GC-MS Analysis of Agarwood from <i>Aquilaria microcarpa</i> in Malaysia

WANG Jun; MEI Wenli; WANG Hao; WANG Yali; DUAN Ruijun; HUANG Shengzhuo; DAI Haofu

doi:10.3969/j.issn.1000-2561.2021.08.034

Chinese Journal of Tropical Crops >

2021 , Vol. 42 >Issue 8: 2377 - 2386

DOI: https://doi.org/10.3969/j.issn.1000-2561.2021.08.034

Post-harvest Treatment & Quality Safety

GC-MS Analysis of Agarwood from Aquilaria microcarpa in Malaysia

WANG Jun ,
MEI Wenli ,
WANG Hao ,
WANG Yali ,
DUAN Ruijun ,
HUANG Shengzhuo ,
DAI Haofu ^,^*

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Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences / Hainan Engineering Research Center of Agarwood / Hainan Institute for Tropical Agricultural Resources, Haikou, Hainan 571101, China

*DAI Haofu,E-mail： daihaofu@itbb.org.cn。

Received date: 2020-11-09

Request revised date: 2020-12-17

Online published: 2021-09-09

Copyright

Copyright reserved © 2021. Office of Acta Agronomica Sinica All articles published represent the opinions of the authors, and do not reflect the official policy of the Chinese Medical Association or the Editorial Board, unless this is clearly specified.

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Abstract

The volatile components of agarwood from Aquilaria microcarpa in Malaysia were extracted by ether ultrasonic extraction and its chemical constituents were analyzed by gas chromatography-mass spectrometry (GC-MS) and were compared with those from Malaysia such as A. sinensis, A. crassna and A. subintegra. The results indicated that 5,8-dihydroxy-2-(2-phenylethyl)chromone, ent-4(15)-eudesmen-11-ol-1-one, (1β,4αβ,7β,8αβ)-octahydro-7-[1-(hydroxymethyl) ethenyl]-1,8α-dimethylnaphthalen-4α(2H)-ol and 6-methoxy-2-(2-phenylethyl) chromone (the relative percentages were all above 10%) were the main volatile components of A. microcarpa samples. There was a big difference between the agarwood samples from cultivated and wild tree, the relative percentage contents of sesquiterpenoids was significantly higher than that of chromones in the former, while the relative percentage contents of chromones was higher than that of sesquiterpenoids or equivalent in the latter. There were more kinds of sesquiterpenoids and chromones in the former, among which the kinds and the relative percentage contents of sesquiterpenoids were more than chromones. However, the latter were less, among which the kinds of sesquiterpenoids were more than chromones, but the relative percentage was relatively low, while the relative percentage of chromones was higher. In addition, a sesquiterpenoid of 11,13-dihydroxy- 9(10)-ene-8α,12-epoxyemophilane was only detected in the agarwood samples from cultivated samples of A. microcarpa. Moreover, the volatile components of Agarwood samples from A. microcarpa, A. sinensis, A. crassna and A. subintegra were compared and analyzed. The results showed that the kinds and relative percentage contents of compounds from A. microcarpa were significantly different from those of the latter three. There were only two common components in all samples, namely 5,8-dihydroxy-2-(2-phenylethyl) chromone and 6-methoxy- 2-(2-phenylethyl)chromone.

Key words： Aquilaria microcarpa; cultivated agarwood samples; wild agarwood samples; GC-MS analysis

Cite this article

WANG Jun , MEI Wenli , WANG Hao , WANG Yali , DUAN Ruijun , HUANG Shengzhuo , DAI Haofu . GC-MS Analysis of Agarwood from Aquilaria microcarpa in Malaysia[J]. Chinese Journal of Tropical Crops, 2021 , 42(8) : 2377 -2386 . DOI: 10.3969/j.issn.1000-2561.2021.08.034

马来西亚野生分布有5种瑞香科（Thymelaeaceae）沉香属（Aquilaria Lam.）植物,分别为小果沉香（A. microcarpa Baill.）、贝卡利沉香（A. beccariana Tiegh.）、马来沉香（A. malaccensis Lam.）、毛沉香（A. hirta Ridl.）和丝沉香[A. filaria (Oken) Merr.],其中小果沉香和贝卡利沉香主要分布于东马来西亚的砂拉越（Sarawak）地区,而后3种则主要分布于西马来西亚^[1]。据调查与实地考察,近年来,还有从其他国家引种至马来西亚种植栽培的种类主要有白木香[A. sinensis (Lour.) Spreng.]、柯拉斯那沉香（A. crassna Pierre ex Lecomte）和近全缘沉香（A. subintegra Ding Hou）等。

小果沉香被认为是砂拉越地区重要的硬木之一^[2],其为大乔木,高可达40 m,生于海拔低于200 m的低地雨林中,在马来西亚砂拉越州广布^[3]。该种与马来沉香形态特征相近,但其果实较小,雄蕊等于或短于花瓣,可与之区别^[4]。到目前为止,有关其研究主要包括解剖学研究^[2]、栽培生理^[5,6,7]、营养繁殖^[8]、组织培养^[9]、生殖生态学^[10]、生物合成^[11]、分子生物学与代谢组学^[12,13,14]、与微生物互作^{[15,16,17,18,19]}、微生物结香^[20,21]、声波层析成像技术探测沉香^[22]等,而对其相关的化学成分和药理活性的研究报道较少,如接菌结香小果沉香叶片甲醇提取物的抗氧化活性成分^[23];小果沉香茎皮中的酚类化合物（phenolic）^[24];小果沉香茎皮中的黄酮类化合物（flavonoid）^[25];小果沉香结香样品乙醇提取物对金黄色葡萄球菌（Staphylococcus aureus）和奇异变形杆菌（Proteus mirabilis）的抗菌活性实验^[26];酚类物质在小果沉香中作用^[27];小果沉香的接菌结香样品提取物（Kemedangan）抗氧化活性研究^[28]。而有关小果沉香结香样品中化学成分方面未有报道,因此,本文在前期调查取样的基础上,对小果沉香结香样品的挥发性成分进行了GC- MS分析,并与马来西亚产白木香、柯拉斯那沉香和近全缘沉香的结香样品挥发性成分进行了对比分析,旨在为小果沉香的进一步深入研究提供依据。

1 材料与方法

1.1 材料

1.1.1 材料与试剂 9份样品中有6份采自东马来西亚砂拉越州,其中2份购自砂拉越与印度尼西亚边境小镇Serikin市场,2份样品采自西马来西亚柔佛州（Johor）,1份采自森美兰州（Sembilan）,样品信息见表1。S1~S6样品由砂拉越森林局（Sarawak Forest Bureau）Dawend jiwaN（大文吉湾）先生鉴定其基源植物为小果沉香（Aquilaria microcarpa Baill.）,S7~S9由王军博士和马来西亚钟楠贵先生共同依次鉴定其基源植物为白木香[A. sinensis (Lour.) Spreng.]、柯拉斯那沉香（A. crassna Pierre ex Lecomte）和近全缘沉香（A. subintegra Ding Hou）。所有输液法结香技术均为中国热带农业科学院热带生物技术研究所研发的整树结香技术。凭证标本均存放于中国热带农业科学院热带生物技术研究所。

表1 9份沉香样品信息

Tab. 1 Nine agarwood samples information

序号 No.	种类 Species	样品来源 Sample source	采集时间 Collection time	地理坐标 Geographical coordiantes	海拔 Altitude/m	凭证标本 Voucher	备注 Notes
S1	Aquilaria microcarpa	Kampung suba, Bau, Sarawak	2018.7.17	1°50°32°° N 110°33°51°° E	23	201801	野生,砍伤法,结香5年
S2	Aquilaria microcarpa	Sarawak Forest Bureau	2018.7.20	1°50°32°° N 110°33°60°° E	15	201810	栽培,输液法,结香1年
S3	Aquilaria microcarpa	Taman Moyan Jaya, Sarawak	2019.7.20	1°53°01°° N 110°24°61°° E	18	201928	栽培,砍伤法,结香1年
S4	Aquilaria microcarpa	Jalan Ban-Lundu, Sarawak	2019.7.21	1°46°19°° N 110°12°96°° E	35	201931	栽培,输液法,结香2年
S5	Aquilaria microcarpa	Serikin Border Market, Sarawak	2018.7.20	1°19°36°° N 110°00°04°° E	45	201816	野生,自然结香,未知
S6	Aquilaria microcarpa	Serikin Border Market, Sarawak	2018.7.20	1°19°36°° N 110°00°04°° E	36	201817	野生,自然结香,未知
S7	Aquilaria sinensis	Sekoloh Menengah Sevi Memnti, Muar, Johor, Malaysia	2019.7.14	1°92°02°° N 102°67°68°° E	19	201907	栽培,输液法,结香2年
S8	Aquilaria crassna	Bank Rakyat Yong Peng, Muar, Johor, Malaysia	2019.7.15	2°00°73°° N 103°04°81°° E	8	201901	栽培,输液法,结香1年
S9	Aquilaria subintegra	Kampung Terentang, Jasin, Malacca,Sembilan,Malaysia	2019.7.17	1°87°48°° N 102°80°09°° E	18	201918	栽培,输液法,结香2年

使用的试剂均为分析纯。

1.1.2 仪器与设备 Agilent 7820A-Agilent 5977E气质联用仪,美国安捷伦公司;色谱柱为HP-5MS 5% Phenyl Methyl Siloxane（30 mm×0.25 mm× 0.25 μm）弹性石英毛细管柱;EN2062电子秤,上海民侨精密科学仪器有限公司;B5510E-MT超声波清洗仪,美国Bransonic公司。

1.2 方法

1.2.1 样品制备选取6份小果沉香与各1份的白木香、柯拉斯那沉香和近全缘沉香（共9份）体积相同的结香树干,分别去除白木,将沉香和其余部位分开。将沉香块分别粉粹后,再均匀混合,各取20 g,分别加入60 mL乙醚溶剂进行超声提取3次,每次超声30 min,静置5 min后过滤得到乙醚提取液,合并3次提取液,挥干后得到油状提取物。分别取油状提取物适量,配制成1 mg/mL的待测样品溶液。

1.2.2 GC-MS分析条件（1）色谱条件。色谱柱为HP-5MS 5% Phenyl Methyl Siloxane（30 m× 0.25 mm×0.25 μm）弹性石英毛细管柱。升温程序：柱温50 ℃,以5 ℃/min升温至310 ℃,保持10 min。汽化室温度250 ℃;载气为高纯He（99.999%）;载气流量1.0 mL/min;不分流进样;溶剂延迟时间4.0 min。

（2）质谱条件。电子轰击（EI）离子源;电子能量70 eV;离子源温度230 ℃;四极杆温度150 ℃;接口温度280 ℃;发射电流34.6 μA;倍增器电压1612 V;质量扫描范围40~800 m/z。

2 结果与分析

9份沉香样品乙醚提取物的总离子流图见图1,对总离子流图中的各峰经质谱数据系统检索及核对Nist2005和Wiley275标准质谱图,并借鉴前人的鉴定方法^{[29,30,31,32]},结合质谱图与文献数据进行对照,并用峰面积归一化法测定色谱峰在样品中的总相对含量分别为小果沉香S1~S6：67.81%、82.83%、96.44%、96.66%、100.00%、68.49%和白木香（98.87%）、柯拉斯那沉香（100.00%）和近全缘沉香（94.16%）,具体见表2。所检测和鉴定的倍半萜和色酮个数和相对百分含量分别为S1：倍半萜10个（34.39%）、色酮3个（30.93%）,其中含量最大的成分为5,8-二羟基-2-(2-苯乙基)色酮（28.35%）,其次为奇楠愈创木酮（8.23%）;S2：倍半萜19个（63.38%）、色酮4个（18.42%）,其中含量最大的成分为对映-4(15)-桉叶烷-11-醇- 1-酮（20.04%）,其次为5,8-二羟基-2-(2-苯乙基)色酮（13.69%）;S3：倍半萜19个（76.50%）、色酮3个（17.83%）,其中含量最大的成分为对映- 4(15)-桉叶烷-11-醇-1-酮（20.96%）,其次为5,8-二羟基-2-(2-苯乙基)色酮（16.87%）;S4：倍半萜17个（61.67%）、色酮6个（34.99%）,其中含量最大的成分为5,8-二羟基-2-(2-苯乙基)色酮（14.96%）,其次为(1β,4aβ,7β,8aβ)-八氢-7-[1-(羟甲基)乙烯基]-1,8a-二甲基萘-4a(2H)-醇（14.45%）;S5：倍半萜11个（30.51%）、色酮4个（69.49%）,其中含量最大的成分为5,8-二羟基-2-(2-苯乙基)色酮（40.09%）,其次为6-甲氧基-2-(2-苯乙基)色酮（12.40%）;S6：倍半萜6个（21.55%）、色酮4个（39.78%）,其中含量最大的成分为5,8-二羟基- 2-(2-苯乙基)色酮（29.70%）,其次为6,7-二甲氧基-2-(2-苯乙基)色酮（8.43%）;S7：倍半萜10个（34.39%）、色酮7个（60.12%）,其中含量最大的成分为6,7-二甲氧基-2-(2-苯乙基)色酮（39.10%）,其次为瓦伦-1(10),8-二烯-11-醇（9.67%）;S8：倍半萜11个（31.57%）、色酮6个（65.38%）,其中含量最大的成分为6-甲氧基- 2-(2-苯乙基)色酮（22.96%）,其次为2-(2-苯乙基)色酮（21.40%）;S9：倍半萜12个（53.87%）、色酮5个（38.22%）,其中含量最大的成分为6-甲氧基-2-(2-苯乙基)色酮（16.21%）,其次为(+)-9-羟基-芹子-4,11-二烯-14-醛（14.61%）。

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图1 9份沉香样品的GC-MS总离子流图

Fig. 1 GC-MS total iron chart of nine agarwood samples

表2 9份沉香样品乙醚提取物中的特征性化学成分

Tab. 2 Characteristic chemical constituents of the ether extracts of the nine samples of agarwood

序号 No.	化合物 Compounds		分子式 Molecular formula			分子量 M_W			相对百分含量RC/%
序号 No.	化合物 Compounds		分子式 Molecular formula			分子量 M_W			S1			S2			S3			S4			S5			S6			S7			S8			S9
1	benzylacetone 苄基丙酮		C₁₀H₁₂O			148			-			-			-			-			-			-			1.03			3.05			-
2	hydrocoumarin 二氢香豆素		C₉H₈O₂			148			-			-			0.85			-			-			-			-			-			-
3	2,4-di-tert-butylphenol 2,4-二叔丁基苯酚		C₁₄H₂₂O			206			1.36			1.03			-			-			-			3.84			-			-			-
4	β-agarofuran β-沉香呋喃^△		C₁₅H₂₄O			220			-			0.94			0.42			-			-			-			-			-			-
5	(-)-10-epi-γ-eudesmol (-)-10-表-γ-桉叶油醇^△		C₁₅H₂₆O			222			-			6.96			4.32			-			-			-			-			-			-
6	γ-eudesmol γ-桉叶醇^△		C₁₅H₂₆O			222			-			-			0.42			-			-			-			-			-			-
7	(-) jinkoh-eremol 沉香雅槛蓝醇^△		C₁₅H₂₂O₂			222			-			2.51			3.14			-			-			-			1.28			-			-
8	kusunol (valerianol) 枯树醇^△		C₁₅H₂₆O			222			-			2.12			3.01			1.42			-			-			3.95			0.80			-
9	(E)-2-((8R,8aS)-8,8a-dimethyl-3,4,6,7,8,8a-hexahydronaphthalen-2(1H)-ylidene)propanal (E)-2-((8R,8aS)-8,8a-二甲基-3,4,6,7,8,8a-六氢萘-2(1H)-亚基)丙醛^△		C₁₅H₂₂O			218			-			-			-			-			0.80			-			-			-			-
10	(-)-α-costol (-)-α-木香醇^△		C₁₅H₂₄O			220			-			-			-			5.29			1.16			-			-			-			1.68
11	6,7-dimethyl-1,2,3,5,8,8a-hexahydronaphthalene 6,7-二甲基-1,2,3,5,8,8a-六氢萘		C₁₂H₁₈			162			-			-			1.26			-			-			-			-			-			-
12	(S)-4a-methyl-2-(1-methylethyl)-3,4,4a,5,6,7-hexahydronaphthalene (S)-4a-甲基-2-(1-甲基乙基)-3,4,4a,5,6,7-六氢萘		C₁₄H₂₂			190			-			-			-			-			-			-			2.41			-			-
13	α-humulene α-蛇麻烯^△		C₁₅H₂₄			204			-			-			-			-			-			-			-			2.18			-
14	7α-H-9(10)-ene-11,12-epoxy-8-oxoeremophilane 7α-H-9(10)-烯-11,12-环氧-8-氧艾里莫芬烷^△		C₁₅H₂₂O₂			234			-			0.88			2.49			-			3.20			-			2.60			-			-
15	isobaimuxinol 异白木香醇^△		C₁₅H₂₆O₂			238			-			2.21			2.08			-			1.63						-			-			-
16	(-)-guaia-1(10),11-dien-15-ol (-)-愈创木-1(10),11-二烯-15-醇^△		C₁₅H₂₄O			220			-			-			3.47			-			-			-			-			-			-
17	β-eudesmol β-桉叶醇^△		C₁₅H₂₆O			222			-			1.07			1.16			-			-			-			-			-			-
18	baimuxinol 白木香醇^△		C₁₅H₂₆O₂			238			-			-			-			-			1.71			-			-			-			-
19	dihydrokaranone 二氢卡拉酮^△		C₁₅H₂₂O			218			-			-			-			-			-			-			1.72			0.73			-
20	(-)-guaia-1(10),11-dien-15-al (-)-愈创木-1(10),11-二烯-15-醛^△		C₁₅H₂₂O			218			-			1.44			2.09			7.96			-			1.64			1.87			-			4.05
21	rel-(2R,3S)-2,3-dimethyl-2-(3-methyl-2-butenyl)-1-cyclohexanone rel-(2R,3S)-2,3-二甲基-2-(3-甲基-2-丁烯基)-1-环己酮^△		C₁₃H₂₂O			194			-			2.74			-			2.42			1.67			-			1.36			-			-
22	(-)-7β-H-eudesmane-4α,11-diol (-)-7β-H-艾里莫芬-4α,11-二醇^△		C₁₅H₂₈O₂			240			-			-			-			-			1.08			-			-			-			-
23	baimuxinal 白木香醛^△		C₁₅H₂₄O₂			236			-			2.69			13.28			1.33			-			-			8.11			4.10			0.98
24	aristolene epoxide 环氧马兜铃烯^△		C₁₅H₂₄O			220			-			1.35			-			-			-			-			-			-			-
25	valenca-1(10),8-dien-11-ol 瓦伦-1(10),8-二烯-11-醇^△		C₁₅H₂₄O			220			-			3.52			3.42			1.15			-			-			9.67			11.23			2.91
26	2-(4a,8-dimethyl-1,2,3,4,4a,5,6,7-octahydro-naphthalen-2-yl)-prop-2-en-1-ol 2-(4a,8-二甲基-1,2,3,4,4a,5,6,7-八氢萘-2-基)- 丙-2-烯-1-醇^△		C₁₅H₂₄O			220			-			-			2.43			-			-			-			-			-			-
27	4-hydroxy-β-cyclocitral 4-羟基-β-环柠檬醛		C₁₀H₁₆O₂			168			-			-			-			-			-			2.22			-			-			-
28	ent-4(15)-eudesmen-11-ol-1-one 对映-4(15)-桉叶烯-11-醇-1-酮^△		C₁₅H₂₆O₂			238			-			20.04			20.96			-			-			-			-			-			-
29	5-epi-7α,15-dihydroxyacorenol 5-表-7α,15-二羟基菖蒲螺烯醇^△		C₁₅H₂₆O₂			238			-			-			-			0.88			-			-			-			-			-
30	valerenol 缬烯醇^△		C₁₅H₂₄O			220			-			-			0.47			2.60			-			-			-			-			2.73
序号 No.	化合物 Compounds			分子式 Molecular formula			分子量 M_W			相对百分含量RC/%
序号 No.	化合物 Compounds			分子式 Molecular formula			分子量 M_W			S1			S2			S3			S4			S5			S6			S7		S8		S9
31	15-hydroxyacorenone 15-羟基菖蒲螺烯酮^△			C₁₅H₂₄O₂			236			-			1.85			-			-			-			-			-		-		-
32	(1β,4aβ,7β,8aβ)-octahydro-7-[1-(hydroxymethyl)ethenyl]-1,8a-dimethylnaphthalen-4a(2H)-ol (1β,4aβ,7β,8aβ)-八氢-7-[1-(羟甲基)乙烯基]-1,8a-二甲基萘-4a(2H)-醇^△			C₁₅H₂₆O₂			238			-			6.31			5.70			14.45			6.07			2.17			1.87		0.97		-
33	cis-Z-α-bisabolene epoxide 顺式-Z-α-环氧化红没药烯^△			C₁₅H₂₄O			220			-			-			-			-			-			3.49			-		0.78		-
34	4β-methoxy-12-hydroxy-5(6)-ene-zizaane 4β-甲氧基-12-羟基-5(6)-烯-深冬烷^△			C₁₆H₂₆O₂			250			-			-			-			0.78			-			-			-		-		-
35	(3R,3aR,4aS,5R,9aS)-3,5,8-trimethyl-3a,4,4a,5,6,7,9,9a-octahydroazuleno[6,5-b]furan-2(3H)-one (3R,3aR,4aS,5R,9aS)-3,5,8-三甲基-3a,4,4a,5,6,7, 9,9a-八氢薁[6,5-b]呋喃-2(3H)-酮^△			C₁₅H₂₂O₂			234			3.09			1.12			-			-			-			-			-		-		-
36	(-)-2α-hydroxyguaia-1(10),11-dien-15-oic acid (-)-2α-羟基愈创木-1(10),11-二烯-15-酸^△			C₁₅H₂₀O₂			232			-			-			-			2.09			-			-			-		2.63		8.18
37	eremophila-9,11(13)-dien-12-ol艾里莫芬- 9,11(13)-二烯-12-醇^△			C₁₅H₂₄O			220			-			-			-			-			-			1.44			-		-		-
38	(4aR,5S)-1-hydroxy-4a,5-dimethyl-3-(propan-2-ylidene)-4,4a,5,6-tetrahydronaphthalen-2(3H)-one (4aR,5S)-1-羟基-4a,5-二甲基-3-(丙-2-亚基)-4,4a,5,6-四氢萘-2(3H)-酮^△			C₁₅H₂₀O₂			232			-			-			-			-			-			-			-		-		9.80
39	11-hydroxy-valenc-1 (l0)-en-2-one 11-羟基-瓦伦-1(10)-烯-2-酮^△			C₁₅H₂₄O₂			236			-			2.93			4.10			-			-			-			1.96		1.03		-
40	(-)-9-hydroxy-selina-3,11-dien-14-al (-)-9-羟基-芹子-3,11-二烯-14-醛^△			C₁₅H₂₂O₂			234			-			-			-			1.03			-			-			-		-		1.36
41	α-santalol α-檀香醇^△			C₁₅H₂₄O			220			-			-			-			-			-			5.19			-		-		1.96
42	2-(4a,8-dimethyl-2,3,4,4a,5,6-hexahydro- naphthalen-2-yl)-prop-2-en-1-ol 3-(4a,8-二甲基-2,3,4,4a,5,6-六氢-萘-2-基)-丙- 2-烯-1-醇^△			C₁₅H₂₂O			218			-			-			-			-			-			-			-		-		1.72
43	(-)-guaia-1(10),11-diene-15,2-olide (-)-愈创木-1(10),11-二烯-15,2-内酯^△			C₁₅H₂₀O₂			234			-			-			-			1.87			-			-			-		-		-
44	1,5-dimethyl-3-hydroxy-8-(1-methylene-2-hydroxyethyl-1)-bicyclo[4.4.0]dec-5-ene 1,5-二甲基-3-羟基-8-(1-亚甲基-2-羟乙基-1)-二环[4.4.0]癸-5-烯^△			C₁₅H₂₄O₂			236			-			-			-			-			-			-			-		-		3.89
45	11,13-dihydroxy-9(10)-ene-8α,12- epoxyemophilane 11,13-二羟基-9(10)-烯-8α,12-环氧艾里莫芬烷^△			C₁₅H₂₄O₃			252			-			1.23			1.23			0.79			-			-			-		-		-
46	cholic acid胆酸			C₂₄H₄₀O₅			408			-			-			-			-			-			3.51			-		-		-
47	α-santonin α-山道年^△			C₁₅H₁₈O₃			246			1.20			-			-			-			-			-			-		-		-
48	eremophil-9-ene-11,12,13-triol 艾里莫芬-9- 烯-11,12,13-三醇^△			C₁₅H₂₆O₃			254			-			-			2.31			8.98			6.02			3.78			-		-		-
49	xanthinin 苍耳素^△			C₁₇H₂₂O₅			306			1.80			-			-			-			-			-			-		-		-
50	12,15-dioxo-α-selinen 12,15-二氧-α-芹子烯^△			C₁₅H₂₀O₂			232			7.84			1.47			-			2.54			3.61			-			-		1.14		-
51	qinan-guaiane-one 奇楠愈创木酮^△			C₁₅H₂₀O₃			248			8.23			-			-			-			-			-			-		-		-
52	(+)-9-hydroxy-selina-4,11-dien-14-al (+)-9-羟基-芹子-4,11-二烯-14-醛^△			C₁₅H₂₂O₂			234			1.84			-			-			6.09			3.57			-			-		5.98		14.61
53	4,9-dihydroxy-6-methyl-3,10-dimethylene-3a,4,7,8,9,10,11,11a-octahydro-3H-cyclodeca[b]furan-2-one 4,9-二羟基-6-甲基-3,10-二亚甲基- 3a,4,7,8,9,10,11,11a-八氢-3H-环癸[b]呋喃-2-酮^△			C₁₅H₂₀O₄			264			1.18			-			-			-			-			-			-		-		-
序号 No.	化合物 Compounds	分子式 Molecular formula			分子量 M_W			相对百分含量RC/%
序号 No.	化合物 Compounds	分子式 Molecular formula			分子量 M_W			S1			S2			S3			S4			S5			S6			S7			S8		S9
54	confertin 密花豚草素^△	C₁₅H₂₀O₃			248			2.63			-			-			-			-			-			-			-		-
55	2-(2-phenylethyl)chromone 2-(2-苯乙基)色酮^*	C₁₇H₁₄O₂			250			1.20			-			-			0.72			-			-			0.88			21.40		9.62
56	qinan lactone 奇楠内酯^△	C₁₅H₂₆O₂₃			254			5.40			-			-			-			-			-			-			-		-
57	3,7,7-trimethyl-8-(2-methyl-propenyl)-bicyclo [4.2.0]oct-2-ene 3,7,7-三甲基-8-(2-甲基-丙烯基)-二环[4.2.0] 辛-2-烯^△	C₁₅H₂₄			204			1.18			-			-			-			-			-			-			-		-
58	6-methoxy-2-(2-phenylethyl)chromone 6-甲氧基-2-(2-苯乙基)色酮^*	C₁₈H₁₆O₃			280			1.38			2.89			0.52			6.74			12.40			1.65			5.44			22.96		16.21
59	2-[2-(4-methoxyphenyl)ethyl]chromone 2-[2-(4-甲氧基)苯乙基]色酮^*	C₁₈H₁₆O₃			280			-			-			-			-			-			-			1.20			-		2.20
60	5,8-dihydroxy-2-(2-phenylethyl)chromone 5,8-二羟基-2-(2-苯乙基)色酮^*	C₁₇H₁₄O₄			282			28.35			13.69			16.87			14.96			40.09			29.70			8.40			12.15		3.42
61	6-hydroxy-2-(2-phenylethyl)chromone 6-羟基-2-(2-苯乙基)色酮^*	C₁₇H₁₄O₃			266			-			-			-			-			-			-			-			2.49		-
62	squalene鲨烯	C₃₀H₅₀			410			-			-			-			-			-			1.43			-			-		-
63	6-methoxy-2-[2-(3-methoxyphenyl)ethyl] chromone 6-甲氧基-2-[2-(3-甲氧基苯基)乙基]色酮^*	C₁₉H₁₈O₄			310			-			1.04			-			-			-			-			-			-		-
64	6-methoxy-2-[2-(4-methoxyphenyl)ethyl] chromone 6-甲氧基-2-[2-(4-甲氧基苯基)乙基]色酮^*	C₁₉H₁₈O₄			310			-			-			-			3.69			6.94			-			3.12			0.97		-
65	6,7-dimethoxy-2-(2-phenylethyl)chromone 6,7-二甲氧基-2-(2-苯乙基)色酮^*	C₁₉H₁₈O₄			310			-			-			0.44			7.01			10.05			8.43			39.10			5.41		6.77
66	6-methoxy-2-[2-(4-methoxy-3-hydroxylphenyl)ethyl]chromone 6-甲氧基-2-[2-(4-甲氧基-3-羟基苯基)乙基] 色酮^*	C₁₉H₁₈O₅			326			-			0.80			-			1.87			-			-			-			-		-
67	6,7-dimethoxy-2-(4-methoxyphenethyl) chromone 6,7-二甲氧基-2-(4-甲氧基苯乙基)色酮^*	C₂₀H₂₀O₅			340			-			-			-			-			-			-			1.98			-		-
68	stigmasterol 豆甾醇	C₂₉H₄₈O			412			1.13			-			-			-			-			-			0.92			-		0.98
69	γ-sitosterol γ-谷甾醇	C₂₉H₅₀O			414			-			-			-			-			-			-			-			-		1.09
总计								67.81			82.83			96.44			96.66			100			68.49			98.87			100		94.16
倍半萜类化合物								34.39			63.38			76.50			61.67			30.51			21.55			34.39			31.57		53.87
色酮类化合物								30.93			18.42			17.83			34.99			69.49			39.78			60.12			65.38		38.22
其他类化合物								2.49			1.03			2.11			0			0			7.16			4.36			3.05		2.07

	注：^*代表色酮类化合物;^△代表倍半萜类化合物;-代表未检出。
	Notes: ^* stands for chromones; ^△ stands for sesquiterpenes; - stands for not detected.

3 讨论

倍半萜类成分和色酮类成分是沉香中2类特征性成分,也是沉香的主要成分^[33,34,35]。本研究通过GC-MS分析发现,9份沉香样品中倍半萜类和色酮类成分的总相对百分含量为68.49%~ 100.00%。所有样品中只有2种成分为共有成分,即5,8-二羟基-2-(2-苯乙基)色酮和6-甲氧基-2-(2-苯乙基)色酮,其中前者在小果沉香中的相对百分含量均较高,超过10.00%,最高可达40.09%（S5）,而在白木香、柯拉斯那沉香和近全缘沉香均较低,分别为8.40%,12.15%和3.42%。杨峻山等^[36]报道了5,8-二羟基-2-(2-苯乙基)色酮在国产白木香所产沉香中首次分离得到。

杨锦玲等^[37]曾报道越南产柯拉斯那沉香中的黄熟香和吊口香中也有5,8-二羟基-2-(2-苯乙基)色酮,且相对百分含量也较高。另外,邵杭等^[38]在柳叶拟沉香中也分离得到该化合物,并发现其对乙酰胆碱酯酶具有一定的抑制活性,抑制率为38.0±1.0（50 μg/mL）。雷智东^[39]报道该化合物对金黄色葡萄球菌有较强的抑制作用。赵桐^[40]用茉莉酸甲酯（MeJA）气雾成功诱导白木香产生2种色酮,其中包括6-甲氧基-2-(2-苯乙基)色酮,表明该化合物可由人工诱导产生,但目前对其生物活性方面研究未见报道。

6份小果沉香结香样品中挥发性成分变化较大,但野生沉香与栽培沉香结香样品具有较大差别,其中野生样品（S1、S5、S6）中的倍半萜和色酮的种类均较少,二者之和分别为14种、15种和10种,其中倍半萜的相对百分含量较低（<40%）,而色酮的种类少,但相对百分含量较高（>30%）。而栽培样品（S2、S3、S4）中的倍半萜和色酮种类均较多,二者之和分别为23种、22种和23种,其中倍半萜的相对百分含量较高（>60%）,而色酮相对百分含量较低（<40%）。所有小果沉香栽培结香样品中均有白木香醛（S2为2.69%、S3为13.28%、S4为1.33%）和瓦伦萨- 1(10),8-二烯-11-醇（S2为3.52%、S3为3.42%、S4为1.15%）,它们在3种其他种类的沉香中也均含有,其含量分别为（S7为8.11%,9.67%;S8为4.10%,11.23%;S9为0.98%,2.91%）,而在小果沉香野生结香样品中则未检测到。另外,11,13-二羟基-9(10)-烯-8α,12-环氧艾里莫芬烷（S2为1.23%、S3为1.23%、S4为0.79%）只在栽培小果沉香结香样品中检测到。野生的自然结香年限相对较长,色酮类化合物所占比例较大,而栽培沉香结香的年限较短,倍半萜类化合物所占比例较大。杨锦玲等^[41]建立了白木香结香样品的指纹图谱,研究结果发现,倍半萜类化合物以白木香醛为主（相对百分含量为0.83%-27.20%）,色酮类化合物以6,7-二甲氧基-2-(2-苯乙基)色酮为主（相对百分含量为1.40%-31.23%）,5,8-二羟基-2-(2-苯乙基)色酮也有,但含量较低（2.00%- 6.57%）。而本次试验的白木香结香样品中的倍半萜类化合物以瓦伦-1(10),8-二烯-11-醇（9.67%）为主,其次为白木香醛（8.11%）,色酮类化合物以6,7-二甲氧基- 2-(2-苯乙基)色酮为主（39.10%）,其次为5,8-二羟基-2-(2-苯乙基)色酮（8.40%）,与杨锦玲等^[41]的研究结果基本一致。

杨锦玲等^[37]曾报道越南柯拉斯那沉香之红土沉香、黄熟香和吊口香的乙醚提取物GC-MS分析,结果表明,红土沉香和黄熟香均以色酮类成分为主,种类也较多,相对百分含量分别为71.27%（11种）和68.10%（14种）,前者又以2-[2-(4-甲氧基)苯乙基]色酮（28.94%）、2-(2-苯乙基)色酮（22.63%）、2-[2-(3-羟基-甲氧基)苯乙基]色酮（14.27%）为主要成分,后者以6,7-二甲氧基-2-(2-苯乙基)色酮（39.11%）和5,8-二羟基-2-(2-苯乙基)色酮（5.57%）为主。越南吊口沉香以倍半萜类化合物较多,总相对百分含量为21.31%,其中又以白木香醛（16.00%）为主。而本次研究的柯拉斯那沉香的倍半萜类成分以瓦伦-1(10),8-二烯-11-醇（11.23%）为主,色酮类成分以6-甲氧基-2-(2-苯乙基)色酮（22.96%）、2-(2-苯乙基)色酮（21.40%）、5,8-二羟基-2-(2-苯乙基)色酮（12.15%）、6,7-二甲氧基-2-(2-苯乙基)色酮（5.41%）为主,与越南柯拉斯那沉香之黄熟香相近。

Pripdeevech等^[42]采用固相微萃取的方法,研究了近全缘沉香的挥发性成分,结果表明,其相对百分含量最大的成分为十二酸异戊酯（isoamyl dodecanoate）（38.00%）,其次为枯树醇（kusunol）（6.40%）和沉香雅蓝醇（jinkoh-eremol）（5.60%）。Monggoot等^[43]采用4种内生真菌诱导近全缘沉香产生沉香,挥发性成分研究结果表明,含量最高的成分为氧化沉香螺旋醇（oxo-agarospirol）,其相对百分含量均超过50.00%。本次研究结果与此前的研究有较大差异,其中含量最大的成分为6-甲氧基-2-(2-苯乙基)色酮（16.21%）,其次为(+)-9-羟基-芹子-4,11-二烯-14-醛（14.61%）,这可能跟结香方法和提取方法等有关。

综上所述,小果沉香结香样品油脂的化学成分组成以5,8-二羟基-2-(2-苯乙基)色酮为主,野生结香样品与栽培结香样品区别较大,野生样品中的倍半萜和色酮的种类均较少,其中倍半萜的种类多于色酮类化合物,但相对百分含量较低,而色酮的种类少,但相对百分含量却较高。栽培结香样品中的倍半萜和色酮种类均较多,其中倍半萜的种类也多于色酮类化合物,其相对百分含量也较高,色酮的种类较少,其相对百分含量也较低。另外,有一种倍半萜类化学成分,即11,13-二羟基-9(10)-烯-8α,12-环氧艾里莫芬烷只在栽培小果沉香结香样品中检测到。另外,将小果沉香与白木香、柯拉斯那沉香、近全缘沉香结香样品对比分析发现,所有种类和样品中有2种共有成分,即5,8-二羟基-2-(2-苯乙基)色酮和6-甲氧基- 2-(2-苯乙基)色酮。小果沉香与白木香、柯拉斯那沉香、近全缘沉香结香样品的种类与相对百分含量具有较大差异。本研究可以为小果沉香的真伪鉴定及质量标准研究等方面提供重要参考。

References

Publishing order | Descend order by publishing year | Descend order by cited within

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