Therapeutic Monitoring of Plasma Digoxin for COVID-19 Patients Using a Simple UPLC-MS/MS Method

Background:: Cardiovascular diseases (CVD) were reported in 8% - 16% of patients with 2019 coronavirus disease (COVID-19). Digoxin was one of the main drugs to treat CVD. Objective:: The clinician applied for therapeutic drug monitoring (TDM) of digoxin according to the drug usage of the patients to monitor their concentration of digoxin , so as to avoid its toxic and side effects, and provide a theoretical reference for clinical usage of digoxin in patients with COVID-19. Methods:: A method for quantifying digoxin concentration in plasma with ultra performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) was developed . After a simple protein precipitation of plasma with methanol, digoxin and its internal standard (digoxin-d3) were detected in positive ion mode using multiple reaction monitoring .Results: Plasma digoxin in the range of 0.2 - 10 ng/mL had a good linearity. The UPLC-MS/MS method was validated with inter-run accuracies from 91.3% to 107.4% and precisions less than 13%. Nine plasma samples (5 at valley concentration and 4 at follow-up after stopping dosing) from three patients with COVID-19 were tested. The mean plasma digoxin concentration was 0.73 ng/mL (ranged from 0 to 1.31 ng/mL). Digoxin was detected at the concentration of 0.93 ng/mL after stopping drug administration for 14 days. Conclusion:: In this study, we established a simple UPLC-MS/MS method using protein-precipitation to perform TDM of digoxin in patients with COVID-19, and found that about 56% of digoxin plasma concentration was within the treatment window (0.8 - 2.0 ng/mL). Digoxin can be remained in the body for nearly 14 days in severe patients with COVID-19 after stopping dosing.

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P63 UPLC/MS/MS assay for the simultaneous determination of seven antibiotics in human serum – application to pediatric studies

Archives of Disease in Childhood ◽

10.1136/archdischild-2019-esdppp.101 ◽

2019 ◽

Vol 104 (6) ◽

pp. e43.2-e43

Author(s):

S Magreault ◽

O Chaussenery-Lorentz ◽

T Storme ◽

E Jacqz-Aigrain

Keyword(s):

Multiple Reaction Monitoring ◽

Internal Standard ◽

Ultra Performance Liquid Chromatography ◽

Routine Activity ◽

Therapeutic Drug ◽

Pharmacokinetic Studies ◽

Pediatric Dose ◽

Liquid Chromatography Tandem Mass ◽

Sampling Volume ◽

Acquity Uplc

BackgroundAntimicrobials are widely used in children but pediatric dose regimens are not always validated, and PK studies, required to validate dosage, are difficult to conduct in children. Low sampling volume limits the number of PK samples drawn per patient and analytical methods adapted to small volumes are not always available. Due to the wide inter-patient pharmacokinetic (PK) variability in children, particularly neonates, therapeutic drug monitoring is required to adapt dosage to individual patients. In such clinical and analytical context, our aim was to develop a unique, rapid and highly sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) assay to quantify 7 antibiotics (amoxicillin, azithromycin, cefotaxime, ciprofloxacin, meropenem, metronidazole and piperacillin) in low sample volumes (50 µL) for both routine monitoring and pharmacokinetic studies.MethodsAfter protein precipitation by acetonitrile, the antibiotics and their associated deuterated internal standard were separated on a Waters Acquity UPLC HSS T3 (100 mm x 2.1 mm; 1.8 µm). The mobile phases consisted of a gradient of ammonium acetate (pH 2.4; 5mM) and acetonitrile acidified with 0.1% (v/v) formic acid (started ratio of 93:7, v/v), run at 0.5 mL/min flow rate (total run time: 2.75 min). Ions were detected in the turbo-ion-spray-positive and multiple-reaction-monitoring modes.ResultsThis method was linear from 0.1–50 µg/mL. Accuracy and precision were evaluated using Quality Control (2, 10, 35 µg/mL). Validation of the method proved that precision, selectivity and stability were all within the recommended limits.ConclusionThis method has the advantage of a unique, efficient and standardized analytical tool for rapid measurement of 7 antibiotics in low blood volume. It has been successfully applied for routine activity and pharmacokinetic studies in children and neonates.Disclosure(s)Nothing to disclose.

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A Novel UPLC-MS/MS Assay for the Measurement of Linezolid and its Metabolite PNU-142300 in Human Serum and its Application to Patients With Renal Insufficiency

Frontiers in Pharmacology ◽

10.3389/fphar.2021.641872 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yingying Wang ◽

Er-min Gu ◽

Xiaoxiang Du ◽

Ren-ai Xu ◽

Guanyang Lin

Keyword(s):

Liquid Chromatography ◽

Renal Insufficiency ◽

Human Serum ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Gradient Elution ◽

Ultra Performance Liquid Chromatography ◽

Serum Samples ◽

The Matrix ◽

Liquid Chromatography Tandem Mass

The contribution of the metabolites of linezolid to the associated myelosuppression is unknown in patients who are renal impairment. In this research, the purpose of our experiment was to explore and develop a quick and robust ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) assay for the determination of linezolid and its metabolite PNU-142300 in human serum simultaneously. The analytes were prepared using a simple and convenient approach with acetonitrile for protein crash, and then separated from the matrix on a Waters Acquity Ultra performance liquid chromatography (UPLC) BEH C18 (2.1 mm × 50 mm, 1.7 μm) column in a program of gradient elution, where the mobile phase was consisted of water with 0.1% formic acid and acetonitrile, and was placed at 0.40 ml/min flow rate. Multiple reaction monitoring (MRM) was employed and conducted for UPLC-MS/MS detection with ion transitions at m/z 338.01 → 296.03 for linezolid, m/z 369.96 → 327.98 for PNU-142300 and m/z 370.98 → 342.99 for tedizolid (Internal standard, IS), respectively. This method had good linearity respectively in the calibration range of 0.01–20 μg/ml for linezolid, and 0.05–100 μg/ml for PNU-142300. In the intra- and inter-day, the precision of linezolid and PNU-142300 was below 14.2%, and the accuracy in this method was determined to be from −9.7 to 12.8%. In addition, recovery and matrix effect of the analytes were all found to be acceptable, and the analytes during the assay and storage in serum samples were observed to be stable. The novel optimized UPLC-MS/MS assay was also successfully employed to determine the concentration levels of linezolid and PNU-142300 in human serum. The results showed that linezolid-associated myelosuppression occurs more frequently in patients with renal insufficiency, and the metabolite-to-parent concentration ratio of PNU-142300 is predicted to reduce this toxicity of myelosuppression.

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Ultra-Performance Liquid Chromatography Coupled with a Triple Quadrupole Mass Spectrometric Method for the Quantification of Anti-epileptic drugs Methsuximide and Normesuximide in Human Plasma and its Application in a Pharmacokinetic Study

Current Pharmaceutical Analysis ◽

10.2174/1573412917666210211122311 ◽

2021 ◽

Vol 17 ◽

Author(s):

Karthik Rajendran ◽

Karthika Anoop ◽

Krishnaveni Nagappan ◽

Genguchetti Mohan Sekar ◽

Sankham Devendran Rajendran

Keyword(s):

Human Plasma ◽

Pharmacokinetic Study ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Current Method ◽

Ultra Performance Liquid Chromatography ◽

Mass Spectrometric ◽

Therapeutic Drug ◽

Spectrometric Method ◽

Mass Spectrometric Method

Background: Extensive therapeutic drug monitoring needs an analytical method for efficient and sensitive quantification of analytes of interest in clinical pharmacology. Objective: A rapid, robust, sensitive and simple UPLC-MS/MS method to quantify Methsuximide (Ms) and N-desmethyl methsuximide/Normesuximide (MsMET) in human plasma was optimized, developed, and validated for application in a pharmacokinetic study. Method: Reverse phased chromatography was performed using Zorbax SB-C18, 4.6 x 75 mm., 3.5 µm as stationary phase, methanol and 0.1% formic acid (60:40 v/v) as mobile phase which was delivered isocratically at a flow rate of 0.9 mL/min. The sample injection volume was 5 µL. Mass spectrometric quantification of the analytes was performed using positive electrospray ionization as mass interface along with multiple reaction monitoring (MRM) as acquisition mode. Results: The selected mass transition ions for analyte, metabolite and its respective internal standards are as follows, precursor ion (m/z) and product ion (m/z): Ms (204.06 and 119.02), MsMET (190.05 and 119.82), Ms internal standard (MsIS) (209.17 and 124.02), and MsMET internal standard (MsMETIS) (195.09 and 124.16), respectively. The current method was found to be linear for Ms (60.72-5920 ng/mL) and MsMET (60.38-6010 ng/mL) with r2 values not less than 0.999. The mean recoveries of all analytes ranged between 71.37 and 86.38 percentage. Conclusion: This method was validated in accordance with USFDA’s bioanalytical guidelines. This method could be applied for a routine analysis of Ms and MsMET in clinical pharmacological practice.

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Determination of eugenitin in mouse blood by ultra-performance liquid chromatography-tandem mass spectrometry and its application to a pharmacokinetic study

Acta Chromatographica ◽

10.1556/1326.2021.00937 ◽

2021 ◽

Author(s):

Chongliang Lin ◽

Dezhen Song ◽

Haodong Jiang ◽

Lvqi Luo ◽

Xi Bao ◽

...

Keyword(s):

Biological Fluids ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Ultra Performance Liquid Chromatography ◽

Quantitative Detection ◽

Syzygium Aromaticum ◽

Mouse Blood ◽

The Matrix ◽

Liquid Chromatography Tandem Mass

Abstract Eugenitin is a non-volatile chromone derivative which is always found in dried flower buds of Syzygium aromaticum L. (Merr.) & L.M. Perry. Until now, there were no reports about the pharmacokinetics of eugenitin in biological fluids. A UPLC-MS/MS method developed to determine eugenitin in mouse blood. The blood samples were prepared by protein precipitation with acetonitrile. Chrysin (internal standard, IS) and eugenitin were gradient eluted by mobile phase of acetonitrile and water (0.1% formic acid) in a BEH C18 column. The multiple reaction monitoring (MRM) of m/z 221.1→206.0 for eugenitin and m/z 255.1→152.9 for IS with an electrospray ionization (ESI) source was used for quantitative detection. The calibration curve ranged from 0.5 to 500 ng/mL (r > 0.995). The accuracy ranged from 98 to 113%, the precision was less than 12%, and the matrix effect was between 86 and 94%, the recovery was better than 81%. The developed method was successfully used for pharmacokinetics of eugenitin in mice after intravenous (5 mg/kg) and oral (20 mg/kg) administration, and the absolute availability of eugenitin was 12%.

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A Rapid UPLC-MS Method for Quantification of Gomisin D in Rat Plasma and Its Application to a Pharmacokinetic and Bioavailability Study

Molecules ◽

10.3390/molecules24071403 ◽

2019 ◽

Vol 24 (7) ◽

pp. 1403 ◽

Cited By ~ 2

Author(s):

Xiaoyong Zheng ◽

Feng Feng ◽

Xiunan Jiang ◽

Jieying Qiu ◽

Xiaojun Cai ◽

...

Keyword(s):

Multiple Reaction Monitoring ◽

Internal Standard ◽

Ultra Performance Liquid Chromatography ◽

Rat Plasma ◽

Ionization Source ◽

Quality Marker ◽

Liquid Chromatography Tandem Mass ◽

Bioavailability Study ◽

Multiple Reaction Monitoring Mode ◽

Positive Ion

Gomisin D, a lignan compound isolated from Fructus Schisandra, is a potential antidiabetic and anti-Alzheimer’s agent. Recently, gomisin D was used as a quality marker of some traditional Chinese medicine (TCM) formulas. In this study, a rapid ultra-performance liquid chromatography/tandem mass spectrometry method (UPLC-MS/MS) was developed and validated to quantify gomisin D in rat plasma for a pharmacokinetic and bioavailability study. Acetonitrile was used to precipitate plasma proteins. Separations were performed on a BEH C18 column with a gradient mobile phase comprising of acetonitrile and water (0.1% formic acid). An electrospray ionization source was applied and operated in the positive ion mode. The multiple reaction monitoring mode (MRM) was utilized to quantify gomisin D and nomilin (internal standard, IS) using the transitions of m/z 531.2 → 383.1 and m/z 515.3 → 161.0, respectively. The calibration curve was linear over the working range from 1 to 4000 ng/mL (R2 = 0.993). The intra- and interday precision ranged from 1.9% to 12.9%. The extraction recovery of gomisin D was in the range of 79.2–86.3%. The validated UPLC-MS/MS method was then used to obtain the pharmacokinetic characteristics of gomisin D after intravenous (5 mg/kg) and intragastric (50 mg/kg) administration to rats. The bioavailability of gomisin D was 107.6%, indicating that this compound may become a promising intragastrical medication. Our results provided useful information for further preclinical studies on gomisin D.

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An LC-MS/MS Validated Method for Quantification of Chlorzoxazone in Human Plasma and Its Application to a Bioequivalence Study

Journal of Chromatographic Science ◽

10.1093/chromsci/bmz052 ◽

2019 ◽

Vol 57 (8) ◽

pp. 751-757

Author(s):

Jiake He ◽

Ning Li ◽

Jiaqiu Xu ◽

Jing Zhu ◽

Yang Yu ◽

...

Keyword(s):

Human Plasma ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Bioequivalence Study ◽

Therapeutic Drug ◽

Reference Formulation ◽

Chromatography Tandem Mass Spectrometry ◽

Liquid Chromatography Tandem Mass ◽

One Step

Abstract A simple, sensitive, specific, accurate liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for determination of chlorzoxazone in human plasma was developed and validated to evaluate the pharmacokinetic characteristics of chlorzoxazone test or reference formulation. Sample preparation was achieved by one step protein precipitation and dilution with acetontrile. The chromatographic separation was performed at 40°C with a gradient mobile phase (0.3 mL/min) and a Shimadzu VP-ODS C18 analytical column (column size: 150 × 2.0 mm). TSQ quantum access triple-quadrapole MS/MS detection was operated in a negative mode by multiple reaction monitoring. Ion transitions at m/z 168.0→132.1 for chlorzoxazone and m/z 451.3→379.3 for repaglinide (internal standard) were used for the LC-MS/MS analysis. The calibration was linear (r ≥ 0.995) over the tested concentration range of 0.2–20 μg/mL for chlorzoxazone in plasma. Precision, accuracy, recovery, matrix effect and stability for chlorzoxazone were evaluated and were excellent within the range of tested concentrations. This method was successfully applied to a bioequivalence study in 20 healthy Chinese volunteers. This method could also contribute to the personalized medication and therapeutic drug monitoring of chlorzoxazone.

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UPLC-MS/MS Method for the Determination of 14 Compounds in Rat Plasma and Its Application in a Pharmacokinetic Study of Orally Administered Xiaoyao Powder

Molecules ◽

10.3390/molecules23102514 ◽

2018 ◽

Vol 23 (10) ◽

pp. 2514 ◽

Cited By ~ 4

Author(s):

Mingyue Xu ◽

Zhanling Xu ◽

Qingxuan Xu ◽

Hongyue Zhang ◽

Mingyang Liu ◽

...

Keyword(s):

Pharmacokinetic Study ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Ultra Performance Liquid Chromatography ◽

Rat Plasma ◽

Chinese Herbs ◽

Tandem Mass ◽

Ionization Source ◽

Liquid Chromatography Tandem Mass ◽

Negative Ionization

Xiaoyao Powder (XYP), a common Chinese medicine, comprises eight traditional Chinese herbs and has been widely used clinically to treat liver damage and mental disorders. An ultra-performance liquid chromatography–tandem mass spectrometry method was developed to investigate the pharmacokinetics of 14 compounds (albiflorin, paeoniflorin, ferulic acid, senkyunolide I, quercetin, isoliquiritigenin, atractylenolide III, ligustilide, atractylenolide II, liquiritin, liquiritigenin, saikosaponin c, glycyrrhizic acid, and saikosaponin a) in XYP. Naringenin was used as the internal standard. The compounds were separated using an ACQUITY UPLCTM BEH C18 column (1.7 μm, 50 × 2.1 mm) with a mobile phase consisting of acetonitrile and 0.1% formic acid in water at a flow rate of 0.3 mL/min. Detection was performed on a triple-quadrupole tandem mass spectrometer using multiple reaction monitoring and an electrospray ionization source in both positive and negative ionization modes. All calibration curves exhibited good linearity (r2 > 0.9974) over the measured ranges. The intra- and inter-day precisions were within 12%, and the accuracy ranged from 89.93% to 106.64%. Extraction recovery and matrix effect results were satisfactory. The method was successfully applied in a pharmacokinetic study of the 14 compounds in rat plasma after the oral administration of XYP.

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

International Journal of Pharmacy and Pharmaceutical Sciences ◽

10.22159/ijpps.2016v8i9.12918 ◽

2016 ◽

Vol 8 (9) ◽

pp. 194 ◽

Cited By ~ 1

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

Objective: To quantify 4-hydroxycyclophosphamide in cancer patients’ plasma for therapeutic drug monitoring of cyclophosphamide.Methods: 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>139.978, 4-hydroxycyclophosphamide-semicarbazide at m/z 338.011>224.97, and hexamethylphosphoramide as internal standard at m/z 180.17>92.08.Results: 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.Conclusion: 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.

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Comparison of HPLC-DAD and UPLC-MS/MS in monitoring serum concentration of lamotrigine

Current Pharmaceutical Analysis ◽

10.2174/1573412917666210215150712 ◽

2021 ◽

Vol 17 ◽

Author(s):

Xubin Wang ◽

Zhibin Chen ◽

Xiaofan Ke ◽

Yingying Wang ◽

Lufeng Hu ◽

...

Keyword(s):

Liquid Chromatography ◽

High Performance ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Epileptic Seizures ◽

Serum Levels ◽

Ultra Performance Liquid Chromatography ◽

Anti Epileptic Drug ◽

Deming Regression ◽

Liquid Chromatography Tandem Mass

Background: Lamotrigine (LTG) is a broad-spectrum and first-line anti-epileptic drug. To monitor the serum levels of LTG in epileptic seizures patients, high-performance liquid chromatography with diode-array detection (HPLC-DAD) and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) methods were established and compared. Methods: Imatinib was used as the internal standard (IS) for both methods. LTG and IS were detected at 246 nm by HPLC-DAD. In UPLC-MS/MS, LTG and IS positive ion were detected by multiple reaction monitoring (MRM), with m/z of 256/210.9 and 494/394.02, respectively. A total of 37 blood samples from epileptic patients were determined and studied by these two methods. Results: There was an acceptable linearity for the two methods. The concentration range of LTG was 0.59 ~ 22.20 mg/L by HPLC, and 0.28 ~ 23.97 mg/L by UPLC-MS/MS.The Pearson regression coefficient of Deming regression was 0.9653 (95% CI: 0.9332 to 0.9821). Bland–Altman method demonstrated that the concentration of LTG determined by UPLC-MS/MS was 8.3% higher than that determined by HPLC (limits of agreement, -32.0% to +48.6%). Conclusion: There was a significant correlation between the two methods. Both HPLC and UPLC-MS/MS can be used for routine clinical monitoring of LTG.

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Pharmacokinetics of ebeiedinone in mouse blood by UPLC–MS/MS

Acta Chromatographica ◽

10.1556/1326.2019.00680 ◽

2020 ◽

Vol 32 (3) ◽

pp. 194-198

Author(s):

Yongxi Jin ◽

Yuyan Chen ◽

Jiawen Liu ◽

Xi Bao ◽

Yinghao Zhi ◽

...

Keyword(s):

Pharmacokinetic Study ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Ultra Performance Liquid Chromatography ◽

Absolute Bioavailability ◽

Mouse Blood ◽

Limit Of Quantification ◽

Chromatography Tandem Mass Spectrometry ◽

The Matrix ◽

Liquid Chromatography Tandem Mass

An ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was established to determine ebeiedinone in mouse blood, and the pharmacokinetics of ebeiedinone after intravenous (0.5 mg/kg) and oral (2, 4, and 8 mg/kg) administration was studied. Twenty-four mice were randomly divided into 4 groups, 1 group was for intravenous administration (0.5 mg/kg), and other 3 groups were for oral administration (2, 4, and 8 mg/kg), with 6 rats in each group. Yubeinine was used as an internal standard. Multiple reaction monitoring (MRM) mode was used to quantitatively analyzed ebeiedinone m/z 414.4 → 91.1 and the internal standard m/z 430.4 → 412.3 in the electrospray ionization (ESI) positive interface. In the concentration range of 1–2000 ng/mL, the ebeiedinone in the mouse blood was linear (r2 > 0.995), and the lower limit of quantification was 1.0 ng/mL. In the mouse blood, the intra-day precision coefficient of variation (CV) was less than 15%, and the inter-day precision CV was less than 15%. The accuracy ranged from 85.4% to 114.6%, and the average recovery was higher than 61.3%. The matrix effect was between 87.0% and 106.5%. These data met the pharmacokinetic study requirements of ebeiedinone. The UPLC–MS/MS method was sensitive, rapid, and selective and was successfully applied to the pharmacokinetic study of ebeiedinone in mice. The absolute bioavailability of ebeiedinone was 30.6%.

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