Liquid Chromatography/Tandem Mass Spectrometry for the Simultaneous Determination of Ursodiol and its Major Metabolites, Tauroursodeoxycholic Acid and Glycoursodeoxycholic Acid in Human Plasma

A rapid and sensitive method is described for the quantification of ursodiol and its major metabolites glycoursodeoxycholic acid (GUDCA) and tauroursodeoxycholic acid (TUDCA) in human plasma using single internal standard (Ursodeoxycholic Acid d4). Solid phase extraction was performed and chromatographic separation of 5µL injected sample was achieved using Waters Xterra, 5µm column with a mobile phase comprised of methanol and 5 mM ammonium formate with 0.1 % acetic acid ( 70 : 30, v/v ). The mass spectrometer was used in negative ion mode and multiple reactions monitoring using electro spray ionization mode as an interface. The method was fully validated and the calibration curves were linear over the concentration range of 25.9 to 15300.1 ng/mL for ursodiol, 2.7 to 1587.5ng/mLfor tauroursodeoxycholicacid and 25.4 to 15040.9 ng/mL for glycoursodeoxycholic acid. The method was sensitive and specific, with the lower limit of quantification of 25.9, 2.7 and 25.4 ng/ml for ursodiol, tauroursodeoxycholic acid and glycoursodeoxycholic acid respectively. The present method includes a simple and rapid sample preparation with shorter analysis run time and less flow rate compared to previously reported methods. The method was applied successfully for a bioequivalence study in healthy subjects.

Download Full-text

Rapid and Sensitive Method for Quantitative Determination of Lopinavir and Ritonavir in Human Plasma by Liquid Chromatography- Tandem Mass Specrtometry

E-Journal of Chemistry ◽

10.1155/2009/709478 ◽

2009 ◽

Vol 6 (1) ◽

pp. 223-230 ◽

Cited By ~ 7

Author(s):

G. A. Temghare ◽

S. S. Shetye ◽

S. S. Joshi

Keyword(s):

Liquid Chromatography ◽

Human Plasma ◽

Solid Phase ◽

Internal Standard ◽

Therapeutic Monitoring ◽

Tandem Mass ◽

Limit Of Quantification ◽

Liquid Chromatography Tandem Mass ◽

Sensitive Method

A rapid and sensitive liquid chromatography-mass spectrometric (LC-MS-MS) method for the simultaneous determination of lopinavir and ritonavir in human plasma using abacavir as internal standard has been developed and validated. Sample preparation of plasma involved solid phase extraction. Detection was performed using an Applied Biosystems Sciex API 2000 Mass spectrometer. The assay of lopinavir and ritonavir was linear over the range of 50 ng mL-1to 20000 ng mL-1and 20 ng mL-1to 3000 ng mL-1 respectively with a precision of <15% and accuracy in the range of 85-115%. The limit of quantification in plasma for lopinavir and ritonavir was 50 ng mL-1and 20 ng mL-1respectively. The described method has the advantage of being rapid and easy and it could be applied in therapeutic monitoring of these drugs in human plasma

Download Full-text

Liquid Chromatography/Tandem Mass Spectrometry for the Simultaneous Determination of Alverine and its Metabolite, Monohydroxy Alverine, in Human Plasma: Application to a Pharmacokinetic Study

E-Journal of Chemistry ◽

10.1155/2011/109414 ◽

2011 ◽

Vol 8 (1) ◽

pp. 201-211 ◽

Cited By ~ 1

Author(s):

Rahul C. Gavhane ◽

Ketan K. Nerurkar ◽

Ashok M. Kalamkar ◽

Mitesh R. Patil ◽

Satish G. Pingale ◽

...

Keyword(s):

Human Plasma ◽

Pharmacokinetic Study ◽

Solid Phase ◽

Internal Standard ◽

Selected Reaction Monitoring ◽

Limit Of Quantification ◽

Liquid Chromatography Tandem Mass ◽

Assay Precision ◽

Male Subjects

A rapid and sensitive LC-MS-MS method for the determination of alverine (ALV) and its major metabolite, monohydroxy alverine (MHA), in human plasma using imipramine as an internal standard was developed and validated. The analytes were extracted from 0.5 mL aliquots of human plasma by solid phase extraction, using oasis cartridge. Chromatographic separation was carried on Thermo Gold C18 column (50 × 4.6 mm, 5 μ) at 30 °C, with isocratic mobile phase, a flow rate of 0.4 mL/min and a total run time of 3.5 min. Detection and quantification were performed using a mass spectrometer in the selected reaction-monitoring mode with positive electrospray ionization atm/z282.3 → 91.11 for alverine,m/z298.3 → 106.9 for mono-hydroxy-alverine, andm/z281.0 → 86.0 for internal standard (IS) respectively. This assay was linear over a concentration range of 0.060-10 ng/mL with a lower limit of quantification of 0.060 ng/mL for both alverine and monohydroxy alverine. The coefficient of variation for the assay precision were <9.18% and <8.44%, the accuracy were >104.66% and >100.38% for alverine and monohydroxy alverine respectively. This method was successfully applied to a pharmacokinetic study after oral administration of alverine citrate 60 mg capsule in healthy male subjects.

Download Full-text

A Validated High-Performance Liquid Chromatography-Tandem Mass Spectrometric (Lc-Ms/Ms) Method for Simultaneous Determination of R(+)-Ketorolac and S(−)-Ketorolac in Human Plasma and Its Application to a Bioequivalence Study

Chromatography Research International ◽

10.4061/2011/214793 ◽

2011 ◽

Vol 2011 ◽

pp. 1-11 ◽

Cited By ~ 3

Author(s):

Sabyasachi Patri ◽

Anil K. Patni ◽

Sunil S. Iyer ◽

Arshad H. Khuroo ◽

Tausif Monif ◽

...

Keyword(s):

Liquid Chromatography ◽

Human Plasma ◽

Solid Phase ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Bioequivalence Study ◽

Mass Spectrometric ◽

Tandem Mass ◽

Liquid Chromatography Tandem Mass ◽

Tandem Mass Spectrometric

We report a selective, accurate, and reproducible liquid chromatography-tandem mass spectrometric (LC-MS/MS) method that employs solid phase extraction for quantification of ketorolac enantiomers in human plasma. Resolution of R(+)-ketorolac and S(−)-ketorolac was achieved using a Chiral-AGP column and a mobile phase of ammonium formate buffer (10 mM, pH ):acetonitrile (85 : 15, v/v and 70 : 30, v/v) in a gradient time program. S(+)-etodolac was used as the internal standard (IS). Quantification was achieved using a positive electrospray ionization (ESI+) interface under multiple reaction monitoring (MRM) condition. The method was validated over the concentration range of 9.36–1198.69 ng/ml for R(+)-ketorolac and 6.07–776.74 ng/ml for S(−)-ketorolac. Matrix effect was found negligible and the method showed good performances in terms of accuracy (89.6–102.7%) and precision (1.7–6.7%) for both enantiomers. Extraction recoveries of R(+)-ketorolac, S(−)-ketorolac, and S(+)-etodolac were 82.04, 70.94, and 93.90%, respectively. Results of all stability exercises in human plasma were within acceptable limits. The method was successfully applied to a single dose cross over bioequivalence study in healthy human male volunteers. Incurred Sample Reanalysis (ISR) was performed by randomly selecting 10% of total subject samples of the study using Statistical Analysis Software (SAS). Values of 91.1% for R (+)-ketorolac and 83.5% for S(−)-ketorolac indicated good acceptance for ISR.

Download Full-text

Determination of Chloramphenicol in Bovine Milk by Liquid Chromatography/Tandem Mass Spectrometry

Journal of AOAC International ◽

10.1093/jaoac/86.4.703 ◽

2003 ◽

Vol 86 (4) ◽

pp. 703-706 ◽

Cited By ~ 16

Author(s):

Lambert K Sørensen ◽

Tina H Elbæk ◽

Helga Hansen

Keyword(s):

Solid Phase ◽

Atmospheric Pressure Chemical Ionization ◽

Bovine Milk ◽

Negative Ion ◽

Tandem Mass ◽

Liquid Chromatography Tandem Mass ◽

Pressure Chemical Ionization ◽

Pressure Chemical ◽

Negative Ion Mode

Abstract A rapid confirmatory liquid chromatographic/tandem mass spectrometic method was developed for determination of chloramphenicol in bovine milk. Chloramphenicol was extracted directly from milk by solid-phase extraction on a C18 cartridge. The extract was further cleaned up on neutral aluminium oxide. Three transition products were monitored in negative ion mode after atmospheric pressure chemical ionization. The detection capability related to the transition product of lowest abundance was 0.03 μg/kg. The mean recovery was 90% at levels of 0.1–0.2 μg/kg. The relative repeatability standard deviations were 4.3, 3.8, and 2.8% at levels of 0.1, 0.2, and 1.0 μg/kg, respectively.

Download Full-text

Quantification of Amiridine in Human Plasma by High-Performance Liquid Chromatography Coupled with Electrospray Tandem Mass Spectrometry

Chromatography Research International ◽

10.1155/2013/524806 ◽

2013 ◽

Vol 2013 ◽

pp. 1-5

Author(s):

Igor I. Miroshnichenko ◽

Angelina I. Platova

Keyword(s):

Human Plasma ◽

High Performance ◽

Solid Phase ◽

Internal Standard ◽

Mass Spectrometric ◽

Pharmacokinetic Studies ◽

Tandem Mass ◽

Ionization Source ◽

Limit Of Quantification ◽

Tandem Mass Spectrometric

The aim of this study was to develop and validate a high-performance liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for analysis of the amiridine in human plasma. The analyte and internal standard (IS), zolpidem, were extracted from human plasma by solid phase extraction (SPE with SOLA cartridges) and separated on a Zorbax SB-C18 column using methanol and 0.2% formic acid in water as mobile phase. Detection was performed using an electrospray ionization source and mass spectrometric positive multireaction monitoring mode (+MRM) at a voltage capillary of +2000 V. The assay was linear over the concentration range 0.5–200 ng/mL with the lowest limit of quantification (LLOQ) of 0.5 ng/mL. The method also afforded satisfactory results in terms of the sensitivity, specificity, precision (intra- and interday %), accuracy, recovery, and the stability of the analyte under various conditions. The method can be successfully applied to pharmacokinetic studies.

Download Full-text

Quantification of Lumefantrine in Human Plasma Using LC-MS/MS and Its Application to a Bioequivalence Study

Journal of Pharmaceutics ◽

10.1155/2013/437697 ◽

2013 ◽

Vol 2013 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Satish G. Pingale ◽

Kiran V. Mangaonkar

Keyword(s):

Mass Spectrometry ◽

Human Plasma ◽

Analytical Method ◽

Internal Standard ◽

Bioequivalence Study ◽

Mass Spectrometric ◽

Plasma Samples ◽

Limit Of Quantification ◽

Chromatographic Condition ◽

Ionization Mode

An analytical method based on protein precipitation has been developed and validated for analysis of lumefantrine in human plasma. Artesunate was used as an internal standard for lumefantrine. Inertsil ODS column provided chromatographic separation of analytes followed by detection with mass spectrometry. The method involves simple isocratic chromatographic condition and mass spectrometric detection in the positive ionization mode using an API-3000 system. The total run time was 2.5 minutes. The proposed method has been validated with linear range of 200–20000 ng/mL for lumefantrine. The intrarun and interrun precision values are within 6.66% and 5.56%, respectively, for lumefantrine at the lower limit of quantification level. The overall recovery for lumefantrine and artesunate was 93.16% and 91.05%, respectively. This validated method was used successfully for analysis of plasma samples from a bioequivalence study.

Download Full-text

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.

Download Full-text

Determination of Ranitidine in Human Plasma by SPE and ESI-LC-MS/MS for Use in Bioequivalence Studies

ISRN Chromatography ◽

10.1155/2013/484592 ◽

2013 ◽

Vol 2013 ◽

pp. 1-7

Author(s):

Karini B. Bellorio ◽

Maria Isabel R. Alves ◽

Nelson R. Antoniosi Filho

Keyword(s):

Solid Phase Extraction ◽

Human Plasma ◽

Extraction Method ◽

Limit Of Detection ◽

Solid Phase ◽

Internal Standard ◽

Phase Extraction ◽

Good Separation ◽

Limit Of Quantification

A method for determining ranitidine in human plasma by ESI-LC-MS/MS was validated, using propranolol as internal standard. The extraction method used was solid phase extraction (SPE). Chromatographic separation was performed in a Chromolith C18 (50 mm × 4.6 mm i.d.) analytical column, which provided good separation of ranitidine and propranolol peaks with an analysis time of 2.5 minutes. Extraction yields of 94.4% for ranitidine and 89.4% for the internal standard were obtained. The lower limit of quantification (LLOQ) was 3.00 ng/mL, and limit of detection (LOD) was 0.05 ng/mL, with linearity ranging from 3.00 to 500 ng/mL. The results, thus, showed that this method is suitable for application in bioequivalence studies of ranitidine in human plasma.

Download Full-text

Quantification of Niacin and Its Metabolite Nicotinuric Acid in Human Plasma by LC-MS/MS: Application to a Clinical Trial of a Fixed Dose Combination Tablet of Niacin Extended-Release/Simvastatin (500 mg/10 mg) in Healthy Chinese Volunteers

International Journal of Analytical Chemistry ◽

10.1155/2015/212437 ◽

2015 ◽

Vol 2015 ◽

pp. 1-9 ◽

Cited By ~ 2

Author(s):

Pingping Zhang ◽

Yantong Sun ◽

Guobing Shi ◽

Yin Sui ◽

Qiuying Li ◽

...

Keyword(s):

Human Plasma ◽

Extended Release ◽

Internal Standard ◽

Negative Ion ◽

Fixed Dose ◽

Ionization Source ◽

Combination Tablet ◽

Dose Combination ◽

Relative Errors ◽

Negative Ion Mode

Our paper aimed to develop rapid, sensitive, and specific LC-MS/MS method for the quantification of niacin (NA) and its metabolite nicotinuric acid (NUA) in human plasma. Following protein precipitation with acetonitrile, the NA, NUA, and internal standard (5-fluorouracil) were separated on a Zorbax 300SB-C8column (250 mm × 4.6 mm, 5 μm) with a mobile phase consisting of methanol-2 mM ammonium acetate (3 : 97, v/v) at a flow rate of 1 mL/min (split 1 : 1). A tandem mass spectrometer equipped with electrospray ionization source was used as the detector and operated in negative ion mode. The linear concentration ranges of the calibration curves were 5–800 ng/mL for NA and NUA. The intra-assay RSD for quality control (QC) samples were from 5.0% to 8.7% for NA, and 5.5% to 7.6% for NUA. The interassay RSD for QC samples were from 2.8% to 9.4% for NA, and 3.7% to 5.8% for NUA. The relative errors for QC samples were from −2.2% to 2.3% for NA, and −0.6% to 3.2% for NUA. The method was successfully applied to the investigation of the pharmacokinetic profiles of NA, NUA in human after single dose administration of Niacin extended-release/Simvastatin tablet (500 mg/10 mg).

Download Full-text

A Sensitive LC–MS/MS Method for the Determination of Afatinib in Human Plasma and Its Application to a Bioequivalence Study

Journal of Chromatographic Science ◽

10.1093/chromsci/bmab040 ◽

2021 ◽

Author(s):

Xi Luo ◽

Xiu Jin Zhang ◽

Wen Ling Zhu ◽

Jin Ling Yi ◽

Wen Gang Xiong ◽

...

Keyword(s):

Human Plasma ◽

Mobile Phase ◽

High Performance ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Bioequivalence Study ◽

Liquid Chromatography Tandem Mass ◽

Multiple Reaction Monitoring Mode ◽

Ionization Mode

Abstract A high performance liquid chromatography–tandem mass spectrometry assay for the determination of afatinib (AFT) in human plasma was established. A simple sample preparation of protein precipitation was used and separation was achieved on a C18 column by the gradient mixture of mobile Phase A of water (containing 0.1% ammonia) and the mobile Phase B of acetonitrile and water (V:V = 95:5, containing 0.2% ammonia). The multiple reaction monitoring mode was used to monitor the precursor-to-production transitions of m/z 486.2 → m/z 371.4 for AFT and m/z 492.2 → m/z 371.3 for AFT-d6 (internal standard) at positive ionization mode. The calibration curve ranged from 0.100 to 25.0 ng·mL−1 and the correlation coefficient was greater than 0.99. The intra- and inter-batch precision was less than or equal to 10.0%. Accuracy determined at four concentrations was in the range of 92.3–103.3%. In summary, our method was sensitive, simple and reliable for the quantification of AFT and was successfully applied to a bioequivalence study.

Download Full-text