scholarly journals Bioanalytical Method Development and Validation for Determination of Sulfasalazine in Rabbit Plasma by HPLC-UV

2021 ◽  
Vol 33 (7) ◽  
pp. 1692-1698
Author(s):  
S.S. Jadiya ◽  
N. Upmanyu ◽  
S. Arulmozhi ◽  
V. Jain ◽  
S. Sankaran ◽  
...  
Keyword(s):  
High Performance ◽  
Method Development ◽  
High Efficiency ◽  
Internal Standard ◽  
Ultra Violet ◽  
Precipitation Method ◽  
Pharmacokinetic Studies ◽  
Rabbit Plasma ◽  
Bioanalytical Method

In present study, an advanced, simple and a rapid reverse phase high performance liquid chromatography (RP-HPLC) method was developed for the quantitative determination of sulfasalazine in rabbit plasma. Sulfasalazine was separated using Chromatopak C-18 basic peerless (250 mm × 4.6 mm, 5μ) column in an isocratic mode using mobile phase consisting of the mixture of 10mM Ammonium acetate pH adjusted to 4.5 and acetonitrile (70:30 v/v) with a flow rate of about 1.0 mL/min at ambient temperature. An ultra-violet detection of sulfasalazine and the internal standard was carried out at 362 nm. Both sulfasalazine and internal standard (IS, 4-hydroxy benzoate) were extracted from plasma matrices with high efficiency using a simple protein precipitation method. The method was found to be highly selective with no carryover effects. Linearity of sulfasalazine was found with the range of 2.5-100 μg/mL with the value of r2 > 0.995 a correlation coefficient. At all three quality control levels, developed bioanalytical method was found as repeatable and reproducible as well. The average recoveries of sulfasalazine from plasma were in the range of 95.59-97.16%. The bioanalytical samples showed good and acceptable stability of sulfasalazine solution at different storage, packaging and handling conditions. Hence, in conclusion, the validated and developed HPLC-UV method could be effectively utilized for determination of sulfasalazine in pharmacokinetic studies involving novel formulations.

scholarly journals Robust and Sensitive High-Performance Liquid Chromatographic-UV Detection Technique for the Determination of Tigecycline in Rabbit Plasma

2011 ◽  
Vol 94 (3) ◽  
pp. 847-856 ◽  
Author(s):  
Kostas M Zorpas ◽  
Georgia N Valsami ◽  
Evangelos V Vryonis ◽  
Athanasios T Skoutelis ◽  
Helen A Archontaki
Keyword(s):  
High Performance ◽  
Internal Standard ◽  
Pharmacokinetic Profile ◽  
High Performance Liquid Chromatographic ◽  
Hplc Method ◽  
Pharmacokinetic Studies ◽  
Rabbit Plasma ◽  
Elution Time ◽  
Liquid Chromatographic

Abstract An isocratic HPLC method with detection at 248 nm was developed and fully validated for the determination of tigecycline in rabbit plasma. Minocycline was used as an internal standard. A Hypersil BDS RP-C18 column (250 × 4.6 mm, 5 μm particle size) was used with the mobile phase phosphate buffer (pH 7.10, 0.070 M)–acetonitrile (76 + 24, v/v) at a flow rate of 1.0 mL/min. The elution time of tigecycline and minocycline was approximately 8.1 and 9.9 min, respectively. Calibration curves of tigecycline were linear in the concentration range of 0.021–3.15 μg/mL in plasma. The LOD and LOQ in plasma were estimated as 7 and 21 ng/mL, respectively. The intraday and interday precision values of the method were in the range of 5.0–7.1 and 5.6–9.1%, while the corresponding accuracy values were in the ranges of 92.8–111.1 and 97.6–102.3%, respectively. At the LOQ, the intraday precision was 18.7%, while intraday and interday accuracy values were 97.3 and 98.0%, respectively. Robustness of the proposed method was studied using a Plackett-Burman experimental design. A pharmacokinetic profile is presented for confirmation of the applicability of the method to pharmacokinetic studies.

scholarly journals Bioanalytical Method Development and Validation of Veratraldehyde and Its Metabolite Veratric Acid in Rat Plasma: An Application for a Pharmacokinetic Study

Molecules ◽  
2020 ◽  
Vol 25 (12) ◽  
pp. 2800
Author(s):  
Hyun Wook Huh ◽  
Hee-Yong Song ◽  
Young-Guk Na ◽  
Minki Kim ◽  
Mingu Han ◽  
...  
Keyword(s):  
Method Development ◽  
Internal Standard ◽  
Rat Plasma ◽  
Precipitation Method ◽  
Veratric Acid ◽  
Bioanalytical Method ◽  
Positive Mode ◽  
Percutaneous Administration ◽  
The One

A simple, sensitive, and rapid UHPLC-MS/MS method was developed for the simultaneous determination of veratraldehyde and its metabolite veratric acid in rat plasma. Cinnamaldehyde was used as an internal standard (IS) and the one-step protein precipitation method with 0.2% formic acid in acetonitrile (mobile phase B) was used for the sample extraction. Reversed C18 column (YMC-Triart C18 column, 50 mm × 2.0 mm, 1.9 µm) was used for chromatographic separation and was maintained at 30 °C. The total run time was 4.5 min and the electrospray ionization in positive mode was used with the transition m/z 167.07 → 139.00 for veratraldehyde, m/z 183.07 → 139.00 for veratric acid, and m/z 133.00 → 55.00 for IS. The developed method exhibited good linearity (r2  ≥  0.9977), and the lower limits of quantification ranged from 3 to 10 ng/mL for the two analytes. Intra-day precision and accuracy parameters met the criteria (within ±15%) during the validation. The bioanalytical method was applied for the determination of veratraldehyde and veratric acid in rat plasma after oral and percutaneous administration of 300 and 600 mg/kg veratraldehyde. Using the analytical methods established in this study, we can confirm the absorption and metabolism of veratraldehyde in rats for various routes.

Determination of 8-methoxypsoralen in suction-blister fluid and serum by liquid chromatography.

1980 ◽  
Vol 26 (13) ◽  
pp. 1825-1828 ◽  
Author(s):  
M J Herfst ◽  
P M Edelbroek ◽  
F A de Wolff
Keyword(s):  
High Performance ◽  
Internal Standard ◽  
Reversed Phase ◽  
Pharmacokinetic Studies ◽  
Blister Fluid ◽  
Liquid Chromatograph ◽  
Suction Blister ◽  
Drug Concentrations ◽  
Suction Blister Fluid

Abstract A method is described for determination of 8-methoxypsoralen to 0.2 mL of suction-blister fluid or 1 mL of serum from psoriatic patients being treated with this drug. The drug is extracted from the biological matrix at pH 9.0 with a mixture of dichloromethane and light petroleum ether. 5-Methoxypsoralen is used as internal standard. Separation and quantitation are performed on a “high-performance” liquid chromatograph with use of an RP 18 reversed-phase column and detection at 245 nm. Accuracy and precision are good. Some benzodiazepines and their metabolites interfere. The lowest detectable concentration is 10 microgram/L, which means that the method is sufficiently sensitive to measure the drug concentrations in serum and suction-blister fluid for pharmacokinetic studies in patients being treated with a therapeutic dosage.

scholarly journals Influence of hydroxybosentan on determination of bosentan in human plasma

2015 ◽  
Author(s):  
Edyta Gilant ◽  
Katarzyna Buś - Kwaśnik ◽  
Izabela Domel ◽  
Michał Kaza ◽  
Piotr J. Rudzki
Keyword(s):  
Human Plasma ◽  
Maximum Concentration ◽  
High Performance ◽  
Internal Standard ◽  
European Medicines Agency ◽  
Bioanalytical Method ◽  
Elimination Half ◽  
Validation Parameters ◽  
Pharmacologically Active

Bosentan is a drug used in the treatment of pulmonary arterial hypertension. Evaluation of bosentan pharmacokinetics is an important step of the drug development. The aim of our study was to evaluate bioanalytical method reliability by assessing the influence of metabolite on the determination of bosentan in human plasma. In human body, bosentan is converted to three metabolites. Hydroxybosentan is an only pharmacologically active metabolite and its maximum concentration in plasma reaches 5-8% of the bosentan’s maximum concentration. Hydroxybosentan’s elimination half-life was reported to be very similar to that of bosentan (5-6 h) or slightly greater (6-14 h). It was observed that times to reach the maximum concentrations are similar for both compounds. Combination of high-performance liquid chromatography with UV-Vis detection and liquid-liquid extraction enabled to was determine bosentan in human plasma in the range of 50-4000 ng∙mL-1. During the method validation, metabolite back-conversion was studied using human plasma samples spkied with hydroxybosentan to obtain concentration of 500 ng·mL-1. Additionally, the influence of metabolite on extraction of bosentan and the internal standard was assessed. We found that there was no influence of metabolite on determination of bosentan in developed bioanalytical method. The method was validated according to European Medicines Agency (EMA) and Food and Drug Administration (FDA) guidelines, in compliance with the principles of Good Laboratory Practice (GLP). All of the validation parameters met acceptance criteria what confirmed method’s reliability.

scholarly journals Influence of hydroxybosentan on determination of bosentan in human plasma

2015 ◽  
Author(s):  
Edyta Gilant ◽  
Katarzyna Buś - Kwaśnik ◽  
Izabela Domel ◽  
Michał Kaza ◽  
Piotr J. Rudzki
Keyword(s):  
Human Plasma ◽  
Maximum Concentration ◽  
High Performance ◽  
Internal Standard ◽  
European Medicines Agency ◽  
Bioanalytical Method ◽  
Elimination Half ◽  
Validation Parameters ◽  
Pharmacologically Active

Bosentan is a drug used in the treatment of pulmonary arterial hypertension. Evaluation of bosentan pharmacokinetics is an important step of the drug development. The aim of our study was to evaluate bioanalytical method reliability by assessing the influence of metabolite on the determination of bosentan in human plasma. In human body, bosentan is converted to three metabolites. Hydroxybosentan is an only pharmacologically active metabolite and its maximum concentration in plasma reaches 5-8% of the bosentan’s maximum concentration. Hydroxybosentan’s elimination half-life was reported to be very similar to that of bosentan (5-6 h) or slightly greater (6-14 h). It was observed that times to reach the maximum concentrations are similar for both compounds. Combination of high-performance liquid chromatography with UV-Vis detection and liquid-liquid extraction enabled to was determine bosentan in human plasma in the range of 50-4000 ng∙mL-1. During the method validation, metabolite back-conversion was studied using human plasma samples spkied with hydroxybosentan to obtain concentration of 500 ng·mL-1. Additionally, the influence of metabolite on extraction of bosentan and the internal standard was assessed. We found that there was no influence of metabolite on determination of bosentan in developed bioanalytical method. The method was validated according to European Medicines Agency (EMA) and Food and Drug Administration (FDA) guidelines, in compliance with the principles of Good Laboratory Practice (GLP). All of the validation parameters met acceptance criteria what confirmed method’s reliability.

Bioanalytical Method Development of Atenolol Enantiomers: Stereoselective Behavior in Rabbit Plasma by RP-UFLC Method

2019 ◽  
Vol 16 ◽  
Author(s):  
Charan Raju C ◽  
B M Gurupadayya ◽  
Prachi Raikar
Keyword(s):  
Method Development ◽  
Pharmacokinetic Studies ◽  
Pharmacokinetic Data ◽  
Rabbit Plasma ◽  
Bioanalytical Method ◽  
Sodium Hydrogen Phosphate ◽  
Method Development And Validation ◽  
Rate Of Recovery ◽  
Fast Liquid Chromatography ◽  
Development And Validation

Objective: The objective of the method was to develop a new, simple and reliable enantioselective Reverse Phase- Ultra Fast Liquid Chromatography (RP-UFLC) method for the separation of Atenolol enantiomers. Comprehensive study was performed by extending the work to pharmacokinetic studies using rabbit plasma. Background: Many methods were reported for enantioseparation of Atenolol enantiomers but, no attempts were made for chiral separation of Atenolol using rabbit plasma. Moreover, pharmacokinetic data to prove the efficiency of particular enantiomers in rabbit plasma was not studied. Method: In the present examination, the binary RP-UFLC technique was developed on Phenomenex® Lux cellulose i5 segment (150×4.6 mm, 5µ) using di-sodium hydrogen phosphate buffer (pH 6.8): acetonitrile (35:65 v/v) as the mobile phase. Results: The elution of Atenolol was observed at 225 nm with a stream rate of 1 mL.min-1. The described technique offered a linear relationship with a regression coefficient of r2= 0.997 and 0.996 for (R) and (S)-enantiomer respectively between the concentration range of 2-10 ng.mL-1. Atenolol enantiomers were detected at a retention time (tR) of 2.7 min and 3.10 min R and S-enantiomer respectively. The rate of recovery of both Atenolol enantiomers was observed to be (R) 98.18% and (S) 100.45% individually. USFDA guidelines May 2018, were systematically followed for bioanalytical method development and validation. Conclusion: The developed technique was applied for the separation of Atenolol enantiomers and for the pharmacokinetic determination of Atenolol enantiomers in rabbit plasma.

Determination of Midodrine and its Active Metabolite Desglymidodrine by LC-MS/MS and its Application in Human Pharmacokinetic Studies

2019 ◽  
Vol 15 (4) ◽  
pp. 355-362
Author(s):  
S.T. Narenderan ◽  
S.N. Meyyanathan ◽  
B. Babu ◽  
Karthik Yamjala ◽  
S.J. Ashwini
Keyword(s):  
Active Metabolite ◽  
Internal Standard ◽  
Precipitation Method ◽  
Pharmacokinetic Studies ◽  
Stability Parameters ◽  
Liquid Chromatography Tandem Mass ◽  
Human Pharmacokinetic ◽  
Tandem Mass Spectroscopy ◽  
Linear Concentration Range

Background: Midodrine (MD) is a prodrug which is converted into Desglymidodrine (DMD) after oral administration. </P><P> Objective: The aim of the present study is to develop and validate a precise, accurate liquid chromatography- tandem mass spectroscopy (LC-MS/MS) method for the separation and detection of Midodrine and Desglymidodrine. Methods: The quantification of prodrug Midodrine (MD) and its active metabolite Desglymidodrine (DMD) in human plasma was performed using simple and economical protein precipitation method. Caffeine was used as an internal standard (IS). LC separation was carried out using Jones C18 column (4.6mm x 150mm, 3µm). Isocratic elution was performed using 10mM ammonium formate (pH 4.0 adjusted with formic acid): methanol 30:70, v/v as a mobile phase, at a constant of flow of 0.5 ml/min. Results: Mass spectrometric detection was carried out at positive electrospray ionization with proton adducts at m/z 255.0˃237.1, 198.1˃180.2 and 195.0˃138.1 for Midodrine, Desglymidodrine and caffeine (IS) respectively, in MRM mode. The method was validated over a linear concentration range of 0.3-110 ng/ml (r2 =0.996 and r2 =0.9988) for both Midodrine and Desglymidodrine. The recovery of Midodrine and Desglymidodrine were found to be 99 ± 0.12%. The precision (intra-day and inter-day) and accuracy studies fulfilled the acceptance criteria. Conclusion: The method shows to be stable for the studied stability parameters and was successfully applied for clinical pharmacokinetic studies.

A Novel LC-MS Method for the Determination of Abiraterone in Rat Plasma and its Application to Pharmacokinetic Studies

2021 ◽  
Vol 17 ◽  
Author(s):  
Linzhi Dai ◽  
Pei Lv ◽  
Yun He ◽  
Xiaoli Wang ◽  
Lili Chen ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Mobile Phase ◽  
High Performance ◽  
Analytical Procedure ◽  
Internal Standard ◽  
Rat Plasma ◽  
Pharmacokinetic Studies ◽  
Quadrupole Mass ◽  
Limit Of Quantitation

Background: High–performance liquid chromatography (HPLC)–ultraviolet (UV) and liquid chromatography (LC)–mass spectrometry (MS)/MS methods have been used to analyse abiraterone (ART); however, a single-quadrupole mass spectrometer with LC-MS systems has never been used to analyse ART. Objective: The study aimed to establish a novel, simple assay of quantitating ART in rat plasma through LC–MS. Method: The analytical procedure involved the extraction of ART and D4-ART (internal standard, IS) from rat plasma through simple protein precipitation. Chromatographic separation was achieved using an isocratic mobile phase (acetonitrile: 5 mM ammonium formate with 0.1% formic acid, 50:50 v/v) at a flow rate of 0.30 mL/min on a Waters XBridge® C18 column with a total run time of 5 min. LC–MS ion transitions monitored were 350.1 and 354.1 for ART and IS, respectively. The method was validated, and the results met acceptance criteria. Results: The lower limit of quantitation achieved was 1 ng/mL, and linearity was 1–8000 ng/mL. The intra- and inter-day precisions were 1.26%–14.20% and 5.49%–13.08%, respectively, in rat plasma.

scholarly journals Nonextractive Procedure and Precolumn Derivatization with 7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole for Trace Determination of Trimetazidine in Plasma by High-Performance Liquid Chromatography with Fluorescence Detection

2008 ◽  
Vol 91 (5) ◽  
pp. 1037-1044 ◽  
Author(s):  
Ibrahim A Darwish ◽  
Ashraf M Mahmoud ◽  
Nasr Y Khalil
Keyword(s):  
Human Plasma ◽  
Fluorescence Detection ◽  
High Performance ◽  
Internal Standard ◽  
High Performance Liquid Chromatographic ◽  
Pharmacokinetic Studies ◽  
Precolumn Derivatization ◽  
Trace Determination ◽  
Relative Standard

Abstract A highly sensitive high-performance liquid chromatographic method with fluorescence detection has been developed and validated in a single laboratory for the trace determination of trimetazidine (TMZ) in human plasma. Fluoxetine (FLX) was used as the internal standard. TMZ and FLX were isolated from plasma by protein precipitation with acetonitrile and derivatized by heating with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole in pH 8 borate buffer at 70C for 30 min. Separations were performed in the isocratic mode on a Nucleosil CN column with the mobile phase acetonitrile10 mM sodium acetate buffer (pH 3.5)methanol (47 + 47 + 6, v/v/v) at a flow rate of 1.0 mL/min. The derivatized samples were excited at 470 nm and monitored at an emission wavelength of 530 nm. Under the optimum chromatographic conditions, a linear relationship with a good correlation coefficient (r 0.9997, n 5) was obtained for the peak area ratio of TMZ to FLX and for TMZ concentrations of 1120 ng/mL. The proposed method has the lowest limits of detection and quantitation reported to date for the determination of TMZ in plasma with values of 0.3 and 0.95 ng/mL, respectively. The values for intra- and interassay precision were satisfactory; the relative standard deviations were 4.04. The accuracy of the method was demonstrated; the recoveries of TMZ from spiked human plasma were 98.13102.83 0.24.04. The method has high throughput because of its simple sample preparation procedure and short run time (&lt;10 min). The results demonstrated that the proposed method would have great value when applied in pharmacokinetic studies for TMZ.

Sign in / Sign up

Export Citation Format

Share Document