Voriconazole In Human Plasma Research Articles

Simultaneous Quantification of Vancomycin, Linezolid and Voriconazole in Human Plasma by UHPLC-MS/MS: Application in Therapeutic Drug Monitoring.

Individual differences challenge the treatment of vancomycin, linezolid and voriconazole in severe infections. This study aimed to build a simple and economical method for simultaneous determination of the three antibiotics in human plasma by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and provided a reference for therapeutic drug monitoring (TDM) of infected patients. The plasma samples were precipitated by acetonitrile and detected and separated on a shim-pack GIST C18 column following the gradient elution within 5min. Mass quantification was performed on multiple reaction monitoring mode under positive electrospray ionization. The linear ranges of vancomycin, linezolid and voriconazole were 1.00-100.00, 0.10-15.00 and 0.10-20.00μg·mL-1, respectively, with good linearity (R2 > 0.99). The accuracy and precision, matrix effect, extraction recovery and stability were validated, and the results all meet the acceptance criteria of China Food and Drug Administration (CFDA) guidelines. The UHPLC-MS/MS method was established and validated for the simultaneous determination of vancomycin, linezolid and voriconazole in human plasma and successfully applied to routine TDM for individualized treatment.

Simultaneous quantification of venetoclax and voriconazole in human plasma by UHPLC-MS/MS and its application in acute myeloid leukemia patients

Venetoclax, an orally bioavailable BCL‐2 inhibitor, has been regarded as a breakthrough for the treatment of leukemia but has a wide interindividual variability and drug-drug interaction in pharmacokinetics. In this study, a simple and sensitive ultra-high performance liquid chromatography-tandem with mass spectrometry method was established and fully validated to quantify venetoclax and voriconazole simultaneously in human plasma. After protein precipitation, the analytes were separated on a Hypersil GOLD C18 column (3 µm, 2.1 × 50 mm) by gradient elution. The mass detection was operated under multiple reaction monitoring mode at m/z 868.5 →636.2 for venetoclax, 350.0 → 127.0 for voriconazole and 353.0 → 127.0 for voriconazole-D3(Internal Standard). The calibration ranges were 0.1–10 μg/mL for venetoclax and 0.05–10 μg/mL for voriconazole with correlation coefficients (r2)＞0.998. The validated method was successfully applied to the pharmacokinetic study in acute myeloid leukemia patients receiving venetoclax with or without voriconazole. The results suggested that co-treatment with 200 mg q12h voriconazole, the peak concentration of venetoclax (100 mg qd) could be raised to the same level as the 400 mg qd group. However, the trough concentration of venetoclax (100 mg qd) was much higher than that of the 400 mg qd group. Therapeutic drug monitoring might give some guidance for the adjustment of dosing regimens to guarantee the drug efficacy and safety of patients to some extent.

Determination of Voriconazole in Human Plasma by Liquid Chromatography Tandem Mass Spectrometry: Application in Therapeutic Drug Monitoring

Determination of Voriconazole in Human Plasma by Liquid Chromatography Tandem Mass Spectrometry: Application in Therapeutic Drug Monitoring

Rapid Quantitative Detection of Voriconazole in Human Plasma Using Surface-Enhanced Raman Scattering.

There is an increasing demand for rapid detection techniques for monitoring the therapeutic concentration of voriconazole (VRC) in human biological fluids. Herein, a rapid and selective surface-enhanced Raman scatting method for point-of-care determination of VRC in human plasma was developed via a portable Raman spectrometer. This approach has enabled the quantification of the VRC spiked into human plasma at clinical relevant concentrations. A gold nanoparticle solution (Au sol) was used as the SERS substrate, and the agglomerating conditions on its sensitivity were optimized. The method involves the formation of hot spots, and the signal of VRC molecules adsorbed on the surface of the SERS hot spot was amplified by 105. The calibration curve was linear in the range of 0.02-10 ppm, with satisfactory repeatability. The limit of detection was as low as 12.3 ppb. The variation in VRC spectra over time on different substrates demonstrated good reproducibility. Notably, the salting-out extraction method developed in this study was rapid and suitable for the quantitation of drugs in biological samples. Compared with traditional methods, this approach allows for the point-of-care quantification of VRC directly in a complex matrix, which may open up new exciting opportunities for future use of the SERS technique in clinical applications.

Comparison of LC-MS3 and LC-MRM Method for Quantifying Voriconazole and Its Application in Therapeutic Drug Monitoring of Human Plasma.

The TDM of voriconazole which exhibits wide inter-individual variability is indispensable for treatment in clinic. In this study, a method that high-performance liquid chromatography tandem mass spectrometry cubed (HPLC-MS3) is first built and validated to quantify voriconazole in human plasma. The system is composed of Shimadzu Exion LCTM UPLC coupled with a Qtrap 5500 mass spectrometer. The separation of voriconazole is performed on a Poroshell 120 SB-C18 column at a flow rate of 0.8 mL/min remaining 7 min for each sample. The calibration curves are linear in the concentration range of 0.25–20 μg/mL. Intra-day and inter-day accuracies and precisions are within 8.0% at three concentrations, and the recoveries and matrix effect are all within accepted limits. In terms of stability, there is no significant degradation of voriconazole under various conditions. The HPLC-MS3 and HPLC-MRM (multiple reaction monitoring) methods are compared in 42 patients with Passing–Bablok regression and Bland–Altman plots, and the results show no significant difference between the two methods. However, HPLC-MS3 has a higher S/N (signal-to-noise ratio) and response than the MRM. Finally, the HPLC-MS3 assay is successfully applied to monitor the TDM (therapeutic drug monitoring) of voriconazole in human plasma, and this verifies that the dosing guidelines for voriconazole have been well implemented in the clinic and patients have received excellent treatment.

Determination of Voriconazole in Human Plasma Using RP-HPLC/UV-VIS Detection: Method Development and Validation; Subsequently Evaluation of Voriconazole Pharmacokinetic Profile in Pakistani Healthy Male Volunteers

Abstract In this research work, an isocratic, reversed-phase high-performance liquid chromatography-ultraviolet/visible detector method was developed for analysis of voriconazole standard (stock-solution) and in plasma samples. Optimization and validation of the method was carried out as per international guidelines. The method offered a simple liquid–liquid extraction technique, which exhibited best recovery of voriconazole along with fluconazole, i.e., internal standard. Different experimental conditions were tried and ultimately, the best outcomes were accomplished utilizing C-18 Perkin-Elmer® column with particulars of 150 mm length, 4.6 mm inner diameter and 5 μm particle size, protected by an RP-18 Perkin-Elmer® Pre-column guard cartridge with specifications of 10 μm particle size, 30 mm length and 4.6 mm inner diameter, utilizing mobile-phase of acetonitrile-water (ACN: H2O) in proportion of 60: 40 v/v, having a flow rate of 1.5 mL/min, and wavelength of 254 nm. All the analytes were observed to be separated in ≤7 min. A linear calibration curve was obtained at concentration range of 01–10 μg/mL of voriconazole. The correlation coefficient of voriconazole was observed to be 0.999, and average recovery (in percent) was 97.4%, whereas the relative standard deviation value was ≤2%. The lower limit of detection was 0.01 μg/mL, whereas, lower limit of quantification was 0.03 μg/mL, respectively. This developed method provided outstanding results of all validation parameters, i.e., recovery, accuracy, selectivity, precision and reproducibility. The method proposed for voriconazole analysis was applied effectively for further research investigation of voriconazole in human-plasma samples (to assess the pharmacokinetic parameters), pharmaceutical formulations and pharmacokinetic drug–drug interaction’s.

Determination of voriconazole in human plasma by liquid chromatography-tandem mass spectrometry and its application in therapeutic drug monitoring in Chinese patients.

ObjectiveTo develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of voriconazole in human plasma, and to evaluate its application in clinical therapeutic drug monitoring.MethodPlasma samples were obtained from Chinese patients receiving voriconazole, precipitated with methanol (using fluconazole as an internal standard), and then subjected to LC-MS/MS using an SB C18 column with a methanol and water mobile phase at a flow rate of 0.4 mL/minute. Quantification was performed by multiple-reaction monitoring using the precursor and product ion pair m/z 350–280.9 for voriconazole and m/z 307–219.9 for fluconazole.ResultsThe calibration curve was linear over a range of 0.1–10.0 µg/mL (R2 = 0.9995). The inter-day and intra-day relative standard deviations were <7.68% and <8.97%, respectively. Extraction recovery, matrix effect, and stability were also validated. Sixty-eight plasma samples from 42 patients were analyzed, and the voriconazole concentrations in 25 samples (36.8%) were outside the optimal range of 1.5–4.5 µg/mL.ConclusionsWe developed a simple and accurate method of drug monitoring, which could improve the efficacy and prevent adverse reactions of voriconazole.

Determination of total and free voriconazole in human plasma: Application to pharmacokinetic study and therapeutic monitoring

•Voriconazole (VCZ) is highly efficient in the treatment of invasive fungal infections. However, the therapy requires the monitoring of VCZ plasma concentration, due to a narrow therapeutic index and large inter-individual variability of pharmacokinetics of VCZ, which often exhibits nonlinearity. Total plasma concentration of VCZ does not always provide sufficient information, especially in hemato-oncology patients, who often show significant fluctuations in plasma protein concentration. Therefore, the monitoring of free drug seems crucial for a complete understanding of the ongoing antifungal therapy.•The study aimed to develop an HPLC-FLD method for the determination of total and free VCZ in clinical samples.•The analysis of total VCZ comprised the one-step samples preparation by plasma protein precipitation. The free drug was isolated from plasma by ultrafiltration. The non-specific bindings (NSB) between the drug and filter membranes were comprehensively evaluated.•The elaborated method fulfilled the validation requirements for bioanalytical methods. The ultrafiltration technique is recommended for the analysis of free VCZ, but requires a preconditioning step. Ultrafiltration was impacted with adsorption of the drug on the filter membrane, which was as high as 61–82% and was found to decrease after pretreatment of the membranes with 5% Tween 80. The method was applied for the quantification of plasma VCZ in patients undergoing antifungal therapy. Total and free VCZ concentrations ranged from 0.21 to 4.92 μg/mL and from 0.07 to 1.28 μg/mL, respectively; the mean protein binding was 47%.

Content detemination of voriconazole in human plasma by HPLC method and its application

Content detemination of voriconazole in human plasma by HPLC method and its application

Reliable and Easy-To-Use Liquid Chromatography-Tandem Mass Spectrometry Method for Simultaneous Analysis of Fluconazole, Isavuconazole, Itraconazole, Hydroxy-Itraconazole, Posaconazole, and Voriconazole in Human Plasma and Serum.

A fast and easy-to-use liquid chromatography-tandem mass spectrometry method for the determination and quantification of 6 triazoles [fluconazole (FLZ), isavuconazole (ISZ), itraconazole (ITZ), hydroxy-itraconazole (OH-ITZ), posaconazole (PSZ), and voriconazole (VRZ)] in human plasma and serum was developed and validated for therapeutic drug monitoring. Sample preparation was based on protein precipitation with acetonitrile and subsequent centrifugation. Isotope-labeled analogues for each analyte were used as internal standards. Chromatographic separation was achieved using a 50 × 2.1 mm, 1.9 μm polar Hypersil Gold C18 column and mobile phase consisting of 0.1% formic acid/acetonitrile (45%/55%, vol/vol) at a flow rate of 340 μL/min. The triazoles were simultaneously detected using a triple-stage quadrupole mass spectrometer operated in selected reaction monitoring mode with positive heated electrospray ionization within a single runtime of t = 3.00 minutes. Linearity of all azole concentration ranges was verified by the Mandel test and demonstrated for all azoles. All calibration curves were linear and fitted using least squares regression with a weighting factor of the reciprocal concentration. Limits of detection (μg/L/L) were FLZ, 9.3; ISZ, 0.3; ITZ, 0.6; OH-ITZ, 8.6; PSZ, 3.4; and VRZ, 2.1. The lower limits of quantitation (μg/L/liter) were FLZ, 28.3; ISZ, 1.0; ITZ, 1.7; OH-ITZ, 26.2; PSZ, 10.3; and VRZ, 6.3. Intraday and interday precisions ranged from 0.6% to 6.6% for all azoles. Intraday and interday accuracies (%bias) of all analytes were within 10.5%. In addition, we report on a 29-year-old white woman (94 kg body weight) with a history of acute myeloid leukemia who underwent stem cell transplantation. Because of diagnosis of aspergillus pneumonia, antifungal pharmacotherapy was initiated with different application modes and dosages of ISZ, and plasma concentrations were monitored over a time period of 6 months. A precise and highly sensitive liquid chromatography-tandem mass spectrometry method was developed that enables quantification of triazoles in plasma and serum matrix across therapeutically relevant concentration ranges. It was successfully implemented in our therapeutic drug monitoring routine service and is suitable for routine monitoring of antifungal therapy and in severely ill patients.

Therapeutic drug monitoring of vancomycin and voriconazole by liquid chromatography-tandem mass spectrometric method

A reliable liquid chromatography-tandem mass spectrometric(LC-MS/MS) method was developed and validated for the simultaneous determination of vancomycin and voriconazole in human plasma. The analytes and internal standard 10-hydroxycarbazepine were separated at a flow rate of 0.9 mL/min using a Zorbax SB-C18 column (50 mm×4.6 mm, 2.7 μm). Positive ion electrospray ionization was used to detect vancomycin, voriconazole and intrenal standard(IS) 10-hydroxycarbazepine followed by multiple reaction monitoring(MRM) of the transition at m/z 725.5→144.2, 350.3→281.0 and 253.1→208.0, respectively. The total run time for both vancomycin and voriconazole samples was 5 min; 0.30 μg/mL was the lower limit of quantification. The precision of intraday and interday was no more than 12.4%. The method was successfully and resoundingly applied in therapeutic drug monitoring of 156 patients treated with vancomycin and voriconazole.

Ultra-fast quantitation of voriconazole in human plasma by coated blade spray mass spectrometry

Voriconazole is a triazole broad-spectrum antifungal medication often used to treat fungal infections caused by Aspergillus and Fusarium species. One of the main challenges associated with the implementation of this medication is its narrow therapeutic concentration range, demonstrating toxicity at concentrations above 6μg/mL and limited efficacy at concentrations below 2μg/mL. As a result, methodologies which permit the rapid and accurate quantitation of voriconazole in patients are highly desirable. In this work two different approaches based on coated blade spray directly coupled to mass spectrometry (CBS-MS) are introduced; each enabling the quantitation of voriconazole in plasma samples with a simple and fast sample preparation and no chromatographic step. The first approach involves a rapid extraction (1min) of the target analyte from 300μL of human plasma using conventional laboratory vessels (e.g. vial, 96-well plate). Alternatively, the second strategy consists of a 2min extraction from a plasma droplet (10μL) placed on the coated area of the blade. Both procedures were successfully validated and good linearity (R2≥0.998), accuracy (91–122%) and precision (<8%) were attained in the concentration range evaluated (0.1–50μg/mL). Moreover, very good results in terms of relative matrix effects were obtained given that the slopes of the calibration curves constructed in five different plasma lots exhibited relative standard deviation (RSD) values below 7%. Herein we demonstrated that CBS-MS is a technology suitable for the ultra-fast determination of voriconazole in human plasma samples. Indeed, the proposed methodology can be easily used either for routine drug monitoring or for in vitro pharmacokinetic studies in applications where very small sample volumes are available and great temporal resolution is needed.

UPLC-MS/MS determination of voriconazole in human plasma and its application to a pharmacokinetic study.

A sensitive and rapid ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed to determine voriconazole in human plasma. Sample preparation was accomplished through a simple one-step protein precipitation with methanol. Chromatographic separation was carried out on an Acquity UPLC BEH C18 column using an isocratic mobile phase system composed of acetonitrile and water containing 1% formic acid (45:55, v/v) at a flow rate of 0.50 mL/min. Mass spectrometric analysis was performed using a QTrap5500 mass spectrometer coupled with an electrospray ionization source in the positive ion mode. The multiple reaction monitoring transitions of m/z 351.0 → 281.5 and m/z 237.1 → 194.2 were used to quantify voriconazole and carbamazepine (internal standard), respectively. The linearity of this method was found to be within the concentration range of 2.0-1000 ng/mL with a lower limit of quantification of 2.0 ng/mL. Only 1.0 min was needed for an analytical run. This fully validated method was successfully applied to the pharmacokinetic study after oral administration of 200 mg voriconazole to 20 Chinese healthy male volunteers.

Determination of Voriconazole in Human Plasma by HPLC–ESI-MS and Application to Pharmacokinetic Study

A fast, sensitive, high-performance liquid chromatography-tandem mass spectrometry method was developed for the determination of voriconazole in human plasma. Carbamazepine was used as the internal standard and the sample pretreatment involved one-step protein precipitation. Chromatographic separation was conducted on an Ultimate C18 column with a mobile phase consisting of acetonitrile-water (containing 0.1% formic acid; 40:60, v/v) at a flow rate of 0.3 mL/min. The detection of voriconazole was performed on a triple-quadrupole tandem mass spectrometer by multiple reaction monitoring with an electrospray ionization source in the positive mode. The standard curve was linear (r(2) ≥ 0.99) within the concentration range of 2.49-293 ng/mL. The intra-day and inter-day precision values were below 5.3%, and the accuracy was within -4.3-5.7%. The method was applicable to the clinical study of the pharmacokinetics of voriconazole in healthy volunteers following oral administration.

Fast and simple LC–MS/MS method for quantifying plasma voriconazole

BackgroundAn increasing number of publications point to the importance of voriconazole plasma monitoring. Our aim was to develop a fast and simple LC–MS/MS method for the quantification of plasma voriconazole. Methods20μL of plasma was extracted by adding a precipitation reagent containing the internal standard (d3-voriconazole). Analysis was performed on a Quattro Micro tandem mass spectrometer equipped with an Alliance HPLC 2795 separations module. MRM transitions were m/z 350.0→281.4 for voriconazole and m/z 353.0→284.4 for d3-voriconazole. Quantification was done by both linear calibration curves and multiplication of the response ratio by a predefined factor. ResultsA method using protein precipitation as only pretreatment with an analysis time of 2min was developed. Within- and between-run precision, expressed as coefficient of variation, at 6 concentrations ranged from 2.8% to 11.7%. Accuracy, expressed as a percentage of the theoretically added concentration, ranged from 89.0% to 109.0%. Linearity was demonstrated from 0.06mg/L (0.17μM) to 20.0mg/L (57.3μM). ConclusionsThe described method offers a simple and rapid analysis of voriconazole in human plasma for clinical routine. Quantification with a predefined factor permits omitting calibration to each run and thereby reduces workload, costs and turn-around time.

Ultra-performance liquid chromatographic method for measurement of voriconazole in human plasma and oral fluid

A simple, sensitive and selective ultra-performance liquid chromatography method for the determination of voriconazole in plasma and oral fluid was developed and validated. After a liquid-liquid extraction with methyl-tert-butyl ether, the analyte and internal standard were separated on a Hypersil Gold C18 column (2.1 × 100 mm, p.d. 1.9 µm), eluted with a mobile phase composed of thietylammonium phosphate buffer and acetonitrile (70:30, v/v). Total run time was 4 min, total mobile phase consumption of 2.2 mL. Detection was performed with a photodiode array detector with quantitation at 256 nm. Voriconazole concentrations in oral fluid were on average 57.5% (±5.3) of those measured in paired plasma samples.

Quantification of voriconazole in human plasma by high-performance liquid chromatography–electrospray ionization mass spectrometry: application to a bioequivalence study

A sensitive and specific liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) method was developed and validated for the identification and quantification of voriconazole (VRC, CAS 137234-62-9) in human plasma. Following liquid-liquid extraction, VRC and loratadine (internal standard, CAS 79794-75-5) were separated using a mobile phase comprised of methanol: water (0.1% formic acid) = 75:25 v/v on a Shimadzu Shim-pack VP-ODS C18 (150 x 2.0 mm ID, 5 microm) column and analyzed by electrospray ionization mass spectrometry. The chromatographic separation was achieved in less than 6 min. The standard curves were linear (r = 0.9994) over the concentration range of 2-2000 ng/mL for VRC and had good accuracy and precision. Both intra- and inter-batch standard deviations were less than 15%. The method was successfully applied to study the comparative bioavailability of VRC tablets test vs. reference in healthy Chinese volunteers through the statistical comparison of pharmacokinetic parameters obtained with the two formulations.

Fast and Reliable Determination of Voriconazole in Human Plasma by LC-APCI-MS/MS

A fast and reliable liquid chromatography coupled with atmospheric pressure chemical ionization tandem mass spectrometry (LC-APCI-MS/MS) method was developed and validated for the quantification of voriconazole in human plasma. The proposed method was validated in a linear range of 50-10,000 ng/ml, and the total run time was 1.5 min. This method was successfully used to support routine therapeutic drug monitoring of voriconazole.

Determination of voriconazole in human serum and plasma by micellar electrokinetic chromatography

The micellar electrokinetic capillary chromatography (MEKC) separation and analysis of voriconazole and UK 115794 (internal standard) were examined and an assay for determination of voriconazole in human plasma and serum was developed. The MEKC medium comprises a 2:15 (v/v) mixture of methanol and a pH 9.3 buffer composed of 5 mM Na 2B 4O 7, 7 mM Na 2HPO 4 and 54 mM SDS. Sample preparation is based upon liquid/liquid extraction with ethylacetate and dichloromethane (75%/25%) at physiological pH. Using this approach with 250 μl serum or plasma and reconstitution of the dried extract into 100 μl of a buffer composed of 0.5 mM Na 2B 4O 7 and 0.7 mM Na 2HPO 4 (pH 9.3), the detection and quantitation limits were determined to be 0.1 and 0.2 μg/ml, respectively, a sensitivity that is suitable for therapeutic drug monitoring of voriconazole (provisional therapeutic range: 1–6 μg/ml) in human plasma and serum samples. The method was validated and compared to an HPLC method, showing excellent agreement between the two for a set of 91 samples that stemmed from patients being treated with voriconazole. The MEKC assay is also demonstrated to be suitable to explore pharmacokinetic data of voriconazole.

Determination of voriconazole in human plasma and saliva using high-performance liquid chromatography with fluorescence detection

Voriconazole is a widely used triazole antifungal agent with a broad spectrum including Aspergillus species. A simple, sensitive and selective high-performance liquid chromatography method for the determination of voriconazole in human plasma and saliva was developed. Drug and internal standard (UK-115 794) were extracted from alkaline plasma and saliva with n-hexane–ethyl acetate (3:1, v/v) and analyzed on a Luna C 18 column with fluorimetric detection set at excitation and emission wavelengths of 254 and 372 nm, respectively. The calibration curve was linear through the range of 0.1–10 μg/ml using a 0.3 ml sample volume. The intra- and inter-day precisions were all below 6.1% for plasma and below 9.1% for saliva. Accuracies ranged from 94 to 109% for both matrices. Mean recovery was 86 ± 4% for voriconazole. The method showed acceptable values for precision, recovery and sensitivity and is well suited for routine analysis work and for pharmacokinetic studies.