Application Guide—Coupling Thin-Layer Chromatography with Mass Spectrometry and Flash Chromatography for Intelligent and Rapid Purification

The synthesis, purification, and final confirmation of compounds have always posed significant challenges. From ensuring the successful synthesis of a product to developing a flash purification method and ultimately confirming the identity of the fractions, this lengthy workflow can be time-consuming and repetitive. By establishing a completely new workflow, AIS enables a rapid and effortless process—from reaction monitoring through flash purification to fraction identification and confirmation.

Application Guide—Coupling Thin-Layer Chromatography with Mass Spectrometry and Flash Chromatography for Intelligent and Rapid Purification

2023-11-14

The synthesis, purification, and final confirmation of compounds always pose challenges. From ensuring the successful synthesis of a product to establishing... Flash Purification methods, followed by final confirmation of the fractions. ID The lengthy workflow can be time-consuming and repetitive. AIS By establishing a brand-new workflow, it’s possible to quickly and easily transition from reaction monitoring to... Flash Purification and fraction ID The confirmation process includes the following key steps:

Step 1： Use AIS Plate Express TLC Thin-layer chromatography-mass spectrometry interface and expression CMS Monitoring with a small desktop mass spectrometer Suzuki React, then proceed. TLC/CMS Analysis.

Step 2： TLC To Flash , by automatically from the mobile phone TLC The method for developing board images, using Interchim puriFlash XS520 Flash Chromatography Genius ^® Purify the compound using the system.

Step 3： Through CMS and As soon as possible ，30s Internal identification fraction ID 。

Experimental instruments

Experimental Instruments and Conditions

Sample mixture, including molecular weight, structure, and reactivity.

Experimental method

Step 1: Monitor the reaction using thin-layer chromatography.

Bromoaniline and phenylboronic acid in ethanol, using a palladium catalyst (Figure). 1 ) React at room temperature 2 Hour. 2 Hours later, take a sample at TLC Mark the spots on the plate and develop them in a solvent system consisting of dichloromethane and cyclohexane. Then, via... TLC / CMS Monitor the reaction -Plate Express TLC With expression® CMS Coupled with a small desktop mass spectrometer, from TLC Directly obtain mass spectrometry information from the plate to identify the sample (Figure). 2 ).

Figure 2 After climbing the board TLC The spot aligns with the red laser dot. Press the button, TLC Begin extracting the sample and transferring it to the mass spectrometer for analysis. The mass spectrometry data are shown in the figure above.

Step 2： TLC To Flash : Method conversion

Via the mobile app — Achieve TLC To Flash Method conversion: Upload thin-layer chromatography separation information to... Flash In the system, optimal separation conditions are provided for fraction collection. PuriFlash ^® The system undergoes purification.

Take a photograph of the thin-layer chromatography plate. Enter detailed information such as the title, mark the solvent front and the product of interest, and specify the purification amount. 400mg ) and the solvent used (cyclohexane: 10% And dichloromethane: 90% ). The data is sent via Bluetooth to puriFlash® System, Genius The software automatically helps build the method.

Genius Displayed on the software TLC Data, the system based on TLC The data recommends the optimal gradient purification method. After the run is completed, TLC Board and General UV Curve. The screen displays the total after running. UV Spectrum (Figure 6 ), using the touchscreen function, you can locate the peaks in the fraction for subsequent fraction confirmation analysis.

Figure 3 After the run is completed TLC Board and General UV Curve. The screen displays the total after running. UV Using the touchscreen function, you can locate the peaks in the fraction to facilitate the next step of fraction confirmation analysis.

Step 3 Fraction ID <30 Get it in seconds

Through UV Collect the fractions and confirm the fractions. ID To ensure that the target substance has been eluted, you can take a sample of each compound’s fraction and analyze it by... ASAP/CMS Conduct analysis.

Through As soon as possible and expression® CMS To perform fraction confirmation, simply dip the probe into the fraction collection tube to collect a sample, then insert it directly into the mass spectrometer. 30s The result can be obtained immediately and is consistent with the expected outcome.

Figure 4 Atmospheric Solid Analysis Probe (ASAP) Immerse it into one of the identified fraction collection tubes, then insert it into the mass spectrometer for analysis.

Summary

Flash Intelligent purification, expression ^® CMS 、 TLC Thin-layer chromatography-mass spectrometry interface and As soon as possible Combined with direct injection analysis probes, a simple sampling technique, intuitive software, and advanced detection technologies, this system streamlines your workflow, enabling you to easily and confidently synthesize, identify, and purify your compounds. Thanks to an intuitive workflow, you can be assured of:

l Use TLC and expression ^® CMS Identify the thin-layer chromatography spots of interest and use them for purification;

l Through PuriFlash ^® Purification system and Genius Methods for establishing and optimizing software;

l Use As soon as possible Direct injection analysis probe and expression ^® CMS A small desktop mass spectrometer is used for fraction confirmation.

Keywords:

Application Guide—Using the expression CMS Compact Benchtop Mass Spectrometer to Monitor and Optimize Flow Chemistry Reactions

Application Guide—PuriFlash® 5.250 + iELSD Enables High-Throughput Purification of Active Pharmaceutical Ingredients!