New functions of sepabean software: recommendation of elution gradient in normal phase separation

Qian Meiyuan, Qiu Wenjun and Xu Bo application technology research center are the most commonly used separation methods for the separation and purification of organic small molecule samples In the normal phase separation mode, the fixed phase with strong polarity and the mobile phase with weak polarity are used as the combination, and the samples and impurities are separated by the difference of the distribution ratio of sample molecules in these two phases Thin layer liquid chromatography Chromatography (hereinafter referred to as TLC) is a common method for organic synthesis personnel to explore the separation conditions when conducting the positive phase separation experiment Based on the mobility of the sample on the TLC plate, the corresponding specific shift value RF is calculated Then, according to the differences of RF values of each component in the sample and the sample loading amount, the researchers determine the type of flash column needed by experience, and the subsequent separation practice In the experiment, the gradient parameters of elution were optimized continuously, and finally the separation and preparation of samples were completed The above experimental process is a typical work with strong experience dependence, which requires rich practical experience of researchers For most of the new experimental rookies just in contact with organic synthesis and flash preparative chromatography, the "routine" in this process is quite profound So, is there a simple and easy to learn, easy to quickly start the experimental device? The answer is yes Today, I'd like to introduce sepabean ® machine, an intelligent rapid liquid chromatography system from Changzhou Santai technology In the latest version of sepabean control software, the function of automatic recommendation of separation method is integrated The suitable separation column can be automatically recommended according to the sample quantity and TLC information of the sample, and the optimized elution gradient conditions can be automatically generated, which greatly simplifies the experimental process and improves the work efficiency of the majority of researchers The following is just an example to briefly introduce the implementation of the above functions First, prepare the standard sample: weigh 5.0g of methyl benzoate, 10.0g of dimethyl phthalate and 10.0g of diethyl phthalate, mix the above three samples and dissolve them in 20ml of ethyl acetate, add n-hexane after the sample is fully dissolved, and dilute them to 100ml Then, the sample spot plate is made on the TLC plate, the developing agent is petroleum ether / ethyl acetate (PE: EA = 10:1), the spot plate results as shown in Figure 1 are obtained, and the RF value of each sample component is calculated Figure 1 TLC spot plate results of mixture samples Next, input TLC information of samples in sepabean control software, select the developer and proportion used, and input RF value of each component of samples, as shown in Figure 2 Figure 2 Input TLC information of samples in sepabean control software Figure 3 After the TLC information of samples is input, enter the next step of separation setting, as shown in Figure 3 It can be seen that the elution gradient has been automatically generated by software After inputting the sample loading amount of the sample, the recommended separation column will appear in the selection menu of the separation column, and the user can decide whether to choose from it according to the nature of the sample For the samples with low separation difficulty, the separation column recommended by the software can meet the requirements Next, after setting the parameters such as flow rate, detection wavelength, collection method, collection rack type and collection volume, the separation experiment can be started Table 1 shows the parameter settings of this preparation and purification experiment Table 1 Sepabean ® machine chromatographic column 12g sepaflash ® e series normal phase silica gel column (amorphous silica gel, packing particle size 40-63 μ m, pore diameter 60 μ m, order No.: s-8101-0012) detection wavelength 254 nm; 280 nm mobile phase solvent A: n-hexane; solvent B: flow rate of ethyl acetate 30 ml / min, injection volume 0.5 ml (125 mg) Elution gradient (according to the RF value after TLC point plate of the input sample, generated after automatic optimization by instrument control software) column volume (CV) solvent B (%) 0.4.7 3 4.7 8 9.4 13 12.5 Figure 4 Flash preparation and separation map of mixture sample Figure 4 shows the separation spectrum of this flash preparation and purification experiment It can be seen from it that the three components in the sample have been baseline separated, which shows that the elution gradient parameters generated automatically by the instrument control software are relatively appropriate Users only need one sample separation experiment to obtain satisfactory results, and no need to carry out tedious separation condition optimization work, thus greatly improving the work efficiency It can save a lot of valuable time and reduce solvent consumption Sepabean ® machine introduced by Changzhou Santai Technology Co., Ltd is an intelligent rapid liquid chromatography system, which has the characteristics of convenient operation, easy to use, built-in chemical knowledge database, self-learning and safe information sharing, and has applied for a series of invention or utility model patents In order to make the instrument more intelligent and easy to use, we will also develop a series of new functions, please look forward to! 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