Box-Behnken Design响应面法优化银杏叶总黄酮柱层析纯化工艺

doi:10.3976/j.issn.1002-4026.2022.06.002

Abstract

Abstract:

The study’s aim was to obtain Ginkgo biloba extract with a high mass fraction of total flavonoids. On the basis of single-factor test, the sample concentration, ethanol volume fraction, eluate volume, and elution flow rate were used as the investigation factors and the mass fraction and yield of total flavonoids extracted from G. biloba were used as the investigation indicators. Box-Behnken design (BBD) and response surface method were used to optimize the column chromatography process for purifying the total flavonoids extracted from G. biloba. Optimal purification was achieved by using a sample concentration of 3.1 mg/mL, ethanol volume fraction of 71%, eluate volume of 2 BV (BV is the column void volume), and elution flow rate of 2 BV/h. Under the optimal conditions, three batches of verification experiments were conducted. The average mass fraction of total flavonoids in the G. biloba extract reached 35.77%, and the yield was 93.30%. The mass fraction of total lactones in the G. biloba extract reached the standards of The Pharmacopoeia of the People's Republic of China. Using BBD and response surface method for optimizing the column chromatography process for the purification of total flavonoids is scientific, reasonable, and feasible.This study can be used as a reference for the industrial production of total flavonoids.

Key words: Box-Behnken Design and response surface method, macroporous resin, column chromatography, total flavonoids from Ginkgo biloba, purification

CLC Number:

R284.2

BI Yan-yan, MA Qing-wen, XU Hui, LIU Jian, LONG Rui, TANG Wei-ming, GUO Tian-tian, HAO Da-wei. Optimization of the column chromatographic purification of total flavonoids extracted from Ginkgo biloba using Box-Behnken design and response surface method[J].Shandong Science, 2022, 35(6): 8-15.

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Table 1

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References 15

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差异来源	自由度	R_综
差异来源	自由度	平方和	均方	F值	P值
模型	14	1 683.99	120.29	134.47	<0.000 1
A	1	65.10	65.10	72.78	<0.000 1
B	1	13.89	13.89	15.53	0.001 5
C	1	1.72	1.72	1.92	0.187 5
D	1	2.92	2.92	3.26	0.092 3
AB	1	4.80	4.80	5.36	0.036 3
AC	1	0.10	0.10	0.11	0.740 1
AD	1	1.27	1.27	1.41	0.254 0
BC	1	7.21	7.21	8.06	0.013 1
BD	1	0.49	0.49	0.55	0.471 5
CD	1	7.70	7.70	8.61	0.010 9
A²	1	465.73	465.73	520.66	<0.000 1
B²	1	1 074.99	1 074.99	1201.78	<0.000 1
C²	1	531.23	531.23	593.89	<0.000 1
D²	1	273.61	273.61	305.88	<0.000 1
残差	14	12.52	0.89
失拟项	10	10.44	1.04	2.01	0.261 9
纯误差	4	2.08	0.52
总差	28	1 696.52

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Optimization of the column chromatographic purification of total flavonoids extracted from Ginkgo biloba using Box-Behnken design and response surface method

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