*2.1. Reagents*

*n*-Decanol, *<sup>n</sup>*-decyl-β-<sup>d</sup>-glucopyranoside, Amberlite XAD-2 polymeric resin (20–60 mesh), butylated hydroxyanisole (BHA), butylated hydroxy toluene (BHT), ascorbic acid, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,4,6-tri(2-pyridyl)- *s*-triazine (TPTZ) and saturated *n*-alkanes mixture (C7–C30), were purchased from Sigma-Aldrich Corp. (St. Louis, MO, USA). Authentic volatile chemicals were purchased from commercial sources (Sigma-Aldrich and Wako Pure Chemical Industries, Ltd., Osaka, Japan). The other reagents used were of analytical grade and were purchased from commercial sources.

## *2.2. Plant Materials*

*M. tricuspidata* fruits were collected in late October 2017 at a fully mature stage from plants cultivated in a farm located in Milyang district, Gyeongsangnam-do, Republic of Korea. A voucher specimen has been deposited at the Herbarium of Department of Food Science and Technology, College of Agricultural Life Science, Chonbuk National University. The fruit was freeze-dried for 4 day. The samples were powdered and stored in a freezer (−20 ◦C) until use.

#### *2.3. Isolation of Steam-Distilled Essential Oil*

A powdered sample (100 g) and distilled water (2 L) were placed ina3Lround flask. The essential oil was isolated by means of simultaneous steam distillation and extraction at atmospheric pressure in a modified Likens–Nickerson type apparatus using *n*-pentane-diethyl ether (1:1) containing *n*-decanol (950 μg) as an internal standard for2h[32]. After the isolated oil was dried over anhydrous sodium sulfate for 12 h, the solvent was concentrated to a volume of 0.5 mL using a Vigreaux column at 40 ◦C and thereafter was evaporated o ff under a stream of nitrogen. The resulting residue was redissolved in 1 mL of *n*-pentane-diethyl ether (1:1) and subjected to gas chromatography (GC) and GC–mass spectrometry (GC–MS) analysis.

#### *2.4. Isolation of Free Volatiles and Glycosidically Bound Volatiles*

The powdered sample (100 g) was homogenized with 300 mL of methanol for 1 min in a Waring blender. The homogenate was centrifuged at 4500 × g for 20 min. The residue was homogenized with 300 mL of methanol followed by centrifugation as above. The supernatant was combined and the solvent was concentrated to remove methanol under reduced pressure at 40 ◦C. The residue was dissolved in 100 mL of distilled water and was passed through a previously preactivated (with methanol) Amberlite XAD-2 (20–60 mesh) adsorbent column (5 × 35 cm) at a flow rate of 3 mL/min according to a previously reported method [33]. After the column was washed with 1.5 L of distilled water, free volatiles (FV) and glycosidically bound volatile (GBV) fraction was isolated by sequentially eluting with each 1 L of *n*-pentane:diethyl ether (1:1) and methanol, respectively. The FV fraction was dried over anhydrous sodium sulfate for 12 h and filtered through filter paper. The filtrate was concentrated to remove solvent under reduced pressure at 40 ◦C. The resulting residue was redissolved in 1 mL of *n*-pentane-diethyl ether (1:1). The methanol eluate designated as GBV was concentrated under reduced pressure to dryness at 40 ◦C. After residue was redissolved in 50 mL of 0.1 M citrate-phosphate bu ffer (pH 4.8), the aqueous layer was washed triplicate with each 50 mL of *n*-pentane:diethyl ether (1:1) to remove remaining free volatiles and added *<sup>n</sup>*-decyl-β-<sup>d</sup>-glucopyranoside (1900 μg) as an internal standard. The GBF was hydrolyzed by *Aspergillus niger* cellulase (80 mg, 24 U as β-glucosidase) at 37 ◦C for 36 h with gentle shaking. The liberated aglycones were isolated by liquid-liquid extraction using ethyl acetate (50 mL × 3). After the liberated glycosidically bound aglycone fractioin (GBAF) was dried over anhydrous sodium sulfate for 12 h, the solvent was evaporated using rotary evaporator at 40 ◦C. The resulting residue was dissolved in ethyl acetate. The extracts prepared were stored at −20 ◦C until use.

#### *2.5. Gas Chromatography (GC) and GC–Mass Spectrometry (GC–MS) Analysis*

GC analysis was performed on a Hewlett-Packard model 6890 series gas chromatograph, with a flame ionization detector (FID), a split ratio of 1:30 using Agilent J&W DB-5MS fused silica capillary column (30 m × 0.32 mm, i.d., 0.25 μm film thickness, Santa Clara, CA, USA) and Agilent J&W Supelcowax 10 fused silica capillary column (30 m × 0.32 mm, i.d., 0.25 μm film thickness). The column temperatures were programmed from 50 ◦C to 230 ◦C at 2 ◦C/min and then kept constant at 230 ◦C for 20 min. The injector and detector temperatures were 250 ◦C, respectively. The carrier gas was nitrogen, at a flow rate of 1.0 mL/min. Peak areas were measured by electronic integration and the concentrations of volatile compounds were expressed as *n*-decanol equivalent (assuming response factor of all analytes was 1.0). The concentrations are to be considered only relative values as recovery after extraction and calibration factors related to the standard were not determined [34,35].

The GC–MS analysis was performed on an Agilent Technologies 7890A GC and 5975C mass selective detector operating in the EI mode at 70 eV, fitted with a DB-5MS fused silica capillary column (30 m × 0.25, i.d., 0.25 μm film thickness) and Supelcowax 10 fused silica capillary column (30 m × 0.32 mm, i.d., 0.25 μm film thickness), respectively. Both column temperatures were programmed from 50 ◦C to 230 ◦C at 2 ◦C per minute and then kept constant at 230 ◦C for 20 min. The injector and ion source temperatures were 250 ◦C. The carrier gas was helium at a flow rate of 1.0 mL/min. Identification of the compounds was achieved by comparing their retention times with those of authentic standards and mass spectral data in Wiley7n,1 database (Hewlett-Packard, Palo Alto, CA, USA), and NIST (National Institute of Standards and Technology, USDA) Webbook, and reported retention indices in the literatures [36]. Retention indices of each compound was calculated by a homologous series of saturated *n*-alkanes (C7–C30) (concentration of 1000 μg/mL in *n*-hexane) under the same conditions [37]. All compounds identified based on comparisons of only mass spectral data were listed as tentatively identified.

#### *2.6. Determination of Total Phenolic Content*

Total phenol content of the sample was measured according to the method described by Chandra et al. [38] with some modifications. Briefly, 20 μL of each fraction (at concentration of 1000 μg/<sup>1</sup> mL methanol) was mixed with 50% Folin–Ciocalteu phenol reagen<sup>t</sup> (20 μL) in 96-well plates. After 5 min, 1 N sodium carbonate solution (20 μL) was added to the mixture and distilled water was added to adjust the final volume to 200 μL. After incubation at room temperature (RT) in the dark for 30 min, the absorbance of test sample against a blank was measured at 725 nm using a VersaMax enzyme-linked immunosorbent assay (ELISA) microplate reader (Molecular Devices, LLC, San Jose, CA, USA). Total phenol content was calculated based on a calibration curve of gallic acid. The results were expressed as mg gallic acid equivalent (mg GAE)/g.
