Rapid Analytical Method Development and Validation of RP-HPLC Method for the Simultaneous Estimation of Exemestane and Genistein with Specific Application in Lipid-Based Nanoformulations

Abstract

Exemestane (EXE), an irreversible aromatase inhibitor, is employed as a therapy for hormone-dependent breast cancer. Several studies have also established the budding effects of genistein (GEN) in various types of cancer such as breast, prostate, as well as skin due to its feeble estrogenic and anti-estrogenic properties. Considering the promising benefits of GEN, it was combined with EXE to accomplish superior therapeutic efficiency with fewer side effects. The quantification of the exact concentration of EXE and GEN when delivered as a combination would be required for which HPLC method was developed and validated. For this purpose, the C18 ODS column having dimensions of 150 × 4.6 mm, 5 μm, using mobile phase A as methanol:water (35:15, v/v), with formic acid (0.01%), and B as acetonitrile (in the ratio of A:B−−30:70 v/v) at a flow rate of 1 mL/min was commonly used. The Box− Behnken design was chosen as our experimental model, and the interactions among the independent and dependent variables were analyzed. Parameters like linearity, system suitability, specificity, precision (intra-and interday), robustness, ruggedness, LOD (limit of detection), and LOQ (limit of quantification) were selected for the validation of our proposed method. EXE and GEN were eluted individually at 245 and 270.5 nm, respectively, while both of the agents were determined simultaneously at 256 nm, showing retention time as 2.10 and 1.67 min, respectively, and the calibration plot was observed to be linear in the range of 5−110 μg/mL. Hence, the method that we developed and validated was found to be suitable for the identification of both the drugs simultaneously in combination and in our in-house-developed nanoformulation.

Novel Chitosan-Coated Liposomes Coloaded with Exemestane and Genistein for an Effective Breast Cancer Therapy

“…EXE and GEN when analyzed concomitantly demonstrated separation at retention times of 2.10 and 1.67 min, respectively. Additionally, the drug encapsulated in the prepared liposome formulation was also determined using the high-performance liquid chromatography (HPLC) technique, indicating that no significant changes in retention times were observed, assuring the selectivity and high specificity of the developed method …”

Quantification of rifampicin loaded into inhaled polymeric nanoparticles by reversed phase high-performance liquid chromatography in pulmonary nonphagocytic cellular uptake

“…The percentage of recovery was considered acceptable when the value obtained was within the range of 90–110% and the standard error (SE) was less than 2%. 20…”

“…The percentage of recovery was considered acceptable when the value obtained was within the range of 90-110% and the standard error (SE) was less than 2%. 20 3.4.5 Stress stability assays. The stress stability tests of RIF samples containing a known concentration of RIF were exposed to different shock factors (photosensitivity, oxidation, and catalyzed hydrolysis by acids and/or bases) at 25 °C and protected from light (except for samples destined to study the photosensitivity of RIF) for 24 h. Briey, a RIF sample in PBS (pH 7.0) was exposed under visible light for 24 h to evaluate photosensitivity.…”

Development and Validation of a Robust RP-HPLC Method for the Simultaneous Analysis of Exemestane and Thymoquinone and Its Application to Lipid-Based Nanoformulations

“…This was evaluated by six replicate injections of EXE and THY, wherein RT, theoretical plates, peak area, and tailing factor of both analytes at 243 nm were considered for estimating the results. As per the guidelines set by the US-FDA, the relative standard deviation (% RSD) for RT and peak area should be less than 2%, , for the tailing factor, it should not exceed by two, while it should be more than 2000 ( N > 2000) for the theoretical plates of the column …”