Abstract
Objective: To develop a simple, accurate and precise reverse-phase high-performance liquid chromatography (RP-HPLC) method and subsequently validate for the simultaneous estimation of praziquantel (PZQ) and pyrantel pamoate (PP) in the pharmaceutical dosage form.
 Methods: The chromatographic separation was achieved on Phenomenex Luna C18 column (250 mm × 4.6 mm, 5 μm) as stationary phase maintained at an ambient temperature with a mobile phase comprising of water: acetonitrile (20: 80) at a flow rate of 1.0 ml/min and UV detection at 220 nm.
 Results: The retention time of PZQ and PP was found to be 3.897 min and 1.697 min respectively. The method was validated in terms of specificity, accuracy, precision, linearity and robustness as per ICH guidelines. Linearity was obtained in the concentration range of 20–60 μg/ml for both PZQ and PP with correlation coefficients of 0.987 and 0.998 respectively. The accuracy of the method was determined using a recovery test and found as 98.44 % to 100.35 %. All parameters are found to be within the acceptable limit.
 Conclusion: The developed RP-HPLC method was simple, rapid, accurate, precise for the simultaneous estimation of PZQ and PP in bulk and tablet dosage form.
Highlights
Helminth infections, which are influenced by parasites, affect more than one billion people in the world
Various mobile phases containing methanol, water, acetonitrile, ammonium acetate buffer and water with triethanolamine in different ratios were tried with different flow rates
In the growing era of international competition for maintaining the products standard in high commercial and market value, development and validation of analytical method became mandatory Analytical method development is the process of demonstrating whether an analytical method is acceptable for use in the workplace to quantify the concentration of the subsequent sample
Summary
Helminth infections, which are influenced by parasites, affect more than one billion people in the world. PZQ is (11 b RS)-2-(cyclohexylcarbonyl)-1,2,3,6,7,11bhexahydro-4H pyrazino [2,1-a] isoquinoline-4-one, shown in fig. 1a, is an anthelmintic effective against flatworms It is very slightly soluble in water, freely soluble in chloroform and alcohol. Molecular weight of PZQ is 312.4 and the empirical formula is C9H24N2O2. 1b acts as a depolarizing neuromuscular blocking agent causing sudden contraction, followed by paralysis of the helminths. PP chemically designated as 1,4,5,6-tetrahydro-1-methyl-2-[(E)-2(2-thienyl)vinyl] pyrimidine hydrogen; 4,4'-Methylenebis(3hydroxynaphthalene-2-caroxylate) shown in fig. It is freely soluble in ethanol, methanol, acetone, dimethyl sulfoxide and slightly soluble in DMF. The molecular weight is 594.7 and the empirical formula is C34H30N2O6S
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