Title:RP-UFLC based Bioanalytical Method Development, Optimization, and
Validation for the Estimation of Isradipine in Rabbit Serum
Volume: 19
Issue: 5
Author(s): Debashish Ghose, Suryakanta Swain*, Chinam Niranjan Patra and Bikash Ranjan Jena
Affiliation:
- School of Pharmacy and Paramedical Sciences, K.K. University, Nalanda, 803115, Bihar, India
Keywords:
Critical process parameters, critical analytical attributes, critical method parameters, pharmacokinetics, stability, isradipine.
Abstract:
Introduction: The objective of this study is to provide a rapid, sensitive, consistent, and costeffective
method for quantifying isradipine using ultra-fast liquid chromatography.
Methods: Quality by Design principles will form the basis of this approach, grounded on response surface
analysis. Shimadzu liquid chromatographic system equipped with a photodiode array detector and LC
solution software was used to conduct the RP-UFLC method development and validation. An ODS C18
(250 x 4.6 mm; 5 μm) UFLC column was used to complete the analysis. The RSM methodology utilized
a central composite design to perform the optimization studies.
Results: The mobile phase ratio and flow rate were considered crucial method parameters, as well as the
peak area, retention time, and USP plate were considered critical analytical attributes. The optimal conditions
for chromatographic separation were followed using 80% acetonitrile and water (20% v/v) as mobile
phase, a 1 mL/min flow rate, an injection volume of 20 μL, 40°C of column oven temperature, and maximum
absorption at λmax 254 nm using graphical optimization technique. When examining concentrations
between 5 and 150 ng/mL, linearity was observed with an R2 of 0.999. The method created was validated
by employing stability testing per the recommendations provided by ICH Q2 (R1). The analysis of blood
serum was modified so that it could be used to examine the pharmacokinetic parameters.
Conclusion: As a result, high accuracy, precision, sensitivity, linearity, and robustness were established
for predicting the amount of isradipine present in its freeze-dried nano-formulations.
