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The HPLC set up is composed of two main parts, a mobile phase and a stationary phase. The stationary phase is a solid packed in a column, usually
consisting of a silica gel while the mobile phase is a liquid solvent mixture. The mobile phase is pumped up the column where separation takes place
followed by detection.
There are several volumes in the column such as the column volume, bed volume, elution volume, retention volume and void volume to name a
few.
The HPLC column void volume denoted Vm or V0 is in simple terms the volume of the mobile phase in the column. It is the part of a fraction that
when added to the volume of the stationary phase makes up a whole fraction or 100% volume.
The HPLC column void volume is considered in the presence of the stationary phase particles, meaning that if there is no stationary phase, then the
void volume cannot be calculated. The HPLC column void volume is also roughly the minimum amount of the solvent that has to be pumped through
the column before any sample that is introduced by injection can be detected.
Related: Steps for HPLC Method Development
Determination of the HPLC column void volume is mainly approximate. If unknown, the volume is determined by the injection of an unretained
standard that usually has minimal retention in any given phase. This means that its role is almost inert relative to the components of the mobile
phase. A product of this unretained standard and the flow rate of the system gives the column void volume of that particular system. It should be
noted that the packed volume of the column and the flow rate can also be utilized to the same effect in the calculation of the volume.
To determine the various volumes in High-Performance Liquid Chromatography system, the dimensions of the capillary column that holds the mobile
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5/20/2019 HPLC Column Void Volume : Pharmaceutical Guidelines
HPLC column void volume and the flow rate are useful in the calculation of a value known as the column void time, otherwise known as to. Knowing
this value is helpful in the determination of other critical chromatographic values such as the resolution and the separation factor of a compound.
Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widely-read
pharmaceutical blog since 2008. Sign-up for the free email updates for your daily dose of pharmaceutical tips.
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