Ultra fast liquid chromatographyProminenece UFLC

Rapid analysis has always been used as an important technical topic, but with the rapid popularization of LCMS, improving cost-effectiveness has become an urgent issue and an important factor in accelerating the process.
Rapid analysis refers to shortening the analysis time, usually achieved by shortening the chromatographic column or increasing the flow rate of the mobile phase. However, using commonly used packed columns with a particle size of 5 μ m, the separation performance will significantly decrease using any method, making rapid separation meaningless.
Therefore, the following two methods have been developed:
• Use small-sized fillers
This method will make rapid separation possible. However, due to the increase in flow resistance inside the chromatographic column, the pressure on various components of the instrument and the chromatographic column will also significantly increase.
• High temperature separation
Higher temperatures can accelerate the diffusion of substances and reduce the flow resistance in the chromatographic column, making rapid separation possible. High temperature analysis is effective in reducing column pressure, but it should be noted that high temperature can cause degradation of the chromatographic column and lead to sample decomposition, which should be taken into account when applied.
In order to solve the problem of back pressure, specialized instruments with improved pressure bearing capacity have been developed, which are usually relatively expensive. However, these specialized instruments have to make compromises in terms of injection performance and sensitivity. Moreover, packed columns with overly fine fillers have high hardware requirements, and the development of analytical methods using such columns sacrifices generality. Therefore, if this instrument is used for routine HPLC analysis, its efficiency will decrease.
Prominence supports fast separation
Prominence uses standard components to meet both traditional and high-speed analysis needs of researchers, and can support operating temperatures up to 85 ℃, making rapid analysis possible at lower pressures. At the same time, the analytical performance and flux level demonstrated by the instrument are rare even in more expensive instrument configurations.
Prominence UFLC supports rapid analysis through the following features:
The small column tube and small volume flow pool reduce the peak broadening effect outside the column.
Use a Shim pack XR-ODS chromatographic column with a particle size of 2.2 μ m.
Shim pack XR-ODS is a new generation of rapid analytical chromatography column that considers the issue of filler refinement and cleverly finds a balance between separation efficiency and pressure, which can improve or maintain separation efficiency while shortening analysis time.

Chromatographic conditions; column: described above, mobile phase: water/acetonitrile (3/7,v/v), flow rate: described above, temperature: 40 ℃, detection: absorbance at 245 nm, sample volume: 4μL(XR-ODS), 10μL(VP-ODS).
Peaks; 1: actophenone, 2: propiophenone, 3: butyrophenone, 4: balenophenone, 5: hexanophenone, 6: heptanophenone, 7: octanophenone.
The Prominence SIL-20A automatic sampler achieves ultra fast injection of 10 μ l in 10 seconds.

Chromatographic conditions; column: Shim-pack XR-ODS(3mm i.d.×30 mm, 2.2μm), mobile phase: water/acetonitrile(4/6 to 2/8 in 0.4 min, convex gradient), flow rate: 3mL/min, temperature: 80 ℃, detection: absorbance at 245 nm, sample volume: 4μL(each 800 nm ol),.0.1 min delayed injection.
Peaks; 1: actophenone, 2: propiophenone, 3: butyrophenone, 4: balenophenone, 5: hexanophenone, 6: heptanophenone, 7: octanophenone.
Supports high temperature analysis up to 85 ℃.
Temperature and rapid analysis: As the column temperature increases, the theoretical number of trays increases, column efficiency increases, and column pressure decreases.

Chromatographic conditions;
mobile phase:　water/acetonitrile (3/7, v/v),
detection: absorbance at 245 nm.
Peaks: 1: Acetophenone 2: Phenylacetone 3: Phenylbutanone 4: Phenylpentanone 5: Phenylhexanone 6: Phenylheptanone 7: Benzophenone
The effect of temperature on compounds: High temperature analysis is effective in reducing column pressure, but high temperature can cause degradation of chromatographic columns and lead to sample decomposition, which should be taken into account when applied. High temperature analysis is only used as an auxiliary tool for rapid analysis.
A detector with high sampling frequency can achieve fast data acquisition without losing information.
Specialized chromatographic method conversion software makes it easy to convert conventional chromatographic conditions to UFLC conditions, ensuring the universality of analytical methods.

Reproducibility of rapid analysis
It is more difficult to achieve reproducibility of component retention time in rapid separation than in conventional analysis. The reproducibility (RSD) of retention time increases inversely with the square root of time, so more stringent liquid delivery performance is required than conventional analysis. Prominence UFLC can ensure excellent liquid delivery capability with a micro stroke liquid delivery resolution of 3nL/min and a control response of 0.1 seconds during gradient liquid delivery, and can exhibit extremely superior retention time reproducibility.

What is high flux
The goal of rapid liquid chromatography is high throughput, which means that more samples are analyzed daily or hourly. In order to achieve high throughput, it is not only necessary to shorten the analysis time of individual samples, but also to optimize the entire cycle injection and analysis time. The original intention of pursuing speed is not only to shorten the analysis time, but also to shorten the analysis cycle. The sample injection speed of Prominence UFLC is 10 seconds, which greatly shortens the analysis cycle.

Application Examples

UFLC analysis of PTC derived amino acids

There are two detection methods for amino acid analysis. One is the post column derivatization method, which uses ion exchange chromatography column separation and derivatization for further detection. The other is the pre column derivatization method, which uses reverse phase chromatography column separation after derivatization. From the perspective of rapid analysis, the pre column derivatization method is effective. Here, amino acid analysis was performed in a shorter time using Shim pack XR-0DS.
Derive using phenyl isothiocyanate. The post column derivatization method requires 1 hour, while the pre column derivatization method using a conventional chromatographic column requires 25 minutes of analysis, which can be shortened to 4 minutes.

UFLC analysis of pigments

The pigments added to food include synthetic compounds and natural compounds, which have high water solubility and are often analyzed by HPLC. However, when multiple pigments are combined, simultaneous analysis requires a long time. Here is an example of simultaneous analysis of pigments using Shim pack XR-ODS ultra fast LC. The results of simultaneous analysis of 12 pigment components are shown in the following chromatogram. The analysis time required for using conventional LC is 50 minutes, while using ultra fast LC, the analysis is completed in about 6 minutes.

UFLC analysis of polycyclic aromatic hydrocarbons (PAHs)

PAHs are chemical substances used to determine the degree of atmospheric pollution caused by vehicles and other exhaust gases. PAHs have strong carcinogenicity, so atmospheric monitoring of these substances is an important measurement item. PAHs exhibit strong fluorescence and can be analyzed with high sensitivity using a fluorescence detector.
For the analysis of these 10 PAHs, UFLC achieved 10 times faster analysis speed (analysis time of 5 minutes) compared to conventional LC (analysis time of 50 minutes) without loss of separation state.