Application Of HPLC Analysis in Rotary Evaporator

High performance liquid chromatography (HPLC) is also called high pressure liquid chromatography, high speed liquid chromatography, high resolution liquid chromatography and modern column chromatography. It is an efficient and rapid analytical separation technology developed rapidly in the 1970s, and it is an important means of modern separation testing.

High performance liquid chromatography (HPLC) uses liquid as mobile phase, and adopts high-pressure infusion system to pump the mobile phase such as single solvent with different polarities or mixed solvent and buffer solution with different proportions into the chromatographic column with stationary phase. After the components in the column are separated, they enter the detector for detection, so as to realize the analysis of the sample.

The separation mechanism of high performance liquid chromatography is the same as that of conventional column chromatography, but the packing is finer and needs to be pushed by high pressure pump, which has high column efficiency and fast analysis speed. Different from gas chromatography, the mobile phase in liquid chromatography also participates in the separation process of components, and its composition, proportion and pH value can be flexibly adjusted, with various separation modes. In practice, the retention value and selectivity of samples in chromatographic column are adjusted by changing the composition of mobile phase, so that different samples can be separated.

 

Components of HPLC analysis

 

Column: The column is the core component of HPLC system, which is used to separate the compounds in the sample. Common separation columns include reverse phase column, ion exchange column, gel filtration column, etc., and their selection depends on the properties of the compound to be separated and the analysis purpose.

Mobile Phase: Mobile phase is a combination of solvents used in HPLC to pass a sample through a separation column. The mobile phase usually consists of a mixture of solvents, which can be a single solvent or a mixture of multiple solvents in a certain proportion. Choosing the appropriate flow is very important to the separation effect.

Pump: The pump is used to provide the mobile phase and push it through the separation column at a certain pressure. Common pumps include constant pressure pump and gradient pump. Gradient pump can realize linear or nonlinear change of mobile phase composition, so as to obtain better separation effect.

Injector: Injector is used to introduce the sample to be analyzed into HPLC system. The sampler can be a manually operated syringe or an automatic sampler. They usually have the characteristics of accurate volume and good repeatability to ensure accurate sample injection.

Column Oven: The column oven is used to control the temperature of the separation column. By controlling the temperature, the distribution coefficient and separation speed of the sample in the column can be changed, thus optimizing the separation effect.

Detector: The detector is used to detect the separated compound and generate an electrical signal related to its concentration. Common detectors include ultraviolet-visible absorption detector, fluorescence detector and electrochemical detector. The choice of detector depends on the characteristics of the compound to be analyzed and the detection sensitivity requirements.

Data Acquisition System: The data acquisition system is used to receive, record and analyze the signals output by the detector. It can convert the detected signal into parameters such as peak area and retention time, and provide analysis and interpretation of the results.

 

After the solvent is removed by rotary evaporator, the obtained sample is usually concentrated or solid. If you want to further analyze or detect the compounds in the sample, you can analyze them by high performance liquid chromatography (HPLC).

General steps for analyzing samples after rotary evaporation by HPLC

 

1. Sample dissolution: dissolve the sample after rotary evaporation in an appropriate solvent. The solubility of the substance to be analyzed and the requirements of HPLC analysis conditions should be taken into account when selecting solvents.

2. Preparation of standard solution: If quantitative analysis is needed, the standard solution of the compound to be tested can be prepared for subsequent correction and calculation of the analysis results.

3. Carry out HPLC analysis: inject the sample into HPLC system and set up appropriate separation column, mobile phase and detector. According to the purpose of analysis, select appropriate methods and parameters, such as flow rate, temperature and gradient program.

4. Data collection and analysis: the chromatogram of the sample is collected by HPLC system, and the target compound is quantified or identified according to the peak area, retention time or other related parameters. Standard curve method and internal standard method can be used for quantitative analysis.

5. Results Interpretation: According to the analysis results, the compounds in the sample were qualitatively and quantitatively evaluated. According to needs, you can generate reports or record analysis results.

 

The application of rotovap in HPLC analysis has two practical meanings

 

• Sample concentration: In some cases, the concentration of the sample to be analyzed may be too low, so the chemical rotary evaporator can be used to concentrate the sample to increase the detection sensitivity of the analyte in HPLC.
• Solvent removal: In some samples, solvents may interfere with HPLC analysis, so it is necessary to remove or replace the solvents in the samples with solvents suitable for HPLC. The lab rotary evaporator can help to remove the solvent and transform the sample into a form suitable for HPLC analysis.

 

HPLC analysis and rotary evaporator are two commonly used techniques in chemical analysis, and they have some differences and connections in principle and application.

 

Difference

 

Principle: HPLC is based on the principle of liquid chromatography, and the separation and quantitative analysis are realized by the interaction between the stationary phase (column) and the mobile phase (solvent). The evaporator uses the principle of heating and rotation to evaporate the liquid sample quickly and remove the solvent, so as to concentrate the sample or change the solvent environment of the sample.

Application field: HPLC is widely used in chemistry, pharmacy, environment and other fields, and can be used for separation, identification and quantitative analysis of compounds in complex mixtures; Rotary evaporation is mainly used in the process of sample concentration, solvent removal and sample preparation, and is commonly used in organic synthesis, natural product extraction, sample preparation and other fields.

Analysis speed: HPLC analysis usually takes a long time to separate and detect samples, which may take several minutes to several hours; The vacuum rotary evaporator unit can evaporate liquid samples quickly in a short time, usually only taking a few minutes.

 

Connect

 

Sample treatment: Rotary evaporator equipment is often used to concentrate samples or remove solvents during sample preparation to make samples suitable for further analysis methods, such as HPLC. The effect and accuracy of HPLC analysis can be improved by concentrating the sample and removing the solvent by rotary evaporator.

Solvent selection: In HPLC analysis, the choice of solvent is very important to the separation effect; The rotary evaporator can help to remove the solvent from the sample and transform the sample into a form suitable for HPLC analysis. Therefore, the rotary evaporator can provide satisfactory solvent conditions for HPLC analysis.

 

We provide manual lifting rotovap and electric lifting rotovap, please contact us sales@achievechem.com to customized your own rotary evaporator machine.