Abstract
rapid high-performance liquid chromatography (HPLC) method for the quantification of vancomycin in mouse plasma samples was developed and validated. Norvancomycin was used as the internal standard. Chromatographic separation was achieved on a Vydac C18 column (4.6×50 mm, 3 μm particle size) and the detection was made at 214 nm. A gradient elution was programmed with the mobile phases of 0.1% v/v trifluoroacetic acid (A) and 95:5 v/v acetonitrile: 0.1% TFA (B) and a flow rate of 1 ml/min. The total run time was 15 min. The calibration curve was linear over the range of 0.1-20 μg/ml, with a correlation coefficient (r) higher than 0.997 and the lower limit of quantitation (LLOQ) of 0.1 μg /ml. The intra-day accuracy values were between 90 and 112% and the inter-day ones ranged from 96 to 104%. Precision values ranged from 1.7 to 9.5% for intra-day and 6.3 to 9.4% for inter-day. Stability studies under normal laboratory conditions without significant loss of the drug. The assay was successfully applied to the pharmacokinetics and bio-distribution study of novel formulations of vancomycin in mice.
Highlights
Vancomycin (Figure 1) was first isolated as “compound 05865” in 1953 by Edmund Kornfield of Eli Lilly and Co, from a dirt sample from Borneo that contained the organism, Streptomyces orientalis [1]
Representative chromatograms of mouse blank plasma, mouse blank plasma spiked with 5 μg/ml concentration of norvancomycin and mouse plasma spiked with 10 μg/ml vancomycin and 5 μg/ml norvancomycin were shown in (Figures 2a-c), respectively
Given the fact that the minimum inhibitory concentration (MIC) of vancomycin is in the 1-4 μg/ml range, the sensitivity of our method is satisfactory for experimental and clinical pharmacokinetic investigations
Summary
Vancomycin (Figure 1) was first isolated as “compound 05865” in 1953 by Edmund Kornfield of Eli Lilly and Co, from a dirt sample from Borneo that contained the organism, Streptomyces orientalis [1]. Immune enzymatic techniques have been widely used clinically owing to their simplicity and high speed, several drawbacks associated with these techniques hinder their usage in research studies involving pharmacokinetics of vancomycin [5,6]. These drawbacks have led to the development of various chromatographic techniques for the accurate and precise determination of vancomycin in biological fluids. Vancomycin has been quantified in tissues [8,14] Some of these methods provide high sensitivity and specificity, tedious extraction techniques and expensive technology are a major hindrance for their practical implementation. A simple HPLC method with a routine extraction procedure is desirable
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