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

Er, White; Carroll, MA; Je, Zarembo

doi:10.7164/antibiotics.30.811

Cited by 49 publications

(7 citation statements)

References 7 publications

(7 reference statements)

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“…The lipid solubility and UV absorbance of CAP make it a prime candidate for reversed-phase HPLC analysis. One method has been described in which HPLC was used to quantitate intermediates in CAP synthesis (24); more recently, reversed-phase HPLC has been applied (26). Initial evaluation of this method revealed a significant interference between CAP and theophylline.…”

Section: Resultsmentioning

confidence: 99%

High-Performance Liquid Chromatographic Assay of Chloramphenicol in Serum

Koup¹,

Brodsky

Lau

et al. 1978

Antimicrob Agents Chemother

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A new method for the analysis of serum chloramphenicol by reversed-phase, high-performance liquid chromatography (HPLC) is described. The method involves a preliminary extraction of 0.1 ml of serum with ethyl acetate containing an internal standard, chromatography with a reversed-phase C18 microparticulate column with an acetonitrile-acetate buffer mobile phase, and detection by measuring UV absorbance at 270 nm. Assay performance was compared with an existing microbiological assay. The HPLC method demonstrated both increased precision and increased sensitivity. The specificity of the HPLC method was also evaluated. The new method presents an alternative approach to the analysis of clinical specimens.

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Section: Resultsmentioning

confidence: 99%

High-Performance Liquid Chromatographic Assay of Chloramphenicol in Serum

Koup¹,

Brodsky

Lau

et al. 1978

Antimicrob Agents Chemother

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“…Microparticulate reverse-phase columns are widely used for the HPLC analysis of ,-lactam antibiotics (2,3,9,10). Ion pair chromatography is employed with reverse-phase columns when dealing with a compound which is in an ionic form in the mobile phase and has to be ion paired to be retained on the column.…”

Section: Discussionmentioning

confidence: 99%

High-pressure liquid chromatographic analysis of aztreonam in sera and urine

Pilkiewicz¹,

Remsburg²,

Fisher³

et al. 1983

Antimicrob Agents Chemother

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Aztreonam (SQ 26,776) is a new synthetic monocyclic P-lactam antibiotic which is specifically active against aerobic gram-negative bacteria. High-pressure liquid chromatographic (HPLC) systems were developed for the quantitative analysis of aztreonam in human, monkey, rat, mouse, and rabbit sera and urine. The HPLC conditions employed for these analyses were a ,uBondapak C18 column, a mobile phase made up of 0.005 M tetrabutylammonium hydrogen sulfate at pH 3.0 and acetonitrile or methanol, UV detection at 293 nm, and a flow rate of 2.0 ml/min. For human sera and urine, the mobile phase was 80%o 0.005 M tetrabutylammonium hydrogen sulfate-0.005M (NH4) 2SO4 and 20% acetonitrile (vol/vol). For the range of sera and urine, HPLC analyses were shown to have excellent detector linearity of aztreonam over a concentration range of 1.0 mg/ml to 0.5 ,ug/ml. Correlation coefficients for plots of aztreonam peak area versus its concentration were -0.990. The detection limit of aztreonam was 1.0 jig/ml in sera and 5.0 ,ug/ml in urine. HPLC and microbiological assays of aztreonam in human sera and urine were in good agreement.Aztreonam (SQ 26,776) (Fig. 1) is a totally synthetic monocyclic ,-lactam antibiotic specifically active against aerobic gram-negative bacteria (6). It is a member of the monobactams, naturally occurring 3-lactams produced by bacteria (7). This compound is currently undergoing clinical development, and human pharmacology data have been reported in detail (4,5).The assay of antibiotics in body fluids has traditionally been performed by microbiological assay, but recently high-pressure liquid chromatography (HPLC) has been extensively employed as an alternative procedure (8,11 3.9 mm; length, 3.0 cm). The guard column was packed with Bondapak C18 on Corasil. For studies with both chromatographic systems, the following conditions were employed: a UV detector setting of 293 nm, a solvent flow rate of 2.0 ml/min, and a chart speed of 0.5 cm/min. Quantitation of aztreonam was performed by an external standard method of calculation, and peak area measurements were used.Aztreonam. Aztreonam, prepared in our laboratories, was used as either the dipolar ion form shown in Fig. 1 or the disodium or dipotassium salt formed from the free diacid.

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“…However, the availability of small-particle size packings for reversed-phase chromatography suggests that very high chromatographic efficiencies and shorter analysis times can be expected with this procedure (8,9). The most recent papers deal with anion-exchange chromatography (5), reversed-phase chromatography (61, and NMR spectroscopy (7).…”

Section: Referencesmentioning

confidence: 99%