Hyoscine Butylbromide Analysis by British Pharmacopoeia Method and 1260 Infinity II Prime LC

Importance of the Topic

The accurate quantification of hyoscine butylbromide and its impurities is essential for ensuring the safety, efficacy, and regulatory compliance of this antispasmodic pharmaceutical. Reliable analytical methods support product quality control, batch release, and adherence to pharmacopeial standards.

Objectives and Study Overview

This study demonstrates the implementation of the British Pharmacopoeia method 2.2.29 for hyoscine butylbromide analysis using the Agilent 1260 Infinity II Prime LC system. The aim was to verify method performance, including resolution, precision, and operating pressure, when separating the API and specified impurities A and B.

Methodology and Instrumentation

Chromatographic separation was carried out on an Agilent ZORBAX StableBond SBAQ column (4.6 mm × 100 mm, 1.8 µm) with a binary gradient of perchloric acid solution and acetonitrile. Key parameters:

• Mobile Phase A: 0.2% perchloric acid in acetonitrile/water (5:95, v/v)
• Mobile Phase B: 0.2% perchloric acid in water/acetonitrile (30:70, v/v)
• Gradient: 91→15% A over 10 min, total run time 15 min
• Flow rate: 2.5 mL/min, column temperature: 50 °C, injection volume: 2 µL
• Detection: UV at 210 nm

Instrumentation comprised the Agilent 1260 Infinity II Flexible Pump, Multisampler, Multicolumn Thermostat, Diode Array Detector HS, and OpenLab CDS Workstation v2.3.0.
Sample preparation included test solution and serial dilutions to generate Reference Solutions A and B, containing known levels of impurities for system suitability and quantitation.

Main Results and Discussion

Blank injections confirmed baseline stability. Reference Solution A and B injections established retention times: impurity B at 1.74 min, impurity A at 2.04 min, and hyoscine butylbromide at 5.75 min. System suitability criteria were met, yielding a resolution >3.6 between impurities A and B (minimum required 2.0). Precision testing (six replicates of Reference Solution B) showed retention time RSD <0.1% for all peaks. The maximum operating pressure was 680 bar, below the 800 bar limit of the 1260 Infinity II system, demonstrating robust performance under sub-2 µm UHPLC conditions.

Benefits and Practical Applications

The method provides fast, high-resolution separation within a 15-minute run, enabling high throughput in quality control laboratories. The precision and low pressure requirements ensure reproducible results without excessive instrument strain, supporting routine batch testing of hyoscine butylbromide pharmaceuticals.

Future Trends and Potential Applications

Adaptation of this UHPLC approach to other quaternary ammonium compounds can streamline impurity profiling across related drug classes. Integration with mass spectrometric detection could enhance specificity and low-level impurity identification. Further miniaturization and automation will improve sample throughput and data quality in pharmaceutical analytics.

Conclusion

The Agilent 1260 Infinity II Prime LC effectively meets British Pharmacopoeia requirements for hyoscine butylbromide analysis. It delivers sharp, well-resolved peaks, high precision, and operates reliably under high pressure, making it an ideal choice for pharmaceutical QC applications.

References

1. Hyoscine Butylbromide. British Pharmacopoeia 2.2.29, 2019.