scholarly journals Full validation of curcumin analytical method by LC-MS/MS points out that the degree of hemolysis in plasma affects the quantitation: application in dog plasma study

2019 ◽  
Vol 62 (4) ◽  
Author(s):  
Mariana Dolores ◽  
Alma Villaseñor ◽  
Alma Revilla Vázquez ◽  
Helgi Jung ◽  
Victor Hugo Santiago Rios ◽  
...  
Keyword(s):  
Internal Standard ◽  
Daily Practice ◽  
Reaction Monitoring ◽  
Selective Method ◽  
Clopidogrel Bisulfate ◽  
Dog Plasma ◽  
Relative Standard ◽  
Molecular Reaction ◽  
Acquity Uplc

Abstract. Curcumin has gained great attention in the last decades due to its fascinating properties for humans, such as anti-inflammatory or as cytotoxic against cancer. These effects are also claimed for pets such as cats and dogs, where curcumin administration is a daily practice routine. However, curcumin presents poor oral bioavailability, driving scientists to look for new delivery systems. In the last decades, several analytical methods for the quantification of curcumin in plasma have been published. To our knowledge there are no published reports on the effect of the level of hemolysis in the determination of this compound. In the present paper, a highly specific, sensitive and selective method is presented using Molecular Reaction Monitoring (SRM) using positive ionization (ESI+) mode. Curcumin and clopidogrel bisulfate – used as internal standard (IS) – were separated on an Acquity UPLC BECH Shield RP 18 column (1.7µm, 2.1 X 100mm) with 0.1% formic acid in acetonitrile and water in proportion of 60:40 (v/v). The analyte transitions were 369.3→177.06 m/z for curcumin and 322→212.05 m/z for IS. The method was fully validated and showed good linearity (R2 ≥ 0.999) over the range of 3-160 ng/mL. The Relative Standard Deviation (RSD) were less than 6% for intra-and inter-day analysis and recovery spanned 85-95%. We proved that the degree of hemolysis impaired curcumin quantitation. This method was applied to test curcumin bioavailability in both a mucoadhesive nanocapsule formulation and traditional capsules in dogs that attended routine veterinary consultation.Resumen. La curcumina ha ganado gran atención en las últimas décadas debido a sus propiedades terapéuticas para los humanos, como antiinflamatorio o citotóxico contra el cáncer. Estos efectos también se observan en pequeñas especies como gatos y perros, donde la administración de curcumina se ha vuelto una alternativa. Sin embargo, la curcumina presenta una baja biodisponibilidad oral, lo que impulsa a los científicos a buscar nuevos sistemas de administración. En las últimas décadas, se han publicado varios métodos analíticos para la cuantificación de curcumina en plasma. Actualmente, no hay informes publicados sobre el efecto del grado de hemólisis en la determinación de este compuesto. En este trabajo se desarrolló un método específico, sensible y selectivo utilizando el Monitoreo de reacción seleccionado (SRM) en modo de ionización positiva (ESI +). La curcumina y el bisulfato de clopidogrel, utilizado como patrón interno (IS), se separaron en una columna Acquity UPLC BECH Shield RP 18 (1,7 μm, 2,1 X 100 mm) con ácido fórmico al 0,1% en acetonitrilo y agua a una proporción de 60:40 (v/v). Las transiciones de los analitos fueron 369.3 → 177.06 m/z para curcumina y 322 → 212.05 m/z para IS. El método fue validado y demostró ser lineal (r2 ≥ 0.999) en el rango de 3-160 ng/mL. La desviación relativa estándar (RSD) fue inferior al 6% para el análisis intra e interdía y el porcentaje de recuperación fue 85-95%. Se descubrió que el grado de hemólisis afecta la cuantificación de curcumina. El método desarrollado se aplicó para evaluar la biodisponibilidad de curcumina tanto en una formulación de nanocápsulas mucoadhesivas como en cápsulas tradicionales en perros que asistieron a consultas veterinarias de rutina.

ICH/FDA Guidelines-Compliant Validated Stability-Indicating HPLC-UV Method for the Determination of Axitinib in Bulk and Dosage Forms

2020 ◽  
Vol 16 (8) ◽  
pp. 1106-1112
Author(s):  
Ibrahim A. Darwish ◽  
Nasr Y. Khalil ◽  
Mohammad AlZeer
Keyword(s):  
High Performance ◽  
Kinase Inhibitor ◽  
Degradation Products ◽  
Internal Standard ◽  
Dosage Forms ◽  
Uv Detector ◽  
Stability Indicating ◽  
Therapeutic Benefits ◽  
Relative Standard

Background: Axitinib (AXT) is a member of the new generation of the kinase inhibitor indicated for the treatment of advanced renal cell carcinoma. Its therapeutic benefits depend on assuring the good-quality of its dosage forms in terms of content and stability of the pharmaceutically active ingredient. Objective: This study was devoted to the development of a simple, sensitive and accurate stabilityindicating high-performance liquid chromatographic method with ultraviolet detection (HPLC-UV) for the determination of AXT in its bulk and dosage forms. Methods: Waters HPLC system was used. The chromatographic separation of AXT, internal standard (olaparib), and degradation products were performed on the Nucleosil CN column (250 × 4.6 mm, 5 μm). The mobile phase consisted of water:acetonitrile:methanol (40:40:20, v/v/v) with a flow rate of 1 ml/min, and the UV detector was set at 225 nm. AXT was subjected to different accelerated stress conditions and the degradation products, when any, were completely resolved from the intact AXT. Results: The method was linear (r = 0.9998) in the concentration range of 5-50 μg/ml. The limits of detection and quantitation were 0.85 and 2.57 μg/ml, respectively. The accuracy of the method, measured as recovery, was in the range of 98.0-103.6% with relative standard deviations in the range of 0.06-3.43%. The results of stability testing revealed that AXT was mostly stable in neutral and oxidative conditions; however, it was unstable in alkaline and acidic conditions. The kinetics of degradation were studied, and the kinetic rate constants were determined. The proposed method was successfully applied for the determination of AXT in bulk drug and dosage forms. Conclusions: A stability-indicating HPLC-UV method was developed and validated for assessing AXT stability in its bulk and dosage forms. The method met the regulatory requirements of the International Conference on Harmonization (ICH) and the Food and Drug Administration (FDA). The results demonstrated that the method would have great value when applied in quality control and stability studies for AXT.

Development and Validation of a New Method for the Determination of Anti-hepatitis C Agent Simeprevir in Human Plasma using HPLC with Fluorescence Detection

2020 ◽  
Vol 16 (4) ◽  
pp. 428-435
Author(s):  
Ahmed F.A. Youssef ◽  
Yousry M. Issa ◽  
Kareem M. Nabil
Keyword(s):  
Hepatitis C ◽  
Human Plasma ◽  
Fluorescence Detection ◽  
Internal Standard ◽  
Protein Precipitation ◽  
Assay Method ◽  
Fluorescence Detector ◽  
Limit Of Quantification ◽  
Relative Standard

Background: Simeprevir is one of the recently discovered drugs for treating hepatitis C which is one of the major diseases across the globe. Objective: The present study involves the development of a new and unique High-Performance Liquid Chromatography (HPLC) method using fluorescence detection for the determination of simeprevir (SIM) in human plasma. Methods: Two methods of extractions were tested, protein precipitation using acetonitrile and liquidliquid extraction. A 25 mM dipotassium hydrogen orthophosphate (pH 7.0)/ACN (50/50; v/v), was used as mobile phase and C18 reversed phase column as the stationary phase. The chromatographic conditions were optimized and the concentration of simeprevir was determined by using the fluorescence detector. Cyclobenzaprine was used as an internal standard. Results: Recovery of the assay method based on protein precipitation was up to 100%. Intra-day and inter-day accuracies range from 92.30 to 107.80%, with Relative Standard Deviation (RSD) range 1.65-8.02%. The present method was successfully applied to a pharmacokinetic study where SIM was administered as a single dose of 150 mg SIM/capsule (Olysio®) to healthy individuals. Conclusion: This method exhibits high sensitivity with a low limit of quantification 10 ng mL-1, good selectivity using fluorescence detection, wide linear application range 10-3000 ng mL-1, good recovery and highly precise and validation results. The developed method can be applied in routine analysis for real samples.

scholarly journals Quantitative Determination of Azithromycin in Human Plasma by Liquid Chromatography–Mass Spectrometry and its Application in Pharmackokinetic Study

2012 ◽  
Vol 11 (1) ◽  
pp. 55-63 ◽  
Author(s):  
Maizbha Uddin Ahmed ◽  
Mohammad Safiqul Islam ◽  
Tasmin Ara Sultana ◽  
AGM Mostofa ◽  
Muhammad Shahdaat Bin Sayeed ◽  
...  
Keyword(s):  
Mobile Phase ◽  
Solid Phase ◽  
Internal Standard ◽  
Extraction Process ◽  
Serum Samples ◽  
Limit Of Quantification ◽  
Relative Standard ◽  
Linear Concentration Range ◽  
Linear Concentration

Azithromycin is an effective and well-known antimicrobial agent. In the present study, a simple, sensitive and specific LC/MS/MS method has been developed and validated for the quantification of Azithromycin in  human serum samples using Clarithromycin as internal standard. Azithromycin was extracted from biological matrix  by using solid phase extraction process. The chromatographic separation was performed on Luna C18 (3 ?, 2x150   mm) column with a mobile phase consisting of 35 mM ammonium acetate buffer (mobile phase-A) and acetonitrile  and methanol in ratio of 90:10 ( as mobile phase-B) at a flow rate of 0.25 mL/min. The method was validated over a  linear concentration range of 0.5?50.0 ng/mL and limit of quantification (LOQ) was 0.5 ng/mL with a coefficient of  correlation (r2) = 0.9998. The intra-day and inter-day precision expressed as relative standard deviation were 1.64% – 8.43% and 2.32% – 9.92%, respectively. The average recovery of azithromycin from serum was 98.11%. The method  was successfully applied to a pharmacokinetic study after oral administration of Azithromycin 200 mg/5 ml suspension in healthy Bangladeshi volunteers. DOI: http://dx.doi.org/10.3329/dujps.v11i1.12488 Dhaka Univ. J. Pharm. Sci. 11(1): 55-63, 2012 (June)

scholarly journals Determination of 1,3-Dichloropropanol in Soy Sauce and Related Products by Headspace Gas Chromatography with Mass Spectrometric Detection: Interlaboratory Study

2005 ◽  
Vol 88 (5) ◽  
pp. 1404-1412 ◽  
Author(s):  
Sarah Hasnip ◽  
Colin Crews ◽  
Nicholas Potter ◽  
Paul Brereton ◽  
Henri Diserens ◽  
...  
Keyword(s):  
Gas Chromatography ◽  
Standard Deviation ◽  
Relative Standard Deviation ◽  
Internal Standard ◽  
Mass Spectrometric ◽  
Soy Sauce ◽  
Headspace Gas Chromatography ◽  
Relative Standard ◽  
Headspace Gas

Abstract An interlaboratory study was performed to evaluate the effectiveness of a headspace gas chromatography (GC) method for the determination of 1,3-dichloro-propan-2-ol (1,3-DCP) in soy sauce and related products at levels above 5 ng/g. The test portion is mixed with an internal standard (d5-1,3-DCP) and ammonium sulfate in a sealed headspace vial. After achieving equilibrium, the headspace is sampled either by gas-tight syringe or solid-phase microextraction (SPME) and analyzed by GC with mass spectrometric detection. 1,3-DCP is detected in the selected-ion mode (monitoring m/z 79 and 81 for 1,3-DCP and m/z 82 for the deuterated internal standard) and quantified by measurement against standards. Test materials comprising soy, dark soy, mushroom soy, and teriyaki sauces, both spiked and naturally contaminated, were sent to 9 laboratories in Europe, Japan, and the United States; of these, 5 used SPME and 4 used syringe headspace analysis. Test portions were spiked at 5.0, 10.0, 20.0, 100.0, and 500.0 ng/g. The average recovery for spiked blank samples was 108% (ranging from 96–130%). Based on results for spiked samples (blind pairs at 5, 10, 20, 100, and 500 ng/g) as well as a naturally contaminated sample (split-level pair at 27 and 29 ng/g), the relative standard deviation for repeatability (RSDr) ranged from 2.9–23.2%. The relative standard deviation for reproducibility (RSDR) ranged from 20.9–35.3%, and HorRat values of between 1.0 and 1.6 were obtained.

Improved Codistillation Method for Determination of Carbon Tetrachloride, Ethylene Dichloride, and Ethylene Dibromide in Grain and Grain-Based Products

1985 ◽  
Vol 68 (4) ◽  
pp. 759-762
Author(s):  
Jonathan W Devries ◽  
Paul A Larson ◽  
Raymond H Bowers ◽  
Joyce A Keating ◽  
James M Broge ◽  
...  
Keyword(s):  
Carbon Tetrachloride ◽  
Internal Standard ◽  
Ethylene Dichloride ◽  
Standard Mixture ◽  
Ethylene Dibromide ◽  
Acetone Water ◽  
Relative Standard ◽  
The Common ◽  
Hexane Solution

Abstract A method is described for the determination of the common fumigants carbon tetrachloride (CC14), ethylene dichloride (EDC), and ethylene dibromide (EDB) in grain and grain-based products. A properly prepared sample is mixed with water and hexane, an internal standard mixture of 1,2-dichloropropane (DCP) and 1,2-dibromopropane (DBP) is added, and the fumigants are codistilled with the hexane into an appropriate receiver. After the hexane solution is dried over sodium sulfate, the quantities of fumigants present are quantitated on a gas chromatograph (GC) equipped with an electron capture detector (ECD). For the matrices investigated, the relative standard deviation of the method was 6.0,9.7, and 23.1% for CC14, EDC, and EDB, respectively. Recoveries of added fumigants were 107, 95, and 101%, respectively. Comparison with an acetone-water soak extraction method gave a correlation of 0.967 between methods for EDB with odds of a difference between methods of 35%.

Gas Chromatographic-Thermal Energy Analysis Method for Determination of Volatile N-Nitrosamines in Baby Bottle Rubber Nipples: Collaborative Study

1987 ◽  
Vol 70 (1) ◽  
pp. 64-68 ◽  
Author(s):  
J Ian Gray ◽  
Michael A Stachiw
Keyword(s):  
Thermal Energy ◽  
Energy Analysis ◽  
Collaborative Study ◽  
Internal Standard ◽  
Analytical Techniques ◽  
The United States ◽  
Baby Bottle ◽  
Relative Standard ◽  
Better Than

Abstract A collaborative study was conducted on the U.S. Food and Drug Administration (FDA) dichloromethane extraction method for determining volatile N-nitrosamines in baby bottle rubber nipples. Following dichloromethane extraction, A'-nitrosamines were determined by gas chromatography-thermal energy analysis. Six pairs of blind duplicate rubber nipple samples representing 6 lots were analyzed by 11 collaborating laboratories. All samples were portions taken from equilibrated composites of cut-up rubber nipples obtained from manufacturers in the United States. Recoveries of the internal standard (N-nitrosodipropylamine) at approximately 20 ppb ranged from 10 to 120%. Reproducibility relative standard deviations (RSDJ were between 35 and 45% for N-nitrosamine levels from 10 to 20 ppb. However, when data from laboratories with recoveries less than 75% were excluded (this is now specified in the method), RSD„ values were between 11 and 32% for N-nitrosamine levels from 6 to 26 ppb. Values were consistent with or better than those reported for other analytical techniques designed to quantitate trace contaminants at the low ppb level, e.g., afiatoxin in foods. The method has been adopted official first action for the quantitation of volatile N-nitrosamines in baby bottle rubber nipples.

scholarly journals Determination of Fleroxacin in Pure and Tablet Forms by Liquid Chromatography and Derivative UV-Spectrophotometry

2003 ◽  
Vol 86 (2) ◽  
pp. 229-235 ◽  
Author(s):  
Dorota Kowalczuk ◽  
Hanna Hopkała
Keyword(s):  
Mobile Phase ◽  
Internal Standard ◽  
Spectrophotometric Assay ◽  
Uv Spectrophotometry ◽  
Aminobenzoic Acid ◽  
Tetrabutylammonium Hydroxide ◽  
Relative Standard ◽  
Liquid Chromatographic ◽  
Derivative Uv Spectrophotometry

Abstract Derivative UV-spectrophotometric and liquid chromatographic (LC) methods for fleroxacin determination were validated. In the spectrophotometric assay, first-, second-, third-, and fourth-order measurements were applied with the use of peak–zero and peak–peak techniques. The linear correlation between amplitude of the peak and concentration of the examined drug ranged from 2.0 to 12.0 μg/mL. An isocratic LC analysis was performed on a Purospher ODS column with an acidic mobile phase containing tetrabutylammonium hydroxide. Measurements were made at a wavelength of 285 nm with 4-aminobenzoic acid (PABA) as internal standard. The calibration curve was linear (r = 0.9999) in the studied range of concentration (1.0–10.0 μg/mL). The accuracy (mean recovery, about 100%), precision (relative standard deviation <1%), selectivity, and sensitivity of the elaborated methods were satisfactory.

scholarly journals Determination of Tianeptine in Tablets by High-Performance Liquid Chromatography with Fluorescence Detection

2007 ◽  
Vol 90 (3) ◽  
pp. 720-724
Author(s):  
Sevgi Tatar Ulu
Keyword(s):  
Lower Limit ◽  
High Performance ◽  
Limit Of Detection ◽  
Internal Standard ◽  
Linear Calibration ◽  
Fluorescence Detector ◽  
Relative Standard ◽  
Validation Parameters ◽  
Limit Of Quantitation

Abstract A sensitive and selective high-performance liquid chromatographic method has been developed for the determination of tianeptine (Tia) in tablets. The method is based on derivatization of Tia with 4-chloro-7-nitrobenzofurazan (NBD-Cl). A mobile phase consisting of acetonitrile10 mM orthophosphoric acid (pH 2.5; 77 + 23) was used at a flow rate of 1 mL/min on a C18 column. The Tia-NBD derivative was monitored using a fluorescence detector, with emission set at 520 nm and excitation at 458 nm. Gabapentin was selected as an internal standard. Linear calibration graphs were obtained in the concentration range of 45300 ng/mL. The lower limit of detection (LOD) was 10 ng/mL at a signal-to-noise ratio of 4. The lower limit of quantitation (LOQ) was 45 ng/mL. The relative standard values for intra- and interday precision were <0.46 and <0.57%, respectively. The recovery of the drug samples ranged between 98.89 and 99.85%. No chromatographic interference from the tablet excipients was found. The proposed method was validated in terms of precision, robustness, recovery, LOD, and LOQ. All the validation parameters were within the acceptance range. The proposed method was applied for the determination of Tia in commercially available tablets. The results were compared with those obtained by an ultraviolet spectrophotometric method using t- and F-tests.

scholarly journals Simultaneous Determination of Levocetirizine and Pseudoephedrine in Dog Plasma by Liquid Chromatography-Mass Spectrometry in the Presence of Dextrocetirizine

2012 ◽  
Vol 15 (4) ◽  
pp. 519 ◽  
Author(s):  
Jae Kuk Ryu ◽  
Sun Dong Yoo
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Oral Administration ◽  
Mobile Phase ◽  
Internal Standard ◽  
Liquid Chromatography Mass Spectrometry ◽  
Chiral Column ◽  
Accuracy And Precision ◽  
Dog Plasma

Purpose. This study describes the development of a rapid and sensitive LC-ESI-MS assay for simultaneous enantioselective determination of levocetirizine and pseudoephedrine in dog plasma in the presence of dextrocetirizine. Methods. Separations were achieved on an Ultron ES-OVM chiral column using the mobile phase consisting of 10 mM aqueous NH4OAc (pH 6.6) and acetonitrile (9:1 v/v). Results. The retention times of pseudoephedrine, dextrocetirizine, levocetirizine and diazepam (internal standard) were 5.2, 8.3, 9.6 and 11.6 min, respectively, and the total run time was less than 15 min. The assay was validated to demonstrate the linearity, accuracy and precision, recovery and stability. The calibration curves were linear over the concentration range from 1 – 200 ng/mL for levocetirizine and from 5 – 1000 ng/mL for pseudoephedrine. Conclusions. The developed assay was successfully applied to a pharmacokinetic study after oral administration of the racemic cetirizine (0.5 mg/kg, or 0.25 mg/kg as levocetirizine) and pseudoephedrine (12 mg/kg) in the dog. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

scholarly journals Simultaneous Determination of Cholecalciferol (Vitamin D3) and Ergocalciferol (Vitamin D2) in Foods by Selected Reaction Monitoring

2009 ◽  
Vol 92 (2) ◽  
pp. 511-517 ◽  
Author(s):  
Gianluca Dimartino
Keyword(s):  
Vitamin D3 ◽  
Ion Trap ◽  
Atmospheric Pressure Chemical Ionization ◽  
Internal Standard ◽  
Selected Reaction Monitoring ◽  
Vitamin D2 ◽  
Reaction Monitoring ◽  
Ionization Source ◽  
Relative Standard ◽  
Pressure Chemical Ionization

Abstract Cholecalciferol (vitamin D3) and ergocalciferol (vitamin D2) were determined simultaneously by selected reaction monitoring (SRM) mass spectrometry for different food matrixes. A small amount of starting sample was saponified and extracted before injection into a linear ion trap mass spectrometer equipped with an atmospheric pressure chemical ionization source. Dihydrotachysterol, which is absent from food and has a structure similar to that of vitamins D3 and D2, was used as an internal standard. Calibration curves for the 2 vitamins showed linearity with R2 values of 0.9999 and 0.9989 for vitamins D3 and D2, respectively. Limits of detection for vitamins D3 and D2 were 0.5 ng/g (1.3 pmol/g) and 1.75 ng/g (4.4 pmol/g) and limits of quantitation were 1.25 ng/g (3.24 pmol/g), and 3.75 ng/g (9.45 pmol/g), respectively. Accuracy and precision of the method were tested with the infant formula reference standard of the National Institute of Standards and Technology, which showed a relative standard deviation of 6. Recoveries ranged from 95 to 105. Several food products were tested with AOAC Method 982.29, which is currently in use for vitamins D3 and D2, and results were comparable within 6.

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