Fast Analysis of Zidovudine by UHPLC Using a Syncronis C18 1.7 μm Column

Importance of the Topic

Zidovudine is a cornerstone antiretroviral drug in HIV/AIDS therapy, and its rapid, reliable quantitation is essential for pharmaceutical quality control and regulatory compliance. Employing ultra-high performance liquid chromatography (UHPLC) with sub-2 µm particle columns can dramatically reduce analysis times and solvent usage while preserving chromatographic integrity.

Objectives and Study Overview

This application note describes the adaptation of the United States Pharmacopeia (USP) monograph for zidovudine analysis onto a UHPLC platform. The primary goal was to leverage a Syncronis C18 column packed with 1.7 µm particles to accelerate the established method and evaluate its performance against pharmacopoeial standards.

Methodology and Instrumentation

Sample preparation:

• Prepared a 1000 µg/mL zidovudine stock in methanol and diluted to 100 µg/mL with mobile phase (20:80 methanol/water, v/v).

Chromatographic conditions:

• Column: Syncronis C18, 1.7 µm, 50 × 2.1 mm
• Column temperature: 25 °C
• Flow rate: 0.8 mL/min
• Injection volume: 0.5 µL
• Detection: UV at 265 nm
• Mobile phase: methanol/water (20:80, v/v)

Instrumentation used:

• Thermo Scientific Accela UHPLC system
• Thermo Scientific ChromQuest 5.0 software

Main Results and Discussion

Scaling down the USP method from a 5 µm, 250 × 4.0 mm column to the Syncronis C18 1.7 µm, 50 × 2.1 mm column using a transfer calculator achieved an eightfold reduction in run time (from 26 min to ~2.42 min). Six replicate injections yielded:

• Retention time RSD: 0.05%
• Tailing factor RSD: 0.30%
• Column efficiency: ~118 500 theoretical plates
• Peak area RSD: 0.79%

All metrics met USP acceptance criteria (≤2% RSD), confirming method robustness and repeatability.

Benefits and Practical Applications

The optimized UHPLC method offers:

• Eightfold faster analysis throughput
• Lower solvent consumption and reduced waste
• High efficiency and reproducible peak shapes
• Full compliance with pharmacopeial standards

These advantages support high-volume quality control in pharmaceutical production and research laboratories.

Future Trends and Opportunities

Emerging developments may include:

• Further miniaturization with sub-1 µm or core-shell particles
• Adoption of green solvents and sustainable chromatography
• Automated, AI-driven method development and optimization
• Integration with high-resolution mass spectrometry for impurity profiling

Conclusion

The transfer of the USP zidovudine assay to a UHPLC format using a Syncronis C18 1.7 µm column successfully reduced run time eightfold while preserving chromatographic performance and meeting regulatory precision requirements. This approach delivers a robust, high-throughput solution for pharmaceutical quality control.

References

1. Thermo Fisher Scientific. Isocratic Method Transfer Calculator. Separated by Experience. Accessed 2011.
2. Caufield WV, Stewart JT. Chromatographia. 2001;54:561–568.
3. World Health Organization. WHO Model List of Essential Medicines. March 2005. http://whqlibdoc.who.int/hq/2005/a87017_eng.pdf.
4. United States Pharmacopeia. Monograph for Zidovudine. USP 29–NF 24, Section m89510.