Quantitation of Antibiotics Using High-Pressure Liquid Chromatography: Tetracycline

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

doi:10.1128/aac.9.5.754

Cited by 37 publications

(10 citation statements)

References 12 publications

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“…If the microbial assay values were compared with those of cephalothin alone, desacetylcephalothin alone, or the sum of cephalothin plus desacetylcephalothin (all determined by the chemical assay), the correlation coefficients were 0.79, 0.84, and 0.80, respectively. This lower correlation coefficient in dogs may be due to the higher concentrations of desacetylcephalothin in the sera of Recently, the HPLC technique has been successfully implemented for rapid, accurate, and specific measurements of tetracycline (13) and amphotericin B (12) in serum and cerebrospinal fluid. The complete methodology involved simple extraction of antibiotics from the samples, followed by separation by HPLC and, finally, detection of the compounds by ultraviolet spectrometry.…”

Section: Resultsmentioning

confidence: 99%

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Quantitation of Antibiotics by High-Pressure Liquid Chromatography: Cephalothin

Nilsson-Ehle¹,

Yoshikawa

Schotz

et al. 1978

Antimicrob Agents Chemother

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A teclpique for quantitative determination of cephalothin and desacetylcephalothin in serum using a method based on high-pressure liquid chromatography is described. Both compounds were quantitatively extracted from serum by using dimethylformamide. After separation of the drugs by reverse-phase chromatography and detection by ultraviolet light absorption at 254 nm, serum concentrations of cephalothin and desacetylcephalothin as low as 1.0 jig/ml were measured.No interfering absorption was found in extracts of serum samples from normal humans or patients receiving a variety of other drugs, including other antimicrobial agents. Serum specimens obtained after an intravenous infusion of cephalothin to twp human subjects and three dogs were assayed by the chemical and microbiological assays. When values, as determined by the chemical method and the microbiological assay in samples from human subjects, were compared, the correlation coefficient (r) was 0.96, whereas, in the dogs, the r value was 0.79. The chemical procedure described in this report for determination of cephalothin and desacetylcephalothin permits future pharnacokinetic investigations of these compounds. This assay is sensitive, specific, accurate, and rapid (approximately 30 min), and appears suitable for routine clinical use.

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Section: Resultsmentioning

confidence: 99%

“…Recently we have successfully applied highpressure liquid chromatography (HPLC) to the quantitation of antibiotics in serum and cerebrospinal fluid (12,13). This technique has advantages over standard microbiological assays with regard to accuracy, specificity, and rapidity.…”

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Quantitation of Antibiotics by High-Pressure Liquid Chromatography: Cephalothin

Nilsson-Ehle¹,

Yoshikawa

Schotz

et al. 1978

Antimicrob Agents Chemother

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“…An HPLC technique for the analysis of tetracycline after extraction from serum has recently been reported by Nilsson-Ehle et al (4). Cooper et al (2) reported on the use of HPLC, employing anion exchange chromatography of cephalothin and deacetylcephalothin after ion pair extraction from human serum.…”

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Rapid Analysis of Cefazolin in Serum by High-Pressure Liquid Chromatography

Wold

1977

Antimicrob Agents Chemother

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A high-pressure liquid chromatography (HPLC) method has been developed for the analysis of cefazolin in serum. Serum was deproteinized by the addition of6% trichloroacetic acid and injected onto a reverse-phase column with a mobile phase of 10 to 15% methanol in 1% aqueous acetic acid. Cefazolin chromatographed without interference from ultraviolet-absorbing components of serum, with a retention time of 3.1 min. Standard curves comparing peak area with concentration prepared from dog or human sera were linear over a range of 1.6 to 200 ug/ml. Results from the HPLC assay were compared with microbiological assays (cylinder plate method) on both standard serum samples and sera from dogs and human subjects receiving intramuscular cefazolin. The HPLC method was somewhat more accurate in comparison with the microbiological assay performed on serum samples of known concentration. The comparison of results from an analysis of serum levels of dogs or human subjects receiving cefazolin indicated that the two methods would lead to identical conclusions concerning pharmacokinetics or the achievement of therapeutic serum levels. The HPLC assay method presents an alternative to conventional microbiological assays, with marked improvement in speed (30 min) and considerable potential for future development.

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“…Several authors have reported on the use of reverse phase methods for the determination of tetracyclines. 7,8,9,10) For reasons discussed in our previous paper1), we prefer to use reverse phase instead of ion exchange chromatography for antibiotics. However, ion exchange high pressure liquid chromatography has also been used successfully for the analysis of cephalosporins,11, 12,13) penicillins 14, 151 and tetracyclines.…”

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Reverse phase high speed liquid chromatography of antibiotics. II. Use of high efficiency small particle columns.

Er¹,

Carroll²,

Je³

1977

J. Antibiot.

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Improved methods for the separation and quantitation of cephalosporins, penicillins, tetracyclines and several miscellaneous antibiotics by reverse phase high speed liquid chromatography are presented. The methods have been improved significantly by the substitution of high efficiency, small particle (-10 pm) reverse phase columns in place of the previously used medium efficiency, pellicular columns. The conditions and procedures described here illustrate that considerable improvements in separation and sensitivity of detection of antibiotics are achieved. Pure compounds, complex mixtures of antibiotics in a variety of dosage forms and fermentation broths are routinely analyzed by the described procedures.In a previous paper" we described conditions for the determination of cephalosporins, penicillins, tetracyclines and other miscellaneous antibiotics by reverse phase high speed liquid chromatography.A second paper 2) dealt with a specific application of this technique to the determination of cephalexin and cephradine in various pharmaceutical formulations and in physiological fluids.

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Quantitation of Antibiotics Using High-Pressure Liquid Chromatography: Tetracycline

Cited by 37 publications

References 12 publications

Quantitation of Antibiotics by High-Pressure Liquid Chromatography: Cephalothin

Quantitation of Antibiotics by High-Pressure Liquid Chromatography: Cephalothin

Rapid Analysis of Cefazolin in Serum by High-Pressure Liquid Chromatography

Reverse phase high speed liquid chromatography of antibiotics. II. Use of high efficiency small particle columns.

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