Study of silica gel regeneration applied on cyclosporine A column chromatography

doi:10.16085/j.issn.1000-6613.2022-1317

Abstract

Abstract:

The industrial production of cyclosporine A involves fungal fermentation, extraction, concentration, crystallization, silica gel column chromatography, decolorization, secondary crystallization and other steps, among which silica gel column chromatography produces a large amount of waste silica gel, which is under great pressure of environmental protection treatment and high cost for enterprises. Based on this, the silica gel regeneration process of cyclosporine A column chromatography was studied in this paper, and the regeneration conditions were optimized by response surface methodology: four column volumes were backwashed with ethanol containing 0.26% polysorbitol, and 2.1 column volumes were forward washed with deionized water containing 0.15% hydrogen peroxide at a flow rate of 5mL/min. The recycled silica gel was pressed out with compressed air and dried until the moisture content was 6%. The yield of the regenerated silica gel can reach more than 75% when used according to the original process, and the yield of the third regenerated silica gel can still reach 75%. This study established a set of feasible cyclosporine purified silica gel regeneration process, which can effectively save factory production cost and reduce environmental pressure.

Key words: cyclosporine A, column chromatography, silica gel regeneration, purification

摘要：

环孢菌素A工业化生产涉及真菌发酵、萃取、浓缩、结晶、硅胶柱层析、脱色、二次结晶等步骤，目前硅胶柱层析步骤的硅胶只能一次使用，现实生产过程产生了大量的废硅胶，环保处理压力大、企业成本高。据此本文开展了环孢菌素A柱层析硅胶再生工艺研究，针对硅胶上吸附的杂质性质，研究了硅胶再生洗脱溶剂系统，通过响应面法优化再生条件，结果表明：选用含有0.26%聚山梨醇的乙醇反向冲洗4个柱体积，再使用双氧水浓度为0.15%的去离子水正向冲洗2.1个柱体积，冲洗流速为5mL/min；使用压缩空气将再生后的硅胶压出，烘干至水分含量为6%。按照本方法得到的再生硅胶按原工艺使用时，环孢菌素A回收率完全达标，并且可以三次重复使用达标。本研究建立了一套切实可行的环孢素纯化硅胶再生工艺，可有效节约工厂生产成本和减轻环境压力。

关键词: 环孢菌素A, 柱层析, 硅胶再生, 纯化

CLC Number:

Q819

HUANG Ye, YAN Xing, WU Qiaowei, CHAI Xiaotao, PAN Gongying, ZHANG Jinfeng, LI Xiangqian. Study of silica gel regeneration applied on cyclosporine A column chromatography[J]. Chemical Industry and Engineering Progress, 2022, 41(S1): 461-468.

黄晔, 言行, 吴巧薇, 柴小涛, 潘恭赢, 张金峰, 李相前. 环孢菌素A柱层析硅胶再生工艺[J]. 化工进展, 2022, 41(S1): 461-468.

Figures/Tables 9

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因素	水平
因素	-1	0	+1
A：乙醇用量/BV	2	3	4
B：聚山梨醇浓度/%	0.1	0.2	0.3
C：双氧水浓度/%	0.1	0.2	0.3
D：去离子水用量/BV	2	3	4

因素	水平
因素	-1	0	+1
A：乙醇用量/BV	2	3	4
B：聚山梨醇浓度/%	0.1	0.2	0.3
C：双氧水浓度/%	0.1	0.2	0.3
D：去离子水用量/BV	2	3	4

来源	平方和	自由度	均方	F值	p值	显著性
合计	1553.21	28	—	—	—
模型	1521.66	14	108.69	48.22	< 0.0001	**
A：乙醇的使用量	286.65	1	286.65	127.18	< 0.0001	**
B：聚山梨醇浓度	93.30	1	93.30	41.39	< 0.0001	**
C：双氧水浓度	113.47	1	113.47	50.34	< 0.0001	**
D：去离子水使用量	18.03	1	18.03	8.00	0.0134	*
AB	37.76	1	37.76	16.75	0.0011	**
AC	0.65	1	0.6480	0.28	0.6002
AD	25.65	1	25.65	11.38	0.0045	**
BC	9.95	1	9.95	4.42	0.0542
BD	221.12	1	221.12	98.11	0.0020	**
CD	164.10	1	164.10	72.81	< 0.0001	**
A²	85.85	1	85.85	38.09	< 0.0001	**
B²	135.43	1	135.43	60.09	< 0.0001	**
C²	393.05	1	393.05	174.39	< 0.0001	**
D²	192.00	1	192.00	85.19	< 0.0001	**
残差	31.55	14	2.25	—	—
失拟项	24.60	10	2.46	1.41	0.3948
纯误差	6.96	4	1.74	—	—

来源	平方和	自由度	均方	F值	p值	显著性
合计	1553.21	28	—	—	—
模型	1521.66	14	108.69	48.22	< 0.0001	**
A：乙醇的使用量	286.65	1	286.65	127.18	< 0.0001	**
B：聚山梨醇浓度	93.30	1	93.30	41.39	< 0.0001	**
C：双氧水浓度	113.47	1	113.47	50.34	< 0.0001	**
D：去离子水使用量	18.03	1	18.03	8.00	0.0134	*
AB	37.76	1	37.76	16.75	0.0011	**
AC	0.65	1	0.6480	0.28	0.6002
AD	25.65	1	25.65	11.38	0.0045	**
BC	9.95	1	9.95	4.42	0.0542
BD	221.12	1	221.12	98.11	0.0020	**
CD	164.10	1	164.10	72.81	< 0.0001	**
A²	85.85	1	85.85	38.09	< 0.0001	**
B²	135.43	1	135.43	60.09	< 0.0001	**
C²	393.05	1	393.05	174.39	< 0.0001	**
D²	192.00	1	192.00	85.19	< 0.0001	**
残差	31.55	14	2.25	—	—
失拟项	24.60	10	2.46	1.41	0.3948
纯误差	6.96	4	1.74	—	—

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