Additive – acids, bases and other chemicals added to the co-solvent portion of the mobile phase to improve the chromatography
Back pressure – pressure after the column at the back pressure regulator, it is also the pressure set by the user to control system pressure
Back pressure regulator (BPR) – instrument module used in SFC to control the back pressure on the system at a user determined set point
Chiral – a compound is considered chiral if it has one or more chiral centers resulting in optical isomers or enantiomers that are “mirror images” of each other
Chiral center – typically a carbon (C) atom within a molecule that has four different bonded functional groups
Chiral columns – specific column chemistries required to separate optical isomers (chiral compounds)
Chromophore – molecular group which absorbs light at a particular frequency
Collection timing delay – the time between when the peak exits the detector and reaches the collection selector valve or solenoid
Column oven – oven used in SFC systems for temperature control and column selection
Column volume – volume inside the column “cylinder” based on length and ID minus the approximate volume of the stationary phase
Conditioning solvent – solvent used with the splitter to enhance signal in the MS detector (QDa)
Co-solvent – in SFC, the organic portion of the mobile phase (solvent B)
Critical point – the specific temperature and pressure point for a fluid above which it exists as a supercritical fluid
Density – mass per volume (grams per liter)
Detector channels – single channels that can be monitored independently and in parallel for detection or collection
Diastereomers – compounds that differ in the optical orientation of two chiral centers
Diffusion coefficient – the rate at which a diffusing substance is transported between opposing systems when there is a concentration difference between them
Diffusivity – in chromatography, the ability of a substance to diffuse into and out of the stationary phase particles of the column based on the diffusion coefficient
Dwell volume – the volume from the point of the gradient mixing to the head of the column in a chromatographic system
Efficiency – chromatographic efficiency is measured by the number of theoretical plates or plate height, a low plate height indicates high efficiency and results when a peak has good retention and narrow width
Enantiomers – compounds that differ in the optical orientation of a single chiral center
Extra-column volume – volume of the system between the injector and the detector without the column; impacts peak quality
Extraction – to separate or obtain a product (extract) from a mixture (or matrix) by force
Focused gradient – shallow gradients generated to optimize the separation of a target compound based on the elution percentage from a scouting gradient
Fraction analysis – qualitative and/or quantitative analysis of the collected fractions
Front pressure – pressure before the column in SFC (system pressure)
Gradient methods – chromatography method in which the mobile phase composition changes over the course of a defined period of time
Gradient slope – the rate of change in organic solvent composition per column volume during a gradient
Hydrophilic – miscible with or soluble in water, polar
Hydrophobic – is not miscible with or soluble in water, non-polar
Inlet pressure – pressure of the CO2 before it enters the pump
Isocratic methods – chromatography methods in which the mobile phase composition remains constant over the course of the separation
Joule-Thomson cooling – the cooling of a real gas or liquid when it expands through a valve or opening
Loading – the amount of sample injected onto the chromatographic system
Loading capacity – the amount (mass) of compound that can be loaded based on the dimensions of the column
Lypophilic – lipid soluble, typically used to indicate low polarity
Make-up solvent – solvent used to aid in collection in SFC systems after the CO2 has evaporated
Mass spec detection (MS) – a detection technique that separates compounds based on mass
Mass-flow control – CO2 pumps that control flow based on mass of CO2 (g/min)
Matrix – the substance containing the target compound or isolate
Matrix effects – detection and chromatographic interferences caused by components in the matrix
Mixed-stream injection – injections are performed into the pre-mixed mobile phase
Mobile-phase – the solvent or solvents used to elute compounds from the stationary phase (column)
Modifier – in SFC this is synonymous with co-solvent or the organic portion of the mobile phase
Modifier-stream injection – injections are performed into the co-solvent (modifier) stream before mixing with CO2
Multi-step purification – when purification is accomplished using multiple methods with differing selectivity, whether using two techniques like LC with SFC, or different columns achiral with chiral
Non-polar – describes compounds or solvents that have low polarity, or are hydrophobic and have high LogP values
Normal-phase liquid chromatography (NPLC) – a separation technique in which the mobile phase is typically non-polar and the separation occurs on a stationary phase that is polar
Open-bed collection – collection systems that deposit fractions into tubes or bottles placed in racks in a bed open to the atmosphere
Orthogonal – describes mutually independent or well separated capabilities that span the entire capability range
Phase separation – when a single substance or composition of substances separate into two or more distinct phases
Photodiode array detection (PDA) – a two dimensional UV detection technique where one axis shows time and the other shows UV spectral scans
Polar – describes compounds or solvents that have high polarity, or are hydrophilic and have low LogP values
Pressure drop – the drop in pressure across the column measured by the difference between the front pressure and the back pressure
Productivity – the rate at which the final product can be generated or purified
Purity – a measure of how pure a fraction is based on analytical analysis
Recovery – a measure of the amount of product collected compared to the amount injected or the amount of starting material
Resolution – a measure of the width of two peaks relative to the distance between those peaks
Reversed-phase chromatography (RPLC) – a widely used separation technique in which the mobile phase is polar and the separation occurs on a stationary phase that is non-polar
Scale-up – transferring an analytical scale method to a preparative scale method
Shallow gradient – chromatography method in which the rate of change of organic solvent per column volume is low
Solvating capacity – the amount of compound that can be dissolved per volume of solvent
Split ratio – the ratio of the total preparative flow that is directed to the detectors by the splitter
Splitter – device used send a small ratio of the preparative flow stream to the detectors, accompanied by make-up or conditioning solvent to enhance signal
Stationary phase – the active particles in the column where chromatographic separation occurs
Supercritical fluid – the result when a fluid is above its critical pressure and critical temperature and there is no longer an interface between liquid and gas phases.
System pressure – typically stated as the pump pressure in liquid chromatography, same as front pressure in SFC
Throughput – amount of sample processed and collected per unit time
Van Deemter curves - A plot of plate height vs. average linear velocity of the mobile phase
Viscosity – The degree to which a fluid resists flow under an applied force
Volume-flow control – CO2 pumps that control flow based on volume of CO2 (mL/min)
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q. J. Runco, L. Subbarao, R. Chen, Qualitative and Quantitative Analysis of ß-carotene using UPC2, Application Note, January 2013, 720004550EN.
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u. J. Runco, J. McCauley, Chiral Purification of Volatile Flavors and Fragrances by SFC, Application Note, August 2014, 720005150EN.
v. C.J. Hudalla, A. Tarafder, J. Jablonski, R. Roshchin, K. Fountain, M. Patel, M. Hardink.
w. For more information of useful Application Notes and References, please visit www.waters.com and put the reference numbers above in the search box.
x. T. Yan, F. Riley, UPC2 Strategy for Scaling SFC Methods: Applications for Preparative Chromatography, Application Note, May 2014, 720005064EN.
Beginner's Guide to Preparative SFC
Preparative SFC Enabling Technology
Preparative SFC Method Development
Preparative SFC Principles of Scaling in Prep SFC
Preparative SFC Glossary