Application case of sepaflash series C18 reversed phase purification column -- Separation and purification of a pharmaceutical synthesis intermediate

In the synthesis process of drug intermediates, it is usually difficult for the applied technology research center to directly obtain the target compounds that meet the purity requirements, which requires the separation and purification of the synthetic products The commonly used purification methods include distillation, extraction, precipitation, filtration, column chromatography and so on Column chromatography (also known as chromatography) is more and more widely used in the separation and purification of various compounds due to its high efficiency, high product purity, easy automation and other characteristics In the classification of column chromatography, according to the polarity of stationary phase and mobile phase, it can be simply divided into two categories: positive phase separation and reverse phase separation Among them, the separation mode that the polarity of stationary phase is stronger than that of mobile phase is called positive phase separation mode Because of historical reasons, this separation mode first appeared, so it is named With the development of application technology, the range of application of reversed phase separation mode has greatly exceeded the normal phase separation mode In the reverse phase separation mode, the most commonly used stationary phases include C18, C8, C4, etc This paper introduces the application of C18 stationary phase in the separation and purification of a pharmaceutical synthesis intermediate Experimental part 1 Sample information the sample used in this paper is from the last step synthesis reaction of a drug intermediate, and the reaction formula is shown in Figure 1 Fig 1 The structural formula of the possible impurities in the sample is shown in Fig 2 Figure 2 Structural formula of possible impurities 2 Preliminary analysis of sample the preliminary purity analysis of the sample was carried out by HPLC (see Figure 3) The result shows that the purity of the sample is 52%, and the content of main impurities is 46% According to the analysis of HPLC chromatogram, the target products and impurities have been separated by baseline, and the separation degree is high, so the reverse phase separation method is used for separation and purification in the future, and it can be considered to increase the sample loading to shorten the preparation and purification process and improve the work efficiency Fig 3 HPLC analysis spectrum of the sample Fig 3 The flash preparation and purification experimental parameters of the sample are shown in Table 1, and the flash preparation and purification spectrum is shown in Fig 4 Table 1 Sepabean ® machine t chromatographic column 330g sepabelash ® bonded series C18 reversed-phase purification column (spherical silica gel, 20-45 μ m, 100 μ m, order No.: sw-5222-330-sp) detection wavelength 220 nm; 254 nm mobile phase solvent A: water; solvent B: methanol flow rate 50 ml / min injection volume 5ml (1.6g) elution gradient time (min) Solvent B (%) 0.30 53 75 63 75 73 90 Fig 4 Flash preparation and purification spectrum of sample 4 Purity analysis of purified product After the prepared sample is evaporated and dried, the purity analysis is carried out by HPLC (see Fig 5) The result shows that the purity is 99.75%, which meets the purity standard of the target product Figure 5 Results and discussion of HPLC analysis spectrum of purified product in this case, the sample was separated and purified by C18 reversed-phase purification column According to the analysis results of the sample under HPLC conditions, combined with sepabean ® machine t produced by Santai Technology The automatic recommendation function of elution gradient in its own control software can easily and quickly convert the analytical conditions of HPLC into the elution conditions of flash preparation chromatography In addition, the high sample loading of 330g column also makes it possible to obtain the target product that meets the purity requirements in one injection, which undoubtedly greatly improves the experimental efficiency Sepabean ® machine t produced by Santai technology and sepaflash ® series C18 reversed-phase purification 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 the ordering information of the flash purification column used with it, please visit CBG online store.