scholarly journals A review on bioanalytical method development and pre-method validation concepts using SPE and LLE process by LC-MS/MS method

2021 ◽  
Vol 12 (2) ◽  
pp. 1353-1359
Author(s):  
Arun Kumar V ◽  
Vijey Aanandhi M ◽  
Gandhimathi R ◽  
Sumithra M
Keyword(s):  
Method Validation ◽  
Analytical Method ◽  
Method Development ◽  
Biological Fluids ◽  
Internal Standard ◽  
Pharmacokinetic Studies ◽  
Detection Techniques ◽  
Extraction Procedures ◽  
Highly Sensitive ◽  
Pharmacokinetic Properties

The review article provides guidance for the development of a Bio-Analytical Method intended for the estimation of drugs in biological fluids. The development of a suitable analytical method for the identification, isolation and quantification of different drugs and or their metabolites from biological fluids is an essential and challenging component of pharmacokinetic studies. The uses of detection techniques that are highly sensitive and specific for the quantification of drugs and or metabolites in biological fluids are preferred. To collect the information on physiochemical, pharmacokinetic, chromatographic and extraction procedures of the drug/Metabolites from available sources, e.g. Medline, Journals, Analytical abstract, Physician Desk Reference, Library etc. Summarize the physiochemical, pharmacokinetic properties, chromatographic and extraction procedures. Select the equipment according to the availability and sensitivity. Select the initial chromatographic conditions and also identify the extraction. The selection of Internal standard should be similar to the analyte. Prepare the required solutions and stock dilutions at the required concentration. Tune the instrument with suitable solutions and optimize parameters for Q1/Q3 ions. Condition the system and column with the selected mobile phase. Perform pre-method validation experiments. Select the best weighing factors for the standards. Compile all the chromatograms and raw data and archive since the development of a Bio-Analytical method for a given drug is so essential to start with, this guidance for approaching and conclude for a suitable method that later can be validated. It is important to note that this is intended to identify minimal criteria for producing consistent and comparable data. This procedure is applicable to the methods that are to be developed for the estimation of drugs and metabolites from biological fluids using any chromatographic devices in the Bio- Analytical Department.

scholarly journals Bio analytical method development and validation for Ezetimibe and Pitavastain and its applications to pharmacokinetic studies in Rabbit plasma by using LCMS/MS

2020 ◽  
Vol 11 (4) ◽  
pp. 7854-7862
Author(s):  
Potturi Ramadevi ◽  
Kantipudi Rambabu
Keyword(s):  
Method Development ◽  
Extraction Process ◽  
Pharmacokinetic Studies ◽  
Rabbit Plasma ◽  
Highly Sensitive ◽  
The Us ◽  
Positive Mode ◽  
Method Development And Validation ◽  
Development And Validation ◽  
Preparation Techniques

For the gradation of Ezetimibe and Pitavastain in rabbit plasma, a highly sensitive and simple LC-MS/MS assay was developed and witnessed. The chromatographic conditions are isocratic with a waters symmetry C18 (150 x 4.6 mm, 3.5) column in isocratic mode. The detection was carried out using a mobile phase of 0.1 percent formic acid and 60:40 acetonitrile, and the detection was carried out using MS in a positive mode of electrospray ionisation. The valid approach was checked with a linear range of 10-200 ng/ml Ezetimibe and 2-40 ng/ml Pitavastain. The intraday and interday precision values were found to be within reasonable limits. The liquid extraction process is used to remove these drugs from rabbit plasma. And these drugs have been shown to be stable in freeze-thaw, autosampler, and benchtop tests in the future. The fluid chromatography coupled mass spectrometry strategy was approved by the US Food and Drug Administration for quantification of Ezetimibe and Pitavastain in rabbit plasma using D4–ezetimibe and D4–pitavastain as within norms using LC-MS consolidated with quadrupole spectrometer by electro shower ionisation process. The aim of this study is to evaluate the applicability of this approach to ezetimibe and pitavastain at different evaluation levels while taking into account various factors such as instrument stability, precision, and accuracy, sample preparation techniques, instrument calibration, recovery, and matrix effect by using Ezetimibe and Pitavastain, as well as their internal guidelines.

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 DEVELOPMENT AND VALIDATION OF LC-MS/MS METHOD FOR ESTIMATION OF UROCARB IN HUMAN PLASMA

2019 ◽  
pp. 125-130
Author(s):  
IRYNA DRAPAK ◽  
BORYS ZIMENKOVSKY ◽  
LINA PEREKHODA ◽  
SERGIY KOVALENKO ◽  
LILIYA LOGOYDA
Keyword(s):  
Formic Acid ◽  
Human Plasma ◽  
Internal Standard ◽  
Sample Volume ◽  
Pharmacokinetic Studies ◽  
Coefficients Of Variation ◽  
Highly Sensitive ◽  
Regulatory Guidelines ◽  
Initial Content ◽  
Development And Validation

Objective: The present study was aimed to develop a rapid, specific and sensitive method based on LC-MS/MS method was developed for the determination of urocarb using etomidate as an internal standard. Methods: Chromatography was achieved on Discovery C18, 50 × 2.1 mm, 5 μm column. Samples were chromatographed in a gradient mode (eluent A (acetonitrile-water–formic acid, 5: 95: 0.1 v/v), eluent B (acetonitrile–formic acid, 100: 0.1 v/v)). The initial content of the eluent B of 8%, which increases linearly to 1.0 min to 100%, is maintained up to 1.5 min and returned to the original 8% to 1.51 min. The mobile phase was delivered at a flow rate of 0.400 ml/min into the mass spectrometer ESI chamber. The sample volume was 4 μl. Results: The total chromatographic run time was 2.0 min and the elution of urocarb and IS (etomidate) occurred at ~1.53 and 1.67 min, respectively. A linear response function was established at 1-100 ng/ml for urocarb and etomidate in human plasma. The % mean recovery for urocarb in LQC, MQC and HQC was 104.1 %, 100.0 % and 97.4 %. The lowest concentration with the RSD<20% was taken as LLOQ and was found to be 1.03 ng/ml for urocarb. The within-run coefficients of variation ranged between 0.271 % and 0.478 % for urocarb. The within-run percentages of nominal concentrations ranged between 99.12 % and 100.21 % for urocarb. The between-run coefficients of variation ranged between 0.388 % and 0.601 % for urocarb. The between-run percentages of nominal concentrations ranged between 98.78 % and 101.11 % for urocarb. Conclusion: A highly sensitive, specific, reproducible, rapid and high-throughput LC-MS/MS assay was developed and validated to quantify urocarb in human plasma as per the regulatory guidelines. Due to the sensitivity of the developed method, it can be applied to the monitoring of plasma levels in the analysis of drug in preclinical and clinical pharmacokinetic studies. All the parameters and results were found within the acceptance limit as given in the validation protocol.

scholarly journals ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF AMLODIPINE IN HUMAN PLASMA USING LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY/MASS SPECTROMETRY

2018 ◽  
Vol 11 (7) ◽  
pp. 393
Author(s):  
Akiful Haque M ◽  
Shanthi Priya D K ◽  
Dibyalochan Mohanty ◽  
Vasudha Bakshi ◽  
Narender Boggula
Keyword(s):  
Mass Spectrometry ◽  
Human Plasma ◽  
Analytical Method ◽  
High Performance ◽  
Method Development ◽  
Solid Phase ◽  
Internal Standard ◽  
High Performance Liquid Chromatographic ◽  
Economic Method ◽  
Positive Ion

Objective: The objective of the present investigation was to develop a novel, simple, and economic method for the estimation of amlodipine in positive ion mode in human plasma using amlodipine maleate d4 as an internal standard.Methods: The chromatographic separation was performed on Zorbax SB, C18, 50 mm*4.6 mm, and 3.5 mm. The mobile phase was prepared with a mixture of 5 mm ammonium acetate in 0.1% formic acid: High performance liquid chromatographic (HPLC) grade methanol:HPLC grade acetonitrile (40:30:30) that run isocratically at the flow rate of 0.700 ml/min and run time at 2.50 min.Results: The analytical method is valid for the estimation of amlodipine, in human plasma over a range of 0.100 ng/ml–9.990 ng/ml with the detection of amlodipine m/z - 409.10 (parent) and 238.00 (product), and internal standard Amlodipine Maleate d4 m/z - 413.20 (parent), and 238.00 (product) in positive ion mode. The results of carryover test, matrix effect, linearity, precision and accuracy, stabilities, dilution integrity, and run size evaluation test presented in this report are within the acceptance range.Conclusion: A sensitive method for the separation and determination of amlodipine in plasma has been developed based on solid-phase extraction with disposable extraction cartridges in combination with LC and mass spectrophotometers (MS/MS).

scholarly journals METHODS OF METOPROLOL ANALYSIS IN DRUGS AND BIOLOGICAL FLUIDS: REVIEW AND SUGGESTIONS

2020 ◽  
Vol 5 (2) ◽  
pp. 88-97
Author(s):  
M. M. Horyn ◽  
L. S. Logoyda
Keyword(s):  
Analytical Method ◽  
High Speed ◽  
High Performance ◽  
Method Development ◽  
High Efficiency ◽  
Biological Fluids ◽  
Dosage Forms ◽  
Pharmaceutical Chemistry ◽  
Components Separation ◽  
Analysis Methods

Background. Analytical method is increasingly implemented into fundamental pharmaceutical chemistry and analysis, considering their high sensitivity, accuracy, specificity and expressiveness. Objective. Metoprolol’s analytical method development was the research goal. Methods. The sources were world recognized journals (1990-2019) and key words used as filter were “metoprolol”, “spectrophotometry” “high-performance liquid chromatography, HPLC”, “quantitative analysis”, “validation”. Results. Chromatographic methods of analysis have the highest specificity and objectivity and allow qualitative and quantitative determination of Active Pharmaceutic Ingredient (API) in combined dosage forms and biological fluids without prior components separation. The main disadvantage of the described API analysis methods is long terms from the beginning of chromatography to API release and specific solvents used as the mobile phase in HPLC. New methods development and selection such chromatographic conditions that provide high speed and high efficiency at lower pressure of the system are essential. Also, the reduction of analysis time is achieved by simplifying the conditions for sample preparation. Conclusions. Analysts are constantly working on developing new analysis methods and their optimization in order to save time and consumables, which also ensures the efficiency of the developed method. There is no monograph on the substance or dosage forms of metoprolol in SPhU. Therefore, some of the developed methods should be suggested for the SPhU monograph, which is important for ensuring pharmacopoeial quality control of medicines in Ukraine.

scholarly journals BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF TIZANIDINE IN K2EDTA HUMAN PLASMA BY USING LC-MS/MS

2019 ◽  
Vol 8 (5) ◽  
Author(s):  
Dr. R.S. Bhadauria ◽  
Dr. D.B. Joshi
Keyword(s):  
Human Plasma ◽  
Method Validation ◽  
Method Development ◽  
Excitatory Amino Acid ◽  
Muscle Tone ◽  
Biological Fluids ◽  
The Body ◽  
Bioanalytical Method Validation ◽  
Spinal Interneurones ◽  
Bioanalytical Method

Bioanalytical method Validation employed for quantitative determination of drug and their metabolites in biological fluids. Comprises all criteria determining data quality, such as selectivity, accuracy, precision, recovery and senstivity. The main purpose of method validation is to demonstrate that a specific Bioanalytical method can reliably determine the concentration of drug in study sample with high degre of confidence. Validation does not means that method is perfect, but validation means method has met a set of criteria to ensure that it is reliable and consistent. Tizanidine is a central alpha 2 adrenergic agonist –inhibits release of excitatory amino acid in the spinal interneurones. It may facilitate the inhibitory transmitter glycine as well. It inhibits polysyneptic reflexes reduce muscle tone and frequency of muscle spasms without reducing muscle strength. Following oral administration, tizanidine is essentially completely absorbed .The absolute oral bioavailability of tizanidine is approximately 40%, due to extensive first-pass hepatic metabolism. Tizanidine is extensively distributed throughout the body with a mean steady state volume of distribution of 2.4 L/kg following intravenous administration in healthy adult volunteers .tizanidine is approximately 30% bound to plasma proteins. Keywords: Bioanalytical method Validation, LC-MS/MS, Human Plasma, Tizanidine, HPLC.

Key Aspects of Analytical Method Development and Validation

2019 ◽  
Vol 31 (1) ◽  
pp. 32-39
Author(s):  
Suman Shrivastava ◽  
Pooja Deshpande ◽  
S. J. Daharwal
Keyword(s):  
Method Validation ◽  
Analytical Method ◽  
Method Development ◽  
Limit Of Detection ◽  
Limit Of Quantification ◽  
Ich Guidelines ◽  
Validation Parameters ◽  
Method Development And Validation ◽  
Key Aspects ◽  
Development And Validation

Development of a method is crucial for discovery, development, and analysis of medicines in the pharmaceutical formulation. Method validation could also be thought to be one in all the foremost well-known areas in analytical chemistry as is reproduced within the substantial variety of articles submitted and presented in peer review journals every year. Validation of an analytical procedure is to demonstrate that it's appropriate for its intended purpose. Results from method validation are often wont to decide the quality, reliability and consistency of analytical results. Analytical methods need to be validated or revalidated. This review describes general approach towards validation process and validation parameters to be considered during validation of an analytical method. It also refers to various regulatory requirements like WHO, USFDA, EMEA, ICH, ISO/IEC. The parameters described here are according to ICH guidelines which include accuracy, precision, specificity, limit of detection, limit of quantification, linearity range and robustness.

scholarly journals A UPLC-MS/MS METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF SOFOSBUVIR FROM HUMAN PLASMA

2016 ◽  
Vol 9 (1) ◽  
pp. 30 ◽  
Author(s):  
Darshan Bhatt ◽  
B. Rajkamal
Keyword(s):  
Chromatographic Separation ◽  
Method Development ◽  
Internal Standard ◽  
Ultra Performance Liquid Chromatography ◽  
Pharmacokinetic Studies ◽  
Solution Stability ◽  
Tandem Mass ◽  
Stability Studies ◽  
Liquid Chromatography Tandem Mass ◽  
Acceptance Limits

Objective: The present work aimed to develop a simple, rapid, specific and precise ultra-performance liquid chromatography-tandem mass spectrophotometric (LC–MS/MS) validated method for quantification of sofosbuvir and internal standard (ISTD) Sofosbuvir-d3 in human plasma.Methods: Samples prepared by employing liquid-liquid extraction (LLE) using 2.5 ml of ethyl acetate. Chromatographic separation was achieved on Gemini 5µ C18, 50 x 4.6 mm column using a mixture of 0.1% (v/v) formic acid in water to methanol at a ratio of 30:70 v/v as the mobile phase. The flow rate was 0.50 ml/min. The LC eluent was split, and approximately 0.1 ml/min was introduced into Tandem mass spectrometer using turbo Ion Spray interface at 325 °C. Quantitation was performed by transitions of 428.35/279.26 (m/z) for sofosbuvir and 431.38/282.37 (m/z) for sofosbuvir-d3.Results: The concentrations of ten working standards showed linearity between 4.063 to 8000.010ng/ml (r2 ≥ 0.9985). Chromatographic separation was achieved within 2 min. The average extraction recoveries of three quality control concentrations were 75.36% for sofosbuvir and were within the acceptance limits. The coefficient of variation was ≤15% for intra-and inter-batch assays. The %CV of ruggedness ranges 0.35% and 3.09%. The % stability of short term and long term stock solution stability studies was found to be 97.25% and 98.81% respectively.Conclusion: The results obtained for specificity, linearity, accuracy, precision, ruggedness and stability studies were within the acceptance limits. Thus the validated economical method was applied for pharmacokinetic studies of sofosbuvir.

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