Identification of chemical constituents from fruit of Antidesma bunius by GC-MS and HPLC-DAD-ESI-MS

Abstract

Antidesma bunius is an edible berry fruit with many benefits, such as natural antimicrobials, anticancer, natural dyes, etc. However, data on chemical content in the fruit is still limited. The purpose of this research is to identify volatile compounds of Antidesma bunius fruit. We extracted and analyzed the A. bunius fruit’s chemical content using GC-MS and HPLC–DAD-ESI-MS methods. Forty-nine compounds representing 99.91% of the total chromatogram’s relative peak area were detected. Antidesma bunius is rich in 5-hydroxymethylfurfural (HMF) and ten other compounds with relative peak area >1%, such as furaldehyde, citric acid and others. We also found 109 compounds tentatively identified through HPLC–DAD-ESI-MS. Antidesma bunius contained HMF, several volatile compounds, organic acid, long-chain fatty acid, and photochromic compound.

Keywords:
Antidesma bunius; berry; bignay, GC-MS; HPLC–DAD-ESI -MS; volatile

1 Introduction

Antidesma bunius belongs to the Phyllantaceae family. The fruit is included in the type of berries. Antidesma bunius comes from east and south Asia. This plant has several other names, such as bignay, bignai, bugnay, Chinese laurel, Queensland cherry, wild cherry, currant tree, salamander-tree. In Indonesia, this fruit is known as the Buni or Huni fruit (Lizardo et al., 2015).

In Indonesia, Buni fruit is usually consumed directly when ripe or processed into local foods such as Rujak. The acidic taste and the striking color become an attraction for consumption. Besides its refreshing taste, Buni fruit is known to have many health benefits and can be applied to other aspects.

Antidesma bunius has many benefits for human health. Almost all parts of Antidesma bunius have bioactive compounds that can be used as a source of medicine. Buni fruit contains antioxidant compounds, anticancer, anti-inflammation, anti-diabetic, anti-hyperlipidemia and can also be used as an anti-parasitic agent (Chowtivannakul et al., 2016; Jorjong et al., 2015; Lizardo et al., 2015).

Health benefits and also applications of Buni fruit have been widely studied. However, data regarding the content of chemical compounds in the fruit are still limited. The research that has been carried out only covers the characterization of phytochemical content, polyphenol compounds and anthocyanin, but its chemical constituents remain incompletely investigated. Furthermore, no studies have examined the content of volatile compounds and essential oils in the fruit of Antidesma bunius. (Hardinasinta et al., 2020; Jorjong et al., 2015).

Thus, comprehensive profiling of the phytochemicals of Antidesma bunius fruit using sensitive tools is necessary. Therefore, this study aims to analyze volatile compounds in the fruit of Antidesma bunius by using high-performance liquid chromatography-diode array detector-hyphenated with tandem mass spectrometry (HPLC–DAD–ESI-MS) and gas chromatography with tandem mass spectrometry (GC-MS) as a powerful analytical technique.

2. Methods

2.1 Plant materials

Fresh ripe fruit of Antidesma bunius was manually harvested in Sumedang, West Java Province, Indonesia during March 2019. The fruit was maintained at -18 ^oC immediately after collection until they were used.

2.2 Extraction

The fresh ripe fruit of Antidesma bunius (200 g) was crushed and then extracted using 1 L of 80% methanol in the darkroom temperature overnight. The extraction process was repeated three times to ensure exhaustive extraction. The supernatant is then filtered using filter paper, and the resulting filtrate is evaporated using a rotary evaporator at a temperature of 45 ^oC.

2.3 Gas Chromatography-Mass Spectrometry

GCMS analyses were performed on a GC-2010 GC coupled with a GCMS-QP2010 Ultra mass spectrometer. The column was a Rxi-5ms fused-silica capillary column (30.0 m Length x 0.25 mm ID x 0.25 μm Thickness) with helium as the carrier gas and was run at a constant pressure of 37.1 psi. The injection was conducted using a split-less mode at an injector temperature of 250 °C. The oven temperature was ramped from 50 to 300 °C (10 min hold) at a rate of 3 °C/min. The oven temperature was held at 300 °C for 6 min following each analysis. The total run time for each sample was approximately 25 min. The GCMS interface temperature was set to 250 °C. MS mode was used during analytical scanning from 40-700 atomic mass units (amu). The ion source temperature was set to 200 °C. The blank was injected first, followed by the sample injection. The chromatograms obtained from the total ion current (TIC) were integrated without any correction for co-eluting peaks, and the results were expressed as total abundance. TIC peaks and chromatograms were analyzed using Agilent MSD ChemStation G1701DA software (version D 02.00, CA, USA). All peaks were identified based on mass spectral matching (≥ 90%) from both the National Institute of Standards and Technology (NIST) and Wiley libraries. Only compounds with 90% or greater spectral matching accuracy were reported.

2.4 HPLC–DAD-ESI-MS analysis

Separation of phenolic compounds from Antidesma bunius extract was performed on an Agilent 1200 series Rapid Resolution LC (Agilent Technologies, Santa Clara, CA). This instrument was equipped with an Agilent Zorbax C18 column (4.6 x 150 mm, 5 µm) from Agilent Technologies. Acidified water (0.5% acetic acid, v/v) and acetonitrile were used as mobile phases A and B, respectively. The gradient was programmed as follows: 0 min, 0% B; 20 min, 20% B; 30 min, 30% B; 40 min, 50% B; 50 min, 75% B; 60 min, 100% B; 62 min 0% B, and finally, the initial conditions were held for 8 min as a re-equilibration step. The flow rate was set at 0.80 mL/min throughout the gradient. The flow from the HPLC system into the ESI-MS detector was 0.2 mL/min. The injection volume was 10 µL and the column temperature was maintained at 25 °C.

3 Results and discussion

3.1 Chemical components identified in Antidesma bunius fruit by GC-MS

A methanol extract of Antidesma bunius has a deep purple color. In the analysis of volatile components present in Antidesma bunius extract, a total of fifty compounds were detected (Figure 1). Among these components, 5-hydroxymethylfurfural (5-HMF) was found in the most significant concentrations, representing 47.07% of all volatiles. Other predominant components were furfural, citric acid, furan carboxylic acid, 2,3-dihydro-3,5-dihydroxy-6-methyl-(4H)-pyran-4-one, hexanoic acid, 1,6-Anhydro-.beta.-D-glucofuranose, 5,5'-oxy-Dimethylene-bis(2-Furaldehyde), Linolenic acid and gamma-Sitosterol.

Figure 1
Gas Chromatogram of Volatile Compounds of Antidesma bunius extract.

The dominant compound detected in Antidesma bunius extract was 5-HMF with a relative amount of 47.07% (Table 1). This compound can be found naturally in honey and processed foods such as fruit juice, ketchup, UHT milk, etc. (Saeed et al., 2018). HMF is a compound that can be used as an intermediate for polymers, pharmaceuticals, liquid fuels, and the synthesis of several groups of compounds such as dialdehyde, ethers and other organic compounds. 5-HMF can be produced from hexoses such as fructose through several treatments such as dehydration (Gomes et al., 2015). This study result shows the potential of the Antidesma bunius fruit as a natural source of HMF.

Table 1
Volatile components identified in Antidesma bunius fruit by GC-MS.

No	Retention Time (RT) in minutes	Compounds	Molecular Formula*)	Relative Abundance (%)	MS Identification*)
1	4.534	Furazane, 3-amino-4-iodo-	C2H2IN3O	0.25	MS
2	5.273	Furfural	C5H4O2	5.89	MS
3	8.647	Citric acid	C6H8O7	3.99	MS
4	9.389	5-methyl furfural	C6H6O2	0.44	MS
5	10.009	2,4-Dihydroxy-2,5-dimethyl-3(2H)-furan-3-one	C6H8O4	0.53	MS
6	11.085	2,2,6,6-Tetramethylpiperidine	C9H19N	0.24	MS
7	12.250	Itaconic anhydride	C5H4O3	0.80	MS
8	12.841	2-Methyl-1,3-cyclohexanedione 97%	C7H10O2	0.16	MS
9	12.987	Pentanoic acid, 4-oxo	C5H8O3	0.48	MS
10	14.172	Orcinol	C7H8O2	0.18	MS
11	14.447	Methyl 2-furoate	C6H6O3	2.38	MS
12	15.835	Levoglucosenone	C6H6O3	0.31	MS
13	15.908	Butylacetate	C6H12O2	0.28	MS
14	16.628	6-Methyltridecane	C14H30	0.16	MS
15	17.397	Hydroxydihydromaltol	C6H8O4	6.46	MS
16	17.828	Methacrolein dimer	C8H12O2	0.16	MS
17	18.843	Pyrimidine, 4-chloro-5-ethoxy-2-methyl-	C7H9ClN2O	0.10	MS
18	19.653	2-Methylbutyl propionate	C8H16O2	0.48	MS
19	21.842	5-hydroxymethylfurfural	C6H6O3	47.07	MS
20	22.153	2-Cyclohexenone	C6H8O	2.55	MS
21	22.622	No hit compound	-	0.23	MS
22	24.375	No hit compound	-	0.09	MS
23	24.813	3-Heptanol	C7H16O	0.53	MS
24	25.437	2-Ethyl-3-nitroso-1,3-oxazinane	C6H12N2O2	0.15	MS
25	27.787	Decanoic acid	C10H20O2	0.11	MS
26	28.452	Propanoic acid, 2-methyl-, 2-(hydroxymethyl)-1-propylbutyl ester	C12H24O3	0.34	MS
27	29.607	Dodecanal	C12H24O	0.14	MS
28	30.922	Artemisia ketone	C10H16O	0.08	MS
29	33.640	Levoglucosan	C6H10O5	6.76	MS
30	34.128	2,4-Di-tert-butylphenol	C14H22O	0.80	MS
31	34.528	No hit compound	-	0.14	MS
32	37.793	Scyllo-Inositol	C6H12O6	2.03	MS
33	38.128	Cyclododecanol	C12H24O	0.77	MS
34	43.308	No hit compound	-	0.14	MS
35	49.655	Methyl Hexadecanoate	C17H34O2	0.23	MS
36	50.827	Hexadecanoic acid	C16H32O2	0.79	MS
37	51.798	5,5'-oxy-Dimethylene-bis(2-Furaldehyde)	C12H10O5	4.21	MS
38	55.195	Methyl Linoleate	C19H34O2	0.39	MS
39	55.405	Methyl Linolenate	C19H32O2	0.70	MS
40	56.392	Ambrettolide	C16H28O2	0.86	MS
41	56.612	Linolenic acid	C18H30O2	3.16	MS
42	57.267	Stearic acid	C18H36O2	0.43	MS
43	57.515	No hit compound	-	0.11	MS
44	72.035	No hit compound	-	0.21	MS
45	75.347	Squalene	C30H50	0.19	MS
46	79.457	Farnesol	C15H26O	0.19	MS
47	80.477	Tetraprenol	C20H34O	0.28	MS
48	82.427	Vitamin E	C29H50O2	0.52	MS
49	85.432	Stigmasterol	C29H48O	0.29	MS
50	86.903	Rhamnol	C29H50O	2.23	MS

• ^*
  ) Based on comparison of MS spectrum with database of WILEY229.LIB.

This study also detected 5-HMF derivative compounds, namely 5.5'-oxy-Dimethylene-bis (2-Furaldehyde). This compound is the result of the thermal decomposition of 5-HMF. Formation of 5,5'-oxy-Dimethylene-bis (2-Furaldehyde) compounds occur at temperatures of 100-220°C (Nikolov & Yaylayan, 2011).

One of predominant component detected in Antidesma bunius extract was furfural. This compound is commonly found in all fruits and vegetables and other plant parts that contain lignin and cellulose. Furfural is also a multi-purpose compound such as 5-HMF. Furfural can be applied in the petroleum industry, medicine, and also agriculture as insecticides, pesticides, antiseptics, and can also be used as bio-fuel (Wankasi & Tarawou, 2013).

Citric acid was also detected in Antidesma bunius extract (3.99%). This compound is generally found in citrus fruits. Citric acid provides an acidic taste in some fruits such as lemon, orange, lime, grapefruit. Antidesma bunius fruit also has a strong sour taste. Based on this study results, citric acid may also be a contributor to Antidesma bunius fruit sour taste (Chanukya et al., 2017).

Furancarboxylic acid or 2-furoic acid is a heterocyclic carboxylic acid, consisting of a five-membered aromatic ring and a carboxylic acid group. In the industry, this compound is used as a preservative, acting as a bactericide and fungicide. Antidesma bunius also own this antimicrobial property. Previous research shows the presence of antibacterial and antifungal activity from Antidesma bunius extract and food preservative effects (Lizardo et al., 2015). This study shows that 2-furoic acid compounds can provide antimicrobial effects in Antidesma bunius.

Farnesol is a bioactive compound that is also detected in this study. Farnesol is a natural 15-carbon organic compound which is acyclic sesquiterpene alcohol. Farnesol is present in many essential oils and used in perfumery. Farnesol is also known as a natural pesticide for mites and flavoring ingredients. Farnesol has antibacterial activity and is used as a deodorant in cosmetic products. It has been reported to exhibit anticancer and anti-inflammatory effects and alleviate allergic asthma, gliosis, and edema. These findings support the development of Antidesma bunius as a potential source of the pharmacological agent (Jung et al., 2018; Kromidas et al., 2006).

Another compound detected in this study is Hydroxydihydromaltol (2,3-dihydro-3,5-dihydroxy-6-methyl-(4H)-pyran-4-one). It was identified as a novel potent aroma compound in a dairy product, such as Ryazhenka kefir, and found as odorants in roasted chicory (Preininger et al., 2008). This compound is thought to be formed due to the heating process and can affect processed foods’ antioxidant activity. For example, increased Hydroxydihydromaltol is accompanied by an increase in antioxidant activity in dry prune products (Čechovská et al., 2011).

Hexanoic acid is an organic acid compound found in Antidesma bunius. This compound is also found in other fruit such as noni, pineapple and raspberry (Ferlinahayati et al., 2016; Aprea et al., 2015). Hexanoic acid has anticancer activity and also can improve energy metabolism (Narayanan et al., 2015; Miyamoto et al., 2016). Other studies have shown the anticancer activity of Antidesma bunius extract in Hela cells (Puspitasari & Ulfa, 2009). Hexanoic acid can be one of the compounds that provide anticancer activity from Antidesma bunius.

Another predominant compound detected in Antidesma bunius fruit is 1,6-Anhydro-beta-D-glucofuranose. This compound is anhydrosugar and commonly formed from the results of glucose pyrolysis (Hu et al., 2017). However, this compound is also naturally found in Oak bark, Acacia honey and Punica granatum. (Deryabin & Tolmacheva, 2015; Sangeetha & Vijayalakshmi, 2011).

Linolenic acid is an essential fatty acid belonging to the omega-3 fatty acids group. It is highly concentrated in certain plant oils and fruits (Saini & Keum, 2017). Linolenic acid has been reported to inhibit prostaglandin synthesis, resulting in reduced inflammation and the prevention of certain chronic diseases. It also has anti-carcinogenic, lipid metabolism regulation, anti-inflammatory, anti-obese and antioxidant activities (Yuan et al., 2014).

Gamma-Sitosterol is a phytosterol with structural similarity to cholesterol and can be found in plants, animals, and fungi. This compound has many beneficial effects on human health, such as hypolipidemic agents, anticancer, antioxidant, etc. (Balamurugan et al., 2015). In this study, we detected gamma-sitosterol in Antidesma bunius. This result suggests that Antidesma bunius can be a hypolipidemic agent, as proven in a previous study (Chowtivannakul et al., 2016).

2,4-di-tert-butylphenol is a member of the class of phenols carrying two tert-butyl substituents at positions 2 and 4. It has a role as a bacterial metabolite, an antioxidant, and a marine metabolite. It is an alkylbenzene and a member of phenols. Antioxidants/Stabilizers Fuels and fuel additives Intermediate Processing aids.

This study also detected several compounds that can act as flavoring agents. 2-cyclohexenone has an organoleptic property such as roasted savory green odor, 2-Methylbutyl propionate, and Methyl 2-furoate taste like sweet caramel brown sugar musty.

3.2 Chemical components identified in Antidesma bunius fruit by HPLC–DAD-ESI-MS

Among those identified compounds, several compounds have not been detected in Antidesma bunius fruit before. This study is the first to describe these results. Table 2 shows the list of 109 compounds detected through HPLC–DAD-ESI-MS experiments along with their retention times (tR). Seventy-eight compounds are tentatively identified, and 31 compounds remain unknown (Figure 2). The compounds detected in this work were tentatively characterized by MS data, together with the interpretation of the observed MS/MS spectra compared with those found in the literature. Several public databases were consulted in the identification process, such as ChemSpider (Royal Society of Chemistry, 2020), and PubChem (National Center for Biotechnology Information, 2020).

Table 2
Chemical components identified in Antidesma bunius fruit by LC-MS/MS.

Peak No	Retention Time (RT) in minutes	Compounds	Molecular Weight**)	Molecular Formula**)	Reference MS Identification**)
1	4.74	5-Methoxy salicylic acid	168.041	C8H8O4	MS
2	5.3	Suberic acid	174.088	C8H14O4	MS
3	5.64	Butanedioic acid, 2-(6-hydroxyhexyl)-3-methylene-	230.115	C11H18O5	MS
4	5.68	Methyl 5-[2-(4-methoxy-2-methyl-4- oxobutyl)-5-methyl-1,3-dioxolan-4- yl]pentanoate	316.188	C16H28O6	MS
5	5.69	Lariciresinol 4-O-glucoside	568.215	C26H34O11	MS
6	5.94	Unknown	497.253	C28H31N7O2	MS
7	6.02	Similar to: 1-(Carboxymethyl)cyclohexanecarboxylic Acid; ΔMass: -208.0708 Da)	394.16	C16H22N6O6	MS
8	6.03	1- (Carboxymethyl)cyclohexanecarboxylic Acid	186.088	C9H14O4	MS
9	6.51	Phospocholine conjugate acid	184.073	C5H15NO4P	MS
10	6.51	Similar to: Disperse red 17	390.129	C17H20N4O4	MS
11	6.51	Unknown	590.167	C28H27N6O7P	-
12	6.51	Unknown	796.223	C26H43N10O13P3	-
13	6.51	[Similar to: 2-(3,4- Dihydroxyphenyl)ethyl (1S,4aR,7aR)-1- (β-D-glucopyranosyloxy)-4a-hydroxy-7- methyl-5-oxo-1,4a,5,6,7,7ahexahydrocyclopenta[c]pyran-4- carboxylate; ΔMass: -68.0201 Da]	594.189	C21H37N6O8P3	MS
14	6.51	Malonic acid,isobutylidene-,bimol. cyclic ethylene ester	368.147	C18H24O8	MS
15	6.53	[Similar to: 2-(3,4- Dihydroxyphenyl)ethyl (1S,4aR,7aR)-1- (β-D-glucopyranosyloxy)-4a-hydroxy-7- methyl-5-oxo-1,4a,5,6,7,7ahexahydrocyclopenta[c]pyran-4- carboxylate; ΔMass: 274.1081 Da]	252.061	C8H8N6O4	MS
16	6.91	Unknown	596.214	C28H33N6O7P	MS
17	6.91	[Similar to: 1-(Carboxymethyl)cyclohexanecarboxylic acid; ΔMass: -414.1466 Da]	600.236	C21H43N6O8P3	MS
18	6.91	Similar to: 1-(Carboxymethyl)cyclohexanecarboxylic Acid; ΔMass: -208.0708 Da)	394.16	C16H22N6O6	MS
19	6.92	[Similar to: 1- (Carboxymethyl)cyclohexanecarboxylic acid; ΔMass: -67.9870 Da]	254.076	C7H14N2O8	MS
20	6.99	1,3,3-trimethylindolino-6'-nitrobenzopyrylospiran	322.131	C19H18N2O3	MS
21	7.67	1,3-Bis(2,4-dimethoxyphenyl)-1,3-propanedione	344.126	C19H20O6	MS
22	7.68	Rubone	374.136	C20H22O7	MS
23	8.03	[Similar to: [7-Hydroxy-1-(4-hydroxy-3, 5-dimethoxyphenyl)-3-(hydroxymethyl)- 6,8-dimethoxy-1,2,3,4-tetrahydro-2- naphthalenyl]methyl pentopyranoside; ΔMass: -11.9636 Da]	564.184	C27H36O12	MS
24	8.19	[Similar to: 1- (Carboxymethyl)cyclohexanecarboxylic acid; ΔMass: -208.0709 Da]	394.16	C16H22N6O6	MS
25	8.91	4-Ethyl-3-oxa-5-thia-4- boratricyclo[5.2.1.0~2,6~]decane	182.093	C9H15BOS	MS
26	10.54	Unknown	524.263	C29H32N8O2	-
27	10.96	Ethyl 5-methoxy-2-[(4- methoxyphenoxy)methyl]-1- benzofuran-3-carboxylate	356.126	C20H20O6	MS
28	10.98	N-[2-(Adamantan-1-yloxy)ethyl]-3,5- Dimethoxybenzamide	359.209	C21H29NO4	MS
29	11	(15Z)-9,12,13-Trihydroxy-15-	330.241	C18H34O5	MS
		octadecenoic acid
30	11.74	Ethyl 1,4-dioxaspiro[4.5]decane-6- Carboxylate	214.12	C11H18O4	MS
31	11.74	[Similar to: 11(Z),14(Z),17(Z)- Eicosatrienoic acid; ΔMass: 136.1262 Da]	170.13	-	MS
32	11.9	Isoindole-1,3-dione, 2-[2-(adamantan1-yloxy)ethyl]-5-methyl-3a,4,7,7atetrahydro	343.214	C21H29NO3	MS
33	12.03	1-Allyl 2-dodecyl 1,2-pyrrolidinedicarboxylate	367.272	C21H37NO4	MS
34	12.6	Unknown	508.268	C29H32N8O	MS
35	12.64	Unknown	547.279	C31H33N9O	MS
36	12.68	[Similar to: Cholic acid; ΔMass: -0.9951 Da]	409.283	C24H35N5O	MS
37	12.85	2-Nitro-1,3-bis(octyloxy)benzene	379.272	C22H37NO4	MS
38	13.02	Unknown	427.293	C24H37N5O2	-
39	13.04	Unknown	367.272	C15H38N5O3P	-
40	13.09	Unknown	397.282	C23H35N5O	-
41	13.19	(11alpha,13E,15S)-11,15-Dihydroxy-N- (2-hydroxyethyl)-9-oxoprost-13-en-1- amide	397.282	C22H39NO5	MS
42	13.55	[Similar to: 3-[2-(2,3-Dihydroxy-5,6,8atrimethyloctahydro-2Hspiro[naphthalene-1,2'-oxiran]-5- yl)ethyl]-5-hydroxy-2(5H)-furanone; ΔMass: -73.0889 Da]	439.239	C18H42N5O5P	MS
43	13.74	[Similar to: 20-Hydroxy- (5Z,8Z,11Z,14Z)-eicosatetraenoic acid; ΔMass: -135.0531 Da]	455.288	C25H37N5O3	MS
44	13.88	(9Z,12Z)-6,8-Dihydroxy-9,12- octadecadienoic acid	312.23	C18H32O4	MS
45	14.09	Unknown	425.278	C24H35N5O2	MS
46	14.21	1-Allyl 2-tridecyl 1,2- Pyrrolidinedicarboxylate	381.288	C22H39NO4	MS
47	14.49	11-Deoxy prostaglandin F2β	338.246	C20H34O4	MS
48	14.55	(11alpha,13E,15S)-11,15-Dihydroxy-N- (2-hydroxyethyl)-9-oxoprost-13-en-1- amide	397.282	C22H39NO5	MS
49	14.56	10-Undecen-1-yl N- (cyclohexylcarbonyl)alaninate	351.277	C21H37NO3	MS
50	14.84	Unknown	455.288	C25H37N5O3	-
51	14.86	Glycocholic acid	465.309	C26H43NO6	MS
52	14.93	Unknown	439.293	C24H41NO6	-
53	15.03	4-Dodecylbenzenesulfonic acid	326.192	C18H30O3S	MS
54	15.11	Unknown	425.278	C24H35N5O2	-
55	15.19	(+/-)12(13)-DiHOME /(Z)-12,13-dihydroxyoctadec-9-enoic acid	314.246	C18H34O4	MS
56	15.21	Unknown	435.298	C26H37N5O	-
57	15.23	Unknown	469.304	C29H43NO2S	-
58	15.23	17-(4-Hydroxyphenyl)-16-oxa-1,6,10,23-tetraazatetracyclo [9.8.6.2~12,15~.0~14,18~]heptacosa12,14,26-triene-19,24-dione	492.273	C28H36N4O4	MS
59	15.33	Similar to: Cholic acid; ΔMass: -0.9947 Da	409.282	C24H35N5O	MS
60	15.33	[Similar to: Cholic acid; ΔMass: - 68.9820 Da]	477.27	C20H40N5O6P	MS
61	15.54	Unknown	395.267	C23H33N5O	-
62	16.25	Unknown	453.273	C18H40N5O6P	-
63	16.33	Unknown	409.283	C24H35N5O	-
64	16.34	OCTOXYNOL-2	294.219	C18H30O3	MS
65	16.41	Dodecyl 6-{[(2-methoxyethoxy)carbonyl]amino}Hexanoate	401.314	C22H43NO5	MS
66	16.49	8-Iso-15-keto-prostaglandin-F2β	352.225	C20H32O5	MS
67	16.55	Pentadecyl N-2-furoylalaninate	393.288	C23H39NO4	MS
68	16.62	Dodecyl sulfate	266.155	C12H25O4S-	MS
69	16.63	Unknown	375.298	C21H37N5O	-
70	16.69	Dodecyl p-toluenesulfonate	340.207	C19H32O3S	MS
71	16.71	2-Nitro-1,3-bis(octyloxy)benzene	379.272	C22H37NO4	MS
72	16.8	2-Nitro-1,3-bis(octyloxy)benzene	379.272	C22H37NO4	MS
73	16.8	N-({(1R,2R)-3-Oxo-2-[(2E)-2-penten-1- yl]cyclopentyl}acetyl)-L-valyl-L-leucine	422.278	C23H38N2O5	MS
74	16.99	3,5,6-Trihydroxyandrostan-17-one	322.214	C19H30O4	MS
75	17.05	Unknown	551.404	C29H53N5O5	-
76	17.06	Unknown	535.351	C31H45N5O3	-
77	17.18	2-Nitro-1,3-bis(octyloxy)benzene	379.272	C22H37NO4	MS
78	17.21	Unknown	425.277	C24H35N5O2	-
79	17.23	Unknown	437.278	C25H35N5O2	-
80	17.33	[Similar to: Methyl 3-{1-[3-hydroxy-4- (hydroxymethyl)tetrahydrofuran-2-yl]- 2,4-dioxo-1,2,3,4-tetrahydropyrimidin5-yl}acrylate; ΔMass: -143.1715 Da]	455.267		MS
81	17.35	[Similar to: 9-Nitrooleate; ΔMass: - 150.0443 Da]	477.285	C25H39N3O6	MS
82	17.53	Cyclo (isoleucylleucylisoleucylleucylleucyl)	565.419	C30H55N5O5	MS
83	17.6	DPPH	395.086	C18H12N5O6	MS
84	17.62	Unknown	596.296	C28H45N4O8P	-
85	17.63	[Similar to: 9-Nitrooleate; ΔMass: - 150.0443 Da]	477.285	C25H39N3O6	MS
86	17.64	[Similar to: 9-Nitrooleate; ΔMass: - 178.0757 Da]	505.317	C21H44N7O5P	MS
88	17.79	Phenyl laurate	276.209	C18H28O2	MS
89	17.81	4-Undecylbenzenesulfonic acid	312.176	C17H28O3S	MS
90	17.9	Unknown	679.412	C32H63N3O8P2	-
91	17.91	Unknown	601.397	C30H56ClN5O5	-
92	18.04	Methyl N- [(benzyloxy)carbonyl] leucylleucylleucinate	505.317	C27H43N3O6	MS
93	18.17	5-(4-Carboxy-3-methylbutyl)-1,4adimethyl-6-methylenedecahydro-1- naphthalenecarboxylic acid	336.23	C20H32O4	MS
94	18.31	OCTOXYNOL-2	294.219	C18H30O3	MS
95	18.34	13(S)-Hydroperoxylinolenic acid	246.199	C18H30O4	MS
96	18.42	Unknown	345.288	C19H39NO4	-
97	18.53	Unknown	407.267	C17H38N5O4P	-
98	18.54	[Similar to: Phosphatidylinositol-1,2- dipalmitoyl; ΔMass: 238.2296 Da]	572.296	C27H41N8O4P	MS
99	18.7	HEXADECYL 4-NITROPHENYL ETHER	363.277	C22H37NO3	MS
100	18.72	17-EPIOXANDROLONE	306.219	C19H30O3	MS
101	18.76	Unknown	533.335	C31H43N5O3	-
102	18.94	16,17-Dihydroxykauran-18-oic acid	290.225	C20H32O4	MS
103	18.99	[Similar to: Phosphatidylinositol-1,2- dipalmitoyl; ΔMass: 329.2087 Da]	481.317	C24H44N5O3P	MS
104	19.05	[Similar to: Sorbitan monooleate; ΔMass: -50.9877 Da]	479.302	C18H43N9O2P2	MS
105	19.26	4-Dodecylbenzenesulfonic acid	326.192	C18H30O3S	MS
106	19.26	Unknown	598.312	C28H47N4O8P	-
107	19.38	Unknown	563.346	C32H45N5O4	-
108	19.51	Laurophenone	260.214	C18H28O	MS
109	19.58	1-{[(2- Aminoethoxy)(hydroxy)phosphoryl]oxy} -3-hydroxy-2-propanyl (4Z)-4- icosenoate	507.333	C25H50NO7P	MS

• ^**
  ) Based on comparison of MS spectrum with database of National Institute of Standards and Technology (NIST)

Figure 2
HPLC-DAD Base Peak Chromatogram (BPC) for Antidesma bunius extract.

5-Methoxysalicylic acid is also known as 2-hydroxy-5-methoxybenzoate. It is an extremely weak basic (nearly neutral) compound (based on its pKa). 5-Methoxysalicylic acid has been detected in herbs, spices and tea. (Dieryckx et al., 2015) However, this study results indicate that 5-Methoxysalicylic acid is also found in fruits such as A. bunius.

In this research, several fatty acid compounds were identified. (15Z)-9,12,13-Trihydroxy-15-octadecenoic acid is a long-chain fatty acid. This compound can be used for various purposes, such as antifoaming agents, coagulating agents, dispersion agents, emulsifiers, flotation agents, foaming agents, viscosity adjustors, etc. Butanedioic acid, 2-(6-hydroxyhexyl)-3-methylene- is a small acid found in Aspergillus tubingensis as its secondary metabolite. This compound has the potential to have bioactivity (Frisvad et al., 2018). However, there was no research to prove this.

Lariciresinol 4-O-glucoside is a lignan, a type of phenylpropanoids. In food, it is found in sesame seeds, flax and Brassica vegetables (Tchoumtchoua et al., 2019). It is also found in the bark and wood of white fir (Abies alba). The lignans are a large group of polyphenols found in plants and one of the major classes of phytoestrogens. It also acts as antioxidants and has potential anti-inflammatory activity (Korkina et al., 2011).

Phosphocholine is the phosphate of choline; and the parent compound of the phosphocholine family. It has a role as an epitope, a hapten, a human metabolite, a mouse metabolite, and an allergen. It is the conjugate acid of a choline phosphate (1-). Phosphocoline was also found in other fruits such as strawberry and Longan (Antunes et al., 2019; Wang et al., 2020).

There is a compound that is identified similar to Disperse red 17. Disperse dyes are a class of water-insoluble dyes that penetrate synthetic fibers and are held in place by physical forces without forming chemical bonds (Al-Etaibi & El-Apasery, 2019). It is used as a colorant, commonly in hair dyeing agents and the textile industry. Disperse red-17 is a red dye that has red powder, soluble in ethanol and acetone. Other than that, 1,3,3-trimethylindolino-6'-nitrobenzopyrylospiran also detected in this research. It is a photochromic compound. It can change their molecular structures by reversible process upon irradiation of lights of two different wavelengths. Several photochromic compounds were known to have the stability of heat (Malinauskiene et al., 2013). These results indicate the potential of Buni fruit as a source of good quality natural colorant.

Rubone was also detected in this research. This compound was also found in another plant such as Malva verticillata and Myrica nagi fruit (Bao et al., 2018; Patel & Prashar, 2020) Several studies have shown that it has vigorous anticancer activity. Rubone and paclitaxel (PTX) combination therapy retarded cancer cell growth, migration and cancer stem-like cells (CSC) population growth (Xiao & Chen, 2015).

This study also found that several compounds belong to the steroid group. 11-Deoxy prostaglandin F2β is an analog of PGF2. 8-Iso-15-keto-prostaglandin-F2β is an isomer of PGF2α of non-enzymatic origin. These results indicate the potential of Antidesma bunius fruit to be used as a functional food.

4 Conclusions

Based on the study results, it can be concluded that the volatile components present in Antidesma bunius extract identified by GC-MS method as many as fifty compounds. Among these components, 5-hydroxymethylfurfural (5-HMF) was the one found in the greatest concentration. Also, there are ten predominant components and several compounds that can be used as flavoring agents. Antidesma bunius also contained several organic acids, long-chain fatty acids, and photochromic compounds. The compounds detected in this study may have biological activities which showed the great potential of Buni fruit to be applied in several aspects such as functional food, food processing and medicine.

Acknowledgements

This research was supported by the Indonesia Endowment Fund for Education (LPDP).

• Practical Application: The study results indicate the possible use of the fruit of A. bunius for food flavoring, antimicrobial and anticancer agents.

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Data availability

Data citations

National Center for Biotechnology Information. (2020). PubChem Rockville, Bethesda: NHI. Retrieved from https://pubchem.ncbi.nlm.nih.gov

Royal Society of Chemistry. (2020). ChemSpider London: Royal Society of Chemistry. Retrieved from http://www.chemspider.com

Publication Dates

• Publication in this collection
  01 Mar 2021
• Date of issue
  2022

History

• Received
  09 Nov 2020
• Accepted
  04 Dec 2020