Easy-Assessment of Levofloxacin and Minocycline in Relevant Biomimetic Media by HPLC-UV Analysis (original) (raw)
Related papers
Indonesian Journal of Chemistry, 2017
To conduct a bioequivalence study for a copy product of levofloxacin (LEV), a simple and validated analytical method was needed, but the previous developed methods were still too complicated. For this reason, a simple and rapid high performance liquid chromatography method was developed and validated for LEV quantification in human plasma. Chromatographic separation was performed under isocratic elution on a Luna Phenomenex® C18 (150 × 4.6 mm, 5 µm) column. The mobile phase was comprised of acetonitrile, methanol, and phosphate buffer 25 mM that adjusted at pH 3.0 (13:7:80 v/v/v) and pumped at a flow rate of 1.5 mL/min. Detection was performed under UV detector at wavelength of 280 nm. Samples were prepared by adding acetonitrile and followed by centrifugation to precipitate plasma protein. Then followed successively by evaporation and reconstitution step. The optimized method meets the requirements of validation parameters which included linearity (r = 0.995), sensitivity (LLOQ and...
An Improved UPLC Method for Rapid Analysis of Levofloxacin in Human Plasma
Chromatographia, 2008
A rapid, specific, and sensitive ultra-performance liquid chromatographic method for analysis of levofloxacin in human plasma has been developed and validated. Plasma samples were spiked with the internal standard (enoxacin) and extracted with 10:1 (v/v) ethyl acetate–isopropanol. UPLC was performed on a 100 × 2.1 mm i.d., 1.7 µm particle, C18 column with 88:12 (v/v) 0.4% triethylamine buffer (pH 3)–acetonitrile as mobile phase, pumped isocratically at a pressure of 11,000 psi (758 bar) and a flow-rate of 0.3 mL min−1. Ultraviolet detection was performed at 300 nm. The retention times of levofloxacin and enoxacin were 3.4 and 2.8 min, respectively, and the run-time was 5 min. Calibration showed that response was a linear function of concentration over the range 0.05–10 µg mL−1 (r 2 ≥ 0.99) and the method was validated over this range for both precision and accuracy. The relative standard deviation was <15% for both intra-day and inter-day assay (n = 5). Levofloxacin and enoxacin were stable in plasma; there was no evidence of degradation during three freeze–thaw cycles, post-preparative stability at 20 °C was ≥24 h, short-term stability at room temperature was ≥6 h, and long-term stability at −70 °C was ≥30 days. The method was successfully used in a study of the bioequivalence of two levofloxacin tablet formulations in healthy volunteers.
Quimica Nova, 2010
A simple, accurate, economical and reproducible UV Spectrophotometric method for estimation of Gabapentin in tablet dosage form has been developed. The method shows maximum absorbance of Gabapentin at 265nm used for the estimation. Gabapentin was found to be linear in the range for 2-10ug/ml. Percentage of Gabapentin present in pharmaceutical dosage form was found to be in the range of 99.11-99.91% respectively. The developed method validated according to ICH guidelines.
Biomedical Chromatography, 2014
A rapid, sensitive and simple high-performance liquid chromatographic assay with ultraviolet detection was developed for the quantification of levofloxacin in microsamples (100 μL) of human plasma. The extraction procedure included a protein precipitation technique and a short chromatographic running time (4.5 min). Analyses were carried out on a Symmetry C 18 column using a mixture of acetonitrile and 0.01 M potassium dihydrogen aqueous solution (pH 3.4; 14:86 v/v) as mobile phase. The method provided specificity and was linear (r ≥ 0.9992) over the concentration range 0.1-12 μg/mL. The average absolute recovery was 93.59%. The intra-and inter-day coefficients of variation were <6%. Additionally, levofloxacin was stable in all evaluations. The usefulness of this method was demonstrated in a pharmacokinetic study of levofloxacin in healthy adult volunteers. The present method offers two main advantages: (a) the use of microsamples reduces the total volume of blood to be collected from patients; and (b) it provides a good cost-effectiveness ratio. It is concluded that the method is rapid, simple, sensitive, economical and suitable for the determination of levofloxacin in human plasma using a small volume of sample.
Journal of Pharmaceutical and Biomedical Analysis, 1997
A rapid high-performance liquid chromatographic (HPLC) method for the determination of levofloxacin in human plasma and urine has been validated. A single-step liquid liquid extraction procedure was used to isolate levofloxacin from the biological matrix prior to quantitative analysis. The compound was separated on an Inertsil C18 reversed-phase HPLC column and quantified by measuring the UV absorbance at 330 nm. The stereospecificity was achieved in the ligand-exchange mode by incorporating chiral reagents directly into the HPLC mobile phase. Ciprofloxacin was used as the internal standard. The method was linear from 0.08 to 5.18 lag ml-~ of levofloxacin in plasma and from 23 to 1464 lag ml ~ in urine. The overall utility of the method is reflected in its high sample throughput and easy adaptability to robotic automation, thus making the procedure suitable for pharmacological and pharmacokinetic studies of levofloxacin.
Journal of Chromatography B, 2004
The aim of this study was to develop a specific and sensitive high-performance liquid chromatographic (HPLC) assay for the determination of levofloxacin in human plasma, bronchoalveolar lavage and bone tissues. The sample extraction was based on a fully automated liquid-solid extraction with an OASIS cartridge. The method used ultraviolet detection set at a wavelength of 299 nm and a separation with a Supelcosil ABZ+ column. The assay has been found linear over the concentration range 0.25-25 g/ml for levofloxacin in plasma, 1-6 g/ml in bronchoalveolar lavage and 0.5-10 g/g for bone tissues and it provided good validation data for accuracy and precision. The assay will be applied to determine the penetration of levofloxacin in human bronchoalveolar lavage (BAL) and bone tissues during pharmacokinetic steady state.
An attempt was made to develop a single, rapid, specific, and sensitive gradient reversed-phase ultra-performance liquid chromatographic method for quantitative analysis of levofloxacin. The single method thus developed is applied for the quantification of levofloxacin both in aqueous humour as well as pharmaceutical dosage forms (i.e., tablets and eye drops). The newly developed method is applicable for pharmacokinetic studies of eye formulations. The chromatographic separation of levofloxacin was achieved on a Waters Acquity HSS T-3 column (100 x 2.1 mm, 1.8 microm) within a short run-time of 5 min. The method was validated according to the ICH guidelines with respect to system suitability, linearity, limit of quantitation and detection, precision, accuracy, robustness, and specificity. Forced degradation studies were also performed in levofloxacin bulk drug samples to demonstrate the stability-indicating power of the developed ultra-performance liquid chromatography method. The developed method was then successfully applied for the ocular pharmacokinetic study of levofloxacin eye formulations and assay of levofloxacin pharmaceutical dosage form.
Biomedical Chromatography, 2019
Levofloxacin, pefloxacin, ciprofloxacin and moxifloxacin are four fluoroquinolones used in the treatment of serious bacterial infections. A concentration-dependent antibacterial activity of fluoroquinolones is well known. Therefore, therapeutic drug monitoring in daily clinical practice is warranted to assure the therapies efficacy and prevent bacterial resistance. The purpose of the present study was to develop a method using high pressure liquid chromatography with ultraviolet detector for simultaneous quantification of these four fluoroquinolones in human plasma. 50µl of plasma was precipitated by 200µl of methanol using gatifloxacin as internal standard. The chromatographic separation was performed on a Kinetex XB-C18 column using a mobile phase composed of a mixture of orthophosphoric acid 0.4% (v/v), acetonitrile and methanol at a flow rate of 1.2 ml/min. Dual UV wavelength mode was used, with levofloxacin and moxifloxacin monitored at 293 nm, and pefloxacin and ciprofloxacin monitored at 280 nm. The calibration was linear over the range of 0.125-25 mg/L for levofloxacin, 0.1-20 mg/L for moxifloxacin and 0.05-10mg/L for both pefloxacin and ciprofloxacin. Inter-and intra-day truness and precisions were less than 13% for all the compounds under study. The proposed method was simple, reliable, cost-effective and suitable for therapeutic drug monitoring or pharmacokinetics studies.
Separations
A simple, selective, rapid, sensitive and less costly green automated solid phase extraction bio-analytical high-performance liquid chromatographic-based technique with fluorescence detection (Aut-SPE-BA-HPLC-FL) for the quantification of levofloxacin in human serum samples has been developed and validated. The serum samples were loaded into the chromatographic system without prior treatment and then injected into short (20 mm × 4.6 mm, 20 µm) protein-coated (PC) µBondapak CN (µBCN) silica pre-column (PC-µBCN-pre-column). Levofloxacin was retained and pre-concentrated on the head of the PC-µBCN-pre-column, while proteins and other polar components were eliminated using phosphate buffer saline (PBS), pH 7.4, as the first mobile phase in the extraction step. Levofloxacin is then transferred to the analytical column; ZORBAX Eclipse XDB-C18 (150 mm × 46 mm, 5 µm), through the aid of a column-switching valve technique, on-throughs the elution mode using the second mobile phase containing...
2021
This study aims at the development of a validated reverse phase-high performance liquid chromatography (RP-HPLC) method for the simultaneous quantification of levofloxacin (LVX) and clarithromycin (CAM). Eclipse X DB C18 column (5 µm, 4.6 × 250 mm) was used as a stationary phase, and monobasic potassium phosphate (KH 2 PO 4) buffer (0.035 M): acetonitrile (75 : 25) was used as an isocratic mobile phase. The pH of the mobile phase was adjusted to 4.5 with the help of diluted orthophosphoric acid. The diode array detector (DAD) was operated at 205 nm and 294 nm for the detection of CAM and LVX, respectively. Limit of detection (LOD) values for CAM and LVX were 1.22 µg/mL and 0.79 µg/mL, respectively. Limit of quantification (LOQ) values for CAM and LVX were 4.08 µg/mL and 2.64 µg/mL, respectively. The method was precise as % relative standard deviation (% RSD) values were less than 2% for all types of precision. This method was successfully applied to determine the percentage contents of both drugs in the pharmaceutical preparation.