scholarly journals Development and Validation of Rapid LC-MS with Electrospray Ionization for the Quantification of Pramipexole in Human Plasma

2014 ◽  
Vol 37 ◽  
pp. 1-15 ◽  
Author(s):  
Petikam Lavudu ◽  
Avula Prameela Rani ◽  
Chandra Bala Sekharan
Keyword(s):  
Human Plasma ◽  
Internal Standard ◽  
Stability Studies ◽  
Acceptable Accuracy ◽  
Liquid Liquid Extraction ◽  
Accuracy And Precision ◽  
Spiked Plasma ◽  
The Mean ◽  
Development And Validation

A rapid, accurate and precise LC-MS method is described for the quantitative determination of pramipexole in human plasma matrix using ropinirole as internal standard. Pramipexole and ropinirole were extracted from plasma by liquid-liquid extraction technique. The method was validated over the concentration range of 100-2514 pg/mL. The method was found to have acceptable accuracy, precision, linearity and selectivity. The mean extraction recovery from spiked plasma samples was in the range of 79.415-87.00 %. The intra-day accuracy of the assay ranged from 98.924 to 112.236 % and intra-day precision ranged from 3.489 to 6.756 %. Inter-day accuracy and precision results for quality control samples ranged between 100.340 and 107.443% of nominal and precision is observed to be 3.970-5.714 %. The pramipexole was found to be stable after several stability studies. The proposed method yielded a quick, simple and reliable protocol for estimating pramipexole concentrations in human plasma.

Determination of Ibuprofen in Human Plasma and Urine by Gas Chromatography/Mass Spectrometry

2014 ◽  
Vol 97 (2) ◽  
pp. 415-420 ◽  
Author(s):  
Bilal Yilmaz ◽  
Ali Fuat Erdem
Keyword(s):  
Human Plasma ◽  
Internal Standard ◽  
Gas Chromatography Mass Spectrometry ◽  
Oral Tablet ◽  
Liquid Liquid Extraction ◽  
Human Plasma And Urine ◽  
The Mean ◽  
Interday Precision ◽  
Better Than

Abstract This paper describes a GC/MS method for the determination of ibuprofen in human plasma and urine. Ibuprofen and internal standard naproxen were extractedfrom plasma and urine by using a liquid–liquid extraction method. Derivatization was carried outusing N-methyl-N-(trimethylsilyl) trifluoroacetamide. Calibration curves were linear over the concentration range of 0.05–5.0 and 0.1–10.0 μg/mL for plasma and urine, respectively. Intraday and interday precision (RSD) values for ibuprofen in plasma and urine were less than 6.31%, and accuracy (relative error) was better than 12.00%. The mean recovery of ibuprofen was 89.53% for plasma and 93.73% for urine. TheLOD was 0.015 and 0.03 μg/mL and the LOQ was 0.05 and 0.1 μg/mL for plasma and urine, respectively. The method was successfully applied to blood samples from three healthy male volunteers who had been given an oral tablet of 600 mg ibuprofen.

SENSITIVE BIO ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF VENETOCLAX IN HUMAN PLASMA BY LC-ESI-MS/MS

INDIAN DRUGS ◽  
2020 ◽  
Vol 57 (06) ◽  
pp. 32-38
Author(s):  
H. Potluri ◽  
Keyword(s):  
Human Plasma ◽  
Method Development ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Liquid Liquid Extraction ◽  
Liquid Chromatography Tandem Mass ◽  
Positive Mode ◽  
Method Development And Validation ◽  
Development And Validation

A specific and sensitive method of liquid chromatography–tandem mass spectrometry was demonstrated for the experimental determination of venetoclax in human plasma utilising venetoclax-D8 as an internal standard. The column Xbridge C18, 50 × 4.6mm, 5 µm was used for attaining chromatographic separation by utilising 10mM ammonium formate and methanol as isocratic mobile phase in the composition ratio of 20:80 (V/V). The flow-rate selected was 0.7ml/min. Venetoclax and venetoclax-D8 are identified in multiple reaction monitoring (MRM) positive mode with proton adducts at m/z 869.53 →553.21 and m/z 877.14 → 553.23, respectively. For the successful extraction of drug as well as internal standard, liquid-liquid extraction technique was efficiently utilised. The developed technique was established in a linear concentration range of 5.0-5000.0 pg/ml along with correlation coefficient (r2) of 0.9994. Intra and inter-day precisions were found to be 0.7 to 1.90% and 0.7 to 2.0 % for venetoclax and venetoclax-D8, respectively. Accuracy was found to be within 98.6 to 101.99% and 99.17 to 101.14 % for venetoclax and venetoclax-D8, respectively. It was observed that throughout the bench top studies, post-operative stability studies and freeze-thawing cycles, venetoclax retained stability.

scholarly journals Development and Validation of a LC/MS/MS Method for the Determination of Duloxetine in Human Plasma and Its Application to Pharmacokinetic Study

2012 ◽  
Vol 9 (2) ◽  
pp. 899-911 ◽  
Author(s):  
D. Chandrapal Reddy ◽  
A. T. Bapuji ◽  
V. Surayanarayana Rao ◽  
V. Himabindu ◽  
D. Rama Raju ◽  
...  
Keyword(s):  
Human Plasma ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Pharmacokinetic Parameters ◽  
Liquid Liquid Extraction ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Edta Plasma ◽  
Development And Validation

A selective, high sensitive and high throughput liquid chromatography-tandem mass spectrometry (LC-ESI-MS/MS) method has been developed and validated for the chromatographic separation and quantitation of duloxetine in human EDTA plasma using fluoxetine (IS) as an internal standard. Analyte and IS were extracted from human plasma by liquid-liquid extraction using MTBE-n Hexane (80:20).The eluted samples were chromatographed on X-terra RP8 (50 mmx4.6 mm, 5 μm particle size) column by using mixture of 30 mM ammonium formate (pH-5.0±0.05) and acetonitrile as an isocratic mobile phase at a flow rate of 0.40 mL/min and analyzed by mass spectrometer in the multiple reaction monitoring (MRM) using the respective m/z 298.08→154.0 for duloxetine and 310.02→148.07 for IS. The linearity of the response/ concentration curve was established in human plasma over the concentration range 0.100-100.017 ng/mL. The lower detection limit (LOD,S/N>3) was 0.04 ng/mL and the lower limit of quantization (LOQ,S/N>10) was 0.100 ng/mL. This LC-MS/MS method was validated with Intra-batch and Inter-batch precision of 5.21-7.02. The Intra-batch and Inter-batch accuracy was 97.14-103.50 respectively. Recovery of duloxetine in human plasma is 80.31% and ISTD recovery is 81.09%. The main pharmacokinetic parameters were Tmax(hr) = (7.25±1.581), Cmax(ng/mL) (44.594±18.599), AUC0→t, = (984.702±526.502) and AUC0→∞, (1027.147±572.790) respectively.

Development and Validation of a Liquid Chromatography/Mass Spectrometry Method for the Simultaneous Quantitation of Rosuvastatin and Ezetimibe in Human Plasma

2013 ◽  
Vol 96 (2) ◽  
pp. 307-312 ◽  
Author(s):  
Susheel John Varghese ◽  
Ravi Thengungal Kochupappy
Keyword(s):  
Human Plasma ◽  
Internal Standard ◽  
Liquid Chromatography Mass Spectrometry ◽  
Selected Ion Monitoring ◽  
Spectrometry Method ◽  
Esi Ms ◽  
Standard Curve ◽  
Liquid Liquid Extraction ◽  
Development And Validation

Abstract A simple, rapid, and sensitive LC/electrospray ionization (ESI)-MS method was developed and validated for the simultaneous determination of rosuvastatin (ROS) and ezetimibe (EZE) in human plasma. Following liquid–liquid extraction, the analytes and an internal standard, atorvastatin (ATO), were separated using an isocratic mobile phase comprising 0.1% (v/v) formic acid–methanol (20 + 80, v/v) on an RP-C18 column. Detection was performed on a mass spectrometer by selected ion monitoring using their respective [M-H]– ions, m/z 480 for ROS, m/z 408 for EZE, and m/z 557 for ATO. For both analytes, the method was linear in the range of 0.1 to 10 ng/mL. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. A run time of 4 min made it possible to determine many plasma samples/day. The validated LC/ESI-MS method can be used to study pharmacokinetics, bioavailability, and bioequivalence of combined dosage forms of ROS and EZE.

scholarly journals Bioanalytical Method Development and Validation of Estimation of Nimorazole by RP-HPLC in Human Plasma

2019 ◽  
Vol 12 (4) ◽  
pp. 7-12
Author(s):  
Nilam B Jadhav ◽  
Savita S Yadav ◽  
Kuldeep Singh Gusain
Keyword(s):  
Human Plasma ◽  
Method Development ◽  
Therapeutic Drug ◽  
Main Principle ◽  
Liquid Liquid Extraction ◽  
Rp Hplc ◽  
Method Development And Validation ◽  
The Mean ◽  
Development And Validation

Nimorazole is an antiprotozoal medicine used to treat infections caused by protozoa in the stomach, intestines, or genital areas. The main principle of this study was to develop RP-HPLC technique for the quantitative determination of Nimorazole in human plasma. The separation was accomplished by the isocratic method by using column C18 (Thermosil ODS), detection wavelength was 294 nm. The analyte was extracted in acetonitrile by liquid-liquid extraction. Acetonitrile: water in the ratio 30:70 was used as mobile phase for estimation of the drug in human plasma with a flow rate of 0.8 mL/min at a detection wavelength of 294 nm. Retention time was found to be 7.933 ± 0.23 min.The developed method was found to be linear over the concentration range of 60-360 μg/mL, with a correlation coefficient of 0.9991. The LOD and LOQ were found to be 10 μg/mL and 40 μg/mL, respectively. The method ensure for Precision and % RSD was found to be less than 2 % and the mean % recovery was found to be 99.58%. This method was effectively and favourably applied to the plasma samples and it seems to be appropriate tool for regular therapeutic drug monitoring of anti-infective drugs.

scholarly journals Development and Validation of Novel HPLC Bioanalytical Analysis Method for Acalabrutinib: An Anticancer Drug in Human Plasma

2020 ◽  
Vol 32 (10) ◽  
pp. 2606-2610
Author(s):  
G. Atchutarama Krishna ◽  
P. Srinivasarao ◽  
T. Benarji Patrudu ◽  
R. Chidanandaswamy
Keyword(s):  
Human Plasma ◽  
Orthophosphoric Acid ◽  
Internal Standard ◽  
Hplc Method ◽  
Linear Calibration ◽  
Liquid Liquid Extraction ◽  
Ich Guidelines ◽  
Relative Standard ◽  
Development And Validation

The aim of the work is to develop and validate the bioanalytical RP-HPLC method for determination of acalabrutinib in plasma with nifedipine drug as internal standard. Liquid-liquid extraction with diethyl ether and methanol in the ratio of 50:50 (v/v) was used for the extraction of drugs from the biological matrix. The optimized chromatography conditions consist of methanol, acetonitrile and 0.1% orthophosphoric acid in the ratio of 45:35:20 (v/v) as a mobile phase with KNAUER Eurospher II C18 Column (250 × 4.6 mm, 5μ) as stationary phase. Isocratic elution with 0.9 mL flow separates acalabrutinib at 4.6 min and nifedipine at 6.8 min. The method was validated as per ICH guidelines and linear calibration curve was obtained for the peak area ratio of acalabrutinib and nifedipine compound across a range of 50-3000 ng/mL. Greater than 90% recoveries were obtained for acalabrutinib. The relative standard deviation (%RSD) was found to be < 5% for precision studies. Hence, the method was found to be suitable for the analysis of acalabrutinib in spiked human plasma and is used for the pharmacokinetic study

scholarly journals Determination of nimodipine in plasma by HPLC-MS/MS and pharmacokinetic application

2010 ◽  
Vol 46 (4) ◽  
pp. 665-677 ◽  
Author(s):  
Demétrius Fernandes do Nascimento ◽  
Manoel Odorico de Moraes ◽  
Fernando Antônio Frota Bezerra ◽  
Andréa Vieira Pontes ◽  
Célia Regina Amaral Uchoa ◽  
...  
Keyword(s):  
Human Plasma ◽  
Internal Standard ◽  
Pharmacokinetic Profile ◽  
Plasma Samples ◽  
Linear Calibration ◽  
Limit Of Quantification ◽  
Standard Curve ◽  
Liquid Liquid Extraction ◽  
Highly Sensitive

To develop and validate a rapid, specific and highly sensitive method to quantify nimodipine in human plasma using dibucaine as the internal standard (IS). The analyte and IS were extracted from plasma samples by liquid-liquid extraction using hexane-ethyl acetate (1:1 v/v). The chromatographic separation was performed on a Varian® Polaris C18 analytical column (3 μm, 50 x 2.0 mm) and pre-column SecurityguardTM C18 (4.0 x 3.0 mm) with a mobile phase of Acetonitrile-Ammonium acetate 0.02 ml/L (80:20v/v). The method had a chromatographic run time of 4.5 min and linear calibration curve over the range of 0.1- 40 ng/mL (r > 0.9938). The limit of quantification was 100 pg/mL. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. This validated method was successfully applied in determining the pharmacokinetic profile of nimodipine tablets of 30 mg administered to 24 healthy volunteers. The proposed method of analysis provided a sensitive and specific assay for nimodipine determination in human plasma. The time for the determination of one plasma sample was 4.5 min. This method is suitable for the analysis of nimodipine in human plasma samples collected for pharmacokinetic, bioavailability or bioequivalence studies in humans.

scholarly journals DEVELOPMENT AND VALIDATION OF A TLC DENSITOMETRIC METHOD FOR DETERMINATION OF GLIMEPIRIDE IN TABLETS

2014 ◽  
Vol 16 (1) ◽  
pp. 11-15
Author(s):  
Yuni Retnaningtyas ◽  
Lestyo Wulandiri ◽  
Gabriella F Punu
Keyword(s):  
Pharmaceutical Analysis ◽  
Data Recovery ◽  
Antidiabetic Drug ◽  
Intermediate Precision ◽  
Quantification Limit ◽  
Accuracy And Precision ◽  
Quality Control Testing ◽  
The Mean ◽  
Development And Validation

A simple and valid TLC method has been developed for the determination of glimepiride in tablet formulation. After extraction of the analyte with a mixture of methanol and ammonia 0,2M (1:1, v/v), the extracts were spotted on precoated TLC silica gel F254 plates, which were developed with a mixture of toluene:methanol:ethyl acetate (75:20:5, v/v/v). Quantitative evaluation was performed by measuring the absorbance reflectance of the analyte spots at 238 nm. The method was validated for specificity, linearity, accuracy and precision. Good linearity was achieved in the concentration range 100–800 ng/spot. The RSD of repeatability and intermediate precision were found to be less than 2%, whereas the mean of the recovery data was 100-101%. The detection limit and quantification limit were 22 and 74 ng/spot, respectively. The method is specific, linear, precise, and accurate; it can be used for the routine quality control testing of marketed formulations.Keywords: glimepiride, TLC densitometric, validation of pharmaceutical methods, pharmaceutical analysis, antidiabetic drug Sebuah metode Kromatografi Lapis Tipis (KLT) yang sederhana dan valid telah dikembangkan untuk penentuan glimepiride dalam sediaan tablet. Setelah analit dalam sampel diekstraksi dengan campuran metanol dan amonia 0,2 M (1:1,v/v), ekstrak yang terlihat pada lempeng silika gel F254, yang dikembangkan dengan campuran toluen : metanol : etil asetat (75:20:5, v/v/v). Evaluasi kuantitatif dilakukan dengan mengukur reflektansi absorbansi noda analit pada panjang gelombang 238 nm.Metode ini divalidasi meliputi spesifisitas, linearitas, akurasi dan presisi.Linearitas yang baik dicapai pada rentang konsentrasi 100-800 ng / spot. RSD pengulangan dan presisi intermediate menunjukkan nilai kurang dari 2 %, sedangkan rata-rata data recovery adalah 100-101 % .Batas deteksi dan batas kuantifikasi adalah 22 dan 74 ng / noda.Metode ini spesifik, linear, tepat, dan akurat, bisa digunakan untuk pengujian kontrol kualitas rutin tablet glimepirid dipasarkan. Kata Kunci: glimepiride, TLC densitometric, validation of pharmaceutical methods, pharmaceutical analysis, antidiabetic drug

Development and Validation of an RP-HPLC Method for Determination of Atorvastatin and its Hydroxyl Metabolites in Human Plasma

2020 ◽  
Vol 16 (3) ◽  
pp. 238-245
Author(s):  
Dagmara Sowińska ◽  
Alicja Pogorzelska ◽  
Marlena Rakicka ◽  
Justyna Sznura ◽  
Justyna Janowska ◽  
...  
Keyword(s):  
Human Plasma ◽  
Internal Standard ◽  
Hplc Method ◽  
Active Metabolites ◽  
Peak Separation ◽  
Rp Hplc ◽  
Increased Risk ◽  
Relative Standard ◽  
Development And Validation

Background: Atorvastatin (AT) belongs to cholesterol-lowering agents, commonly used in patients with an increased risk of cardiovascular disease. The drug, as well as its hydroxyl metabolites, exhibit pharmacological activity, and their plasma levels may be helpful in the assessment of the therapeutic effectiveness. Objective: Development and validation of a fast and reproducible RP-HPLC method with UV detection for the simultaneous determination of atorvastatin and its active metabolites, para-hydroxy-atorvastatin (p-OH-AT) and ortho-hydroxy-atorvastatin (o-OH-AT) in human plasma. Methods: Optimal conditions of chromatographic separation of the analytes, as well as rosuvastatin, chosen as an internal standard, were studied. The absorbance of the compounds was measured at λ=248 nm. Validation of the method was performed. The usefulness of the method was confirmed for determination of the analytes in plasma of patients treated with the drug. Results: Total peak separation was achieved at LiChrospher 100 RP-18 column with a mobile phase composed of methanol and water (1:1,v:v) and a flow rate of 1.2 ml/min. The method was linear in the ranges of 0.025 - 1.0 μg/ml for AT, o-OH-AT and p-OH-AT. Intra- and inter-assay precision expressed as relative standard deviation was ≤13% for AT, ≤12% for p-OH-AT and ≤11% for o-OH-AT. Intraand inter-day accuracy of the method, expressed as a relative error, was ≤15%. Conclusion: The elaborated HPLC method is specific, repeatable, reproducible, adequately accurate and precise and fulfills the validation requirements for the bioanalytical method. The method was successfully applied for analysis of atorvastatin and its o-hydroxy metabolite in plasma of patients treated with the drug.

Simultaneous Determination of Tramadol and its Metabolite in Human Plasma by GC/MS

2015 ◽  
Vol 98 (1) ◽  
pp. 56-61 ◽  
Author(s):  
Bılal Yılmaz ◽  
Alı Fuat Erdem
Keyword(s):  
Simultaneous Determination ◽  
Human Plasma ◽  
Room Temperature ◽  
Internal Standard ◽  
Stability Studies ◽  
Freeze Thaw ◽  
Calibration Curves ◽  
Interday Precision ◽  
Intravenous Formulation

Abstract A simple and sensitive GC/MS method for the determination of tramadol and its metabolite (O-desmethyltramadol) in human plasma was developed and validated. Medazepam was used as an internal standard. The calibration curves were linear (r = 0.999) over tramadol and O-desmethyltramadol concentrations ranging from 10 to 200 ng/mL and 7.5 to 300 ng/mL, respectively. The method had an accuracy of &gt;95% and intra- and interday precision (RSD%) of ≤4.83% and ≤4.68% for tramadol and O-desmethyltramadol, respectively. The extraction recoveries were 97.6 ± 1.21% and 96.3 ± 1.66%for tramadol and O-desmethyltramadol, respectively. The LOQ using 0.5 mL human plasma was 10 ng/mL for tramadol and 7.5 ng/mL for O-desmethyltramadol. Stability studies showed that tramadol and O-desmethyltramadol were stable in human plasma after 8 h incubation at room temperature or after 1 week storage at –20°C with three freeze-thaw cycles. Also, this method was successfully applied to six patients who had been given an intravenous formulation of 100 mg tramadol with Cmax results of 2018.1 ± 687.8 and 96.1 ±22.7 ng/mL for tramadol and O-desmethyltramadol, respectively.

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