scholarly journals Validation of Cell-Based Assay for Quantification of Sesamol Uptake and Its Application for Measuring Target Exposure

Molecules ◽  
2019 ◽  
Vol 24 (19) ◽  
pp. 3522 ◽  
Author(s):  
Srisongkram ◽  
Weerapreeyakul
Keyword(s):  
Control Sample ◽  
High Sensitivity ◽  
Pharmacological Action ◽  
Target Cells ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Target Exposure ◽  
Relative Standard ◽  
Site Of Action ◽  
High Repeatability

The intracellular drug concentration is needed for determined target exposure at the site of action regarding its pharmacological action and adverse effects. Sesamol is an antiproliferative molecule from Sesamum indicum with promising health benefits. We present a method for measuring the intracellular sesamol content using reverse-phase HPLC with a UV diode array in melanoma cells. Sesamol was completely resolved by isocratic elution (4.152 ± 0.008 min) with methanol/water (70%, v/v) through a 30 °C, 5-µm C-18 column and detection at 297 nm. The present assay offers high sensitivity, fast elution, and an accurate and linear nominal concentration range of 10–1000 ng/mL (R2 = 0.9972). The % accuracy of the sesamol quality control sample was −3.36% to 1.50% (bias) with a 0.84% to 5.28% relative standard deviation (RSD), representing high repeatability and high reproducibility. The % recovery was 94.80% to 99.29%, which determined that there was no loss of sesamol content during the sample preparation. The validated method was applied to monitor intracellular sesamol concentration after treatment from 5 min to 24 h. The remaining intracellular sesamol content was correlated with its antiproliferative effect (R2 = 0.9483). In conclusion, this assay demonstrated low manipulation, quick elution, and high sensitivity, precision, accuracy, and recovery, and it was successfully applied to the quantification of sesamol in target cells.

scholarly journals Solid-Phase Extraction and Reverse-Phase HPLC: Application to Study the Urinary Excretion Pattern of Benzophenone-3 and its Metabolite 2,4-Dihydroxybenzophenone in Human Urine

2008 ◽  
Vol 3 ◽  
pp. ACI.S396 ◽  
Author(s):  
Helena Gonzalez ◽  
Carl-Eric Jacobson ◽  
Ann-Marie Wennberg ◽  
Olle Larkö ◽  
Anne Farbrot
Keyword(s):  
Solid Phase Extraction ◽  
Urinary Excretion ◽  
Human Urine ◽  
Solid Phase ◽  
Whole Body ◽  
Phase Extraction ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Excretion Pattern ◽  
Relative Standard

Background Benzophenone-3 (BZ-3) is a common ultraviolet (UV) absorbing compound in sunscreens. It is the most bioavailable species of all UV-absorbing compounds after topical application and can be found in plasma and urine. Objectives The aim of this study was to develop a reverse-phase high performance liquid chromatography (HPLC) method for determining the amounts BZ-3 and its metabolite 2,4-dihydroxybenzophenone (DHB) in human urine. The method had to be suitable for handling a large number of samples. It also had to be rapid and simple, but still sensitive, accurate and reproducible. The assay was applied to study the urinary excretion pattern after repeated whole-body applications of a commercial sunscreen, containing 4% BZ-3, to 25 healthy volunteers. Methods Each sample was analyzed with regard to both conjugated/non-conjugated BZ-3 and conjugated/non-conjugated DHB, since both BZ-3 and DHB are extensively conjugated in the body. Solid-phase extraction (SPE) with C8 columns was followed by reverse-phase HPLC. For separation a Genesis C18 column was used with an acethonitrile-water mobile phase and the UV-detector was set at 287 nm. Results The assay was linear r 2 > 0.99, with detection limits for BZ-3 and DHB of 0.01 µmol L-1 and 0.16 µmol L-1 respectively. Relative standard deviation (RSD) was less than 10% for BZ-3 and less than 13% for DHB. The excretion pattern varied among the human volunteers; we discerned different patterns among the individuals. Conclusions The reverse-phase HPLC assay and extraction procedures developed are suitable for use when a large number of samples need to be analyzed and the method fulfilled our objectives. The differences in excretion pattern may be due to differences in enzyme activity but further studies, especially about genetic polymorphism, need to be performed to verify this finding.

scholarly journals FORMULATION OF SOLID DOSAGE FORM CONTAINING CLOPIDOGREL AND CILOSTAZOL AND ITS HPLC ANALYSIS

2017 ◽  
Vol 9 (6) ◽  
pp. 12
Author(s):  
Asha Thomas ◽  
Suraj Bhosale ◽  
Rabindra Nanda
Keyword(s):  
Dosage Form ◽  
Hplc Method ◽  
Forced Degradation ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Stability Indicating ◽  
Solid Dosage Form ◽  
Solid Dosage ◽  
Relative Standard ◽  
Verify Now

Objective: The verify now P2Y12 assay suggested that addition of cilostazol to clopidogrel proves to be efficious in the treatment of patients with cardiovascular disease (CVD). Based on these findings, an attempt has been made to formulate solid dosage form containing the two drugs at the recommended concentrations and develop and validate reverse phase high-performance liquid chromatography (HPLC) method for their simultaneous estimation.Methods: A combined tablet dosage form containing cilostazol (100 mg) and clopidogrel (75 mg) was formulated by direct compression method. A reverse phase HPLC method using C8 column, employing 0.025M phosphate buffer: methanol: acetonitrile (20:40:40% v/v) as mobile phase at a flow rate of 1 ml/min with ultraviolet (UV) detection at 237 nm was developed and validated as per International Council on Harmonisation (ICH) guidelines.Results: The prepared powder blend showed excellent flow properties and formulated tablet passed the standard tests for tablets. The tablets were suitably analyzed by the reverse phase HPLC method with a retention time (RT) of 3.82 and 7.72 min for cilostazol and clopidogrel respectively. The method exhibited linearity (10-100mg/ml for cilostazol and 7.5-75mg/ml for clopidogrel) with r2= 0.999 for both drugs respectively. The recoveries of cilostazol and clopidogrel were 98.97% and 98.94% respectively. The relative standard deviation (RSD) was<2% indicating good method precision. The stability indicating properties evaluated by forced degradation studies showed good separation of the drugs from their degradation products.Conclusion: A simple, precise, robust, stability-indicating HPLC method was developed for simultaneous assay of cilostazol and clopidogrel in prepared tablet formulation and validated as per ICH guidelines. This method can be employed for the analysis and stability studies of solid dosage forms containing the two drugs.

A micromethod for quantitative determination of iodoamino acids in thyroglobulin

1997 ◽  
Vol 153 (1) ◽  
pp. 99-104 ◽  
Author(s):  
N Baudry ◽  
B Mallet ◽  
P J Lejeune ◽  
L Vinet ◽  
J L Franc
Keyword(s):  
Acetic Acid ◽  
Enzymatic Hydrolysis ◽  
Mobile Phase ◽  
High Sensitivity ◽  
The Other ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Spectrophotometric Detection ◽  
Hydrolysis Of

Abstract We describe a new method for quantification of iodoamino acids after enzymatic hydrolysis of thyroglobulin. The procedure involves separation of monoiodotyrosine (MIT), diiodotyrosine, tri-iodothyronine and thyroxine by reverse phase HPLC with a Vydac C18 stationary phase and a mobile phase of water–acetonitrile–acetic acid. The separation is monitored by sensitive spectrophotometric detection through a 96-well microplate system based on the catalytic Sandell–Kolthoff reaction of iodide on the oxidation of arsenic(III) by cerium(IV). This new microassay is particularly convenient because of its high sensitivity and its rapidity (less than 2 h). It can detect 1 pmol MIT and 0·5 pmol of the other three iodoamino acids with a recovery higher than 96%. Moreover, the 96-well microplate system allows many samples to be tested simultaneously and avoids the use of radiolabeled iodine. Journal of Endocrinology (1997) 153, 99–104

Atomic absorption determination of Hg (II), Cd (II), and As (III) in natural and waste water after preliminary group preconcentration

2019 ◽  
Vol 85 (2) ◽  
pp. 12-16
Author(s):  
I. V. Saunina ◽  
E. N. Gribanov ◽  
E. R. Oskotskaya
Keyword(s):  
Waste Water ◽  
Atomic Absorption ◽  
High Sensitivity ◽  
Distribution Coefficients ◽  
Neutral Medium ◽  
Atomic Absorption Determination ◽  
Absorption Determination ◽  
Relative Standard ◽  
Preliminary Group

The sorption of Hg (II), Cd (II), and As (III) by natural aluminosilicate is studied. It is shown that the mineral absorbs those toxicants in a rather wide pH range, quantitative extraction of analytes being achieved in a neutral or close to neutral medium (pH values range within 7.0 - 8.0; 6.3 - 7.5; 7.4 - 8.5 for Hg (II), As (III), and Cd (II), respectively). The effect of the time of phase contact on the degree of extraction of elements is shown. The sorption capacity of the mineral in optimal conditions of the medium acidity (0.06 mmol/g for mercury, 0.31 mmol/g for cadmium, and 0.52 mmol/g for arsenic) is determined. The distribution coefficients attain values of aboutnX 103-nX 104. A new combined method for determination of Hg (II), Cd (II), and As (III) in natural and waste water is developed and tested. The method consists in a preliminary group sorption concentration of the analytes by aluminosilicate, desorption of the analytes from the surface of the mineral and their subsequent atomic absorption determination. The correctness of the method is verified in analysis of spiked samples. The method is easy to use and exhibits high sensitivity, reproducibility and accuracy of analyte determination. The relative standard deviation does not exceed 0.13. Economic availability and possibility of using domestic sorption materials are the important advantages of the proposed procedure which can be used in the practice of laboratories monitoring the quality and safety of environmental objects.

Understanding Process Variables and their Interactions for Maximizing Production of Artemisinin Derivative Artemether (Anti-Malarial Drug) Through Cunninghamella echinulata var elegans at 5 L Bioreactor Level

2019 ◽  
Vol 15 (4) ◽  
pp. 442-452
Author(s):  
Kashyap Kumar Dubey ◽  
Punit Kumar
Keyword(s):  
Experimental Finding ◽  
Culture Broth ◽  
Quadratic Model ◽  
Optimum Conditions ◽  
Reverse Phase Hplc ◽  
Polar Solvents ◽  
Reverse Phase ◽  
Experimental Conditions ◽  
Cunninghamella Echinulata ◽  
Life Threatening

Background: Malaria is one of the life threatening diseases which is caused by Plasmodium sp. of protozoa and uses Anopheles mosquitos as vector. Plasmodium vivax and Plasmodium falciparum are common form of malaria parasite. Artemisinin is reported for its antimalarial activities and Artemether which is a methyl ether derivative of Artemisinin, has been found effective against P. falciparum. Methods: In the present study, bioconversion of Artemisinin into Artemether was carried out experimentally and the statistical tools like experimental factorial design and Response Surface Methodology were used to find optimal conditions (concentration of Artemisinin, age of inoculum, temperature & pH) using Cunninghamella echinulata var. elegans. Experimental conditions for maximum product recovery from culture broth were also optimized using various polar and non-polar solvents for extraction. Artemether purity was analyzed by reverse-phase HPLC. Experimental data was fitted in a quadratic model and effect of various parameters was analyzed. Results: It was found that bioconversion of Artemisinin into Artemether is growth associated process. It was observed that molasses used as carbon source supported production of Artemether to 3.4g/L. The biomass and oxygen are key element affecting of bioconversion of Artemisinin into Artemether such as higher dissolved oxygen reduced the Artemether bioconversion. The highest bioconversion of Artemisinin into Artemether was obtained at temperature 25.5oC, 5g/L concentration of Artemisinin, at age of inoculum of 44.5 h and at pH 6.0. Model suggested the highest bioconversion of Artemisinin into Artemether was 54% at shake flask level which was near about experimental finding. An optimal condition for bioconversion was also analyzed and 64% bioconversion was obtained in 5L bioreactor. Conclusion: The outcomes of the study provided optimum conditions for bioconversion of Artemisinin into Artemether.

scholarly journals Development of TLC Chromatographic-Densitometric Procedure for Qualitative and Quantitative Analysis of Ceftobiprole

Processes ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 708
Author(s):  
Żaneta Binert-Kusztal ◽  
Małgorzata Starek ◽  
Joanna Żandarek ◽  
Monika Dąbrowska
Keyword(s):  
High Sensitivity ◽  
Biologically Active ◽  
Fifth Generation ◽  
Ich Guidelines ◽  
Relative Standard ◽  
Lipophilicity Parameter ◽  
Alkaline Environments ◽  
Acetic Acid Water ◽  
Cephalosporin Antibiotics ◽  
The Stability

Currently, there is still a need for broad-spectrum antibiotics. The new cephalosporin antibiotics include, among others, ceftobiprole, a fifth-generation gram-positive cephalosporin, active against Staphylococcus aureus methicillin agonist (MRSA). The main focus of the work was to optimize the conditions of ceftobiprole qualitative determination and to validate the developed procedure according to ICH guidelines. As a result of the optimization process, HPTLC Cellulose chromatographic plates as a stationary phase and a mixture consisting of ethanol:2-propanol: glacial acetic acid: water (4:4:1:3, v/v/v/v) as a mobile phase were chosen. The densitometric detection was carried out at maximum absorbance of ceftobiprole (λ = 232 nm). Next, the validation process of the developed procedure was carried out. The relative standard deviation (RSD) for precision was less than 1.65%, which proves the high compatibility of the results, as well as the LOD = 0.0257 µg/spot and LOQ = 0.0779 µg/spot values, which also confirm the high sensitivity of the procedure. The usefulness of the developed method for the stability studies of ceftobiprole was analyzed. Study was carried out under stress conditions, i.e., acid and alkaline environments, exposure to radiation imitating sunlight and high temperature (40–60 °C). It was found that cefotbiprole is unstable in an alkaline environment and during exposure to UV-VIS radiation. Moreover, the lipophilicity parameter, as a main physicochemical property of the biologically active compound, was determined using experimental and computational methods.

scholarly journals Orphan Designation and Cisplatin/Hyaluronan Complex in an Intracavitary Film for Malignant Mesothelioma

Pharmaceutics ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 362
Author(s):  
Sabrina Banella ◽  
Eride Quarta ◽  
Paolo Colombo ◽  
Fabio Sonvico ◽  
Antonella Pagnoni ◽  
...  
Keyword(s):  
Sodium Hyaluronate ◽  
Chloride Ions ◽  
Complete Resection ◽  
Size Exclusion ◽  
European Medicines Agency ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Pharmaceutical Development ◽  
Film Forming ◽  
Exclusion Chromatography

Pleural mesothelioma is a lung diffuse tumor, whose complete resection is unlikely. Consequently, metastases reappear where the primary tumor was removed. This paper illustrates the orphan medicine designation procedure of an intracavitary cisplatin film and related pharmaceutical development aspects requested by the European Medicines Agency (EMA) in its Scientific Advice. Since cisplatin pharmacokinetics from the implanted film in sheep resulted substantially modified compared to intravenous administration, the formation of a cisplatin/hyaluronan complex had been hypothesized. Here, the interaction between sodium hyaluronate (NaHA) and cisplatin (CisPt) was demonstrated. Size exclusion chromatography qualitatively evidenced the complex in the film-forming mixture, only showing the NaHA peak. Atomic absorption spectroscopy of the corresponding fraction revealed platinum, confirming the interaction. Reverse phase HPLC quantified about 5% free cisplatin in the film-forming mixture, indirectly meaning that 95% was complexed. Finally, a study of CisPt release from the film assessed how CisPt/NaHA complex affected drug availability. In water, a medium without chloride ions, there was no release and the film remained intact for 48 h and longer, whereas the placebo film dissolved in 15 min. In 0.9% NaCl medium, the film became more soluble, dissolving within 3–4 h. However, cisplatin release was still controlled by the existing complex in solution until chloride ions displaced it. While the film modified its dissolution with aging, CisPt release remained unaffected (90% released in 48 h).

Quantitation of Amiodarone and Desethylamiodarone from Blood Serum and Myocardium Using Reverse Phase HPLC

1987 ◽  
Vol 10 (12) ◽  
pp. 2625-2637 ◽  
Author(s):  
J. Alan Menius ◽  
D. James Schumacher ◽  
Emily A. Hull-ryde ◽  
Cyril Y. Leung ◽  
Robin G. Cummings ◽  
...  
Keyword(s):  
Blood Serum ◽  
Reverse Phase Hplc ◽  
Reverse Phase
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