Development of a liquid chromatography tandem mass spectrometry method for the simultaneous measurement of voriconazole, posaconazole and itraconazole

2017 ◽  
Vol 54 (6) ◽  
pp. 686-695 ◽  
Author(s):  
John M Wadsworth ◽  
Anna M Milan ◽  
James Anson ◽  
Andrew S Davison
Keyword(s):  
Therapeutic Drug Monitoring ◽  
Drug Monitoring ◽  
Matrix Effects ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Gradient Elution ◽  
Turnaround Time ◽  
Therapeutic Drug ◽  
Tandem Mass ◽  
Lower Limits

Background Azole-based antifungals are the first-line therapy for some of the most common mycoses and are now also being used prophylactically to protect immunocompromised patients. However, due to variability in both their metabolism and bioavailability, therapeutic drug monitoring is essential to avoid toxicity but still gain maximum efficacy. Methods Following protein precipitation of serum with acetonitrile, 20  µL of extract was injected onto a 2.1 × 50 mm Waters Atlantis dC18 3  µm column. Detection was via a Waters Quattro Premier XE tandem mass spectrometer operating in ESI-positive mode. Multiple reaction monitoring (MRM) detected two product ions for each compound and one for each isotopically labelled internal standard. Ion suppression, linearity, stability, matrix effects, recovery, imprecision, lower limits of measuring interval and detection were all assessed. Results Optimal chromatographic separation was achieved using gradient elution over 8 minutes. Voriconazole, posaconazole and itraconazole eluted at 1.71, 2.73 and 3.41 min, respectively. The lower limits of measuring interval for all three compounds was 0.1 mg/L. The assay was linear to 10 mg/L for voriconazole (R2 = 0.995) and 5 mg/L for posaconazole (R2 = 0.990) and itraconazole (R2 = 0.991). The assay was both highly accurate and precise with % bias of voriconazole, posaconazole and itraconazole, respectively, when compared with previous NEQAS samples. The intra-assay precision (CV%) was 1.6%, 2.5% and 1.9% for voriconazole, posaconazole and itraconazole, respectively, across the linear range. Conclusion A simple and robust method has been validated for azole antifungal therapeutic drug monitoring. This new assay will result in a greatly improved sample turnaround time and will therefore vastly increase the clinical utility of azole antifungal drug monitoring.

scholarly journals Simultaneous Quantification of Methotrexate and Its Metabolite 7-Hydroxy-Methotrexate in Human Plasma for Therapeutic Drug Monitoring

2019 ◽  
Vol 2019 ◽  
pp. 1-10 ◽  
Author(s):  
Xinxin Ren ◽  
Zhipeng Wang ◽  
Yunlei Yun ◽  
Guangyi Meng ◽  
Xialan Zhang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Therapeutic Drug Monitoring ◽  
Human Plasma ◽  
Drug Monitoring ◽  
Internal Standard ◽  
Gradient Elution ◽  
Therapeutic Drug ◽  
Tandem Mass ◽  
The Matrix ◽  
Analytical Time

Objective. To establish and validate a simple, sensitive, and rapid liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the determination of methotrexate (MTX) and its major metabolite 7-hydroxy-methotrexate (7-OH-MTX) in human plasma. Method. The chromatographic separation was achieved on a Zorbax C18 column (3.5 μm, 2.1 × 100 mm) using a gradient elution with methanol (phase B) and 0.2% formic acid aqueous solution (phase A). The flow rate was 0.3 mL/min with analytical time of 3.5 min. Mass spectrometry detection was performed in a triple-quadruple tandem mass spectrometer under positive ion mode with the following mass transitions: m/z 455.1/308.1 for MTX, 471.0/324.1 for 7-OH-MTX, and 458.2/311.1 for internal standard. The pretreatment procedure was optimized with dilution after one-step protein precipitation. Results. The calibration range of methotrexate and 7-OH-MTX was 5.0-10000.0 ng/mL. The intraday and interday precision and accuracy were less than 15% and within ±15% for both analytes. The recovery for MTX and 7-OH-MTX was more than 90% and the matrix effect ranged from 97.90% to 117.60%. Conclusion. The method was successfully developed and applied to the routine therapeutic drug monitoring of MTX and 7-OH-MTX in human plasma.

An UHPLC-UV Method for Determination of Vancomycin in human serum

2020 ◽  
Vol 16 ◽  
Author(s):  
Fang Fang ◽  
Ning Li ◽  
Chunli Xu ◽  
Rong Tan ◽  
Jihong Yang ◽  
...  
Keyword(s):  
Therapeutic Drug Monitoring ◽  
Human Serum ◽  
Drug Monitoring ◽  
Internal Standard ◽  
Sample Pretreatment ◽  
Gradient Elution ◽  
Therapeutic Drug ◽  
Serum Samples ◽  
Column Temperature ◽  
Limit Of Quantitation

Objective: To develop a rapid ultra-performance liquid chromatographic (UHPLC)-UV method for vancomycin determination in human serum for therapeutic drug monitoring (TDM). Methods: Human serum samples were precipitated with 10% perchloric acid, and the supernatant after centrifugation was analyzed on an ACQUITY UHPLC BEH C18 column (2.1 × 50mm, 1.7 μm) via gradient elution with a flow rate at 0.3 mL/min. The mobile phase consisted of acetonitrile and 0.005M KH2PO4 buffer (containing 0.1% triethylamine, pH 3.4). The detection wavelength was set at 210 nm, and the column temperature was set at 40. The total runtime was 6.0 min per analysis. Results: After comprehensive validation, the method was applied to determine the concentration of vancomycin in human serum. The chromatographic peaks of vancomycin and internal standard were not interfered by endogenous matrixes. The retention time (RT) of vancomycin was 1.91 min, while the internal standard was 1.58 min. The good linearity range of vancomycin concentration was 2.5-120 μg/mL (R2>0.999). The lower limit of quantitation (LLOQ) was 2.5 μg/mL. The precision at three quality control (QC) levels (including LLOQ) was restricted within 85-115%. The extraction recovery rate of QC samples (4.0, 20.0, 60.0 μg/mL) were 101.16%、97.70%、94.90%, respectively. Inter- and intra-day precision was less than 8% (RSD). Stability tests under different storage conditions were satisfactory. In patients, the concentration of vancomycin ranged from 7.30 to 89.12 μg/mL determined by the fully validated method. Conclusion: The simple, rapid sample pretreatment procedures and short analysis time made this UHPLC-UV method suitable for therapeutic drug monitoring (TDM) of vancomycin.

A Validated LC-MS/MS Method for the Determination of Mezlocillin in Plasma: an Adapted Method for Therapeutic Drug Monitoring in Children

2020 ◽  
Vol 16 ◽  
Author(s):  
Bo-Hao Tang ◽  
Min Kan ◽  
Xin-Mei Yang ◽  
Rong-Hua Wang ◽  
Hai-Yan Shi ◽  
...  
Keyword(s):  
Therapeutic Drug Monitoring ◽  
Trough Concentration ◽  
Drug Monitoring ◽  
Respiratory Infections ◽  
Internal Standard ◽  
Gradient Elution ◽  
Therapeutic Drug ◽  
Limit Of Quantification ◽  
Liquid Chromatography Tandem Mass

Background: Mezlocillin is off-label used for the treatment of respiratory infections in children. Therapeutic drug monitoring (TDM) data are also limited in children. A sensitive Liquid chromatography tandem mass spectrometry (LC–MS/MS) method adapted to children was developed and validated for the determination of mezlocillin plasma concentration in present study. Method: Mezlocillin, extracted from a volume of 50 μL plasma using acetonitrile, was analyzed on an online LC–MS/MS system with an Agilent 1290 Infinity UHPLC (Agilent Technologies, CA, USA) coupled to an AB SCIEX QTRAP 6500PLUS MS/MS (AB Sciex, Framingham, MA, USA) with ceftiofur as an internal standard. HPLC separation was performed on a C18 column with ultrapure water and acetonitrile as gradient elution at a flow rate of 0.4 mL/min at 30o C. Analyst TM Version 1.5.2 (Applied Biosystems) was used for data acquisition. The total chromatographic run time was 1.6 min. Results: LC/MS/MS method used for TDM of mezlocillin in children was developed and validated. This assay has a lower limit of quantification of 0.025 μg/mL for mezlocillin with 50 μL plasma. Good linearity was achieved for mezlocillin over the range 0.025 to 20 μg /mL. The acceptance criteria were met in all cases. Among 36 patients aged 0.16-1.63 years old, only one patient had detectable trough concentration higher than 1 μg/mL. Conclusion: LC-MS/MS method with 50 μL plasma developed in our study was successfully applied to TDM of mezlocillin in children. The high variability of trough concentration highlighted TDM is important to optimize mezlocillin therapy in children.

scholarly journals ANALYSIS OF 4-HYDROXYCYCLOPHOSPHAMIDE IN CANCER PATIENTS PLASMA FOR THERAPEUTIC DRUG MONITORING OF CYCLOPHOSPHAMIDE

2016 ◽  
Vol 8 (9) ◽  
pp. 194 ◽  
Author(s):  
Yahdiana Harahap ◽  
Christian Samuel ◽  
Rizka Andalusia ◽  
Nadia Farhanah Syafhan
Keyword(s):  
Therapeutic Drug Monitoring ◽  
Cancer Patients ◽  
Drug Monitoring ◽  
High Performance ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Therapeutic Drug ◽  
Mass Detection ◽  
Column Temperature ◽  
Liquid Chromatography Tandem Mass

<p><strong>Objective</strong><strong>:</strong><strong> </strong>To quantify 4-hydroxycyclophosphamide in cancer patients’ plasma for therapeutic drug monitoring of cyclophosphamide.<strong></strong></p><p><strong>Methods</strong><strong>:</strong><strong> </strong>The blood was collected at 0.5 and 1 h after administration of chemotherapy. Prior to analysis, 4-OHCP in plasma was derivatized with semicarbazide HCl, then was extracted using 4 ml ethyl acetate and finally was determined by Ultra High-Performance Liquid Chromatography–tandem mass spectrometry. Chromatographic separation was conducted using waters acquity BEH C18 column (1.7 μm; 50 mm x 2.1 mm). The mobile phase consisted of formic acid 0.1% and methanol (50: 50, v/v), column temperature 30 °C and flow rate of 0.3 ml/min. Mass detection was performed on waters xevo TQD equipped with an electrospray ionization (ESI) source at positive ion mode in the multiple reaction monitoring (MRM). Cyclophosphamide was detected at m/z 260.968&gt;139.978, 4-hydroxycyclophosphamide-semicarbazide at m/z 338.011&gt;224.97, and hexamethylphosphoramide as internal standard at m/z 180.17&gt;92.08.</p><p><strong>Results</strong><strong>:</strong><strong> </strong>The method was linear in the range of 5–1000 ng/ml for cyclophosphamide and also for 4-hydroxycyclophosphamide. The results showed that the level of 4-OHCP in 39 cancer patients was in the range of 5.02 ng/ml to 832.44 ng/ml.<strong></strong></p><p><strong>Conclusion: </strong>4-hydroxycyclophosphamide was detected on 39 patient samples with the lowest level of 5.40ng/ml and the highest level was 832.44 ng/ml. This can be a parameter that the regiment of cyclophosphamide was effective.</p>

scholarly journals Direct analysis in real time-mass spectrometry for rapid quantification of five anti-arrhythmic drugs in human serum: application to therapeutic drug monitoring

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yuzhou Gui ◽  
Youli Lu ◽  
Shuijun Li ◽  
Mengqi Zhang ◽  
Xiaokun Duan ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Therapeutic Drug Monitoring ◽  
Tandem Mass Spectrometry ◽  
Human Serum ◽  
Real Time ◽  
Drug Monitoring ◽  
Multiple Reaction Monitoring ◽  
Direct Analysis ◽  
Therapeutic Drug ◽  
Tandem Mass

Abstract Therapeutic drug monitoring (TDM) is necessary for the optimal administration of anti-arrhythmic drugs in the treatment of heart arrhythmia. The present study aimed to develop and validate a direct analysis in real time tandem mass spectrometry (DART–MS/MS) method for the rapid and simultaneous determination of five anti-arrhythmic drugs (metoprolol, diltiazem, amiodarone, propafenone, and verapamil) and one metabolite (5-hydroxy(OH)-propafenone) in human serum. After the addition of isotope-labeled internal standards and protein precipitation with acetonitrile, anti-arrhythmic drugs were ionized by DART in positive mode followed by multiple reaction monitoring (MRM) detection. The use of DART–MS/MS avoided the need for chromatographic separation and allowed rapid and ultrahigh throughput analysis of anti-arrhythmic drugs in a total run time of 30 s per sample. The DART–MS/MS method yielded satisfactory linearity (R2 ≥ 0.9906), accuracy (86.1–109.9%), and precision (≤ 14.3%) with minimal effect of biological matrixes. The method was successfully applied to analyzing 30 clinical TDM samples. The relative error (RE) of the concentrations obtained by DART–MS/MS and liquid chromatography-tandem mass spectrometry (LC–MS/MS) was within ± 13%. This work highlights the potential usefulness of DART for the rapid quantitative analysis of anti-arrhythmic drugs in human serum and gives rapid feedback in the clinical TDM practices.

scholarly journals UPLC-MS/MS Method for Simultaneous Determination of 14 Antimicrobials in Human Plasma and Cerebrospinal Fluid: Application to Therapeutic Drug Monitoring

2022 ◽  
Vol 2022 ◽  
pp. 1-7
Author(s):  
Huiting Sun ◽  
Han Xing ◽  
Xueke Tian ◽  
Xiaojian Zhang ◽  
Jing Yang ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Therapeutic Drug Monitoring ◽  
Human Plasma ◽  
Drug Monitoring ◽  
Quantitative Methods ◽  
Multiple Reaction Monitoring ◽  
Gradient Elution ◽  
Therapeutic Drug ◽  
Limit Of Quantification ◽  
Acquity Uplc

Pharmacokinetics/pharmacodynamics is the foundation for guiding the rational application of antibiotics in clinical practice, so it is necessary to establish quantitative methods for accurate drug concentration determination. This study aimed to develop a rapid and simple ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for simultaneous quantification of 14 antibiotics (amikacin, etimicin, ceftazidime, cefepime, cefoperazone, ceftriaxone, daptomycin, latamoxef, linezolid, meropenem, biapenem, ampicillin, norvancomycin, and vancomycin) in human plasma and cerebrospinal fluid. Antibiotics were chromatographically separated on a Waters ACQUITY UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) via gradient elution within 3 minutes and were monitored using positive ion fitted with multiple reaction monitoring. The lower limit of quantification was 0.05–2.0 μg·mL−1. The method was verified according to the FDA bioanalysis method validation guidelines, which showed excellent accuracy (from 86.75% to 110.85%) and precision (from 0.46% to 10.97%). At last, this method was successfully applied to therapeutic drug monitoring in 113 patients under antibiotics treatment.

scholarly journals Validation and Application of an HPLC-UV Method for Routine Therapeutic Drug Monitoring of Cefiderocol

Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 242
Author(s):  
Julia Zimmer ◽  
Anka C. Röhr ◽  
Stefan Kluge ◽  
Jonas Faller ◽  
Otto R. Frey ◽  
...  
Keyword(s):  
Therapeutic Drug Monitoring ◽  
Critically Ill ◽  
Drug Monitoring ◽  
Critically Ill Patients ◽  
High Performance ◽  
Internal Standard ◽  
Gradient Elution ◽  
Resistant Bacteria ◽  
Therapeutic Drug ◽  
Serum Samples

Cefiderocol is a new siderophore cephalosporin approved for the treatment of multidrug resistant bacteria including activity against carbapenem-resistant Enterobacterales and Pseudomonas aeruginosa. As cephalosporins are known for their high pharmacokinetic variability in critically ill patients, cefiderocol therapeutic drug monitoring might become a valuable tool. Therefore, we aimed to develop and validate a simple, rapid, cost-effective high performance liquid chromatography (HPLC) method for the quantification of cefiderocol in serum. Samples were treated for protein precipitation followed by chromatographic separation on a reverse phase column (HPLC C-18) with gradient elution of the mobile phase. Cefiderocol was detected via UV absorption and quantification was performed with the internal standard (metronidazole) method. The calibration range showed linearity from 4 to 160 mg/L. The intra and interday precision was less than 10% with a recovery rate of 81%. The method was successfully used for the analysis of subsequent serum samples of critically ill patients and showed good performance in monitoring serum levels and optimizing antibiotic therapy.

scholarly journals Sensitive and Rapid Quantification of Busulfan in Small Plasma Volumes by Liquid Chromatography–Electrospray Mass Spectrometry

2001 ◽  
Vol 47 (8) ◽  
pp. 1437-1442 ◽  
Author(s):  
Thomas E Mürdter ◽  
Janet Coller ◽  
Alexander Claviez ◽  
Frank Schönberger ◽  
Ute Hofmann ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Therapeutic Drug Monitoring ◽  
Drug Monitoring ◽  
Internal Standard ◽  
Therapeutic Drug ◽  
High Dose ◽  
Selected Ion Monitoring ◽  
Hematopoietic Stem ◽  
Adults And Children ◽  
Liquid Chromatographic

Abstract Background: High-dose busulfan is widely used in conditioning regimens before hematopoietic stem cell transplantation in both adults and children. Large interindividual variability in pharmacokinetics after oral administration has been reported; therefore, therapeutic drug monitoring of busulfan may decrease the incidence of drug-related toxicity (for example, hepatic venoocclusive disease) and may also improve therapeutic efficacy. Methods: Busulfan concentrations were quantified using 200 μL of plasma and liquid–liquid extraction with diethyl ether after the addition of [2H8]busulfan as the internal standard. Separation and detection of busulfan and [2H8]busulfan were achieved with a LUNA C8 column (5 μm; 150 × 2 mm i.d.) at 30 °C, a HP 1100 liquid chromatography system, and a HP 1100 single-quadrupole mass spectrometer. Busulfan and [2H8]busulfan were detected as ammonium adducts in selected-ion monitoring mode at m/z 264.2 and 272.2, respectively. Results: The calibration curve was linear at 5–2000 μg/L busulfan. Intra- and interassay imprecision (CV) and bias were both &lt;11%. The limits of detection and quantification were 2 and 5 μg/L, respectively. Extraction recovery of busulfan was &gt;87%. Analysis of pharmacokinetics in four patients receiving high-dose busulfan indicated that minimum busulfan concentrations before the next dose were 405–603 μg/L, with no interference observed. Conclusions: The new rapid and sensitive liquid chromatographic–mass spectrometric assay is an appropriate method for quantification of busulfan in human plasma, making therapeutic drug monitoring of busulfan faster and easier in clinical practice.

scholarly journals Multiplex Liquid Chromatography-Tandem Mass Spectrometry Assay for Simultaneous Therapeutic Drug Monitoring of Ribavirin, Boceprevir, and Telaprevir

2013 ◽  
Vol 57 (7) ◽  
pp. 3147-3158 ◽  
Author(s):  
Manel Aouri ◽  
Darius Moradpour ◽  
Matthias Cavassini ◽  
Thomas Mercier ◽  
Thierry Buclin ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Hepatitis C ◽  
Therapeutic Drug Monitoring ◽  
Tandem Mass Spectrometry ◽  
Antiviral Therapy ◽  
Drug Monitoring ◽  
Reversed Phase ◽  
Therapeutic Drug ◽  
Tandem Mass

ABSTRACTNew directly acting antivirals (DAAs) that inhibit hepatitis C virus (HCV) replication are increasingly used for the treatment of chronic hepatitis C. A marked pharmacokinetic variability and a high potential for drug-drug interactions between DAAs and numerous drug classes have been identified. In addition, ribavirin (RBV), commonly associated with hemolytic anemia, often requires dose adjustment, advocating for therapeutic drug monitoring (TDM) in patients under combined antiviral therapy. However, an assay for the simultaneous analysis of RBV and DAAs constitutes an analytical challenge because of the large differences in polarity among these drugs, ranging from hydrophilic (RBV) to highly lipophilic (telaprevir [TVR]). Moreover, TVR is characterized by erratic behavior on standard octadecyl-based reversed-phase column chromatography and must be separated from VRT-127394, its inactive C-21 epimer metabolite. We have developed a convenient assay employing simple plasma protein precipitation, followed by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) for the simultaneous determination of levels of RBV, boceprevir, and TVR, as well as its metabolite VRT-127394, in plasma. This new, simple, rapid, and robust HPLC-MS/MS assay offers an efficient method of real-time TDM aimed at maximizing efficacy while minimizing the toxicity of antiviral therapy.

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