Skip to main content
NCBI home page
PeerJ logoLink to PeerJ
. 2021 Nov 24;9:e12513. doi: 10.7717/peerj.12513

Chemical profiling of root bark extract from Oplopanax elatus and its in vitro biotransformation by human intestinal microbiota

Jin-Yi Wan 1,#, Jing-Xuan Wan 1,#, Shilei Wang 2, Xiaolu Wang 1, Wenqian Guo 1, Han Ma 1, Yuqi Wu 1, Chong-Zhi Wang 3,✉, Lian-Wen Qi 2, Ping Li 2, Haiqiang Yao 1,✉, Chun-Su Yuan 3
Editor: Rogerio Sotelo-Mundo
PMCID: PMC8627129  PMID: 34900430

Abstract

Oplopanax elatus (Nakai) Nakai, in the Araliaceae family, has been used in traditional Chinese medicine (TCM) to treat diseases as an adaptogen for thousands of years. This study established an ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) method to identify chemical components and biotransformation metabolites of root bark extract from O. elatus. A total of 18 compounds were characterized in O. elatus extract, and 62 metabolites by human intestinal microbiota were detected. Two polyynes, falcarindiol and oplopandiol were recognized as the main components of O. elatus, whose metabolites are further illustrated. Several metabolic pathways were proposed to generate the detected metabolites, including methylation, hydrogenation, demethylation, dehydroxylation, and hydroxylation. These findings indicated that intestinal microbiota might play an essential role in mediating the bioactivity of O. elatus.

Keywords: Oplopanax elatus, Intestinal microbiota, UPLC-Q-TOF/MS, Metabolic profiles, Biotransformation

Introduction

Oplopanax elatus (Nakai) Nakai is the plant of genus Oplopanax, which belongs to the Araliaceae family. It is mainly distributed in northeast China, Korea and far east of Russia (Dou et al., 2009; Yang et al., 2010). As a traditional medicinal plant, O. elatus is being utilized as a ginseng-like herbal medicine and has been long used as an adaptogen to treat arthritis, diabetes mellitus, rheumatism, neurasthenia, and cardiovascular diseases (Dai et al., 2016; Eom et al., 2017; Knispel et al., 2013; Moon et al., 2013; Panossian et al., 2021). Previous studies have identified several components derived from O. elatus, such as the lignans, saponins, phenolic glycosides, and polyynes (Huang et al., 2010; Shao et al., 2016). To date, polyynes have been chiefly reported with high contents in the root of O. elatus (Huang et al., 2014a). Increasing attention has been paid to two main polyynes facarindiol (FAD) and oplopandiol (OPD), because of their significant anti-tumor activities (Purup, Larsen & Christensen, 2009; Qiao et al., 2017; Sun et al., 2016). However, most studies remain focused on the pharmacological and chemical constituents of O. elatus, while its metabolic profiles are rather obscured.

It is widely known that human beings live in symbiotics with coevolutionary microbiota (Thursby & Juge, 2017; Yang & Lao, 2019). The human gastrointestinal tract is the primary habitat for trillions of microbes. The gut microbiota serves metabolic functions crucial for the human host (Bäckhed et al., 2004; Chen et al., 2016; Rajilić-Stojanović & de Vos, 2014) and influences the biofunctions (Barko et al., 2018; Defois et al., 2018; Pagliari et al., 2017). Like most herbal medicines, O. elatus products are orally administered. The multiple constituents of O. elatus are typically brought into contact with intestinal bacteria and subsequently transformed in the digestive tract (Gao et al., 2018; Huang et al., 2014b; Koppel, Maini Rekdal & Balskus, 2017; Shikov et al., 2014; Teschke et al., 2015). However, existing reports did not address intestinal microflora’s biotransformed metabolites of O. elatus. Therefore, elucidating how gut microbes treat these complex components may contribute to a complete understanding of the metabolic profiles and biological activities of O. elatus.

Recently, various analytical platforms are typically applied to identify metabolic profiles in the complex extracts of TCMs. Most notably, ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) is one of the powerful analytical tools (Jin et al., 2018; Wu et al., 2019; Yang et al., 2016). With the newly developed chromatographic technique, the UPLC system allows significant improvements in the resolution, analysis speed, and reduction of solvent waste (Chekmeneva et al., 2018; Du et al., 2017; Wang et al., 2008). Meanwhile, high-resolution Q-TOF/MS can give more specific and accurate mass information on characteristic molecular ions and fragment ions, providing a reliable basis for the qualitative analysis of complex samples (Li et al., 2019; Lou et al., 2015; Wewer et al., 2011). Based on these characteristics, UPLC-Q-TOF/MS was ultimately selected for fast identification of constituents in O. elatus.

In the present study, we focused on the metabolic behavior of human intestinal microflora on O. elatus. A highly selective and sensitive UPLC-Q-TOF/MS method was established to characterize the chemical and metabolic profiles of O. elatus. Furthermore, the proposed metabolic pathways were also summarized. This work will provide a better understanding for exploring the bioactivities of O. elatus in vivo.

Materials & methods

Materials and reagents

The general anaerobic medium for bacteria culture was obtained from Shanghai Kayon Biological Technology Co. Ltd. (Shanghai, China). Formic acid and HPLC-grade acetonitrile were purchased from Merck (Darmstadt, Germany). Deionized water (18 MΩ·cm) was supplied with a Millipore Milli-Q water system (Milford, MA, USA). All other reagents were from standard commercial sources and of analytical purity.

Preparation of Oplopanax elatus extract

Root bark of O. elatus was obtained from Benxi city (Liaoning, China). The voucher samples were deposited at the Tang Center for Herbal Medicine Research at the University of Chicago (Chicago, IL, USA). The air-dried root bark of O. elatus was pulverized into powder and sieved through an 80-mesh screen. Eight g of the powder were extracted twice by heat-reflux with 70% ethanol for 2 h. The combined extract was evaporated under vacuum and lyophilized with a yield of 28%. The samples were stored at 4 °C until use.

Preparation of human intestinal microflora

The Institutional Review Board approved the present study protocol at the University of Chicago (IRB protocol number: 12536). Fresh fecal samples were collected from six healthy adult volunteers (male, aged 20–55, non-smokers without antibiotic consumption for more than 6 months, and written consent was obtained). All the fecal samples were mixed for analysis. A total of five g of samples were homogenized in 30 ml cold physiological saline, and centrifuged at 13,000 rpm for 10 min to obtain the resulting fecal supernatant.

Incubation of sample in intestinal bacteria

Two microliters of the fecal supernatant were added with eight ml anaerobic dilution medium containing five mg of O. elatus extract, which were then anaerobically incubated at 37 °C for 24 h in an anaerobic workstation (Electrotek, UK). The reaction mixtures were extracted three times with water-saturated n-butanol. All the n-butanol layers were mixed and dried under a nitrogen stream and then dissolved in one ml methanol. The solutions were centrifuged at 13,000 rpm for 10 min for analysis.

UPLC-Q-TOF/MS analysis

Data were collected as previously described (Wang et al., 2020). The Agilent 1290 Series UPLC system (Agilent Technologies, Santa Clara, CA, USA) was applied to perform the chromatographic analysis, and a binary pump, an online degasser, an auto plate-sampler, and a thermostatically controlled column compartment were also equipped for this system. The separation was carried out on UPLC ACQUITY HSS C8 column (2.1 mm × 100 mm × 1.7 μm, Waters) with a constant flow rate of 0.4 mL/min, and the column temperature was kept at 40 °C. A gradient mobile phase system of 0.1% formic acid in water (phase A) and acetonitrile (phase B) was applied as follows: 5% B at 0–1 min, 5–20% B at 1–18 min, 20–30% B at 18–27 min, 30–35% B at 27–32 min, 35–60% B at 32–40 min, 60–95% B at 40–50 min, 95% B at 50–53 min, 95–5% B at 53–55 min. The injection volume of samples was set at two μL for MS mode and five μL for MS/MS mode.

The Agilent 6545 Q-TOF-MS system with a Dual electrospray ionization source was used to conduct the detection. Nitrogen (purity > 99.999%) served as a sheath gas and drying gas, and the flow velocities were set at 11 and 8 L/min. The temperatures of sheath gas and drying gas were set at 350 and 320 °C respectively. Positive and negative ion modes were both employed in this study. The other parameters were set as follows: nebulizer pressure, 35 psig; voltage, 3,500 V; fragmentor voltage, 175 V; mass range, m/z 100–1,700; data acquisition rate, 1.5 scans/s; MS/MS spectra collision energy, 50 eV (Wang et al., 2020).

Data analysis

Mass data were analyzed by the Agilent MassHunter Workstation software (Version B.06.01), based on the accurate measurements of m/z values with online databases (MassBank, etc.), to screen probable compounds. The empirical molecular formula was deduced by comparing the theoretical mass of molecular ions at the mass accuracy of less than five ppm.

Results

Optimization of UPLC-Q-TOF/MS conditions

To obtain the chromatograms with better resolution and higher baseline stability of O. elatus extract and its primary metabolites, multiple mobile phases such as acetonitrile-water and methanol-water were detected. Acetonitrile-water was applied as the solvent, for its stronger separation ability, shorter retention time, and lower column pressure. Additionally, 0.1% formic acid added in the water as mobile phase adducts may help to achieve higher response and better peak sensitivity (Tao et al., 2016). Therefore, the optimal solvent system consisting of acetonitrile-water (0.1% formic acid), which remarkably enhanced the efficiency of ionization and satisfactory sensitivity, was ultimately selected as mobile phase with a gradient elution.

In addition, the factors related to MS performance, including ionization mode and collision energy, were further improved. The positive ion mode was ultimately employed to gain comprehensive data for structural characterization and metabolite assignment with much lower background noise. The collision energy was optimized to obtain the higher ionization efficiency and relative abundance of precursor and product ions.

Chemical profiling of O. elatus extract

In total, 18 ingredients of O. elatus were detected in this study, and their chemical structures are shown in Fig. 1. There are six types of compounds, including nine polyynes, three lignans, one phenylpropanoid, two sesquiterpenes, one triterpenoid, and two fatty acids. The total ion chromatogram (TIC) of O. elatus extract is shown in Fig. 2A in the positive ion mode by UPLC-Q-TOF-MS. Table 1 shows the detailed information, including retention time, signal intensity, molecular formula, calculated and experimental mass m/z, ppm error, and fragment ions of these 18 components (Schymanski et al., 2014; Wang et al., 2020).

Figure 1. The chemical structures of bioactive compounds detected in Oplopanax elatus extract.

Figure 1

Open in a new tab

(A) Polyynes; (B) Lignans; (C) Phenylpropanoid; (D) Sesquiterpenes; (E) Triterpenoid; (F) Fatty acids.

Figure 2. UPLC-TOF/MS profiles of O. elatus extract in the positive ion mode.

Figure 2

Open in a new tab

(A) Total ion chromatogram (TIC) of O. elatus extract. (B) TIC of blank sample including dilution medium and human fecal microflora. (C) TIC of biotransformed O. elatus sample by intestinal bacteria.

Table 1. UPLC-Q-TOF/MS data of bioactive components of O. elatus extract in the positive ion mode (Dou et al., 2009; Shikov et al., 2014; Wang et al., 2020).

No. Compound Formula tR (min) Signal intensity (×105) [M+H]+ or [M+Na]+ Fragment ions in the positive mode with the energy 50 V CID
m/z Calc m/z Diff (ppm)
1 4-(3-hydroxyprop-1-en-1-yl)-2,6-dimethoxyphenyl β-D-glucopyranoside C17H24O9 5.24 2.86 ± 0.82 373.1495 373.1493 −0.51 211.1526[M-glc+H]+, 373.1495[M+H]+
2 4’,7-epoxy-4,9,9’-trihydroxy-3,3’-dimethoxy-5’,8-lignan-4,9-bis[O-β-D-glucopyranoside] C32H44O16 10.92 1.74 ± 0.34 707.2521 707.2522 0.08 545.1981[M-glc+Na]+, 707.2521[M+Na]+
3 isolariciresinol 3-O-β-D-glucopyranoside C26H34O11 14.14 0.55 ± 0.17 545.1997 545.1993 −0.70 383.1428[M-glc+Na]+, 545.1997[M+Na]+
4 5-methoxylariciresinol 4-O-β-D-glucopyranoside C27H36O12 15.04 0.89 ± 0.30 575.2112 575.2099 −2.36 412.1434[M-C10H11O2+Na]+, 250.0381[M-C10H11O2-glc+Na]+, 575.2112[M+Na]+
5 2-decenoic acid C10H18O2 23.41 5.62 ± 1.74 171.1378 171.1380 0.92 55.9342[M-HCOOH-C5H10+H]+, 171.1378[M+H]+
6 oploxyne B C18H30O4 36.91 2.73 ± 0.36 311.2212 311.2217 1.57 107.0527[M-H2O-C11H22O2+H]+, 79.0547[M-H2O-C11H22O2-C2H4+H]+, 311.2212[M+H]+
7 oploxyne A C17H26O3 37.62 2.62 ± 0.44 279.1957 279.1955 −0.82 107.0485 [M-H2O-C10H18O+H]+, 79.0545[M-H2O-C10H18O-C2H4+H]+, 279.1957[M+H]+
8 oplopantriol B C18H28O3 37.89 1.28 ± 1.07 293.2110 293.2111 0.41 107.0491[M-H2O-C11H20O+H]+, 79.0538[M-H2O-C11H20O-C2H4+H]+, 293.2110[M+H]+
9 9,17-octadecadiene-12,14-diyne-1,11,16-triol,1-acetate C20H28O4 38.63 1.57 ± 1.21 333.2048 333.2060 3.72 105.0700[M-H2O-C13H22O2+H]+, 79.0546[M-H2O-C13H22O2-C2H2+H]+, 333.2048 [M+H]+
10 oplopandiol acetate C20H30O4 39.18 3.93 ± 1.41 335.2215 335.2217 0.56 107.0503[M-H2O-C13H22O2+H]+, 79.0545[M-H2O-C13H22O2-C2H4+H]+, 335.2215[M+H]+
11 6,9-octadedicenoic acid C18H32O2 39.46 5.13 ± 1.95 281.2470 281.2475 1.81 65.0396[M-CH3COOH-C11H20+H]+, 281.2470[M+H]+
12 falcarindiol C17H24O2 40.76 101.47 ± 12.16 261.1848 261.1849 0.41 105.0713[M-H2O-C10H18+H]+, 79.0548[M-H2O-C10H18-C2H2+H]+, 261.1848 [M+H]+
13 oplopandiol C17H26O2 41.23 119.24 ± 9.28 263.2010 263.2006 −1.69 107.0459[M-H2O-C10H18+H]+, 79.0563[M-H2O-C10H18-C2H4+H]+, 263.2010[M+H]+
14 falcarinol C17H24O 42.85 62.85 ± 7.26 245.1899 245.1900 0.38 105.0699[M-C10H18+H]+, 79.0556[M-C10H18-C2H2+H]+, 245.1899 [M+H]+
15 oplopantriol A C18H26O3 43.31 67.83 ± 4.40 291.1956 291.1955 0.44 105.0659[M-H2O-C11H20O+H]+, 79.0567[M-H2O-C11H20O-C2H2+H]+, 291.1956 [M+H]+
16 curcumene C15H22 44.92 4.37 ± 2.22 203.1797 203.1794 −1.35 134.1063[M-C5H9+H]+, 65.0382[M-2×C5H9+H]+, 203.1797[M+H]+
17 muurolene C15H24 45.27 8.27 ± 3.82 205.1950 205.1951 0.38 65.0550[M-2×C5H9+H]+, 205.1950[M+H]+
18 oleanolic acid C30H48O3 46.88 10.83 ± 3.28 479.3508 479.3496 −2.70 231.1715[M-C16H24O2+Na]+, 479.3508[M+Na]+
Open in a new tab

Polyynes have been found as the main constituents in the root of O. elatus (Yang et al., 2014). Among them, falcarindiol and oplopandiol were determined to have very high contents in the air-dried root bark. As shown in Table 1, polyynes exhibit the same elemental composition and similar MS/MS behaviors, with the characteristic fragment ions at m/z 79.05 in the positive ion mode.

For example, the typical protonated molecular ion [M+H]+ of FAD was observed at m/z 261.1848 in the mass spectrum. The fragment ion at m/z 105.0713 was formed by the losses of H2O and C10H18, with m/z 79.0548 by further loss of C2H2. OPD was identified by the protonated molecular ion [M+H]+ at m/z 263.2010 compared with calculated m/z 263.2006. The fragment ion at m/z 107.0459 was produced by the losses of H2O and C10H18, and m/z 79.0563 was formed by further loss of C2H4.

In addition, phenylpropanoid compound 4-(3-hydroxyprop-1-en-1-yl)-2,6-dimethoxyphenyl β-D-glucopyranoside was determined to be C17H24O9 at m/z 373.1495 ([M+H]+, C17H24O+9; calculated as 373.1493). The neutral loss of 1 × Glc moiety formed fragment ion at m/z 211.1526. Similarly, three lignans were determined by the neutral loss of 1 × Glc moiety in the [M+Na]+ mode.

Detection and identification of metabolites of O. elatus extract

The control sample was prepared in parallel, which used in the dilution medium and human fecal microflora, as shown in Fig. 2B. The biotransformed O. elatus sample by intestinal bacteria is shown in Fig. 2C. Samples were incubated, pretreated, and analyzed under the same conditions as mentioned in “Incubation of sample in intestinal bacteria”. The potential metabolites were detected from the TIC of the transformed O. elatus sample compared to the control group. All the metabolites were further confirmed by the extracted ion chromatograms (EICs) and their MS/MS corresponding fragments. A total of 62 metabolites were identified by UPLC-Q-TOF-MS in the positive mode. Table 2 shows the retention time, signal intensity, experimental and calculated mass m/z, difference between m/z and calculated m/z in ppm, and fragment ions in the MS/MS stage of these 62 metabolites (M1-M62). All these metabolites could not be observed or only in trace amounts in control samples (Schymanski et al., 2014).

Table 2. UPLC-Q-TOF/MS data of metabolites detected from the biotransformed O. elatus sample in the positive ion mode.

No. Description Formula tR
(min)		 Signal intensity (×105) [M+H]+ or [M+Na]+ Fragment ions in the positive mode with the energy 50 V CID
m/z Calc m/z Diff (ppm)
M1 deglycosylation product of 4-(3-hydroxyprop-1-en-1-yl)-2,6-dimethoxyphenyl β-D-glucopyranoside C11H14O4 9.90 2.83 ± 0.32 211.0962 211.0965 1.36 92.0582[M-C3H5O-OCH3×2+H]+, 211.0962[M+H]+
M2 acetylization product of 9,17-octadecadiene-12,14-diyne-1,11,16-triol,1-acetate C22H30O5 16.55 16.27 ± 2.46 375.2170 375.2166 −1.07 79.0544[M-C13H24O3-C2H2+H]+, 375.2170[M+H]+
M3 deglycosylation product of 4’,7-epoxy-4,9,9’-trihydroxy-3,3’-dimethoxy-5’,8-lignan-4,9-bis[O-β-D-glucopyranoside] C20H24O6 20.06 11.37 ± 2.01 383.1470 383.1465 −1.36 188.1603 [M-C10H11O4+Na]+, 383.1470 [M+Na]+
M4 deglycosylation product of isolariciresinol 3-O-β-D-glucopyranoside C20H24O6 20.08 11.32 ± 0.98 383.1464 383.1465 0.30 167.4648 [M-C10H12O3-2×H2O+Na]+, 383.1464[M+Na]+
M5 deglycosylation product of 5-methoxylariciresinol 4-O-β-D-glucopyranoside C21H26O7 22.65 14.28 ± 1.42 413.1578 413.1571 −1.86 231.8773 [M-C9H10O4+Na]+, 413.1578 [M+Na]+
M6 hydroxylation product of oplopandiol acetate C20H30O5 30.58 0.72 ± 0.43 351.2153 351.2166 3.71 107.0840 [M-H2O-C13H22O3+H]+, 79.0547[M-H2O-C13H22O3-C2H4+H]+, 351.2153[M+H]+
M7 hydroxylation product of oploxyne B C18H30O5 30.64 3.02 ± 0.55 327.2163 327.2166 0.92 107.0851 [M-H2O-C11H22O3+H]+, 79.0543 [M-H2O-C11H22O3 -C2H4+H]+, 327.2163[M+H]+
M8 acetylization product of oploxyne B C20H32O5 31.64 1.82 ± 0.63 353.2321 353.2323 0.43 107.0847 [M-H2O-C13H24O3+H]+, 79.0545[M-H2O-C13H24O3-C2H4+H]+, 353.2321[M+H]+
M9 hydroxylation product of curcumene C15H22O 32.61 42.47 ± 4.29 219.1745 219.1743 0.73 63.0237[M-2×C5H9-H2O+H]+, 219.1745 [M+H]+
M10 hydroxylation product of 9,17-octadecadiene-12,14-diyne-1,11,16-triol,1-acetate C20H28O5 34.08 0.98 ± 0.26 349.1994 349.2010 4.45 105.0700[M-H2O-C13H22O3+H]+, 79.0573[M-H2O-C13H22O3-C2H2+H]+, 349.1994 [M+H]+
M11 demethylation product of oploxyne B C17H28O4 34.24 15.83 ± 3.11 297.2061 297.2060 −0.22 107.0508 [M-H2O-C10H20O2+H]+, 79.0554[M-H2O-C10H20O2-C2H4+H]+, 297.2061[M+H]+
M12 dehydroxylation product of 2-decenoic acid C10H18O 34.27 1.36±0.42 155.1431 155.1430 −0.38 56.9427[M-CHO-C5H10+H]+, 155.1431[M+H]+
M13 hydroxylation product of oplopantriol A C18H26O4 34.90 1.54 ± 0.72 307.1902 307.1904 0.61 105.0682[M-H2O-C11H20O2+H]+, 79.0539 [M-H2O-C11H20O2-C2H2+H]+, 307.1902[M+H]+
M14 dehydroxylation product of oplopantriol A C18 H26O2 34.92 52.73 ± 3.40 275.2003 275.2006 0.94 105.0698[M-H2O-C11H20+H]+, 79.0541[M-H2O-C11H20-C2H2+H]+, 275.2003[M+H]+
M15 methylation product of falcarindiol C18H26O2 34.98 49.23 ± 2.49 275.2008 275.2006 −0.89 105.0704[M-H2O-C11H20+H]+,79.0548[M-H2O-C11H20-C2H2+H]+, 275.2008[M+H]+
M16 dehydroxylation product of oplopandiol acetate C20H30O3 35.25 37.62 ± 4.21 319.2256 319.2268 3.68 107.0525 [M-H2O-C13H22O+H]+, 79.0538[M-H2O-C13H22O-C2H4+H]+, 319.2256[M+H]+
M17 hydrogenation product of falcarindiol C17H26O2 35.89 1.36 ± 0.77 263.2008 263.2006 −0.93 105.0695[M-H2O-C10H20+H]+,79.0535[M-H2O-C10H20-C2H2+H]+, 263.2008[M+H]+
M18 acetylization product of oplopantriol B C20H30O4 37.06 12.48 ± 2.41 335.2202 335.2217 4.45 107.0855 [M-H2O-C13H22O2+H]+,79.0543[M-H2O-C13H22O2-C2H4+H]+,335.2202[M+H]+
M19 hydrogenation product of oploxyne A C18H32O4 37.63 7.37 ± 2.49 313.2375 313.2373 −0.53 107.0575[M-H2O-C11H24O2+H]+, 79.0541[M-H2O-C11H24O2-C2H4+H]+, 313.2375[M+H]+
M20 hydroxylation product of muurolene C15H24O 37.76 13.47 ± 3.57 221.1901 221.1900 −0.49 65.0374[M-2×C5H9-H2O+H]+, 221.1901[M+H]+
M21 hydrogenation product of 9,17-octadecadiene-12,14-diyne-1,11,16-triol,1-acetate C20H30O4 37.77 17.38 ± 4.76 335.2204 335.2217 3.85 105.0694[M-H2O-C13H24O2+H]+, 79.0543[M-H2O-C13H24O2-C2H2+H]+, 335.2204 [M+H]+
M22 acetylization product of oplopantriol A C20H28O4 37.90 2.55 ± 0.77 333.2047 333.2060 4.02 105.0705[M-H2O-C13H22O2+H]+, 79.0524[M-H2O-C13H22O2-C2H2+H]+, 333.2047[M+H]+
M23 hydrogenation product of oplopandiol acetate C20H32O4 37.91 12.42 ± 3.28 337.2358 337.2373 4.57 79.0538[M-C13H26O3-C2H4+H]+, 337.2358[M+H]+
M24 hydroxylation product of 6,9-octadedicenoic acid C18H32O3 37.92 17.52 ± 2.71 297.2423 297.2424 0.41 77.0379[M-CH3COOH-C10H20O+H]+, 297.2423[M+H]+
M25 hydrogenation product of oplopantriol B C18 H30O3 38.22 24.52 ± 3.98 295.2263 295.2268 1.60 79.0557[M-C11H24O2-C2H4+H]+, 295.2263[M+H]+
M26 demethylation product of falcarindiol C16H22O2 38.59 2.26 ± 0.74 247.1689 247.1693 1.45 105.0686[M-H2O-C9H16+H]+, 79.0528[M-H2O-C9H16-C2H2+H]+, 247.1689[M+H]+
M27 hydroxylation product of oplopandiol C17H26O3 38.64 50.12 ± 4.62 279.1958 279.1955 −1.18 107.0496[M-H2O-C10H18O+H]+, 79.0541[M-H2O-C10H18O-C2H4+H]+, 279.1958[M+H]+
M28 acetylization product of oplopandiol acetate C22H32O5 39.27 4.92 ± 0.44 377.2326 377.2323 −0.93 79.0543[M-C15H26O4-C2H4+H]+, 377.2326[M+H]+
M29 hydroxylation product of falcarinol C17H24O2 39.54 4.39 ± 0.58 261.1851 261.1849 −0.74 79.0538[M-C10H20O-C2H2+H]+, 261.1851 [M+H]+
M30 hydroxylation product of oploxyne A C17H26O4 39.60 2.74 ± 0.94 295.1902 295.1904 0.63 79.0557[M-C10H20O3-C2H4+H]+, 295.1902[M+H]+
M31 demethylation product of falcarinol C16H22O 39.61 1.02 ± 0.57 231.1744 231.1743 −0.25 79.0544[M-C9H18-C2H2+H]+, 231.1744[M+H]+
M32 demethylation product of oplopantriol B C17H26O3 39.69 33.79 ± 4.95 279.1965 279.1955 −3.70 79.0542[M-C10H20O2-C2H4+H]+, 279.1965[M+H]+
M33 hydrogenation product of oploxyne A C17H28O3 40.06 10.32 ± 1.45 281.2113 281.2111 −0.64 79.0541[M-C10H22O2-C2H4+H]+, 281.2113[M+H]+
M34 hydroxylation product of oplopantriol B C18H28O4 40.07 5.28 ± 1.82 309.2062 309.2060 −0.53 79.0560[M-C11H22O3-C2H4+H]+, 309.2062[M+H]+
M35 demethoxy product of oploxyne B C17H28O3 40.07 9.24 ± 1.23 281.2112 281.2111 −0.28 79.0541[M -C10H22O2-C2H4+H]+, 281.2112[M+H]+
M36 hydrogenation product of oplopantriol A C18 H28O3 40.09 2.83 ± 0.75 293.2109 293.2111 0.76 79.0538 [M-C11H24O2-C2H2+H]+, 293.2109[M+H]+
M37 demethylation product of oleanolic acid C29H46O3 40.18 0.54 ± 0.26 443.3524 443.3520 −0.97 165.0912[M-C16H24O2-2×CH3+H]+, 443.3524[M+H]+
M38 demethylation product of curcumene C14H20 40.18 2.42 ± 0.29 189.1636 189.1638 0.94 51.0229[M-2×C5H9+H]+, 189.1636[M+H]+
M39 dehydroxylation product of oploxyne A C17H26O2 40.29 0.51 ± 0.23 263.2005 263.2006 0.22 79.0552[M-C10H20O-C2H4+H]+, 263.2005[M+H]+
M40 demethylation product of oplopandiol C16H24O2 40.48 7.12 ± 0.44 249.1851 249.1849 −0.78 107.0863[M-H2O-C9H14+H]+, 79.0543[M-H2O-C9H14-C2H4+H]+, 249.1851[M+H]
M41 hydrogenation product of falcarinol C17H26O 40.50 22.15 ± 2.35 247.2053 247.2056 1.39 79.0528[M-C10H22-C2H2+H]+, 247.2053 [M+H]+
M42 methylation product of oplopandiol acetate C21H32O4 40.51 1.10 ± 0.77 349.2373 349.2373 0.10 79.0541[M-C14H26O3-C2H4+H]+, 349.2373[M+H]+
M43 dehydroxylation product of oplopandiol C17H26O 40.53 20.63 ± 3.86 247.2060 247.2056 −1.45 107.0513[M-C10H20+H]+, 79.0528[M-C10H20-C2H4+H]+, 247.2060[M+H]+
M44 dehydroxylation product of falcarinol C17H24 40.53 4.76 ± 1.42 229.1953 229.1951 −0.98 77.0398[M-C9H18-C2H2+H]+, 229.1953 [M+H]+
M45 demethylation product of muurolene C14H22 40.55 4.97 ± 0.99 191.1796 197.1794 −0.91 53.0384[M-2×C5H9+H]+, 191.1796 [M+H]+
M46 dehydroxylation product of falcarindiol C17H24O 41.18 67.22 ± 3.74 245.1903 245.1900 −1.26 105.0692[M-C10H20+H]+, 79.0521[M-C10H20-C2H2+H]+, 245.1903[M+H]+
M47 methylation product of oplopantriol A C19H28O3 41.41 0.52 ± 0.28 305.2111 305.2113 −0.59 79.0547[M-C12H24O2-C2H2+H]+, 305.2111 [M+H]+
M48 demethylation product of oplopantriol A C17H24O3 41.42 53.27 ± 3.42 277.1800 277.1798 −0.65 79.0537[M-C10H20O2-C2H2+H]+, 277.1800[M+H]+
M49 methylation product of oplopandiol C18H28O2 41.43 51.35 ± 2.53 277.2160 277.2162 0.75 107.0871[M-H2O-C11H20+H]+, 79.0547[M-H2O-C11H20-C2H4+H]+, 277.2160[M+H]+
M50 dehydroxylation product of oplopantriol B C18H28O2 41.44 53.87 ± 4.21 277.2164 277.2162 −0.70 79.0541[M-C11H22O-C2H4+H]+, 277.2164[M+H]+
M51 methylation product of falcarinol C18H26O 41.59 3.75 ± 0.89 259.2057 259.2056 −0.23 79.0524[M-C11H22-C2H2+H]+, 259.2057 [M+H]+
M52 methylation product of oplopantriol B C19 H30O3 41.69 1.26 ± 0.82 307.2262 307.2268 1.87 79.0543[M-C12H24O2-C2H4+H]+, 307.2262[M+H]+
M53 methylation product of oploxyne B C19H32O4 41.76 0.37 ± 0.20 325.2371 325.2373 0.73 79.0540[M-C12H26O3 -C2H4+H]+, 325.2371[M+H]+
M54 dehydroxylation product of 9,17-octadecadiene-12,14-diyne-1,11,16-triol,1-acetate C20H28O3 42.15 28.46 ± 1.55 317.2096 317.2111 4.81 79.0537[M-C13H24O2-C2H2+H]+, 317.2096 [M+H]+
M55 dehydroxylation product of oploxyne B C18H30O3 42.16 56.82 ± 2.47 295.2273 295.2268 −1.80 79.0550 [M-C11H24O2 -C2H4+H]+, 295.2273[M+H]+
M56 hydroxylation product of falcarindiol C17H24O3 42.20 79.26 ± 1.34 277.1797 277.1798 0.44 105.0693[M-H2O-C10H18O+H]+,79.0541[M-H2O-C10H18O-C2H2+H]+, 277.1797[M+H]+
M57 methylation product of 9,17-octadecadiene-12,14-diyne-1,11,16-triol,1-acetate C21H30O4 42.52 0.42 ± 0.28 347.2213 347.2217 1.11 79.0541[M-C14H26O3-C2H2+H]+, 347.2213 [M+H]+
M58 hydrogenation product of curcumene C15H24 42.65 70.36 ± 4.29 205.1953 205.1951 −1.09 67.0540[M-2×C5H9+H]+, 205.1953[M+H]+
M59 demethylation product of 6,9-octadedicenoic acid C17H30O2 42.75 0.48 ± 0.32 267.2327 267.2319 −3.17 65.0390[M-CH3COOH-C10H18+H]+, 267.2327[M+H]+
M60 dehydroxylation product of 6,9-octadedicenoic acid C18H32O 43.40 65.77 ± 5.21 265.2536 265.2526 −3.81 69.0688[M-CH3CHO-C11H20+H]+, 265.2536[M+H]+
M61 hydrogenation product of 6,9-octadedicenoic acid C18H34O2 43.50 128.46 ± 8.42 283.2636 283.2632 −1.57 65.0389[M-CH3COOH-C11H22+H]+, 283.2636[M+H]+
M62 acetylization product of oleanolic acid C32H50O4 48.23 2.48 ± 1.63 521.3606 521.3601 −0.94 220.0842[M-C16H24O2-C2H3O+Na]+, 521.3606[M+Na]+
Open in a new tab

Polyynes

A total of 46 metabolites of nine polyynes generated by the transformation of human intestinal microflora were detected and identified. For each polyyne, at least four types of metabolites were identified. Due to the high biological activities, FAD and OPD selected as the representative compounds of polyynes were stated in detail.

The EICs and MS/MS spectrums of metabolites of FAD are shown in Fig. 3. Five metabolites including M15, M17, M26, M46, M56 were detected. M15 was assigned to be the methylation product of FAD with the molecular formula C18H26O2 at m/z 275.2008 ([M+H]+, C18H26O+2; calculated as 275.2006). The fragment ion at m/z 105.0704 was generated by the neutral losses of H2O and C11H20, and m/z 79.0548 was formed by further loss of C2H2. M17 was assigned to be the hydrogenation product of FAD with the characteristic fragment ions at m/z 105.0695 and 79.0535. In addition, metabolites M26, M46, and M56 were assigned as demethylation, dehydroxylation, and hydroxylation products of FAD, respectively.

Figure 3. Metabolites of falcarindiol using UPLC-TOF/MS in the positive ion mode.

Figure 3

Open in a new tab

(A) Extracted ion chromatograms (EICs); (B) MS/MS spectra and structural elucidation.

Figure 4 presents the EICs and MS/MS spectrums of OPD metabolites (M27, M40, M43, and M49). M27 was assigned as the hydroxylation product of OPD, owing to the presence of [M+H]+ at m/z 279.1958. The characteristic fragment ion at m/z 107.0496 was formed by the neutral losses of H2O and C10H18O, and m/z 79.0541 was formed by further loss of C2H4. Similarly, three other metabolites like M40, M43, and M49 were supposed to be the demethylation, dehydroxylation, and methylation products of OPD.

Figure 4. Metabolites of oplopandiol using UPLC-TOF/MS in the positive ion mode.

Figure 4

Open in a new tab

(A) EICs; (B) MS/MS spectra and structural elucidation.

Lignans

M3–M5 were the deglycosylation products of three lignans via the loss of glycose moieties. For example, the parent compound of M3 was determined to be C32H44O16 while M3 was C20H24O6, indicating M3 was the deglycosylation product via the loss of two glucose moieties. M4 and M5 were assigned as the products by losing a glucose moiety from their corresponding parent lignan compounds.

Phenylpropanoids

M1 was identified as the deglycosylation metabolite of phenylpropanoid compound 4-(3-hydroxyprop-1-en-1-yl)-2,6-dimethoxyphenyl β-D-glucopyranoside. The protonated molecular ion [M+H]+ of M1 at m/z 211.0962 was observed in the positive ion mode, providing the molecular formula of C11H14O4.

Others

For two sesquiterpenes, M9, M38, and M58 were identified as the hydroxylation, demethylation, and hydrogenation products of curcumene, while M20 and M45 were the hydroxylation and demethylation product of muurolene, respectively. M37 and M62 were identified as the demethylation and acetylization products of oleanolic acid. In addition, for 2 fatty acids, M12 was the dehydroxylation product of 2-decenoic acid, while M24 and M59-61 were the products of 6,9-octadedicenoic acid.

Proposed metabolic pathways of O. elatus extract

The proposed metabolic pathways of O. elatus extract by human intestinal microflora are presented in Fig. 5. Multiple major metabolite pathways can be observed in this study. The common pathways involved in the biotransformation of O. elatus extract include methylation, demethylation, hydroxylation, dehydroxylation, acetylation, hydrogenation, demethoxylation, and deglycosylation. Among them, polyynes were undoubtedly the most important compounds, as 46 out of 62 metabolites originated from polyynes. By comparing the signal intensity of metabolites, we could find that methylation, dehydroxylation and hydroxylation are major metabolic pathways of polyynes. Moreover, four metabolites of lignans and phenylpropanoid were produced by the loss of glucose. The other metabolites were generated from one triterpenoid and two fatty acids. This indicated that polyynes of O. elatus generated comprehensive biotransformation and were more readily metabolized than other compounds under the same conditions.

Figure 5. The proposed metabolic pathways of O. elatus extract by human intestinal microflora, including (A) Polyynes; (B) Lignans; (C) Phenylpropanoid; (D) Sesquiterpenes; (E) Triterpenoid; (F) Fatty acids. Methylation (1), demethylation (2), hydrogenation (3), hydroxylation (4), dehydroxylation (5), acetylization (6) and demethoxylation (7) were observed in this biotransformation.

Figure 5

Open in a new tab

In summary, the main metabolic pathways of O. elatus refer to hydrolytic and reductive reactions by gut microorganisms. Because of the complexity of active ingredients or constituent concentrations, in vivo exposure, and individual differences, the metabolic profiles of O. elatus might be affected by several factors.

Discussion

In this study, a UPLC-Q-TOF-MS/MS method was developed to screen and identify the chemical composition and metabolites from a traditional Chinese herb, the air-dried root bark of O. elatus. A total of 18 ingredients and 62 metabolites biotransformed by human intestinal microflora were characterized from O. elatus in UPLC-Q-TOF/MS positive ion mode. Two polyynes, falcarindiol and oplopandiol, as the main components of O. elatus and their metabolites by human intestinal microflora are mainly illustrated. It could be noted that the major metabolic pathways of O. elatus refer to methylation, dehydroxylation, and hydroxylation. Studies on the chemical and metabolic profiling of O. elatus by human intestinal microflora will be helpful for the understanding of mechanism research on the active components and further in vivo investigation.

Supplemental Information

Supplemental Information 1. Raw data.

The mass spectrum of Oplopanax elatus (the herbal medication we studied) and its metabolites by human intestinal microbiota, which were detected by an Agilent 6545 Q-TOF-MS system. Mass data including MS and MS/MS information could be analyzed by Agilent MassHunter Workstation software. The software was applied in the screening and identification of the probable compounds, based on the accurate measurements of m/z values with databases.

Click here for additional data file. (1MB, zip)
DOI: 10.7717/peerj.12513/supp-1

Funding Statement

This work was supported by grants from the National Natural Science Foundation of China (81803970), NIH/NCCAM Grants AT004418 and AT005362, Young Talent Promotion Project of China Association of Chinese Medicine (CACM-2019-QNRC2-A01), Young Scientists Development Program of Beijing University of Chinese Medicine, Fundamental Research Funds for the Central Universities (2020-JYB-XJSJJ-003). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Contributor Information

Chong-Zhi Wang, Email: cwang@dacc.uchicago.edu.

Haiqiang Yao, Email: hqyao@bucm.edu.cn.

Additional Information and Declarations

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Jin-Yi Wan performed the experiments, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Jing-Xuan Wan performed the experiments, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Shilei Wang performed the experiments, authored or reviewed drafts of the paper, and approved the final draft.

Xiaolu Wang performed the experiments, authored or reviewed drafts of the paper, and approved the final draft.

Wenqian Guo analyzed the data, authored or reviewed drafts of the paper, and approved the final draft.

Han Ma analyzed the data, authored or reviewed drafts of the paper, and approved the final draft.

Yuqi Wu analyzed the data, authored or reviewed drafts of the paper, and approved the final draft.

Chong-Zhi Wang conceived and designed the experiments, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Lian-Wen Qi performed the experiments, authored or reviewed drafts of the paper, and approved the final draft.

Ping Li analyzed the data, prepared figures and/or tables, and approved the final draft.

Haiqiang Yao conceived and designed the experiments, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Chun-Su Yuan conceived and designed the experiments, prepared figures and/or tables, authored or reviewed drafts of the paper, and approved the final draft.

Human Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

University of Chicago

Data Availability

The following information was supplied regarding data availability:

The raw measurements are available in the Supplementary Files.

References

  • Barko et al. (2018).Barko PC, McMichael MA, Swanson KS, Williams DA. The gastrointestinal microbiome: a review. Journal of Veterinary Internal Medicine. 2018;32(1):9–25. doi: 10.1111/jvim.14875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Bäckhed et al. (2004).Bäckhed F, Ding H, Wang T, Hooper LV, Koh GY, Nagy A, Semenkovich CF, Gordon JI. The gut microbiota as an environmental factor that regulates fat storage. Proceedings of the National Academy of Sciences of the United States of America. 2004;101(44):15718–15723. doi: 10.1073/pnas.0407076101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Chekmeneva et al. (2018).Chekmeneva E, Dos Santos Correia G, Gómez-Romero M, Stamler J, Chan Q, Elliott P, Nicholson JK, Holmes E. Ultra-performance liquid chromatography-high-resolution mass spectrometry and direct infusion-high-resolution mass spectrometry for combined exploratory and targeted metabolic profiling of human urine. Journal of Proteome Research. 2018;17(10):3492–3502. doi: 10.1021/acs.jproteome.8b00413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Chen et al. (2016).Chen F, Wen Q, Jiang J, Li HL, Tan YF, Li YH, Zeng NK. Could the gut microbiota reconcile the oral bioavailability conundrum of traditional herbs? Journal of Ethnopharmacology. 2016;179:253–264. doi: 10.1016/j.jep.2015.12.031. [DOI] [PubMed] [Google Scholar]
  • Dai et al. (2016).Dai SX, Li WX, Han FF, Guo YC, Zheng JJ, Liu JQ, Wang Q, Gao YD, Li GH, Huang JF. In silico identification of anti-cancer compounds and plants from traditional Chinese medicine database. Scientific Reports. 2016;6(1):25462. doi: 10.1038/srep25462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Defois et al. (2018).Defois C, Ratel J, Garrait G, Denis S, Le Goff O, Talvas J, Mosoni P, Engel E, Peyret P. Food chemicals disrupt human gut microbiota activity and impact intestinal homeostasis as revealed by in vitro systems. Scientific Reports. 2018;8(1):11006. doi: 10.1038/s41598-018-29376-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Dou et al. (2009).Dou DQ, Hu XY, Zhao YR, Kang TG, Liu FY, Kuang HX, Smith DC. Studies on the anti-psoriasis constituents of Oplopanax elatus Nakai. Natural Product Research. 2009;23(4):334–342. doi: 10.1080/14786410802075806. [DOI] [PubMed] [Google Scholar]
  • Du et al. (2017).Du LY, Tao JH, Jiang S, Qian DW, Guo JM, Duan JA. Metabolic profiles of the Flos Abelmoschus manihot extract by intestinal bacteria from the normal and CKD model rats based on UPLC-Q-TOF/MS. Biomedical Chromatography. 2017;31(2):e3795. doi: 10.1002/bmc.3795. [DOI] [PubMed] [Google Scholar]
  • Eom et al. (2017).Eom S, Lee J, Kim H, Hyun T. De novo transcriptomic analysis to reveal functional genes involved in triterpenoid saponin biosynthesis in Oplopanax elatus NAKAI. Journal of Applied Botany and Food Quality. 2017;90:18–24. doi: 10.5073/JABFQ.2017.090.004. [DOI] [Google Scholar]
  • Gao et al. (2018).Gao MX, Tang XY, Zhang FX, Yao ZH, Yao XS, Dai Y. Biotransformation and metabolic profile of Xian-Ling-Gu-Bao capsule, a traditional Chinese medicine prescription, with rat intestinal microflora by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry analysis. Biomedical Chromatography. 2018;32(4):e4160. doi: 10.1002/bmc.4160. [DOI] [PubMed] [Google Scholar]
  • Huang et al. (2014a).Huang WH, Shao L, Wang CZ, Yuan CS, Zhou HH. Anticancer activities of polyynes from the root bark of Oplopanax horridus and their acetylated derivatives. Molecules. 2014a;19(5):6142–6162. doi: 10.3390/molecules19056142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Huang et al. (2010).Huang W, Yang J, Zhao J, Wang CZ, Yuan CS, Li SP. Quantitative analysis of six polyynes and one polyene in Oplopanax horridus and Oplopanax elatus by pressurized liquid extraction and on-line SPE-HPLC. Journal of Pharmaceutical and Biomedical Analysis. 2010;53(4):906–910. doi: 10.1016/j.jpba.2010.06.021. [DOI] [PubMed] [Google Scholar]
  • Huang et al. (2014b).Huang WH, Zhang QW, Yuan CS, Wang CZ, Li SP, Zhou HH. Chemical constituents of the plants from the genus Oplopanax. Chemistry & Biodiversity. 2014b;11(2):181–196. doi: 10.1002/cbdv.201200306. [DOI] [PubMed] [Google Scholar]
  • Jin et al. (2018).Jin MM, Zhang WD, Jiang HH, Du YF, Guo W, Cao L, Xu HJ. UPLC-Q-TOF-MS/MS-guided dereplication of Pulsatilla chinensis to identify triterpenoid saponins. Phytochemical Analysis. 2018;29(5):516–527. doi: 10.1002/pca.2762. [DOI] [PubMed] [Google Scholar]
  • Knispel et al. (2013).Knispel N, Ostrozhenkova E, Schramek N, Huber C, Peña-Rodríguez LM, Bonfill M, Palazón J, Wischmann G, Cusidó RM, Eisenreich W. Biosynthesis of panaxynol and panaxydol in Panax ginseng. Molecules. 2013;18(7):7686–7698. doi: 10.3390/molecules18077686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Koppel, Maini Rekdal & Balskus (2017).Koppel N, Maini Rekdal V, Balskus EP. Chemical transformation of xenobiotics by the human gut microbiota. Science. 2017;356(6344):eaag2770. doi: 10.1126/science.aag2770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Li et al. (2019).Li W, Hong B, Li Q, Li Z, Bi K. An integrated serum and urinary metabonomic research of Rhizoma Curcumae-Rhizoma Sparganii drug pair in hysteromyoma rats based on UPLC-Q-TOF-MS analysis. Journal of Ethnopharmacology. 2019;231:374–385. doi: 10.1016/j.jep.2018.11.033. [DOI] [PubMed] [Google Scholar]
  • Lou et al. (2015).Lou Y, Zheng J, Hu H, Lee J, Zeng S. Application of ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry to identify curcumin metabolites produced by human intestinal bacteria. Journal of Chromatography B Analytical Technologies in the Biomedical and Life Sciences. 2015;985:38–47. doi: 10.1016/j.jchromb.2015.01.014. [DOI] [PubMed] [Google Scholar]
  • Moon et al. (2013).Moon HK, Kim YW, Hong YP, Park SY. Improvement of somatic embryogenesis and plantlet conversion in Oplopanax elatus, an endangered medicinal woody plant. Springerplus. 2013;2(1):428. doi: 10.1186/2193-1801-2-428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Pagliari et al. (2017).Pagliari D, Gambassi G, Piccirillo CA, Cianci R. The intricate link among gut “immunological niche,” microbiota, and xenobiotics in intestinal pathology. Mediators of Inflammation. 2017;2017:8390595. doi: 10.1155/2017/8390595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Panossian et al. (2021).Panossian AG, Efferth T, Shikov AN, Pozharitskaya ON, Kuchta K, Mukherjee PK, Banerjee S, Heinrich M, Wu W, Guo DA, Wagner H. Evolution of the adaptogenic concept from traditional use to medical systems: pharmacology of stress- and aging-related diseases. Medicinal Research Reviews. 2021;41(1):630–703. doi: 10.1002/med.21743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Purup, Larsen & Christensen (2009).Purup S, Larsen E, Christensen LP. Differential effects of falcarinol and related aliphatic C(17)-polyacetylenes on intestinal cell proliferation. Journal of Agricultural and Food Chemistry. 2009;57(18):8290–8296. doi: 10.1021/jf901503a. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Qiao et al. (2017).Qiao X, Sun W, Wang C, Zhang L, Li P, Wen X, Yang J, Yuan C. Polyyne-enriched extract from oplopanax elatus significantly ameliorates the progression of colon carcinogenesis in Apc(Min/+) mice. Molecules. 2017;22(10):1593. doi: 10.3390/molecules22101593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Rajilić-Stojanović & de Vos (2014).Rajilić-Stojanović M, de Vos WM. The first 1,000 cultured species of the human gastrointestinal microbiota. FEMS Microbiology Reviews. 2014;38(5):996–1047. doi: 10.1111/1574-6976.12075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Schymanski et al. (2014).Schymanski EL, Jeon J, Gulde R, Fenner K, Ruff M, Singer HP, Hollender J. Identifying small molecules via high resolution mass spectrometry: communicating confidence. Environmental Science & Technology. 2014;48(4):2097–2098. doi: 10.1021/es5002105. [DOI] [PubMed] [Google Scholar]
  • Shao et al. (2016).Shao L, Nie MK, Chen MY, Wang J, Wang CZ, Huang WH, Yuan CS, Zhou HH. Screening and identifying antioxidants from Oplopanax elatus using 2,2’-diphenyl-1-picrylhydrazyl with off-line two-dimensional HPLC coupled with diode array detection and tandem time-of-flight mass spectrometry. Journal of Separation Science. 2016;39(22):4269–4280. doi: 10.1002/jssc.201600838. [DOI] [PubMed] [Google Scholar]
  • Shikov et al. (2014).Shikov AN, Pozharitskaya ON, Makarov VG, Yang WZ, Guo DA. Oplopanax elatus (Nakai) Nakai: chemistry, traditional use and pharmacology. Chinese Journal of Natural Medicines. 2014;12(10):721–729. doi: 10.1016/S1875-5364(14)60111-4. [DOI] [PubMed] [Google Scholar]
  • Sun et al. (2016).Sun W, He YS, Xu LH, Zhang BY, Qi LW, Yang J, Li P, Wen XD. Pharmacokinetic profiles of falcarindiol and oplopandiol in rats after oral administration of polyynes extract of Oplopanax elatus. Chinese Journal of Natural Medicines. 2016;14(9):714–720. doi: 10.1016/S1875-5364(16)30085-1. [DOI] [PubMed] [Google Scholar]
  • Tao et al. (2016).Tao JH, Duan JA, Jiang S, Qian YY, Qian DW. Biotransformation and metabolic profile of buddleoside with human intestinal microflora by ultrahigh-performance liquid chromatography coupled to hybrid linear ion trap/orbitrap mass spectrometer. Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences. 2016;1025:7–15. doi: 10.1016/j.jchromb.2016.04.055. [DOI] [PubMed] [Google Scholar]
  • Teschke et al. (2015).Teschke R, Wolff A, Frenzel C, Eickhoff A, Schulze J. Herbal traditional Chinese medicine and its evidence base in gastrointestinal disorders. World Journal of Gastroenterology. 2015;21(15):4466–4490. doi: 10.3748/wjg.v21.i15.4466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Thursby & Juge (2017).Thursby E, Juge N. Introduction to the human gut microbiota. Biochemical Journal. 2017;474(11):1823–1836. doi: 10.1042/BCJ20160510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Wang et al. (2008).Wang X, Sun W, Sun H, Lv H, Wu Z, Wang P, Liu L, Cao H. Analysis of the constituents in the rat plasma after oral administration of Yin Chen Hao Tang by UPLC/Q-TOF-MS/MS. Journal of Pharmaceutical and Biomedical Analysis. 2008;46(3):477–490. doi: 10.1016/j.jpba.2007.11.014. [DOI] [PubMed] [Google Scholar]
  • Wang et al. (2020).Wang CZ, Wan JY, Wan J, Wang S, Luo Y, Zeng J, Yao H, Zhang CF, Zhang QH, Sawadogo WR, Xu M, Du W, Qi LW, Li P, Yuan CS. Human intestinal microbiota derived metabolism signature from a North American native botanical Oplopanax horridus with UPLC/Q-TOF-MS analysis. Biomedical Chromatography. 2020;34(10):e4911. doi: 10.1002/bmc.4911. [DOI] [PubMed] [Google Scholar]
  • Wewer et al. (2011).Wewer V, Dombrink I, vom Dorp K, Dörmann P. Quantification of sterol lipids in plants by quadrupole time-of-flight mass spectrometry. Journal of Lipid Research. 2011;52(5):1039–1054. doi: 10.1194/jlr.D013987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Wu et al. (2019).Wu Y, Wang P, Yang H, Sui F. UPLC-Q-TOF-MS and UPLC-MS/MS methods for metabolism profiles and pharmacokinetics of major compounds in Xuanmai Ganjie Granules. Biomedical Chromatography. 2019;33(3):e4449. doi: 10.1002/bmc.4449. [DOI] [PubMed] [Google Scholar]
  • Yang et al. (2010).Yang MC, Kwon HC, Kim YJ, Lee KR, Yang HO. Oploxynes A and B, polyacetylenes from the stems of Oplopanax elatus. Journal of Natural Products. 2010;73(5):801–805. doi: 10.1021/np900628j. [DOI] [PubMed] [Google Scholar]
  • Yang & Lao (2019).Yang M, Lao L. Emerging applications of metabolomics in traditional chinese medicine treating hypertension: biomarkers, pathways and more. Frontiers in Pharmacology. 2019;10:158. doi: 10.3389/fphar.2019.00158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Yang et al. (2016).Yang S, Tian M, Yuan L, Deng H, Wang L, Li A, Hou Z, Li Y, Zhang Y. Analysis of E. rutaecarpa Alkaloids constituents in vitro and in vivo by UPLC-Q-TOF-MS combined with diagnostic fragment. Journal of Analytical Methods in Chemistry. 2016;2016:4218967. doi: 10.1155/2016/4218967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • Yang et al. (2014).Yang ZM, Zhao J, Lao KM, Chen XJ, Leong F, Wang CZ, Yuan CS, Li SP. Determination of six polyynes in Oplopanax horridus and Oplopanax elatus using polyethylene glycol modified reversed migration microemulsion electrokinetic chromatography. Electrophoresis. 2014;35(20):2959–2964. doi: 10.1002/elps.201400159. [DOI] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supplemental Information 1. Raw data.

The mass spectrum of Oplopanax elatus (the herbal medication we studied) and its metabolites by human intestinal microbiota, which were detected by an Agilent 6545 Q-TOF-MS system. Mass data including MS and MS/MS information could be analyzed by Agilent MassHunter Workstation software. The software was applied in the screening and identification of the probable compounds, based on the accurate measurements of m/z values with databases.

Click here for additional data file. (1MB, zip)
DOI: 10.7717/peerj.12513/supp-1

Data Availability Statement

The following information was supplied regarding data availability:

The raw measurements are available in the Supplementary Files.

Articles from PeerJ are provided here courtesy of PeerJ, Inc

RESOURCES