红豆杉枝叶中总生物碱的提取及其抗肿瘤活性研究

doi:10.3969/j.issn.0253-2417.2019.06.016

Abstract

Abstract:

The optimum extraction conditions of total alkaloids from Taxus chinensis branches and leaves(TCBL)through enzymolysis treatment and ultrasonic-assisted extraction method were investigated by the single factor experiment and response surface experiment. Then, in vitro antitumor activities of crude total alkaloids extract were studied. The results showed that the optimum process conditions of enzyme and ultrasonic-assisted extraction are ethanol volume fraction of 19%, enzyme solution pH value 3.0, enzyme concentration of 0.10 g/L, liquid-solid ratio 12:1(mL:g), enzymolysis time 52 min, enzymolysis temperature 50℃, ultrasonic power 150 W.The yield of total alkaloid of extracting 1 time was 0.126% under these conditions. Response surface model significance test found that the model P < 0.000 1, which indicated that the difference was very significant, regression equation was effective, and the model had statistical significance.The in vitro antitumor activity of crude total alkaloids extracted from TCBL was studied. The results showed that total alkaloids extract from TCBL had good inhibitory effects on three kinds of cancer cells. Especially, the antitumor activity against human cervical cancer (Hela) cells was the strongest, and its IC₅₀ value was 22.18 mg/L that was similar to that of etoposide (21.2 mg/L). It had better antitumor effect on human breast cancer (MCF7) cells (IC₅₀ 74.95 mg/L). It had a certain inhibitory activity on human hepatocellular carcinoma (HepG2) cells, and its IC₅₀ value was 146.03 mg/L.

Key words: Taxus chinensis branches and leaves, total alkaloids, extraction technology, antitumor activity

CLC Number:

TQ35

Ping JIANG,Yajie ZHAO,Xinye BAO,Fei WANG,Zhanquan DUAN,Maolin WANG. Extraction Technology of Total Alkaloids from Taxus chinensis Branches and Leaves and Their Antitumor Activities[J]. Chemistry and Industry of Forest Products, 2019, 39(6): 115-122.

Figures/Tables 5

Table 1

Fig.1

Table 2

Table 3

Fig.2

References 26

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实验号 No.	pH值¹⁾ pH value	酶质量浓度/(g·L^-1) enzyme mass concentration	乙醇体积分数/% ethanol volume fraction	超声波提取 ultrasonic extraction	总生物碱得率/% total alkaloid yield
1	4	0.0	0	无no	0.062
2	≈7	0.0	20	无no	0.082
3	≈7	0.1	20	无no	0.059
4	4	0.1	0	无no	0.093
5	4	0.1	20	无no	0.121
6	4	0.0	0	有yes	0.065
7	≈7	0.0	20	有yes	0.096
8	≈7	0.1	20	有yes	0.067
9	4	0.1	0	有yes	0.104
10	4	0.1	20	有yes	0.130

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	0.013	14	0.0009	10.50	< 0.0001	**
X₁	4.73×10^-4	1	4.73×10^-4	5.49	0.0344	*
X₂	7.30×10^-5	1	7.30×10^-5	0.85	0.3727
X₃	0.0016	1	0.0016	19.06	0.0006	**
X₄	1.08×10^-4	1	1.08×10^-4	1.25	0.2815
X₁X₂	4.29×10^-5	1	4.29×10^-5	0.50	0.4918
X₁X₃	7.02×10^-4	1	7.02×10^-4	8.16	0.0127	*
X₁X₄	1.00×10^-6	1	1.00×10^-6	0.012	0.9157
X₂ X₃	7.48×10^-5	1	7.48×10^-5	0.87	0.3670
X₂ X₄	9.00×10^-6	1	9.00×10^-6	0.10	0.7512
X₃ X₄	2.50×10^-5	1	2.50×10^-5	0.29	0.5984
X₁²	0.0032	1	0.0032	37.57	< 0.0001	**
X₂²	0.0018	1	0.0018	20.62	0.0005	**
X₃²	0.0047	1	0.0047	54.75	< 0.0001	**
X₄²	0.0047	1	0.0047	54.90	<0.0001	**
残差residual	0.0012	14	8.61×10^-5	3.86	0.1025
失拟项lack of fit	0.0011	10	1.09×10^-4
纯误差pure error	1.13×10^-4	4	2.83×10^-5
总和correlation total	0.0140	28

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Extraction Technology of Total Alkaloids from Taxus chinensis Branches and Leaves and Their Antitumor Activities

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试验号 No.	X₁ 提取时间/min extraction time	X₂ 提取温度/ ℃ extraction temperature	X₃ 超声波功率/W ultrasonic power	X₄ 乙醇体积分数/% volume fraction of ethanol	Y 总生物碱得率/% total alkaloid yield
1	50	40	168	20	0.095
2	60	40	144	20	0.091
3	60	50	120	20	0.061
4	50	40	144	10	0.078
5	40	60	144	20	0.079
6	40	50	168	20	0.055
7	50	50	144	20	0.128
8	50	60	144	30	0.073
9	50	50	144	20	0.122
10	50	60	168	20	0.100
11	50	60	144	10	0.077
12	40	50	144	10	0.081
13	60	50	168	20	0.111
14	40	50	144	30	0.063
15	40	40	144	20	0.093
16	50	40	144	30	0.080
17	50	60	120	20	0.056
18	50	50	144	20	0.115
19	50	50	168	10	0.081
20	50	50	144	20	0.127
21	50	40	120	20	0.068
22	50	50	120	30	0.070
23	50	50	168	30	0.076
24	60	50	144	30	0.068
25	50	50	120	10	0.065
26	50	50	144	20	0.126
27	60	50	144	10	0.084
28	40	50	120	20	0.058
29	60	60	144	20	0.089