, cefaclor, cefamandole, cefazolin, cephalexin, cefoperazone, cephapirin, cefoxitin, and cefotaxime in human plasma. MATERIALS AND METHODSCephalosporin standards were prepared fresh in distilled deionized water or 0.45 M phosphate buffer, pH 6.8 (according to the manufacturers' reconstitution instructions), on the day of analysis. Internal standards were prepared fresh daily in absolute methanol at a concentration approximating the mean of the therapeutic range (Tables 1 and 2 All experiments were performed on antibiotic-spiked human plasma from the hospital blood bank. Plasma (0.3 ml) was combined with an equal volume of ice-cold 70% absolute methanol with internal standard-30% 0.1 M sodium acetate, pH 5.2. This mixture was vortexed for 30 s and incubated at -20°C for 10 min. The specimens were centrifuged at 1,500 x g for 10 min, and 10 ,ul of the resulting supernatant was used for analysis.Chromatography was performed at room temperature sin...

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High-pressure liquid chromatographic method for analysis of cephalosporins

Signs¹,

File²,

Tan³

1984

Antimicrob Agents Chemother

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“…18 This method was extensively used to determine cephalothin concentrations in many studies. [7][8][9][10][11][12][13][14]22,23 However, because the microbiological assay does not discriminate between the active drug (cephalothin) and its active metabolite (acetylcephalothin), it is criticized for some authors. Nevertheless, we have selected it because of its sensibility, simplicity and good correlation with HPLC determination.…”

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“…Nevertheless, we have selected it because of its sensibility, simplicity and good correlation with HPLC determination. 9 Plasma cephalothin disposition curves after intravenous administration were best fitted by an open bicompartmental model in all the cats. This is in agreement with human and horses pharmacokinetic studies.…”

Section: Discussionmentioning

confidence: 99%

“…2,12 Extension of metabolism varies with the species, being of 5-15% in humans, 14 and greater in rabbits, dogs and horses. 5,9,11 The antibacterial activity of desacetylcephalothin is only one-half to onesixteenth that of cephalothin, the lowest value corresponding to Bacillus subtilis ATCC 6633. 15 Cephalothin plasma half-life elimination is lower than 1 hour in most studied species.…”

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confidence: 99%

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Plasma pharmacokinetics, tissue concentrations and urine elimination after cephalothin intravenous administration to cats under surgical conditions

et al. 2015

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Pharmacokinetic profile, tissue concentrations and urine elimination of cephalothin in cats under surgical conditions after a single intravenous dose (30 mg/kg) were studied. . Tissue/plasma concentrations rate was in a range of 0.04 (testicle) to 0.21 (uterus). Cephalothin urine elimination was 66.49% in the first 6 hours after administration. Cephalothin plasma concentrations remained above a minimum inhibitory concentration (MIC)≥1 µg/mL up to 5.5 hours in all the studied cats. However, for MIC≥8 µg/mL (MIC breakpoint) this time is reduced to 2.5 hours. This suggests that proper perioperative prophylactic use of cephalothin in cats requires a dose interval not longer than 2 hours.

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“…The use of high-pressure liquid chromatography (HPLC) as a sensitive and specific method for the analysis of antibiotics, especially B-lactam compounds, is becoming more and more accepted. Recent literature contains an increasing number of papers devoted to the HPLC analysis of ,B-lactam antibiotics including specific assay methods for mecillinam (7), ampicillin (8,12), and cephalothin (9,11), to name a few.…”

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