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BY 4.0 license Open Access Published by De Gruyter Open Access September 25, 2019

Analysis and Biological Evaluation of Arisaema Amuremse Maxim Essential Oil

  • Guiying Li , Yueyao Jiang , Yingjun Li , Tong He , Ying Wang , Tianyi Ji , Wanchen Zhai , Lichun Zhao EMAIL logo and Xiaoping Zhou
From the journal Open Chemistry

Abstract

The chemical composition and antitumor activity of essential oil were compared for four parts of the Arisaema Amuremse Maxim. Chemical constituent of essential oil in every parts of Arisaema Amuremse Maxim was identified by GC-MS analysis systematically. Arisaema Amuremse Maxim had 114 kinds of essential oils. Tubers, petioles, leaves, and fruits were identified 53, 48, 5 and 21 species essential oil, respectively. The antiproliferative activity against Hep2, HCT-116, A549, SW480, HepG2 cell lines had been investigated by MTT assay. The essential oil of tubers, petioles, leaves and fruits all had strong antiproliferative activity. The IC 50 was 19.60μg·mL-1, 17.60μg·mL-1, 23.80μg·mL-1, and 30.23μg·mL-1, respectively. Among them, the essential oil of tubers and petioles had stronger antitumor activity. The essential oil of tubers, petioles, leaves and fruits had strong antiproliferative activity. This study was of great significance to increase the medicinal parts of Arisaema Amuremse Maxim.

1 Introduction

Arisaema Amuremse Maxim, also called aliased corn and snake corn, is traditional Chinese perennial herb medicine, mainly distributed in Heilongjiang, Jilin and Liaoning Provinces of China, as well as in Korea and Russia [1]. It is bitter in taste, pungent, warm, poisonous, and effects on fight swelling [2, 3]. It can also be used to cure the dizziness, epilepsy, anti-tumor, coronary heart disease and inflammation [4, 5, 6]. Currently, there are not many reports on Arisaema Amuremse Maxim chemical components, mainly flavone glycosides, sterols, glycosides and brain esters, etc. As far as the essential oil of this plant was rarely studied, which chemical constituent is more complicated [7]. This essay concentrated on chemical constituent of essential oil. First, we used steam distillation to extract the tubers, petioles, leaves, and fruits of Arisaema Amuremse Maxim and obtained the essential oil. Then the essential oil was extracted by diethyl ether, dried with anhydrous sodium sulfate, and analyzed by GC - MS. Finally, the antineoplastic activities of the essential oils were tested by MTT assay [8, 9, 10, 11, 12, 13, 14]. The Arisaema Amuremse Maxim was adopted and identified by Professor Chendi of the School of Pharmacy of Jilin University in the Changbai Mountain area of Jilin Province in 2016.

2 Experimental section

2.1 Plant material

Arisaema Amuremse Maxim was collected from the Changbai Mountain in Jilin province and was identified by the School of pharmacy Chendi professor of Jilin University.

2.2 Extraction and Identification

100 g dry tubers, petioles, leaves, fruits were put in the different essential oil extractor, Then, 1000 ml distilled water were added to extract for 10 hours. After extraction with diethyl ether and drying with anhydrous sodium sulfate, diethyl ether was evaporated to get essential oils which were all the pale yellow oily special aroma matters.

2.3 GC-MS chromatographic conditions

The GC-MS used for the gas chromatographic analysis consisted of a chromatographic column Agilent hp-5 (0.25mm×30m,0.25μm); the injection temperature was 260⍰ and transmission line temperature was 280⍰, carrier gas was helium, flow rate was 1.0 mL/min, split ratio was 20:1, sample size was 1 μL. Temperature program: column temperature 80⍰ for 3 min, in 10⍰/ min up to 150⍰, not reserved, continued to heat in 5⍰/ min to 240⍰, not reserved, then in 10⍰/ min to 280⍰, keeping 10 min. Mass spectrometer conditions: EI ionization, electron bombardment energy 70 eV, ion source temperature 230⍰, scanning range from 20 to 800 aμm; multiplier voltage 2141 V solvent delay 3 min, nist 2008 standard mass spectrometry gallery. Using gas chromatography data processing system, data processing by computer, identified the kinds of chemical composition by NIST 2008 standard mass spectrometry gallery. The relative content of each component in the sample was determined by peak area normalization method.

2.4 The antiproliferative activity against Hep2, HCT-116, A549, SW480, HepG2 cell lines has been investigated by MTT assay

2.4.1 Cell culture

DMEM complete medium containing 10% fetal bovine serum, 100 μg·mL-1 penicillin and 100 μg·mL-1 streptomycin. Under 37⍰, 5%CO2, saturated humidity conditions. The medium was changed every other day and once every 3-4 days.

2.4.2 The tumor cells in vitro growth inhibition test (MTT)

Hep2 cells (throat cancer), HCT-116 cells (colon cancer), A 549(lung adenocarcinoma cells), SW480 (colon cancer) and HepG-2 (HCC) cells were digested by 0.25% trypsin-0.01% EDTA solution, suspending in the culture medium and counting. Adjusting the cell density to 5 x104 (a ·mL - 1), added to 96 - well culture plate (100 μL per well). After 24h, the experimental group added 100 μl different concentration of each part of Arisaema Amuremse Maxim into each well. The concentration gradients of volatile oils in the four parts were 62.5 μg·mL-1, 125 μg·mL-1, 250 μg·mL-1, 500 μg·mL-1 and 1000 μg·mL-1 and the positive control group added the same concentration gradient 5 - fluorouracil. The blank control group, cell control group, ethanol solvent control group were set up. Each group had three wells. They were put in the saturated humidity, 37⍰, 5% CO2 incubator to continue culturing for 48 h. The cell morphology changes with the inverted microscope were observed

Before the end of the experiment 20 μL of 5mg·mL-1 MTT were added to each well, removed the cell culture medium with a syringe. Then 150 μL DMSO were added into each well, and shaken on the shaking bed with low speed for 10 min to make crystals dissolve completely. The OD570nm absorbance values were measured by an enzyme-linked immune detector.

The following formula was used to calculate the fruit growth inhibition rate (IR) of tumor cells: IR (%) = (1 - (dosing average OD value - blank group average OD value) / (control group average OD value - blank group average OD value) ×100%) and found the half inhibitory concentration (IC50) by the Origin software.

Ethical approval: The conducted research is not related to either human or animal use.

3 Results and discussion

3.1 Identification of essential oil

Aristides Amusement Maxim had 114 kinds of essential oil as shown in Table 1. The most was located in tubers, up to 53; the second was in petioles, totaling 48; the least was found in leaves, only 5. 2-pentadecanone, ethyl ester and ethyl oleate were more abundant in tubers and petioles. Phytone, hexadecanoic acid methyl ester, methyl oleate, heneicosane were common composition of tubers and fruits. Methyl palmitate and methyl oleate were similar in tubers, leaves, fruits, petioles and Leaves both had 6,10,14-trimethyl-2-pentadecane ketone. Nonadecane was common composition of petioles and fruits.

Table 1

Analysis on chemical constituent of essential oil in every parts of Arisaema Amurense Maxim by GC-MS.

NumberChemical compositionTubersPetiolesLeavesFruits
Relative Contents
1Hexanal0.861
2trimethyl Oxazole0.543
32-Furanmethanol5.501
42,6-dimethyl Pyrazine1.562
52,4,5-trimethyl Thiazole0.171
62,3,5-trimethyl Pyrazine1.988
7Benzeneacetaldehyde1.235
83-ethyl-2,5-dimethyl Pyrazine1.317
92,3-Dimethyl-5-ethylpyrazine0.942
10Linalool(3,7-dimethyl-1,6-Octadien-3-ol)0.516
112-n-Butylfuran0.491
122-Isopropyl-4,5-dimethyl Thiazole0.601
132,3-diethyl-5-methyl Pyrazine0.360
142-ethyl-3-hydroxy-4H-Pyran-4-one0.598
152,5-dimethyl-3-(2-methylpropyl) Pyrazine0.209
165-ethyl-2-methyl-4-propyl Thiazole0.382
174,5-dimethyl-2-(2-methylpropyl) Thiazole0.587
182,5-dimethyl-3-(3-methylbutyl) Pyrazine0.398
19(R)-1,5,5,9-tetramethyl-,Spiro[5.5]undeca-1,8-diene0.264
20(+)-Epi-bicyclosesquiphellandrene0.743
212,6-Di(tert-butyl) benzo-1,4-quinone0.853
22[1S-(1π4π5π]-1,8-dimethyl-4-(1-methylethenyl)- Spiro [4.5] dec-7-ene0.296
232-Tridecanone1.708
242,6-di-tert-butyl-p-cresol (Butylated Hydroxytoluene)1.242
252-Dodecanone0.345
261,3-bis(1,1-dimethylethyl)-2-methoxy-5-methyl Benzene0.601
27(Z, Z)-6, 9-Pentadecadien-1-ol0.705
28(Z, Z)-10,12-Hexadecadienal0.387
29Cyclopentadecanone2.076
302-Pentadecanone8.904 0.543
314,5,5a,6,6a,6b-hexahydro-4,4, 6b-trimethyl-2-(1-methylethenyl)- 2H-Cyclopropa [g] benzofuran0.621
322-Nonanone0.562
33Perhydrofarnesyl acetone⍰(6,10,14-trimethyl-2-Pentadecanone)7.1510.388
34(Z)- 9,17-Octadecadienal1.433
355-heptenyl Benzene1.064
36(E)-1-methoxy-9-Octadecene0.822
372-Heptadecanone0.922
38(E,E)-6,10,14-trimethyl-5,9,13-Pentadecatrien-2-one1.189
39Hexadecanoic acid methyl ester1.33338.62253.452
40Dibutyl phthalate1.488
41Hexadecanoic acid ethyl ester3.6920.405
42[1R-(1R*,3E,7E, 11R*,12R*)]-4,8,12,15,15-pentamethyl-Bicy- clo[9.3.1]pentadeca-3,7-dien-12-ol1.034
43Aromadendrene oxide-(2)0.598
443-cyclohexyl-1-phenyl Propane14.859
45(Z, Z)-9,12-Octadecadienoic acid methyl ester1.4755.402
46(Z) 9-Octadecenoic acid methyl ester0.86612.2760.117
47Oxacycloheptadec-8-en-2-one0.861
489,12-Octadecadienoic acid, ethyl ester5.6193.586
49(Z, Z, Z)- 9,12,15-Octadecatrienoic acid ethyl ester1.515
50Ethyl Oleate1.8390.527
51(Z, Z)- 9,12-Octadecadienoyl chloride0.780
52Heneicosane6.2770.559
53Pentacosane2..601
54Tetradecane0.098
551-(2-nitropropyl)- Cyclohexanol0.149
561H-Cyclopropa[a]naphthalene0.067
572,6,10-trimethyl-Tetradecane0.098
58Pentadecane0.171
59Hexadecane0.770
60Heptadecane2.527
615,6-bis(2,2-dimethylpropylidene)-(E,Z)- Decane2.805
621-chloro-Octadecane1.043
63Octadecane1.755
641-Hexadecanol acetate4.052
656,10,14-trimethyl-2-Pentadecanone0.99746.276
661,2-Benzenedicarboxylic-acid bis(2-methylpropyl) ester1.091
67Nonadecane1.0320.279
6814-Methylpentadecanoic acid methyl ester7.699
697,9-Di-tert-butyl-1-oxaspiro (4, 5) deca-6,9-diene-2,8-dione7.909
70tert-Hexadecanethiol1.851
711,2-Benzenedicarboxylic acid butyl 8-methylnonyl ester1.152
72Eicosane1.012
732,4,4,6,6,8,8-Heptamethyl-2-nonene0.624
745,6,6-Trimethyl-5-(3-oxobut-1-enyl)-1-oxaspiro [2.5] octan-4-one0.599
75Methyl-11,14-eicosadienoate0.570
768-Octadecenoic acid methyl ester0.755
775-dodecyldihydro-2(3H)-Furanone0.470
78Hexadecanoic acid butyl ester2.189
79Docosane0.587
80Behenic alcohol5.025
81Heptacosane4.005
82Butyl 9,12-octadecadienoate0.555
83Tetracosane6.873
84Octacosane4.641
851,2-Benzenedicarboxylic acid diisooctyl ester0.516
86Hexacosane0.463
87Sulfurous acid cyclohexylmethyl pentadecyl ester0.462
889-octyl-Tetracosane0.576
89Hexatriacontane6.734
909-octyl-Hexacosane0.486
911-Hexacosene0.538
9213-dodecyl-Hexacosane5.129
93Nonacosane0.773
94i-Propyl 24-methyl-pentacos-5,9-dienoate5.109
95Triacontane2.544
96Hentriacontane3.285
97Dotriacontane1.230
98Tritriacontane1.701
991-(1-Ethyl-2,3-dimethyl-cyclopent-2-enyl)-ethanone1.602
1003,7,11,15-Tetramethyl-2-hexadecen-1-ol1.224
101Methyl-9-Tetradecenoate0.115
102Methyl tetradecanoate1.539
103Pentadecanoic acid methyl ester0.095
10412-methyl Tetradecanoic acid methyl ester0.111
10514-methyl-Pentadecanoic acid methyl ester0.048
106(Z)- 9-Hexadecenoic acid methyl ester0.915
107Heptadecanoic acid, methyl ester0.519
1089,10,12-trimethoxy Octadecanoic acid methyl ester1.305
1099-Octadecenoic acid methyl ester5.033
11013-octadecenoate7.818
111Octadecanoic acid methyl ester2.262
112Ethyl 9-hexadecenoate0.375
113Methyl 12-hydroxystearate3,005
114Methyl 13-phenyl-tridecanoate1.031

The systematic research of essential oil in Arisaema Amuremse Maxim in this paper was the foundation for further study, which was of great significance to increase the medicinal parts of Arisaema Amuremse Maxim.

3.2 The antiproliferative activity was investigated by MTT assay

The essential oil of tubers, petioles, leaves, and fruits was tested against cancer cell lines, including Hep2, HCT-116, A-549, SW-480, HepG-2 for proliferation and survival by MTT assay. In general, the essential oil of Arisaema Amuremse Maxim all activated on cancer cells effectively. The essential oil of tubers, petioles, leaves and fruits all had strong antiproliferative activity. Among them, the essential oil of tubers and petioles had stronger antitumor activity. The results were presented in Table 2.

Table 2

The antiproliferative activity of essential oil.

IC50 (μg·mL-1)
PartsHep2HCT-116A-549HepG-2SW-480Most Sensitive Cells
Fruits50.47±5.2659.26±8.38105.00±18.2640.14±6.1230.23±4.99SW-480
Tubers26.71±3.8419.60±0.9527.44±1.2319.83±1.5635.46±2.37HCT-116
Leaves51.79±4.9836.75±5.2452.52±6.3223.80±2.77138.10±15.52HepG-2
Petioles23.60±1.9419.52±1.3620.80±7.6417.60±1.9627.72±7.11HepG-2

4 Discussion

In the literature, reports were mainly on the study of essential oil of Arisaema Amuremse Maxim tubers. In this paper, the antitumor activity of essential oil in various parts of Arisaema Amuremse Maxim (including tubers, petioles, leaves and fruits) was studied and compared for the first time.

Arisaema Amuremse Maxim had 114 kinds of essential oil. There were 24 kinds of essential oil of tubers with ingredients over 1%. The highest was three-cyclohexyl - 1-phenyl propane, occupied 14.859%. There were 25 kinds of essential oil of petiole with ingredients over 1%, the highest was 7,9-two tertiary butyl-1-oxygen screw [4.5]-decane-6, 9-diene-2,8-diketone (7.909%) and 14-methyl pentadecane acid methyl ester (7.699%). The highest in leaves was 6,10,14-trimethyl-2-pentadecane ketone (46.276%). The highest in fruits was methyl palmitate (53.452%). There were 5 kinds of essential oil for leaves with ingredients over 1%, the highest was 6, 10, 14-trimethyl-2-pentadecane ketone (46.276%). There were 9 kinds of essential oil of fruits with ingredients over 1%, the highest was methyl palmitate (53.452%).

The essential oil of tubers, petioles, leaves, and fruits was tested against cancer cell lines, including Hep2, HCT-116, A-549, SW-480, HepG-2 for proliferation and survival by MTT assay. Among them, the tubers were the most sensitive to HCT-116 cells, and the IC50 was 19.60±0.95 ;Petioles was the most sensitive to HepG-2 cells, IC50 is 17.60±1.96; Fruits were the most sensitive to SW-480 cells, IC50 was 30.23±4.99; Leaves were most sensitive to HepG-2 cells, IC50 was 23.80±2.77. It can be seen from Table 2 that essential oil of tubers, petioles, leaves, and fruits of Arisaema Amuremse Maxim had a good in vitro inhibition over the above four experimental cells. With the increase of the dose, the inhibition of tumor cells gradually increased. Therefore, the four parts of Arisaema Amuremse Maxim, including petiole, leaf and fruit, could be used in medicine. The medicinal part of Arisaema Amuremse Maxim recorded by the first edition of the 2015 edition of the Chinese Pharmacopoeia was its dried tubers. In addition to the medicinal studies on the tuber parts of Arisaema Amuremse Maxim, the petiole, leaf and fruit parts of Arisaema Amuremse Maxim were also studied in this paper. Therefore, this study was of great significance in increasing the effective medicinal parts of Arisaema Amuremse Maxim.

Figure 1 GC-MS chromatograms showed the essential oil in every part of Arisaema amurenseMaxim: (a) Fruits, (b) Tubers, (c) leaves, (d) Petioles.
Figure 1

GC-MS chromatograms showed the essential oil in every part of Arisaema amurenseMaxim: (a) Fruits, (b) Tubers, (c) leaves, (d) Petioles.

5 Conclusions

The essential oil components of the whole plant of Arisaema Amuremse Maxim were studied systematically in this paper, which would lay the foundation for further research on its chemical composition and increasing the effective medicinal parts of Arisaema Amuremse Maxim.

Acknowledgments

The authors expressed many thanks for the funding and support of Department of Pharmaceutical chemistry, College of Pharmacy, Jilin University, and also tgrateful for the assistance from Dr. Xiaohong Yang.

  1. Conflict of interest: Authors declare no conflict of interest.

References

[1] Li C.N., Liu Y.Y., Li P.S., Shi X.J., Xu T.H. and Liu T. H., Chemical constituents and pharmacological activities of arisaema amurense maxim, Jilin Tradit. Chin. Med., 2015, 35, 293-296.Search in Google Scholar

[2] Huang J., Yi J.H., Liu Y.H., Huang Z.F., Chen Y. and Liu Y.H., Determination of eight nucleosides in rhizoma arisaema, rhizoma pinellie and rhizoma typhonii, Chin. Exp. Tradt. Medi. Formulae., 2013, 19, 59-62.Search in Google Scholar

[3] Du X., Rhizoma arisaematis pharmacological action and clinical application research work, Med. Inform., 2011, 7, 3048.Search in Google Scholar

[4] Tang J.H., Ren Y.L., Liu K.Q., Su J., Zhang Z.L., Rhizoma arisaematis pharmacological action and clinical application research work, Shanxi Tradit. Chin. Med., 2010, 478-479.Search in Google Scholar

[5] LI Y., Luo T.S., Dai X.T., Qian J.F., Antibacterial activity of the extracts from arisaema erubescens (wall.) schott, Dali. Univer., 2014, 13, 9-11.Search in Google Scholar

[6] Han J.T., Advances in pharmacological action and clinical application of araceae, Mod. Medi. Health., 2010, 26, 2487-2488.Search in Google Scholar

[7] Kong D.X., Yang X.H., Dong L., Shi Y.Y., LI X.H., Li G.Y., et al., Study on chemical components of volatile oil in stem of arisaema amuremse maxim by GC-MS, Spec. Econo. Animal. Plant., 2013, 66-69.Search in Google Scholar

[8] Tang J.H., Zhang H.M., Dong Y.C., Yang Z.H., Experimental study on apoptosis of BGC823 Cell in human gastric carcinoma induced by araceae alcohol extraction liquid, Shanxi Tradi. Chin. Medi., 2011, 32, 1421-1422.Search in Google Scholar

[9] Zhang Y., Wang S., Bao Y.R., Meng X.S., The study for activity of araceae extractions resisting lung cancer cell, Chin. Mod. Medi., 2013, 20, 80-83.Search in Google Scholar

[10] Qi X.X., Li H., Du G.J., Preliminary study of rhizome arisaematis on anti-tumor, Henan. Univer., 2014, 33, 85-87.Search in Google Scholar

[11] Yang G.P., Lv X.M., Gan P., Peng F., Bai L., Qian J., The antitumor effect of araceae arisaema extract on S180 sarcoma-bearing mice, Lishi. Medi. Mater., 2011, 21, 752-753.Search in Google Scholar

[12] Yang G.P., Cao H.Z., Li Y., Bai L., Qian J., The inhibitory effect of the fruit extract of araceae arisaema on human leukemia cell line in vitro, Northwest Pharm., 2012, 27, 133-135.Search in Google Scholar

[13] Zhang H.M., Tang J.H., Yang Z.H., Gao Y.Y., Experimental study of alcoholic extract of rhizoma arisaematisinduced apoptosis on human K562 cells, Chin. Drug. Appli. Monit., 2011, 8, 214-234.Search in Google Scholar

[14] Yang X.H., Li G.Y., Wang X.Y., Dong L., Chemical analysis and anti-tumor activity test of the volatile oil from arisaema amurense maxim tubers, Faseb., 2014, 28, 647.10.1096/fasebj.28.1_supplement.647.13Search in Google Scholar

Received: 2018-06-11
Accepted: 2018-09-11
Published Online: 2019-09-25

© 2019 Guiying Li et al., published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 Public License.

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