Development and validation of an HPLC method for the determination of vancomycin in human plasma and its comparison with an immunoassay (PETINIA)

Abstract

Vancomycin (VAN) is among those antibiotics for which therapeutic drug monitoring is highly recommended. For this purpose a reliable method with small sample volume was required for quantification of VAN in human plasma. Therefore, a selective and sensitive method of high performance liquid chromatography was developed and validated. The separation was carried out isocratically by using a mobile phase NH4H2PO4 (50 mM, pH 2.2)–acetonitrile (88:12, v/v) at a flow rate of 0.36 mL/min on a nucleodur C18 column (125 mm × 4.6 mm, 5 µm) with UV detection at 205 nm. Sample preparation was done by deproteination of plasma with 70 % perchloric acid and a liquid/liquid extraction. Validation was performed according to the European Medicines Agency guideline. The method showed linearity over the range of 0.25–60 mg/L with a coefficient of determination r2 ≥ 0.999 and a lower limit of quantification of 0.25 mg/L. No interference was observed in blank plasma samples at the retention time of VAN. The percentage relative recovery and coefficient of variation (CV%) values for accuracy and precision were within the acceptable limits. Stability was proved at room temperature for 24 h, after repeated freeze and thaw cycles and storage at −20 °C for 3 months. A good correlation was observed (r = 0.947) by comparing with the results of an immunoassay (PETINIA, Siemens) in 289 samples. In conclusion the method proved simple, sensitive and cost effective for quantification of VAN in human plasma.

Establishment of an ultrasensitive indirect competitive time-resolved fluoroimmunoassay for vancomycin determination

“…It was found that the IC 50 value was 13.74 ng/L, the IC 20 -IC 80 were 3.43-43.62 ng/L, and the IC 10 reached up to 1.07 ng/L according to the established IC-TRFIA for Van. The LOD value was 1.07 ng/L in the presented IC-TRFIA for Van, which is more sensitive (approximately 60-fold) compared with Kong et al (2017) established IC-ELISA for Van with LOD value of 0.06 ng/mL, and approximately 460-fold, 2.0 × 10 5 -fold and 2.5 × 10 5 -fold compare with Liang et al (2015), Sheng and Zhou (2017) and Usman and Hempel (2016) which were established fluorescence "turn-off" biosensor, two-dimensional liquid chromatography and HPLC for Van with LOD value of 0.46 ng/mL, 0.2 and 0.25 μg/mL respectively. In addition, it was slightly better than Liu, Yan, Hua, and Wang (2013), Hu et al (2018), Zhou et al (2015) and Sheng et al (2016) established IC-TRFIAs for thiacloprid, aflatoxin B 1 , chloramphenicol and diniconazole with LOD value of 1.9, 3.55, 8.0 and 21 ng/L, respectively.…”

Correlation between 61 risk factors and vancomycin blood concentration reached based on real world data

“…Thus, it is clinically necessary to monitor VAN blood concentration. Monitoring methods mainly include high-performance liquid chromatography, radioimmunoassay, and enzyme amplification immunoassay [8]. Since VAN is a time-dependent antibacterial drug, the key to ensuring its efficacy is extending the antibacterial drug infusion time such that the drug concentration is always higher than the target organism's minimum inhibitory concentration [9].…”

Design and development of novel fluorescence-based detection methods for bio-analytes

“…HPLC is a type of chromatographic separation technique that is commonly coupled with UV, [55][56][57] (photo)diode array (DA), [58][59] fluorescence, 60 mass spectrometry (MS) 51,[61][62][63] or electrochemical 64 detector for the identification and quantification of the analyte(s) of interest. Due to its high sensitivity and specificity, HPLC has been considered the gold standard for Van quantification among the different analytical methods.…”

“…58 Especially in complicated biological fluids, HPLC proves to be an accurate and reliable tool to detect Van at concentrations below the therapeutic range which are often unattainable by immunoassays. [55][56] Another advantage of HPLC over immunoassays is the ability to isolate and quantify the degraded products of Van. [55][56] Furthermore, a simultaneous detection of Van and other commonly co-prescribed antibiotics has been performed on HPLC systems which improves the efficiency of analysis.…”

“…[55][56] Another advantage of HPLC over immunoassays is the ability to isolate and quantify the degraded products of Van. [55][56] Furthermore, a simultaneous detection of Van and other commonly co-prescribed antibiotics has been performed on HPLC systems which improves the efficiency of analysis. 53,63 However, traditional HPLC systems have several drawbacks 50,56,[61][62] that limited its widespread use in routine laboratories and prompt ongoing efforts to improve existing HPLC techniques.…”