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Selection of ideal mobile phase and stationary phase is very important to get accurate separation of mixtures or impurities as a whole. There indeed are parameters which are explained in this article, important to be kept in mind while a method development of HPLC method. But as such there is no list of such parameters and their accurate limits can be applied while method development. In this article, there have been mention of certain parameters which are generally looked for, and not exact, but optimal application of such has been discussed. When we talk about stationary phase, parameters which we tend to optimize very often include mostly commonly the pH of the analyte as well the mobile phase used and stability of the column packing material over a range of temperature. Choosing of stationary phase greatly depends on the nature of analyte. Stationary phase will depend and vary simultaneously if analyte is lipophilic or an ionic compound. In order to increase separation and increase mass transfer there have been new addition to column packing material apart from the widely used silica. New advances result into greater column loading, better flow, reduced plate height and reduced back pressure. Leaving alone stationary phase, mobile phase selection is also a task of its own. Very commonly used mobile phase solvents include acetonitrile, methanol, THF, water and buffer of salts like acetate, phosphate, etc. Apart from these traditional choices, there have been use of pressurized hot water and ionic liquids as mobile phases which are termed as “Environmental-friendly” because these solvents are devoid of any organic counterpart or organic modifier in them, which produces harmful fumes on getting heated. Apart from the environmentally friendly mobile phase options, we have some specific chemicals to be used when chemical property of the analyte is specific.
Selection of ideal mobile phase and stationary phase is very important to get accurate separation of mixtures or impurities as a whole. There indeed are parameters which are explained in this article, important to be kept in mind while a method development of HPLC method. But as such there is no list of such parameters and their accurate limits can be applied while method development. In this article, there have been mention of certain parameters which are generally looked for, and not exact, but optimal application of such has been discussed. When we talk about stationary phase, parameters which we tend to optimize very often include mostly commonly the pH of the analyte as well the mobile phase used and stability of the column packing material over a range of temperature. Choosing of stationary phase greatly depends on the nature of analyte. Stationary phase will depend and vary simultaneously if analyte is lipophilic or an ionic compound. In order to increase separation and increase mass transfer there have been new addition to column packing material apart from the widely used silica. New advances result into greater column loading, better flow, reduced plate height and reduced back pressure. Leaving alone stationary phase, mobile phase selection is also a task of its own. Very commonly used mobile phase solvents include acetonitrile, methanol, THF, water and buffer of salts like acetate, phosphate, etc. Apart from these traditional choices, there have been use of pressurized hot water and ionic liquids as mobile phases which are termed as “Environmental-friendly” because these solvents are devoid of any organic counterpart or organic modifier in them, which produces harmful fumes on getting heated. Apart from the environmentally friendly mobile phase options, we have some specific chemicals to be used when chemical property of the analyte is specific.
Tofisopam is an anxiolytic used in treatment of anxiety like disorders, prescribed in the dosage of 50mg to 300mg. As the drug is insoluble in water it is necessary to develop an economical and streamlined liquid chromatography for determination of pure and commercial dosage of Tofisopam. The main objective of the work is to develop and validate an RP-HPLC method for determination of Tofisopam as per ICH guidelines. An RP- HPLC method was devised using 0.1% Orthophosphoric acid in water: methanol in the ration 10:90% v/v and validated for parameters such as system suitability, linearity, limit of detection, limit of quantitation, precision, accuracy, assay, robustness, and ruggedness, as per ICH Guidelines. Tofisopam showed the maximum absorbance at 238nm with good system suitability and linearity was entrenched within the range of 10-60µg/ml, with the regression coefficient of 0.9996. The limit of detection and limit of quantitation were found to be 2.75 µg/ml and 8.855µg/ml respectively. Assay was valuated to be 101%, Accuracy was valuated between 98-103%, and %RSD was less the 2% for precision, robustness and ruggedness, so the method is precise and accurate. The method robustness is demonstrated by insignificant variations in absorption values and mobile phase ratio with deliberate alterations. In accordance with the validation results, the procedure is simple, specific, precise, accurate, robust and suitable for routine analysis of Tofisopam in bulk and commercial products.
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