Reproducible measurement of amphotericin B in serum by high-performance liquid chromatography in alkaline buffer

Leclercq, M; Fouillit, M.; Panteix, G.; Guinet, R

doi:10.1016/0378-4347(85)80058-x

Cited by 15 publications

(8 citation statements)

References 14 publications

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“…Although it is generally accepted 43 that DMF is an excellent solvent for amphotericin B, many workers have suggested DMSO 20,21,23 or DMSOsMeOH 19,24 for preparing the standard solutions. Our experiments on the stability with time of amphotericin B dissolved in selected solvents showed ( Table 3) that the antibiotic is much more stable in DMF than in DMSO.…”

Section: Stability Of Amphotericin B In Selected Solventssmentioning

confidence: 99%

“…15,16 A direct spectrophotometric method for measuring amphotericin B in whole blood and plasma has been also developed, 17 but it seriously suffers from high background absorbance mainly caused by hemoglobin. Several alternative, more specific, high-performance liquid chromatographic methods are also available, [18][19][20][21][22] but their applicability is often limited by interfering endogenous compounds, 22 low extraction efficiency, 18,19 and inadequate reproducibility necessitating use of internal standards. [18][19][20] To improve the performance characteristics, a few solid-phase extraction techniques 23,24 have been introduced recently for removing interference from serum and/or urine extracts prior to liquid chromatography.…”

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confidence: 99%

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Rapid Determination of Amphotericin B in Serum and Urine by Third-Order Derivative Spectrophotometry

Botsoglou

Fletouris

Παπαγεωργίου

et al. 1996

Journal of Pharmaceutical Sciences

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A derivative spectrophotometric method for rapid monitoring of amphotericin B in serum and urine down to 30 ng/mliters is described. Samples are treated with acetonitrile, and amphotericin B is directly quantified in the crude extracts on the basis of the intensity of the peak that appears at 402 nm when the normal absorption spectrum is submitted to third-order derivative processing. Accuracy data suggested recoveries in the range of 84.3-94.9% for serum and 85.6-93.4% for urine. The precision of the method was better than 11.3% for serum and 9.2% for urine when samples contained as low as 29.6 ng/mliters of amphotericin B. Ease of applicability, short analysis time, low cost, and reliability are the main advantages of the method.

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Section: Stability Of Amphotericin B In Selected Solventssmentioning

confidence: 99%

mentioning

confidence: 99%

Rapid Determination of Amphotericin B in Serum and Urine by Third-Order Derivative Spectrophotometry

Botsoglou

Fletouris

Παπαγεωργίου

et al. 1996

Journal of Pharmaceutical Sciences

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“…For determination of free Amphotericin B, this has to be achieved without disturbing or breaking the liposomes present in human plasma. High-pressure liquid chromatography (HPLC) methods were reported for the quantification of Amphotericin B in plasma or serum [6][7][8][9][10][11][12][13][14][15][16][17][18][19][20]. These methods quantified the total amount of Amphotericin B in circulation.…”

Section: Introductionmentioning

confidence: 99%

Determination of free and liposomal Amphotericin B in human plasma by liquid chromatography–mass spectroscopy with solid phase extraction and protein precipitation techniques

Deshpande

Gangrade

M.B.Kekare

et al. 2010

Journal of Chromatography B

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“…Although several other HPLC methods for AMB have been described elsewhere (1,3,4,6,7,9,12), most have required many steps in sample preparation (1) or have a very large coefficient of variation (7). The HPLC method of Granich et al (4) for AMB determination required a solid-phase extraction procedure for elimination of potential bilirubin interference.…”

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confidence: 99%

Rapid determination of amphotericin B levels in serum by high-performance liquid chromatography without interference by bilirubin

Hosotsubo

Takezawa

Taenaka

et al. 1988

Antimicrob Agents Chemother

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A high-performance liquid chromatographic method for the determination of amphotericin B concentrations in human serum without bilirubin interference was developed and compared with a microbiological assay. The high-performance liquid chromatographic assay utilized a reversed-phase trimethyl silica column, simple sample preparation, and visible detection. Reproducibility studies yielded coefficient-of-variation ranges from 1.02 to 2.11% for within-day precision and 2.88 to 4.32% for between-day precision. The correlation coefficient with the microbiological assay was 0.984 for amphotericin B.Amphotericin B (AMB) is a polyene antifungal agent which was isolated from Streptomyces nodosus in 1956 (10) and remains today the drug of choice for the treatment of systemic fungal infections (5, 12). The most frequent and clinically significant side effect of AMB therapy is nephrotoxicity (8). Although toxicity is generally dose dependent, there is considerable interindividual variability among patients. Because of these large individual variations in AMB levels in serum with oral administration, a method is required for frequent and rapid estimation of drug levels which provide adequate antifungal activity but do not produce adverse effects.The concentrations of AMB in serum were previously determined by microbiological assays (2, 11). Although bioassays have been used traditionally for the determination of concentrations of antibiotics, they have limited sensitivity and reproducibility. The additional disadvantages of bioassays are their lack of specificity and the length of time required for sample analysis. High-performance liquid chromatography (HPLC) provides a sensitive and specific alternative to bioassays, with the additional advantages of high precision and rapid turnaround time. We developed a sensitive HPLC assay for the measurement of AMB in serum and compared it with the microbiological assay.The HPLC assay was performed with an LC-6A pump, an SPD-6AV UV-visible variable-wavelength detector, and a C-R4A computing integrator (Shimadzu, Kyoto, Japan). Analysis was performed on a reversed-phase CLC-trimethylsilyl column (5 ,um; 150-by 6.0-mm [inside diameter]; Shimadzu). Samples were injected with a Rheodyne syringeloading sample injector (model 7125; Shimadzu) fitted with a 50-,ul loop. The mobile phase for HPLC analysis consisted of an acetonitrile-10 mM acetate buffer (pH 7.4) mixture (40/60, vol/vol). The mobile phase was degassed and delivered with a flow of 1.0 ml/min at room temperature. The A405 of the column effluent was monitored (0.005 absorbance units full scale).Serum samples used in this study were obtained from patients with systemic fungal infections receiving AMB as well as other antibiotics for which various determinations of * Corresponding author. concentrations in serum had been ordered as part of the clinical management of the patients. Serum standards and controls were prepared by dissolving AMB (Japan Squibb, Tokyo, Japan) in dimethyl sulfoxide and diluting the solution in drug-free pool...

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Reproducible measurement of amphotericin B in serum by high-performance liquid chromatography in alkaline buffer

Cited by 15 publications

References 14 publications

Rapid Determination of Amphotericin B in Serum and Urine by Third-Order Derivative Spectrophotometry

Rapid Determination of Amphotericin B in Serum and Urine by Third-Order Derivative Spectrophotometry

Determination of free and liposomal Amphotericin B in human plasma by liquid chromatography–mass spectroscopy with solid phase extraction and protein precipitation techniques

Rapid determination of amphotericin B levels in serum by high-performance liquid chromatography without interference by bilirubin

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