Development and validation of simultaneous quantification method for gemcitabine and betulinic acid: augmenting...

Area of Science:

• Analytical Chemistry
• Pharmaceutical Sciences
• Nanotechnology

Background:

• Combinatorial cancer therapy using gemcitabine (GEM) and betulinic acid (BET) shows promise.
• Nanotechnology advancements necessitate validated methods for co-loaded drug formulations.
• Accurate quantification of GEM and BET is crucial for therapeutic efficacy and formulation development.

Purpose of the Study:

• To develop and validate a robust, simple, and economical analytical method for simultaneous GEM and BET estimation.
• To establish a reliable quantification technique for GEM-BET co-loaded nano-formulations.
• To demonstrate the method's applicability in assessing nano-formulation characteristics.

Main Methods:

• Reverse-phase high-performance liquid chromatography (RP-HPLC) was employed.
• A mobile phase of 0.1% orthophosphoric acid-acetonitrile was used for detection at 248 nm (GEM) and 210 nm (BET).
• Method validation followed regulatory guidelines, assessing linearity, accuracy, precision, robustness, and stability.

Main Results:

• The RP-HPLC method achieved simultaneous quantification of GEM and BET with distinct retention times (5 min and 13 min, respectively).
• Validated parameters confirmed the method's linearity, accuracy, precision (intra- and inter-day variability <2%), robustness, and stability.
• The method demonstrated specificity, with no matrix interference observed in drug-spiked FBS samples.

Conclusions:

• A validated, economical, and robust RP-HPLC method for simultaneous GEM and BET quantification has been successfully developed.
• This analytical tool is suitable for quality control of GEM-BET formulations and analysis in biological samples.
• The method supports the advancement of GEM-BET nano-formulations for cancer therapy.