蒸汽爆破对杜仲叶水提液活性成分、抗氧化活性及香气组分的影响

doi:10.12302/j.issn.1000-2006.202306002

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南京林业大学学报（自然科学版） ›› 2024, Vol. 48 ›› Issue (3): 245-256.doi: 10.12302/j.issn.1000-2006.202306002

蒸汽爆破对杜仲叶水提液活性成分、抗氧化活性及香气组分的影响

周昀菲¹^,²(), 杜庆鑫¹^,³, 王志勇¹^,³, 王璐¹^,³, 王雁¹^,³, 刘攀峰¹^,³, 孙志强¹^,³^,^*()

1.中国林业科学研究院经济林研究所,河南郑州 450003
2.南京林业大学林草学院,江苏南京 210037
3.经济林种质创新与利用国家林业和草原局重点实验室,国家林业和草原局杜仲工程技术研究中心,河南郑州 450003

收稿日期:2023-06-03 修回日期:2023-08-10 出版日期:2024-05-30 发布日期:2024-06-14
通讯作者: *孙志强(zq_sun@paulownia.ac.cn),研究员。
作者简介:周昀菲(1255623163@qq.com)。
基金资助:
中央级公益性科研院所基本科研业务费专项(CAFYBB2022MB003);郑州市基础研究及应用基础研究专项

Steam explosion impact on the active ingredients, antioxidant activity and aroma components of the aqueous extract from Eucommia ulmoides leaves

ZHOU Yunfei¹^,²(), DU Qingxin¹^,³, WANG Zhiyong¹^,³, WANG Lu¹^,³, WANG Yan¹^,³, LIU Panfeng¹^,³, SUN Zhiqiang¹^,³^,^*()

1. Research Institute of Non-timber Forestry, Chinese Academy of Forestry, Zhengzhou 450003, China
2. College of Forestry and Grassland, Nanjing Forestry University, Nanjing 210037, China
3. Key Laboratory of Non-timber Forest Germplasm Enhancement & Utilization of State Forestry and Grassland Administration, Eucommia Engineering Research Center of State Forestry and Grassland Administration, Zhengzhou 450003, China

Received:2023-06-03 Revised:2023-08-10 Online:2024-05-30 Published:2024-06-14

摘要/Abstract

摘要：

【目的】通过分析蒸汽爆破(汽爆)后杜仲成熟叶片活性成分、抗氧化活性和香气组分变化,探究汽爆辅助制备杜仲叶茶的可行性。【方法】对比两个杜仲品种‘华仲8号’和‘华仲12号’成熟叶片的揉捻和不揉捻处理,采用2因素3水平完全试验设计,以叶片总黄酮、多酚和多糖含量确定最佳汽爆参数;采用紫外分光法和高效液相色谱法(HPLC)测定叶片活性成分,测定DPPH(1,1-二苯基-2-三硝基苯肼)和ABTS[2,2-联氮-双-(3-乙基-苯并噻唑-6-磺酸)二铵盐]自由基清除率并分析叶片抗氧化活性与活性成分间的相关性;采用顶空固相微萃取/气相色谱-质谱联用法(HS-SPME/GC-MS)测定三硝基苯香气组分。【结果】依据总黄酮等含量确定最佳汽爆参数为:蒸汽压强0.45 MPa,稳压时间300 s。该条件下‘华仲8号’不揉捻和‘华仲12号’揉捻叶片处理总黄酮和多酚含量较对照分别提高10.87和21.51倍,多酚含量较对照分别提高1.14和0.87倍,环烯醚萜类等6种活性成分含量均极显著提高(P<0.01)。其中‘华仲8号’不揉捻和‘华仲12号’揉捻处理叶片的京尼平苷酸较对照分别提高了0.88和1.23倍,绿原酸较对照分别提高0.45和0.31倍,没食子酸甲酯较对照分别提高10.41和8.06倍。汽爆后杜仲叶抗氧化活性均极显著提高,‘华仲8号’不揉捻强于‘华仲12号’揉捻处理;IC₅₀值与京尼平苷酸、车叶草苷、绿原酸等含量均呈显著负相关(P<0.05)。叶片挥发性化合物中产生了苯甲醇和β-环柠檬醛等芳香物质,具有花香气味的酮类占比最高,相对含量提高了1.93~3.38倍。【结论】蒸汽爆破处理显著提高了杜仲成熟叶水提液的活性成分水溶率和抗氧化活性,其芳香类物质种类增加且含量提高。鉴于目前国内绿叶杜仲资源量远高于红叶杜仲,本研究所提出的杜仲成熟叶片萎凋后直接蒸汽爆破简化了制茶工艺步骤,有效降低了绿叶杜仲茶生产成本,为规模化的杜仲叶加工利用提供了新思路。

关键词: 杜仲, 活性成分, 抗氧化, 香气组分, 蒸汽爆破

Abstract:

【Objective】 To evaluate the feasibility of manufacturing Eucommia ulmoides leaf tea via steam explosion (SE), the changes of active ingredients, antioxidant activity and aroma components were compared between tea produced using E. ulmoides mature leaves with and without an SE. 【Method】 Treatments based on physically rolling or not rolling the leaves were applied using E. ulmoides ‘Huazhong 8’ and ‘Huazhong 12’ mature leaves. A two-factor, three-level complete experimental design was used to determine the optimal SE parameters according to the contents of total flavonoids, polyphenols and polysaccharides, which were determined by UV spectroscopy. The active ingredients were detected by high-performance liquid chromatography (HPLC). The antioxidant activity of the samples was assessed using 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) and 2,2'-diazo-di(3-ethylbenzothiazoline-6-sulfonic acid) diamine salt (ABTS) radical scavenging assays. Additionally, the correlation between the active ingredients of the samples and antioxidant activity was analyzed. The aroma components were determined by headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS). 【Result】 The optimal SE parameters were a pressure of 0.45 MPa and retention time of 300 s. Under these conditions, the total flavonoid and polyphenol contents of ‘Huazhong 8’ and ‘Huazhong 12’ were increased by 10.87 and 21.51, and 1.14 and 0.87-fold compared with the control, respectively. The contents of six active ingredients increased significantly (P < 0.01), with chlorogenic acid and gallic acid increased by 31.31% and 30.72%, respectively and methyl ester increased by 10.41 and 8.06-fold in non-twisted ‘Huazhong 8’ and twisted ‘Huazhong 12’ compared with the control, respectively. Overall, the antioxidant activity of mature E. ulmoides leaves was enhanced significantly with an SE, in which the antioxidant activity of non-twisted ‘Huazhong 8’ was greater than that of twisted ‘Huazhong 12’. The half-maximal inhibitory concentration (IC₅₀) was negatively correlated with the contents of kynurenine, carnoside, and chlorogenic acid (P<0.05). Aromatic substances such as benzyl alcohol and β-cyclocitral were generated from the volatile compounds. 【Conclusion】 An SE treatment significantly improved the water solubility and antioxidant activity of the active ingredients of the aqueous extract of mature E. ulmoides leaves, with an increase in the type and content of aromatic substances. Because the resource availability of green-leaved E. ulmoides is much greater than that of red-leaved E. ulmoides at present, our results suggest that mature E. ulmoides leaves without a physical twisting treatment prior to an SE would result in a simplified tea-making process and effectively reduce the production cost of E. ulmoides green leaf tea, providing novel ideas for the large-scale processing and utilization of E. ulmoides leaves.

Key words: Eucommia ulmoides, active ingredients, antioxidant, aroma components, steam explosion

中图分类号:

TS272
TS201

周昀菲,杜庆鑫,王志勇,等. 蒸汽爆破对杜仲叶水提液活性成分、抗氧化活性及香气组分的影响[J]. 南京林业大学学报（自然科学版）, 2024, 48(3): 245-256.

ZHOU Yunfei, DU Qingxin, WANG Zhiyong, WANG Lu, WANG Yan, LIU Panfeng, SUN Zhiqiang. Steam explosion impact on the active ingredients, antioxidant activity and aroma components of the aqueous extract from Eucommia ulmoides leaves[J].Journal of Nanjing Forestry University (Natural Science Edition), 2024, 48(3): 245-256.DOI: 10.12302/j.issn.1000-2006.202306002.

图/表 8

表1

表2

表3

Total flavonoid, polyphenol and polysaccharide contents in aqueous extracts of E. ulmoides leaves by steam explosion单位:mg/g"

处理 treatment	总黄酮含量 mass fraction of total flavonoid				多酚含量 mass fraction of polyphenol				多糖含量 mass fraction of polysaccharide
处理 treatment	A1	A2	B1	B2	A1	A2	B1	B2	A1	A2	B1	B2
P₁T₁	10.06±0.02 a	6.09±0.03 b	40.62±0.44 d	37.48±1.04 d	133.05±0.45 b	134.70±2.48 c	101.87±0.17 f	139.98±2.41 c	1.36±0.17 h	16.15±0.11 g	24.73±0.08 c	13.50±0.04 e
P₁T₂	1.09±0.01 a	12.63±0.07 b	38.67±0.73 e	32.72±0.51 e	123.06±0.69 c	117.15±0.97 d	98.30±0.50 g	119.53±1.28 e	18.72±0.07 d	17.97±0.08 f	21.23±0.04 g	8.34±0.03 g
P₁T₃	0.41±0.01 b	40.50±0.03 a	48.13±0.83 b	25.14±1.11 f	59.28±1.65 h	82.56±0.87 f	106.00±1.79 e	69.20±1.11 h	25.09±0.07 b	40.41±0.04 d	24.22±0.02 d	2.37±0.02 i
P₂T₁	0.43±0.00 b	1.24±0.01 d	23.67±0.36 g	58.14±0.70 b	125.89±2.22 c	103.24±1.20 e	89.38±1.14 h	58.43±1.64 i	18.88±0.17 d	15.06±0.10 h	17.21±0.02 i	6.23±0.01 h
P₂T₂	0.45±0.01 b	27.98±0.15 a	45.94±0.51 c	60.17±0.91 a	96.20±2.72 d	163.83±2.05 b	184.16±1.96 a	105.05±1.13 f	16.14±0.06 f	22.66±0.07 e	22.01±0.08 f	20.73±0.02 c
P₂T₃	10.28±0.02 a	41.65±0.03 a	60.10±0.23 a	60.64±0.61 a	146.17±2.34 a	219.19±0.94 a	140.77±1.28 c	125.80±1.34 d	22.96±0.07 c	30.35±0.07 d	36.48±0.08 a	21.05±0.02 b
P₃T₁	16.20±0.02 a	1.04±0.02 d	45.99±0.91 c	50.15±1.17 c	69.28±1.28 e	52.07±0.60 i	104.23±0.46 e	201.40±0.64 a	30.21±0.09 a	10.95±0.04 i	23.38±0.01 e	16.33±0.04 d
P₃T₂	0.42±0.01 b	0.09±0.02 d	32.51±0.32 f	19.17±0.51 g	43.95±1.40 g	62.69±0.78 h	120.78±1.04 d	77.00±0.61 g	10.23±0.07 g	33.14±0.04 c	19.14±0.08 h	8.41±0.01 f
P₃T₃	0.53±0.00 a	4.95±0.01 c	41.58±0.86 d	59.38±1.12 ab	62.69±0.17 f	71.59±0.62 g	172.18±1.56 b	151.56±0.35 b	16.77±0.07 e	42.10±0.06 a	33.90±0.62 b	25.56±0.02 a
CK	3.51±0.02		2.67±0.01		102.39±0.69		75.30±1.34		23.59±0.56		24.85±0.08

表3

表4

图1

表5

表6

表7

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处理 treatment	活性物质 active ingredient	蒸汽压强 steam pressure	稳压时间 pressure time	蒸汽压强× 稳压时间 steam pressure× pressure time	处理 treatment	活性物质 active ingredient	蒸汽压强 steam pressure	稳压时间 pressure time	蒸汽压强× 稳压时间 steam pressure× pressure time
A1	总黄酮	64.750^**	1 140.453^**	999.036^**	B1	多酚	3 588.134^**	476.184^**	1 073.366^**
A2	总黄酮	2 222.454^**	466.554^**	677.750^**	B2	多酚	21 089.124^**	1 693.573^**	4 994.324^**
B1	总黄酮	89 425.000^**	117 6011.054^**	1 058 505.108^**	A1	多糖	3 276.301^**	89 262.470^**	23 672.693^**
B2	总黄酮	362 434.719^**	470 119.172^**	100 781.399^**	A2	多糖	301 349.540^**	72 440.380^**	345 765.439^**
A1	多酚	2 172.081^**	2 904.297^**	1 943.560^**	B1	多糖	12 441.648^**	25 441.630^**	86 482.275^**
A2	多酚	3 086.185^**	1 416.836^**	4 949.769^**	B2	多糖	3 145.486^**	33 5745.974^**	53 452.984^**

处理 treatment	京尼平苷酸 kynurenine glucoside	车叶草苷 chelerythrine	原儿茶酸 protocatechuic acid	绿原酸 chlorogenic acid	儿茶素 catechin	松脂醇二葡萄糖苷 pinoresinol diglucoside	没食子酸甲酯 methyl gallate
A1	12.593 4±0.059 9 D	7.088 1±0.000 2 D	0.899 6±0.000 0 F	11.329 8±0.001 0 D	9.552 7±0.003 6 D	8.464 5±0.001 7 D	1.230 3±0.000 2 D
A2	16.363 9±0.039 8 B	8.637 0±0.000 2 B	1.052 2±0.000 0 B	12.773 9±0.000 3 B	12.315 2±0.000 4 B	9.507 4±0.001 9 B	2.144 5±0.000 1 B
B1	18.519 3±0.003 9 A	10.230 1±0.000 5 A	1.034 2±0.000 2 C	13.621 2±0.000 3 A	13.947 1±0.001 7 A	10.712 9±0.001 6 A	2.700 4±0.000 7 A
B2	15.192 9±0.010 3 C	7.602 3±0.000 8 C	1.009 4±0.000 1 D	12.322 3±0.000 3 C	11.45 66±0.000 8 C	8.870 1±0.000 5 C	1.870 9±0.000 4 C
CK1	8.705 2±0.015 0 E	5.817 8±0.000 2 E	1.150 0±0.000 0 A	9.727 8±0.000 3 E	6.638 3±0.002 5 E	7.595 6±0.000 8 E	0.236 7±0.000 0 E
CK2	8.312 3±0.035 5 F	5.626 2±0.000 6 F	0.954 6±0.000 4 E	9.426 6±0.000 1 F	6.226 2±0.001 2 F	7.062 3±0.001 2 F	0.192 2±0.000 2 F

成分 constituent	DPPH自由基清除能力 DPPH free radicals scavenging	ABTS自由基清除能力 ABTS free radicals scavenging
总黄酮total flavonoids	-0.899^*	-0.865^*
多酚polyphenols	-0.837^*	-0.820^*
多糖polysaccharides	0.075	0.037
京尼平苷酸kyonipin glycosides	-0.913^*	-0.877^*
车叶草苷chelerythrin	-0.904^*	-0.869^*
原儿茶酸protocatechuic acid	-0.874^*	-0.845^*
绿原酸chlorogenic acid	-0.895^*	-0.860^*
儿茶素catechins	-0.899^*	-0.865^*
松脂醇二葡萄糖苷 pinoresinol diglucoside	-0.837^*	-0.820^*
没食子酸甲酯 gallic acid methyl ester	0.075	0.037

类别 type	A2		B1		CK1
类别 type	相对含量/% relative content	种类数 species number	相对含量/% relative content	种类数 species number	相对含量/% relative content	种类数 species number
碳氢类hydrocarbons	11.39	10	13.26	16	11.80	15
杂环类heterocycles	8.83	6	9.88	7	19.24	8
醇类alcohols	15.22	9	4.34	4	9.27	4
醛类aldehydes	10.52	7	12.90	8	3.96	4
酮类ketones	29.05	20	23.32	17	6.63	10
酯类esters	16.33	5	18.02	4	41.71	8
酚类phenols	2.52	3	3.03	5	—	—
其他类others	6.14	8	8.73	8	2.28	3
总计total	100.00	68	93.48	69	94.89	52

化合物编号 compound No.	CAS	化学式 chemical formula	名称 name	香气描述 aroma description	阈值^[33] /(mg·kg^-1) threshold value	A2		B1		CK1
化合物编号 compound No.	CAS	化学式 chemical formula	名称 name	香气描述 aroma description	阈值^[33] /(mg·kg^-1) threshold value	相对含量/% relative content	气味活度值 OAV	相对含量/% relative content	气味活度值 OAV	相对含量/% relative content	气味活度值 OAV
1	100-51-6	C₇H₈O	苯甲醇benzyl alcohol	花香、香水	5.500	8.70	1.58	—	—	—	—
2	124-19-6	C₉H₁₈O	壬醛nonanal	柑橘香、花香、油脂味、栗子香、焦糖香	1.100	1.40	1.27	1.35	1.23	0.59	0.54
3	112-31-2	C₁₀H₂₀O	癸醛decanal	橙皮味	0.100	—	—	0.69	6.90	0.35	3.50
4	4313-3-5	C₇H₁₀O	(E,E)-2,4- 庚二烯醛 (E,E)-2,4- heptadienal	青草味	0.051	5.29	103.73	3.78	74.12	—	—
5	432-25-7	C₁₀H₁₆O	β-环柠檬醛β-cyclocitral	烘烤香	3.000	0.89	0.30	0.99	0.33	—	—
6	121-33-5	C₈H₈O₃	香兰素vanillin	类似香荚兰豆香气	0.500	0.81	1.62	0.45	0.90	—	—
7	23726-93-4	C₁₃H₁₈O	大马士酮damascenone	强烈的玫瑰香	0.002	0.72	360.00	1.14	570.00	0.66	330.00
8	3796-70-1	C₁₃H₂₂O	香叶基丙酮geranylacetone	木兰香气	0.010	2.47	247.00	2.60	260.00	0.74	74.00
9	79-77-6	C₁₃H₂₀O	β-紫罗兰酮β-violetone	花香、紫罗兰香	0.007	3.85	550.00	—	—	1.35	192.86
10	79-77-6	C₁₃H₂₀O	α-紫罗兰酮α-violetone	木香、花香、蜂蜜气味	0.004	3.24	810.00	0.64	160.00	—	—
11	17092-92-1	C₁₁H₁₆O₂	二氢猕猴桃内酯dihydrokiwi lactone	甜桃香	0.500	9.96	19.92	6.29	12.58	1.33	2.66

蒸汽爆破对杜仲叶水提液活性成分、抗氧化活性及香气组分的影响

Steam explosion impact on the active ingredients, antioxidant activity and aroma components of the aqueous extract from Eucommia ulmoides leaves

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