UHPLC Method Development for Analyzing a Once-Daily Tablet Formulation for HIV-1 Infection Treatment

Significance of the Topic

Combination antiviral tablets for HIV-1 therapy require efficient analysis of multiple active compounds and impurities to ensure quality and safety. Ultra high performance liquid chromatography (UHPLC) offers high resolution and speed critical in pharmaceutical quality control.

Objectives and Study Overview

This study aimed to develop a rapid, generic UHPLC method for simultaneous separation and quantitation of the four main active pharmaceutical ingredients in a once-daily Stribild tablet formulation, while also monitoring relevant impurities.

Methods and Instrumentation

• Instrumentation: Thermo Scientific Vanquish UHPLC system with binary pump, autosampler, column compartment, and diode array detector.
• Column: Accucore Vanquish C18+, 1.5 µm, 2.1 × 100 mm.
• Mobile phase: water + 0.1% formic acid (A) and methanol + 0.1% formic acid (B).
• Gradient: 5–90% B in 0–6 min, hold 6–9 min, re-equilibrate to 5% B by 10 min at 0.5 mL/min, 50 °C.
• Detection: DAD at 214 nm and 260 nm; 0.5 µL injections across volumes 0.01–1 µL for calibration.
• Standard preparation: single API stock solutions in methanol or water, mixed to reflect tablet dosage ratios.

Main Results and Discussion

The optimized method resolved all four APIs and at least five related impurities within 14 min, achieving baseline separation and critical resolution of 2.1 between cobicistat and an elvitegravir impurity. Retention time reproducibility was excellent (RSD < 0.008%). Calibration curves over a wide injection range delivered correlation coefficients > 0.9998. Detection wavelengths were optimized using 3D diode array data, yielding limits of detection from 0.02 to 1.02 ng on column and LOQ from 0.06 to 3.41 ng.

Benefits and Practical Applications

This method provides a fast, robust approach for quality control of combination HIV formulations and can be adapted for impurity profiling during drug development and manufacturing quality assurance.

Future Trends and Possibilities

Further integration with mass spectrometric detection may enhance specificity for trace impurities. Extension to other fixed-dose combinations and automation of sample preparation can support high-throughput screening in pharmaceutical laboratories.

Conclusion

The developed UHPLC-DAD method demonstrates rapid, reproducible separation of Stribild components and impurities, offering high sensitivity and linearity. It supports efficient quality control workflows for antiretroviral combination products.

References

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• Thermo Fisher Scientific. Vanquish UHPLC System brochure; 2014.