scholarly journals Development and Validation of UPLC-MS/MS Method for Rapid Simultaneous Determination of Levothyroxine and Liothyronine in Human Serum

2020 ◽  
Vol 10 (3-s) ◽  
pp. 176-181
Author(s):  
Rohit Dutt ◽  
Kailash Chander Malik ◽  
Manoj Karwa ◽  
Gaurav Kumar JAIN
Keyword(s):  
Mass Spectrometry ◽  
Human Serum ◽  
High Performance ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Rapid Separation ◽  
Bioequivalence Testing ◽  
Development And Validation ◽  
Liquid Chromatographic

A simple ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) method was developed and fully validated to simultaneously determine levothyroxine (LT4) and liothyronine (LT3) in human serum. Sample preparation was done through protein precipitation with acetonitrile. HyPURITY C18 column was selected to achieve rapid separation for LT4 and LT3 within 4 min. Electrospray ionization (ESI) under multiple reaction monitoring (MRM) was used to monitor the ion transitions for LT4 (m/z 777.54→731.52), LT3 (m/z 651.64→ 605.65) and internal standard LT4-D3 (m/z 780.53 →734.19), operating in the positive ion mode. The method was proved to be accurate (82.35% to 113.56%) and precise (0.73% to 8.28%) over concentration range of 50.37 ng/ml – 300.13 ng/ml for LT4 and 0.5 ng/ml – 50.37 ng/ml for LT3. The validated method could be applied for pharmacokinetic study or bioequivalence testing of combination products of LT4 and LT3. Keywords: Levothyroxine; Liothyronine; Ultra Performance Liquid Chromatographic; Mass Spectrometry; Human Serum

scholarly journals Development and Validation of an HPLC-MS/MS Method for Rapid Simultaneous Determination of Cefprozil Diastereomers in Human Plasma

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Guodong He ◽  
Liping Mai ◽  
Xipei Wang
Keyword(s):  
Mass Spectrometry ◽  
Antimicrobial Activity ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Reaction Monitoring ◽  
Reverse Phase ◽  
Development And Validation ◽  
Positive Ion ◽  
Sensitive Method

Background. Both cis- and trans-cefprozil have antimicrobial activity, but their potencies are quite different. It is therefore necessary to develop a sensitive method to simultaneously determine both isomers for pharmacokinetic and bioequivalence studies. Methods. An LC-MS/MS method, using stable isotope-labeled cefprozil as the internal standard, was developed and validated. The analytes were extracted from plasma by protein precipitation and separated on a reverse-phase C18 column using a gradient program consisting of 0.5% formic acid and acetonitrile within 4 min. The mass spectrometry acquisition was performed with multiple reaction monitoring in positive ion mode using the respective [M+H]+ ions, m/z 391.2→114.0 for cefprozil and 395.0→114.5 for cefprozil-D4. Results. The calibration curves were linear over the ranges of 0.025–15 μg/mL for cis-cefprozil and 0.014–1.67 μg/mL for trans-cefprozil. The accuracies for the cis and trans isomers of cefprozil were 93.1% and 103.0%, respectively. The intra- and interassay precisions for the QC samples of the isomers were < 14.3%. The intra- and interassay precisions at the LLOQ were < 16.5%. Conclusions. The method was sensitive and reproducible and was applied in a pilot pharmacokinetic study of healthy volunteers.

scholarly journals Development and Validation of Ultra High Performance Liquid Chromatographic (UHPLC) Method for the Determination of Roxithromycin in the Broiler Plasma

2019 ◽  
pp. 1-8
Author(s):  
R. D. Singh ◽  
S. K. Mody ◽  
H. B. Patel ◽  
V. N. Sarvaiya ◽  
B. R. Patel ◽  
...  
Keyword(s):  
High Performance ◽  
Internal Standard ◽  
High Performance Liquid Chromatographic ◽  
Pharmacokinetic Studies ◽  
Uv Detector ◽  
Linear Calibration ◽  
The Mean ◽  
Development And Validation ◽  
Liquid Chromatographic

Aims: The present study was designed to develop and validate the UHPLC method for quantitative determination of roxithromycin, a macrolide antimicrobial drug, in broiler plasma for the application of pharmacokinetic studies. Methodology: UHPLC apparatus comprised of ultraviolet (UV) detector was used in the present study. Chromatographic separation was performed by using reverse phase C18 column. Mobile phase was combination of buffer and 55 acetonitrile in the ratio of 55: 45. Buffer part used was 0.1% trifluoroacetic acid (v/v) having pH of 2.1. Erythromycin was used as an internal standard. Isocratic elution mode was employed with flow rate of 1 ml/min and effluents were monitored at wavelength of 220 nm. Liquid-liquid extraction using ice-cold acetonitrile was performed to extract roxithromycin from plasma samples. The data integration was performed using Chromeleon™ version 6.8 software. Results: The linear calibration curve with a mean correlation coefficient (R2) value of 0.9999 was observed for concentrations ranging from 0.20 to 12.80 µg/ml. At any concentration, accuracy was not found to be less than 90%. The mean extraction recovery (n=5) for concentrations of 0.40 µg/ml was 81.36%. The calculated intraday and interday C.V. % was not more than 7.70% and 9.42%, respectively, at any concentration studied. The specificity of the analysis was reflected by the narrow range of retention time ranging between 6.983 to 7.178 minutes. LOD and LOQ of the method under investigation were calculated as 0.131 and 0.398 µg/ml, respectively. Conclusion: A reliable, reproducible, accurate, precise, specific and sensitive method for analysis of roxithromycin in broiler plasma was developed and validated for application in the pharmacokinetic study of the roxithromycin.

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