A Continuous Batch Chromatography Method And Its Application In The Preparation Of High Purity Fish Oil

In this study,we described an effective,intelligent and flexible method for multi-component mixture intermediate product chromatographic preparation technique,which combines the batch chromatography(BC)with the continuous chromatographic(CC)operation mode.We named it continuous batch chromatography(CBC)and used the CBC system to purify eicosapentaenoic acid ethyl esters(EPA-EE)of crude fish oil to evaluate the proposed process.A multi-component mixture contains at least three parts,which are light impurities(small affinity to the stationary),heavy impurities(large affinity to the stationary)and an intermediate product(a target component between the light impurities and heavy impurities).We have built the HPLC and GC analysis methods of fish oil ethyl esters for the qualitative analysis and quantitative testing of EPA-EE.Meanwhile,we also investigated the physicochemical properties and chromatographic properties of the fish oil,such as the stability experiment,solubility experiment and adsorption isotherm experiment.We determined the stationary phase as 120?-10-ODS-BP by Van-Deemter experiment in three different kind of adsorbent.Through investigating the resolution of the components of fish oil crude in different proportions methanol-water system,we determined the mobile phase as 95% methanol.Finally,we determined the flow rate of zone Ⅱ and Ⅵ was 15 m L/min by H-u experiment and elution experiment.At the CBC experiment process,under the guidance of the separation principle of CBC system,the operation conditions of each function zone were obtained by elution experiments.Finally,The running time of zone I is set at 14-15min;the peak threshold of zone II was 940 m V and a time window at 2min;the running time of zone III and V was 12 min and the transfer time of zone IV was 5.4min;the peak threshold of zone VI was 915 m V and a time window at 10min;the running time of the fraction collector and the pump were 4min and 15 min,respectively.The results showed that when the feed concentration is 500g/L,the CBC separation was able to produce EPA-EE with average purity 97.82%,recovery rate 82.01%,productivity is 24.93mg/cm2 cross-sectional area/min,solvent consumption is 0.83L/g,and the average purity of EPA-EE above 98.5% with the recovery rate 81.17%.At the same chromatographic conditions,compared with batch chromatograph(BC)mode,the productivity of EPA-EE of CBC is 5.48 times than that of BC,while the solvent consumption of BC is 1.27 times than that of CBC.CBC is also superior to BC both in the product purity and recovery rate.Obviously,CBC is superior to BC in the separation and preparation processes.Comparing with other continuous chromatography,such as simulate moving bed(SMB),the CBC operation conditions are easier to determine,which can save a large amount of sample and solvent.This method may be useful for commercial preparation of high pure EPA-EE from crude fish oil.There is potential to use CBC for separation purification of intermediate product from other similar multi-component mixture.