东北红豆杉枝叶中原花色素的提取分离及其糖尿病抑制活性

doi:10.3969/j.issn.0253-2417.2022.06.015

Abstract

Abstract:

The proanthocyanidins were extracted by enzymolysis and ultrasonic-assisted coupling extraction method using Taxus cuspidata branches and leaves(TCBL) as the materials. The optimum extraction process was investigated by the single factor experiment and response surface experiment and a proanthocyanidin compound was obtained by extraction and separation. Then, in vitro antidiabetic activities of the fractions isolated from different solvent extract and the ethyl acetate extract by macroporous adsorption resin were studied. The results showed that the optimum process conditions of enzyme and ultrasonic assisted coupling extraction of proanthocyanidins were 60% ethanol aqueous as solvent, enzyme concentration of 0.15 g/L, liquid-solid ratio of 14∶1(mL∶g), enzymolysis time of 50 min, enzymolysis temperature of 40 ℃, and ultrasonic power of 120 W. The extraction yield of proanthocyanidins for one time was 3.84%(extraction rate was 83.48%) under these conditions. Response surface model significance test found that the P value of model was 0.000 2, indicating that the difference was very obvious, the regression equation was effective, and the model had statistical significance. The ethyl acetate extract was separated and purified by AB-8 macroporous adsorption resin, dextran gel, and silica gel column chromatography to obtain a proanthocyanidin compound, which was identified as a dimeric proanthocyanidins of catechin and robinatinol by TOF-MS structure identification and ultraviolet full-wavelength scanning. The anti-diabetes activities of 7 components(CBFr.1-CBFr.7) isolated from different solvent extracts and ethyl acetate extracts by macroporous adsorption resin were studied. The results showed that ethyl acetate extract had a good inhibitory effect on α-amylase activity and α-glucosidase activity, and the inhibition rates were 83.6% and 63.03%, respectively. The median inhibitory concentration(IC₅₀) values of CBFr.1-CBFr.5 for α-amylase were ranging from 0.036 to 0.043 g/L, and they all had a good inhibitory ability to α-amylase. CBFr.3 showed the best inhibitory activity against α-glucosidase, with an IC₅₀ value of 0.04 g/L, which was lower than that of acarbose(0.063 g/L). Analysis showed that CBFr.1-CBFr.4 had better α-glycosidase enzyme inhibition activity and moderate α-amylase inhibitory activity, which could achieve the effect to lower blood sugar and reduce side effects owing to excessive reduction of α-amylase activity. Therefore, CBFr.1-CBFr.4 could be used as type Ⅱ diabetes inhibitors.

Key words: Taxus cuspidata sieb. et Zucc., proanthocyanidins, extraction technology, antidiabetic activity

CLC Number:

TQ35

Ping JIANG, Jingliang QU, Yuxin JIA, Zhichao SUN, Siying LIU, Fei WANG. Extraction and Separation of Total Proanthocyanidins from Taxus cuspidata sieb.et Zucc. Branches and Leaves and Their Antidiabetic Activity[J]. Chemistry and Industry of Forest Products, 2022, 42(6): 108-116.

Figures/Tables 6

Fig.1

Table 1

Table 2

Fig.2

Table 3

Fig.3

References 17

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方差来源 variance source	平方和 sum of squares	自由度 degree of freedom	均方 mean square	F值 F value	P值 P value	显著性 significant
模型model	1.02	9	1.02	19.90	0.000 2	**
X₁	0.33	1	0.33	58.35	0.000 1	**
X₂	0.065	1	0.065	11.38	0.011 9	*
X₃	0.42	1	0.42	73.54	<0.000 1	**
X₁X₂	0.026	1	0.026	4.50	0.071 6
X₁X₃	0.038	1	0.038	6.68	0.036 2	*
X₂X₃	0.000	1	0.000	0.000	1.000 0
X₁²	0.017	1	0.017	3.05	0.124 1
X₂²	0.053	1	0.053	9.24	0.018 9	*
X₃²	0.078	1	0.078	13.63	0.007 1	**
残差residual	0.040	7	5.692×10^-3
失拟项lack of fit	0.017	3	5.708×10^-3	1.00	0.477 2
纯误差pure error	0.023	4	2.83×10^-5
总和correlation total	1.06	16

萃取物 extract	抑制率inhibition rate/%
萃取物 extract	α-淀粉酶 α-amylase	α-葡萄糖苷酶 α-glucosidase
石油醚petroleum ether	16.45	17.69
二氯甲烷dichloromethane	16.83	16.53
乙酸乙酯ethyl acetate	83.60	63.03
正丁醇n-butyl alcohol	63.03	53.97
水water	13.40	16.32
阿卡波糖acarbose	90.90	85.73

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Extraction and Separation of Total Proanthocyanidins from Taxus cuspidata sieb.et Zucc. Branches and Leaves and Their Antidiabetic Activity

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试验号 No.	X₁ 酶解温度/℃ enzymolysis temperature	X₂ 酶解时间/min enzymolysis time	X₃ 酶质量浓度/(g·L^-1) enzyme concentration	Y 原花色素得率/% proanthocyanidins yield
1	50	50	0.10	3.15
2	40	60	0.10	3.15
3	50	50	0.10	3.31
4	40	50	0.05	3.13
5	50	40	0.05	3.05
6	50	50	0.10	3.19
7	40	40	0.10	3.25
8	60	50	0.05	2.89
9	40	50	0.15	3.82
10	50	60	0.05	2.95
11	50	60	0.15	3.37
12	50	50	0.10	3.17
13	50	50	0.10	3.11
14	60	50	0.15	3.19
15	60	60	0.10	2.61
16	60	40	0.10	3.03
17	50	40	0.15	3.47