Ginseng is native to China and some parts of Korea and has been used in various systems of medicine for centuries.
The herb has purported immune boosting abilities among other properties and thus is used in a wide range of research activities.
It has been used to try treat a range of diseases and disorders from menopausal symptoms to erectile dysfunction, hepatitis C, lowering blood pressure etc.
The root of the Asian ginseng has certain chemical components that are termed ginsenosides. These are believed to be responsible for the medicinal properties.
3 main ginsenosides
A HPLC method listed in the Chinese Pharmacopeia identifies three main ginsenosides:-
This traditional method uses 4.6 mm × 150 mm, 5 μm column and takes around 100 minutes. A reduction of this time frame could aid in increasing throughput and reduce lab costs.
In this study the traditional method was scaled to Agilent Poroshell 120 EC-C18 and Poroshell 120 SB-C18 columns.
For the experiment the Agilent Poroshell 120 2.7 μm columns were packed with porous materials to achieve a faster separation while maintaining the linear velocity of the analyte within the column.
The 1200 Series SL LC system including a binary pump, a high performance autosampler, column compartment with thermostat and a diode array detector (DAD) was used.
The original separation method of ginseng was repeated using an Agilent ZORBAX Eclipse Plus C18, 4.6 mm × 150 mm, 5 μm and an Agilent ZORBAX SB-C18, 4.6 mm × 150 mm, 5 μm column.
Results showed that the new method saved around a third of the original analysis time. The time can be reduced further by increasing the flow rate and decreasing the gradient time.
Since these two parameters are directly proportional, analysis time can be halved when using double the original flow rate (raising it from 0.425 mL/min to 0.85 mL/min).
Three times the original flow rate can further reduce the time taken for analysis.
Further the effectiveness of analysis of the Poroshell column is not compromised with the high flow rates compared to that of columns with1.8 μm packing. A reason for this is the fact that as the flow rate rises to three times the original the pressure from the original 174 bar is also raised to about 485 bar. The Agilent 1260 Infinity LC has a pressure limit of at least 600 bar that makes it suitable for this experiment.
To summarize, there was substantial time saving without compromising resolution when ginsenosides were analyzed from ginseng using Poroshell 120 columns instead of traditional 4.6 X 150mm, 5µm columns.
The less porous 2.7µm particle columns show a similar performance when compared to completely porous sub-2µm columns but at lower pressures.
Faster separations on a UHPLC are allowed by higher flow rates, up to the pressure limit of the column of 600-bar.
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