Higher Sample Loading Capacity, Better Performance – The Application of SepaFlash® Ruby Series High Resolution Cartridges

In the daily separation and purification work of the organic synthesis personnel, the application technology research center usually uses the amorphous silica gel packing with the particle size of 40-63 μ m as the separation medium for the samples which are not difficult to be separated The normal phase separation can meet the requirements of preparation and purification However, for some samples which are difficult to separate, or need to collect the impurities in the samples for the next analysis, the silica gel packing in the positive phase separation column has put forward higher requirements The silica gel packing with smaller particle size and larger specific surface area has obvious advantages Its higher resolution and larger sample load make it a satisfactory solution for customers In this application case, a synthetic drug intermediate was used as a sample to make parallel comparison between sepaflash ® Ruby series high-resolution silica gel column, speaflash ® ordinary silica gel column and products of a famous brand The results show that sepaflash ® is a kind of high-resolution silica gel column Ruby series products have higher column efficiency, which provides powerful guarantee for researchers to complete the separation and purification of complex samples Experiment part 1 Sample information the sample in this case is the intermediate of a synthesis reaction, and the final synthesis reaction formula is shown in Figure 1 Figure 1 Chemical reaction formula of synthetic intermediate sample 2 Preliminary analysis of sample TLC spot plate analysis of the sample The developer system is petroleum ether (PE) and ethyl acetate (EA) (1:1) According to the results of TLC point plate (as shown in Figure 2), the RF value of the target product is 0.4, and there are two near impurity points near the target product, RF values are about 0.5 and 0.6 For the case that the Δ RF of the target product and impurity is less than 0.1, higher requirements are put forward for the subsequent preparation of purified packing Figure 2 Sample spot results on TLC plate 3 Flash preparation and purification of samples In this case, we selected three flash separation columns See Table 1 for specific separation column information Table 1 Product specifications of the separation column used in this case: separation column model: 12g, one famous brand of normal silica gel column: 12g, sepaflash, ordinary normal silica gel column: 12g, sepaflash ruby, high-resolution normal silica gel column, silica gel particle size: 20 – 40 μ m (spherical), 40 - 63 μ m (spherical), 25 μ m (spherical) Silica gel pore size 60޹60޹50޹specific surface area n / a 500 m 2 / g 700 m2 / g recommended sample amount 60 mg - 1.2 g 10 mg - 1 g 10 mg - 3 G recommended flow rate 16.8 - 30 ml / min 30 - 60 ml / min 25 - 50 ml / min According to the TLC point plate information of the sample, calculate the RF value of the target compound and the main impurity point, then input the above data in the corresponding page of the iPad end control software of sepabean ® machine t, and the control software can automatically recommend the optimized elution gradient The flash preparation experimental parameters of the sample are shown in Table 2, and the preparation and purification spectra on different flash separation columns are shown in Figure 3 Table 2 Flash preparation experimental parameter instrument for sample sepabean ® machine t chromatographic column 12 g normal phase silica gel column of a famous brand 12 g sepabelash ® ordinary normal phase separation column (Order No.: sw-5101-012) 12 g sepabelash ® Ruby high resolution normal phase silica gel column (Order No.: sw-2102-012-sp (H)) detection wavelength 280 Nm mobile phase solvent A: n-hexane; solvent B: ethyl acetate flow rate 30 ml / min injection volume 250 mg elution gradient column volume (CV) solvent B (%) 0.53.58.40 12 45 25 45 Figure 3 Analysis of flash preparation and purification spectra of samples on three different separation columns Figure 3 shows that sepaflash ® Ruby high-resolution normal phase silica gel column has a higher resolution for impurities in the samples, and two impurities with very close Rf value have also been separated by baseline, which shows that sepaflash ® Ruby series high-resolution silica gel column has higher column efficiency than other specifications of silica gel column, which is very suitable for the fine purification of sample system with high separation difficulty In addition, on the premise of meeting the requirements of product purity and yield, sepafflash ® Ruby series separation column allows more samples to be loaded at one time, so as to improve the separation and purification efficiency, save a lot of reagents and time, and effectively reduce the cost Sepabean ® machine t produced by Santai technology and sepaflash ® Ruby series high-resolution silica gel column can provide users with fast and efficient sample preparation and purification solutions, and help laboratory researchers easily complete various sample purification tasks To learn more about the detailed specifications of sepabean ® machine t, or to order the flash purification column used with it, please visit CBG online store.