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      Comparison of Microbiological and High-Performance Liquid Chromatographic Methods for Determination of Clarithromycin Levels in Plasma

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          Abstract

          An agar well diffusion bioassay method for determination of clarithromycin in human plasma, using Micrococcus Luteus ATCC 9341 as the assay organism, was compared with a selective high-performance liquid chromatographic (HPLC) method with UV detection. Spiked plasma was used to prepare standard and control samples for both methods. The results of the bioassay analyses with spiked plasma samples were concordant by HPLC methods (R 2 =0.871, P < 0.001).The Bland-Altman method also showed good agreement between the results of two methods. HPLC demonstrated an improved precision (0.88-19.86% versus 4.51-26.78%) and accuracy (99.27-103.42 % versus 78.52-131.19 %), compared to those of the bioassay method. The range of linearity obtained by both methods (from 62.5 to 3000 ng/ml for HPLC and from 250 to 3000 ng/ml for the bioassay) includes the range of concentrations of clarithromycin which are considered clinically relevant. However, comparison between HPLC and microbiological assays after oral administration of clarithromycin in healthy volunteers indicated significant differences between the two methods in mean plasma concentration–time profiles. The Bland-Altman method revealed no agreement between the two methods, which can be explained by the presence of active metabolites of clarithromycin in plasma.

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          Most cited references30

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          Statistical methods for assessing agreement between two methods of clinical measurement.

          In clinical measurement comparison of a new measurement technique with an established one is often needed to see whether they agree sufficiently for the new to replace the old. Such investigations are often analysed inappropriately, notably by using correlation coefficients. The use of correlation is misleading. An alternative approach, based on graphical techniques and simple calculations, is described, together with the relation between this analysis and the assessment of repeatability.
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            Analysis of macrolide antibiotics.

            The following macrolide antibiotics have been covered in this review: erythromycin and its related substances, azithromycin, clarithromycin, dirithromycin, roxithromycin, flurithromycin, josamycin, rokitamycin, kitasamycin, mycinamycin, mirosamycin, oleandomycin, rosaramicin, spiramycin and tylosin. The application of various thin-layer chromatography, paper chromatography, gas chromatography, high-performance liquid chromatography and capillary zone electrophoresis procedures for their analysis are described. These techniques have been applied to the separation and quantitative analysis of the macrolides in fermentation media, purity assessment of raw materials, assay of pharmaceutical dosage forms and the measurement of clinically useful macrolide antibiotics in biological samples such as blood, plasma, serum, urine and tissues. Data relating to the chromatographic behaviour of some macrolide antibiotics as well as the various detection methods used, such as bioautography, UV spectrophotometry, fluorometry, electrochemical detection, chemiluminescence and mass spectrometry techniques are also included.
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              An HPLC assay and a microbiological assay to determine levofloxacin in soft tissue, bone, bile and serum.

              A simple, specific and sensitive HPLC assay for levofloxacin in serum, bile, soft tissue and bone was evaluated and validated. The samples were prepared by protein precipitation with acids and methanol, which yielded high recoveries (for serum and bile>98% and for bone and soft tissue>90%). The compounds were separated on a reversed phase column with an acidic mobile phase containing triethylamine. The eluate was monitored by fluorescence detection. The HPLC assay is linear over the usable concentration range (0.1-40 microg/ml) and it provides good validation data for accuracy and precision. Although comparison of HPLC results to the results of a microbiological assay showed congruent results (regression coefficients>0.967). HPLC should be the method of choice for determination of levofloxacin in biological matrices.
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                Author and article information

                Journal
                Iran J Pharm Res
                Iran J Pharm Res
                IJPR
                Iranian Journal of Pharmaceutical Research : IJPR
                Shaheed Beheshti University of Medical Sciences (Tehran, Iran )
                1735-0328
                1726-6890
                Winter 2010
                : 9
                : 1
                : 27-35
                Affiliations
                [a ] Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
                [b ] Drug Applied Research Center, Tabriz University of Medical Sciences,Tabriz, Iran.
                Author notes
                [* ]Corresponding author: E-mail: lotfipoor@tbzmed.ac.ir
                Article
                ijpr-09-27
                3862973
                24363703
                24bcb4ad-02cd-40f0-aa37-9611a358c8de
                © 2010 by School of Pharmacy, Shaheed Beheshti University of Medical Sciences and Health Services

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License, ( http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : January 2009
                : May 2009
                Categories
                Original Article

                clarithromycin,microbiological assay,high-performance liquid chromatographyplasma,bioassay method

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