Development and validation of a rapid HPLC method for the determination of ascorbic acid, phenylephrine, paracetamol and caffeine using a monolithic column P. Go to our Instructions for using Copyright Clearance Center page for details. Authors contributing to RSC publications journal articles, books or book chapters do not need to formally request permission to reproduce material contained in this article provided that the correct acknowledgement is given with the reproduced material.
Published online May The aim of this study was to develop a simple, rapid and sensitive reverse phase high performance liquid chromatography RP-HPLC method for quantification of sirolimus SRL in pharmaceutical dosage forms.
The chromatographic system employs isocratic elution using a Knauer- C18, 5 mm, 4. Mobile phase consisting of acetonitril and ammonium acetate buffer set at flow rate 1.
The analyte was detected and quantified at nm using ultraviolet detector. The method was validated as per ICH guidelines. For all quality control QC standards in intraday and interday assay, accuracy and precision range were Samples were stable during preparation and analysis procedure.
Therefore the rapid and sensitive developed method can be used for the routine analysis of sirolimus such as dissolution and stability assays of pre- and post-marketed dosage forms.
It was first identified as an antifungal agent produced by the Streptomyces hygroscopicus bacterium and was subsequently demonstrated to be a potent immunosuppressive agent that was approved by the U.
Food and Drug Administration FDA to be Method development hplc thesis beside cyclosporine or tacrolimus in kidney transplantation. In contrast to tacrolimus and cyclosporine, which inhibit the production of cytokines, sirolimus binds to the FK peptidyl-prolyl cis-trans isomerase binding protein which modulates the activity of the mammalian Target Of Rapamycin mTOR.
The mTOR inhibits Interleukinmediated signal transduction, resulting in cell cycle stop in the G1-S phase and prevents cell cycle progression and proliferation, hence blocks the response of T- and B-cell activation by cytokines.
These pharmacological properties allow rapamycin not only to be a promising immunosuppressant with the absence of nephrotoxicity but also to be a possible chemotherapeutic agent against many types of solid tumor.
SRL is a white to off-white powder, insoluble in water, very slightly soluble in hexane and petroleum ether, soluble in methanol, diethyl ether, and N, N-dimethylformamide DMF and freely soluble in benzyl alcohol, chloroform, acetone, and acetonitrile ACN.
As stated by the current International Conference on Harmonization ICH guideline, accuracy, precision, linearity and stability of test methods are some of the analytical parameters which require assays validation.
The immunoassay is not commercially available, and there is an acute need for an accurate, rapid, and simple chromatographic assay to determine SRL in drug development procedure and also post marketing investigations. Double-distilled water was used during the entire HPLC procedure.
Analytical grade ammonium acetate, sodium lauryl sulphate SLS and sodium hydroxide were obtained from Merck Darmstadt, Germany. System Gold software was used for data acquisition and system Gold nouveau software was used for data reporting and analysis. For the preliminary analysis different columns in combination with different solvent systems were tried out.
Analytical column used for analysis was Knauer ODS 5 mm, 4. SRL was detected by UV absorption at nm. One ml of obtained solution was diluted in ml volumetric flask with sodium lauryl sulphate 0. Linearity The linearity of an analytical method indicates its ability to obtain the response directly proportional to the concentration of the analyte in the sample within a definite range.
Each set of standards was injected into the column from lowest to highest concentration. LOD is the lowest concentration of the analyte detected by the method and LOQ is the minimum measurable concentration.
The first method relies on visual evaluation method, second, signal to noise ratio method the signal-to-noise ratio of 3: Accuracy and Precision Three quality control samples with concentrations at three levels low, medium and high within calibration range were used in triplicates to determine the accuracy and precision of the method.
The mean measured concentrations for all prepared samples were considered for intraday and interday accuracy and precision evaluation.
In all sample stability experiments, the results were compared with the sirolimus initial concentrations.
Statistical analysis using least square regression indicated excellent linearity for SRL in the mentioned range. Concentration curves for SRL had a mean slope, intercept and r2 of0.Reversed Phase HPLC. Reversed phase HPLC is one of the most utilized modes of chromatography. This comprehensive module will provide you with the knowledge and understanding you need to apply this method, design or improve mobile phases through solvent choice or pH adjustment, and which parameters can be altered to change selectivity and retention and improve resolution.
Full-Text Paper (PDF): BASIC SKILLS TRAINING GUIDE -HPLC method development and validation- an overview. Trang, Hung Khiem, "Development of HPLC methods for the determination of water-soluble vitamins in pharmaceuticals and fortified food products" ().
All Theses. An HPLC method for the separation of glycerol oxidation products, namely glyceraldehyde, dihydroxyacetone, mesoxalic, tartronic, and glycolic and glyceric acids on an ion-exchange 8% cross-linked calcium sulfonated divinylbenzene-styrene resin column was developed and validated.
A reliable and reproducible reversed-phase high performance liquid chromatography (RP-HPLC) was developed for the quantitative determination of Quetiapine Fumarate from marketed bulk tablets.
Introduction to method validation Vicki Barwick LGC Overview • What is method validation? • Why is method validation necessary? When do you validate a method? • During method development • Before using any method for samples – verify own ability to match published data.