Nikhil Gupta*
A drug includes all medicines intended for internal or external use for or in the diagnosis, treatment, mitigation or prevention of disease or disorder in human beings or animals, and manufactured exclusively in accordance with the formulae mentioned in authoritative books. (1) Pharmaceutical analysis is a branch of chemistry involving a process of identification, determination, quantification, purification and separation of components in a mixture or determination of chemical structure of compounds. There are two main types of analysis – Qualitative and Quantitative analysis. Qualitative analysis is performed to establish composition of a substance. It is done to determine the presence of a compound or substance in a given sample or not. The various qualitative tests are detection of evolved gas, limit tests, color change reactions, determination of melting point and boiling point, mass spectroscopy, determination of nuclear half-life etc. Quantitative analysis techniques are mainly used to determine the amount or concentration of analyte in a sample and expressed as a numerical value in appropriate units. These techniques are based on suitable chemical reaction and either measuring the amount of reagent added to complete the reaction or measuring the amount of reaction product obtained the characteristic movement of a substance through a defined medium under controlled conditions, electrical measurement or measurement of spectroscopic properties of the compound. (2)
MATERIALS AND METHODS
|
Reagent |
Specifications |
|
Potassium dihydrogen orthophosphate (Merck – HPLC grade) |
Mol. Formula – KH2PO4 Mol. Weight – 136.09 |
|
Orthophosphoric acid (Merck – HPLC grade) |
Mol. Formula – H3PO4 Mol. Weight – 98 |
|
Ammonium acetate (Merck-GR) |
Mol. Formula-CH3COONH4 Mol. Weight-77.0825 |
|
Methanol (Merck hplc grade) |
Mol. Formula – CH3OH Mol. Weight – 32.04 |
|
Acetonitrile (Merck – HPLC grade) |
Mol. Formula – CH3CN Mol. Weight – 41.05 |
|
Water |
Milli Q Grade |
METHODS
Preparation of buffer (0.01N KH2Po4)
Accurately weighed 1.36gm of potassium dihydrogen Ortho phosphate in a 1000ml of Volumetric flask add about 900ml of milli-Q water and degas in a sonicator and finally make up the volume with water and pH adjusted to 5.4 with dil. OPA
Standard Preparation:
Accurately Weighed and transferred 15mg of levosulpiride and 8mg of Esomeprazole working Standards into a 10ml clean dry volumetric flask, add 3/4th volume of diluent, sonicated for 5 minutes and make up to the final volume with diluents. 1ml from the above two stock solutions was taken into a 10ml volumetric flask and made up to 10ml.
Sample Preparation:
5 tablets were weighed and powdered and transferred into a 50mL volumetric flask, 35mL of diluent added and sonicated for 25 min, further the volume made up with diluent and filtered. From the filtered solution 0.2 ml was pipeted out into a 10 ml volumetric flask and made upto 10ml with diluent.
Linearity:
Linearity solutions are prepared such that 0.25ml, 0.5ml, 0.75ml, 1ml, 1.25ml, 1.5ml from the Stock solutions of Esomeprazole and Levosulpiride are taken in to 6 different volumetric flasks and diluted to 10ml with diluents to get 20ppm, 40ppm,60ppm, 80ppm, 100ppm, 120ppm of Esomeprazole and 37.5ppm, 75ppm, 112.5ppm 150ppm, 187.5ppm, 225ppm of Levosulpiride.
Standard Preparation:
Accurately Weighed and transferred 15mg of levosulpiride and 8mg of Esomeprazole working Standards into a 10ml clean dry volumetric flask, add 3/4th volume of diluent, sonicated for 5 minutes and make up to the final volume with diluents. 1ml from the above two stock solutions was taken into a 10ml volumetric flask and made up to 10ml.
Sample Preparation:
5 tablets were weighed and powdered and transferred into a 50mL volumetric flask, 35mL of diluent added and sonicated for 25 min, further the volume made up with diluent and filtered. From the filtered solution 0.2 ml was pipeted out into a 10 ml volumetric flask and made upto 10ml with diluent.
Accuracy
Standard Preparation:
Accurately Weighed and transferred 15mg of levosulpiride and 8mg of Esomeprazole working Standards into a 10ml clean dry volumetric flask, add 3/4th volume of diluent, sonicated for 5 minutes and make up to the final volume with diluents. 1ml from the above two stock solutions was taken into a 10ml volumetric flask and made up to 10ml.
Sample preparation
50%: 5 tablets were weighed and calculate the average weight of each tablet then 750mg tablet powder was transferred into a 50mL volumetric flask, 30mL of diluent added and sonicated for 25 min, further the volume made up with diluent and filtered. From the filtered solution 0.2ml was pipeted out into a 10 ml volumetric flask and made up to 10ml with diluent.
100%: 5 tablets were weighed and calculate the average weight of each tablet then 1500mg tablet powder was transferred into a 50mL volumetric flask, 30mL of diluent added and sonicated for 25 min, further the volume made up with diluent and filtered. From the filtered solution 0.2ml was pipeted out into a 10 ml volumetric flask and made up to 10ml with diluent.
150%: 5 tablets were weighed and calculate the average weight of each tablet then 2250mg tablet powder was transferred into a 50mL volumetric flask, 30mL of diluent added and sonicated for 25 min, further the volume made up with diluent and filtered. From the filtered solution 0.2ml was pipeted out into a 10 ml volumetric flask and made up to 10ml with diluent.
Method Development
Many trials were done by changing columns and Mobile phases and were reported below.
Trial 1
This trial was run through ods 250 column with mobile phase composition of 55:45 Buffer and Acetonitrile, Flow rate set at 1ml/min.
Fig. 1 Trial chromatogram 1 Observation: resolution is not good
Trial 2
This trial was run through ODS 250mm column with mobile phase composition of 50:50 A water and Acetonitrile, Flow rate set at 1ml/min.
10.5281/zenodo.17503005