scholarly journals Simple and Robust Analysis of Cefuroxime in Human Plasma by LC-MS/MS: Application to a Bioequivalence Study

2014 ◽  
Vol 2014 ◽  
pp. 1-7
Author(s):  
Xingjiang Hu ◽  
Mingzhu Huang ◽  
Jian Liu ◽  
Junchun Chen ◽  
Jianzhong Shentu
Keyword(s):  
Human Plasma ◽  
Protein Separation ◽  
Extraction Procedure ◽  
Internal Standard ◽  
Bioequivalence Study ◽  
Clinical Analysis ◽  
Cefuroxime Axetil ◽  
Precipitate Protein ◽  
Male Subjects ◽  
Chinese Male

A simple, robust LC-MS/MS assay for quantifying cefuroxime in human plasma was developed. Cefuroxime and tazobactam, as internal standard (IS), were extracted from human plasma by methanol to precipitate protein. Separation was achieved on a Zorbax SB-Aq (4.6×250 mm, 5 μm) column under isocratic conditions. The calibration curve was linear in the concentration range of 0.0525–21.0 μg/mL (r=0.9998). The accuracy was higher than 90.92%, while the intra- and interday precision were less than 6.26%. The extraction procedure provides recovery ranged from 89.44% to 92.32%, for both analyte and IS. Finally, the method was successfully applied to a bioequivalence study of a single 500 mg dose of cefuroxime axetil in 22 healthy Chinese male subjects under fasting condition. Bioequivalence was determined by calculating 90% Cls for the ratios ofCmax, AUC0-t, and AUC0-∞values for the test and reference products, using logarithmic transformed data. The 90% Cls for the ratios ofCmax(91.4%~104.2%), AUC0-t(97.4%~110.9%), and AUC0-∞(97.6%~111.1%) values were within the predetermined range. It was concluded that the two formulations (test for capsule, reference for tablet) analyzed were bioequivalent in terms of rate and extent of absorption and the method met the principle of quick and easy clinical analysis.

scholarly journals LC-MS-MS Method for Determination of Metolazone in Human Plasma

2008 ◽  
Vol 5 (3) ◽  
pp. 634-640 ◽  
Author(s):  
Shikha M. N. Roy ◽  
Kiran V. Mangaonkar ◽  
Santosh. M. Yetal ◽  
Santosh. S. Joshi
Keyword(s):  
Human Plasma ◽  
High Performance ◽  
Extraction Procedure ◽  
Internal Standard ◽  
Bioequivalence Study ◽  
Specific Method ◽  
Simple Liquid ◽  
Limit Of Quantification ◽  
Liquid Liquid Extraction

A rapid, sensitive and specific method for quantification of metolazone in human plasma using metaxalone as internal standard is described. Sample preparation involved a simple liquid-liquid extraction procedure. The extract was analyzed by high performance liquid chromatography coupled to electrospray tandem mass spectrometry (LC–MS–MS). Chromatography was performed isocratically on a 5 μm C18analytical column (50 mm × 4.6 mmi.d.) with buffer–acetonitrile 20:80 (v/v) as mobile phase. The response to metolazone was a linear function of concentration over the range 1.00 to 2000.00 ng mL-1. The lower limit of quantification in plasma was 1.0 ng mL-1. The method was successfully applied in a bioequivalence study of a metolazone formulation after administration as a single oral dose.

scholarly journals A sensitive and reliable RP-HPLC method for the detection of cimetidine – a H2 receptor antagonist in human plasma

2019 ◽  
Vol 8 (3) ◽  
pp. 671-674
Keyword(s):  
Human Plasma ◽  
High Performance ◽  
Internal Standard ◽  
Bioequivalence Study ◽  
High Concentration ◽  
H2 Receptor Antagonist ◽  
Chromatography Method ◽  
Thaw Cycle ◽  
Limit Of Quantitation ◽  
The Mean

Bioanalytical methods for bioequivalence studies require high sensibility and rapidity due to the large number of samples and the low plasma concentration of drugs. The present study aimed to develop and validate a high-performance liquid chromatography method to quantify cimetidine (CMT) in human plasma and to apply it in a bioequivalence study. Spiked plasma of 500 µl (l, m and h concentration) was used for the assay. The HPLC injection volume was 20μl of the reconstitute sample where, 2 ml of ethyl acetate used for extraction purposes. Cimetidine was prepared separately for low (80 ng/ml), medium (2000 ng/ml) and high (3600 ng/ml) concentrations and internal standard (ranitidine) concentration was 3000 ng/ml. Freeze thawing and long terms stability were conducted at -25º c. The individual calibration curve for spiked standards was linear with R2= 0.99. The inaccuracy values for QC samples were within 15% of the actual value and not more than 20% for the LOQ. The limit of quantitation (LOQ) was 40 ng/ml, which was also the lowest concentration of cimetidine that was quantitated with the variability of 5.9%. The within day precision and between day precision for LOQ were 10.8 and 5.9 respectively. The retention time for the analyte was 4.1-4.5 minutes during the within a day and between day results. The mean % inaccuracy values for low, medium and high concentration were 6.8, 5.6 and 7.8 respectively for within day and 2.4, 6.1 and 7.9 respectively for between days. The within day and between day % inaccuracy for LOQ concentration was 12.4 and 5.5 respectively. The mean recoveries for low, medium and high concentration of cimetidine were 80.2, 70.9 and 74.2. The overall mean recovery for cimetidine was 75.1%. The maximum inaccuracy for freeze thaw cycle and long term stability samples for low, medium and high was found with CV less than 15% for all concentrations, indicating that cimetidine is stable. The developed method was precise and accurate and was suitable to be applied for the bioequivalence study of cimetidine.

scholarly journals Bioanalytical Method Development and Validation of Memantine in Human Plasma by High Performance Liquid Chromatography with Tandem Mass Spectrometry: Application to Bioequivalence Study

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Ravi Kumar Konda ◽  
B. R. Challa ◽  
Babu Rao Chandu ◽  
Kothapalli B. Chandrasekhar
Keyword(s):  
Human Plasma ◽  
High Performance ◽  
Method Development ◽  
Dynamic Range ◽  
Quantitative Estimation ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Bioequivalence Study ◽  
Healthy Human ◽  
Development And Validation

A simple, sensitive, and rapid HPLC-MS/MS method was developed and validated for quantitative estimation of memantine in human plasma. Chromatography was performed on Zorbax SB-C18(4.6×75 mm, 3.5 μm) column. Memantine (ME) and internal standard Memantine-d6(MED6) were extracted by using liquid-liquid extraction and analyzed by LC-ESI-MS/MS using multiple-reaction monitoring (MRM) mode. The assay exhibited a linear dynamic range of 50.00–50000.00 pg/ml for ME in human plasma. This method demonstrated an intra- and interday precision within the range of 2.1–3.7 and 1.4–7.8%, respectively. Further intra- and interday accuracy was within the range of 95.6–99.8 and 95.7–99.1% correspondingly. The mean recovery of ME and MED6 was86.07±6.87and80.31±5.70%, respectively. The described method was successfully employed in bioequivalence study of ME in Indian male healthy human volunteers under fasting conditions.

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

2011 ◽  
Vol 8 (1) ◽  
pp. 201-211 ◽  
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.

Purification of a basic somatomedin, from human plasma Cohn fraction IV-1, with physicochemical and radioimmunoassay similarity to somatomedin-C and insulin-like growth factor

1979 ◽  
Vol 57 (11) ◽  
pp. 1289-1298 ◽  
Author(s):  
R. Marvin Bala ◽  
B. Bhaumick
Keyword(s):  
Growth Factor ◽  
Human Plasma ◽  
Protein Separation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Extraction Procedure ◽  
Molecular Size ◽  
Protein Band ◽  
Insulin Like Growth Factor ◽  
Igf I

A basic somatomedin (SM) was purified from human plasma Cohn fraction IV-1 using an initial acid–ethanol–acetone extraction procedure followed by alternating molecular size or charge protein separation techniques. The final recovery of SM bioactivity was approximately 2% of that present in the starting Cohn fraction. The purified SM has an approximate molecular weight of 7500, pI 8.6, 4000 SM bioactivity units per milligram (as measured by a hypophysectomized rat bioassay) and a parallel approximately equipotent radioimmunoassay dose–response curve to SM-C and insulin-like growth factor-I (IGF-I). Sodium dodecyl sulfate – polyacrylamide gel electrophoresis of this purified SM revealed a single protein band. The preliminary determination of the amino acid sequence of the N terminus suggested that this SM preparation was over 75% pure and the first five N-terminal amino acids were identical with those of IGF-I.

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

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
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.

DEVELOPMENT OF LIQUID-LIQUID EXTRACTION PROCEDURE AND ITS APPLICATION FOR QUANTIFICATION OF ERLOTINIB IN SPIKED HUMAN PLASMA USING RP-HPLC WITH UV DETECTION

INDIAN DRUGS ◽  
2016 ◽  
Vol 53 (06) ◽  
pp. 46-50
Author(s):  
A Faizee ◽  
◽  
S. S Sonawane ◽  
A. S. Patil ◽  
S. J Kshirsagar ◽  
...  
Keyword(s):  
Human Plasma ◽  
Ammonium Acetate ◽  
Extraction Procedure ◽  
Internal Standard ◽  
Hplc Method ◽  
Liquid Extraction ◽  
Calibration Model ◽  
Spiked Human Plasma ◽  
Liquid Liquid Extraction ◽  
Rp Hplc

A simple, rapid and accurate RP-HPLC method was developed and validated for the quantification of Erlotinib in spiked human plasma using liquid-liquid extraction. Sufficient recovery was obtained when drug and internal standard (Nabumetone) were extracted using ethyl acetate and 1N NaOH. Chromatographic separation was performed on C18 Phenomenex Hyperclone column (250 × 4.6 mm, 5 μm) using mobile phase acetonitrile: 20 mM ammonium acetate buffer pH 4.6 (60:40%,V/V). Flow rate was kept constant at 1 mL/min and detection was carried out at 331 nm. Calibration curve was found to be linear in the range of 100-3200 ng/mL. During the calibration experiments, it was found that heteroscedasticity can be minimized using weighted regression calibration model with weighing factor of 1/x2.

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

2012 ◽  
Vol 9 (3) ◽  
pp. 1605-1612
Author(s):  
M. Ganesan ◽  
S. Nanjundan ◽  
S. Viswanathan ◽  
G. Uma
Keyword(s):  
Human Plasma ◽  
Solid Phase ◽  
Internal Standard ◽  
Bioequivalence Study ◽  
Negative Ion ◽  
Limit Of Quantification ◽  
Tauroursodeoxycholic Acid ◽  
Liquid Chromatography Tandem Mass ◽  
Glycoursodeoxycholic Acid ◽  
Negative Ion Mode

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.

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

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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