Fingerprint Profiling of Extracts from Danshen (Salvia miltiorrhiza) Using Sub-2 μm Superficially Porous LC Columns

Význam tématu

Fingerprint profiling of traditional Chinese medicine extracts such as Danshen (Salvia miltiorrhiza) is essential for comprehensive quality assessment and authentication. Complex herbal matrices contain numerous bioactive compounds, making single-analyte testing insufficient. Chromatographic fingerprints capture the entire chemical profile, enabling differentiation between genuine and adulterated samples and ensuring batch consistency.

Cíle a přehled studie / článku

This application note describes the transfer of Chinese Pharmacopeia (CHP) HPLC fingerprint methods for total phenolic acids and tanshinones to an Agilent UHPLC platform using sub-2 µm superficially porous Poroshell 120 columns. The aim is to boost peak capacity and reduce analysis time while adhering to pharmacopeial fingerprint requirements.

Použitá metodika a instrumentace

UHPLC conditions and instrumentation:

• Agilent 1290 Infinity system: binary pump, autosampler, thermostatted column compartment, diode array detector.
• Columns: Agilent InfinityLab Poroshell 120 EC-C18, 2.1×100 mm and 2.1×150 mm, 1.9 µm superficially porous particles.
• Mobile phases: (A) water with phosphoric acid (0.026% for tanshinones; 0.05% for phenolic acids), (B) acetonitrile.
• Gradient programs optimized for each extract; flow rates 0.33–0.42 mL/min; column temperatures 25–30 °C; detection UV at 270–286 nm.
• Sample prep: total phenolic acids dissolved in water (1 mg/mL); tanshinones in methanol (1 mg/mL).

Hlavní výsledky a diskuse

• Longer 150 mm columns delivered peak capacities of 126–131, compared to 91–98 for 100 mm columns, enhancing resolution of complex profiles.
• Key compounds—rosmarinic acid, lithospermic acid B, cryptotanshinone, tanshinone IIA—were clearly resolved and matched pharmacopeial standards.
• Superficially porous particles maintained high efficiency under UHPLC pressures (up to 835 bar) and shortened analysis times.
• Conditional peak capacity was calculated as nc=(tR,n–tR,1)/w, where w is the average peak width at 4σ height.

Přínosy a praktické využití metody

The optimized UHPLC fingerprint method offers:

• Comprehensive chromatographic profiles for robust quality control of Salvia extracts.
• Faster throughput aligning with pharmaceutical QA/QC workflows.
• Regulatory compliance with existing pharmacopeial fingerprint standards.

Budoucí trendy a možnosti využití

Potential future directions include:

• Coupling with mass spectrometry for structural elucidation and quantitation.
• Development of ultrafast gradients and automation for higher sample throughput.
• Application of chemometric and multivariate data analysis for fingerprint interpretation and batch comparison.
• Extension of superficially porous column technology to other herbal medicines and nutraceuticals.

Závěr

Sub-2 µm superficially porous Poroshell 120 columns significantly improve the fingerprint profiling of Danshen extracts by increasing peak capacity and reducing run times. This UHPLC approach strengthens the reliability and efficiency of traditional Chinese medicine quality control.

Reference

1. TsaI-Hui Lin, Ching-Liang Hsieh, Lin Hsieh. Chinese Medicine 5:22 (2010).
2. China Pharmacopoeia Commission. Salvia Total Phenolic Acids and Tanshinones; China Pharmacopoeia 2015, p. 397.