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Article

UPLC-QTOF-ESI-MS/MS-Based Comparative Study of Phytochemicals in Sapindus mukorossi

by
Neil Patrick Uy
1,
Hak-Dong Lee
1,2,
Jajung Ku
3,
Kyung Choi
4 and
Sanghyun Lee
1,2,*
1
Department of Plant Science and Technology, Chung-Ang University, Anseong 17546, Republic of Korea
2
Natural Product Institute of Science and Technology, Anseong 17546, Republic of Korea
3
Forest Bioresources Department, National Institute of Forest Science, Suwon 16631, Republic of Korea
4
Division of Forest Biodiversity, Korea National Arboretum, Pocheon 11186, Republic of Korea
*
Author to whom correspondence should be addressed.
Horticulturae 2025, 11(6), 682; https://doi.org/10.3390/horticulturae11060682 (registering DOI)
Submission received: 30 May 2025 / Revised: 11 June 2025 / Accepted: 12 June 2025 / Published: 13 June 2025
(This article belongs to the Special Issue Plant Tissue and Organ Cultures for Crop Improvement in Omics Era: 2nd Edition)
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Abstract

:
This study examined the phytochemical compositions of ten Sapindus mukorossi samples from two regions in Korea: Suwon and Daegu. The Folin–Ciocalteu method was used to calculate the total polyphenol content (TPC). Among all extracts tested, leaf samples from Suwon and Daegu (SLE and DLE) exhibited the highest TPC at 2.70 and 2.90 mg tannic acid equivalent/g extract. Similarly, a modified aluminum chloride colorimetric test was used to determine the total flavonoid content (TFC). Similar results were obtained, with SLE and DLE having TFC values of 40.71 and 41.07 mg quercetin equivalent/g extract, respectively. Liquid chromatography with tandem mass spectrometry was used to detect 13 compounds, whereas high-performance liquid chromatography was used to quantify the prominent compounds: rutin, nicotiflorin, and narcissin. Among these, rutin was the most abundant, especially in SLE and DLE (54.37 and 70.21 mg/g, respectively). Furthermore, rutin significantly contributed to the total content of these samples at 78.31 and 85.44 mg/g, respectively. There were significant variations in the distribution of these compounds across different parts of the plant. These findings highlight the importance of S. mukorossi as a source of natural bioactive chemicals and pave the way for further research into its potential applications in healthcare products.

Graphical Abstract

1. Introduction

The soap nut tree, also known as Sapindus mukrossi is a deciduous tree belonging to the Sapindaceae family [1]. This species is native to South-Central and Southeast China and has been introduced to various regions, including India, Japan, Korea, and Nepal [2]. In recent years, there has been increasing emphasis on exploring plant-derived bioactive compounds as natural alternatives to synthetic antioxidants and therapeutic agents [3]. This global trend aligns with the growing consumer preference for sustainable and eco-friendly products in pharmaceuticals, nutraceuticals, and cosmetics. S. mukorossi, with its diverse phytochemical profile, represents an untapped resource to address these demands. It has been a staple in Traditional Chinese Medicine for centuries [4], highlighting its potential as a source of bioactive compounds with therapeutic value. Recent studies have shown that S. mukorossi possesses antipyretic, anti-inflammatory, antioxidant, and analgesic properties [5].
Beyond these medicinal applications, S. mukorossi has been traditionally used in the aforementioned countries for centuries mainly due to its abstergent properties [6]. This is attributed to the abundance of saponins in the pericarps of its fruit. Saponins are the primary phytochemicals found in soap nuts [7]. These medicinally important phytochemicals play a key role in plant defense against bacteria [8]. Furthermore, saponins are used in industries that manufacture detergents and shampoos due to their surfactant properties [9].
Although the saponins of S. mukorossi are well-documented, less attention has been paid to its other bioactive constituents, particularly flavonoids. These secondary metabolites exhibit antioxidant, anti-inflammatory, and therapeutic properties [10]. Their relevance to health and industry underscores the importance of further exploration into the phytochemical composition of S. mukorossi. Despite the extensive traditional and industrial use of this plant, detailed studies on the flavonoid glycosides present in various parts of the plant are lacking [11]. Previous research has primarily focused on saponin content, leaving significant gaps in the profiling and quantification of other bioactive compounds [12]. Additionally, variations in these phytochemicals across different plant parts (e.g., leaves, fruits, and seeds) have not been comprehensively examined. Addressing these gaps is essential to fully elucidate the bioactive potential of S. mukorossi.
Flavonoid glycosides, such as rutin, nicotiflorin, and narcissin, are bioactive compounds with potent antioxidant properties [13,14,15]. They play crucial roles in reducing oxidative stress and have potential applications in pharmaceuticals, nutraceuticals, and cosmetics. Investigating these specific compounds allows for a deeper understanding of the therapeutic potential of S. mukorossi and paves the way for its future applications in various industries.
This study aimed to address these gaps by extracting and analyzing the compounds present in various parts of S. mukorossi using colorimetric assays and ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-ESI-MS/MS), quantifying the identified flavonoid glycosides using high-performance liquid chromatography (HPLC), and comparing the phytochemical content across different plant parts to identify patterns and potential applications.

2. Materials and Methods

2.1. Plant Materials

The S. mukorossi samples were acquired from the Korea National Arboretum, Pocheon, Republic of Korea. The samples were named according to the plant part used, and their geographical locations were the National Institute of Forest Science, Suwon, and Kyungpook National University, Daegu, Republic of Korea (Figure 1). The samples from Suwon, Republic of Korea, are referred to as leaf (SLE), stem (SST), fruit receptacle (SFR), flesh (SPF), and seed (SSE), whereas the samples from Daegu, Republic of Korea, are referred to as leaf (DLE), stem (DST), fruit receptacle (DFR), flesh (DPF), and seed (DSE).

2.2. Instruments and Reagents

Chromatographic analysis was performed using a Waters Alliance e2695 separation module (Milford, MA, USA) equipped with a Waters 2998 photodiode array (PDA) detector, quaternary pump, and autosampler. The HPLC solvents (water and acetonitrile (ACN)) were purchased from Honeywell (Burdick and Jackson, Muskegon, MI, USA), and trifluoroacetic acid (TFA) was purchased from J. T. Baker (Phillipsburg, PA, USA). Standard compounds rutin (1), nicotiflorin (2), and narcissin (3) were procured from the Natural Product Institute of Science and Technology (www.nist.re.kr; accessed on 12 January 2025), Anseong, Republic of Korea (Figure 2).

2.3. Crude Extraction

Ten grams of homogenized air-dried samples were combined with 200 mL of 95% ethanol. The mixture was then refluxed for 5 h at 80 °C three times. The extracts were filtered and subsequently concentrated in a vacuum with a rotary evaporator (Eyela, Tokyo, Japan) at 55 °C to obtain concentrated EtOH extracts. The extracts were set aside until further use.

2.4. Total Polyphenol Content Assay

The TPC of the ten S. mukorossi extracts was determined using a previously described method with minimal changes [16]. First, a stock solution (2.5 mg/mL) of each sample was serially diluted to achieve three concentrations (2.5–0.63 mg/mL). Next, 60 μL of each sample was placed in the 96-well microtiter plate. Then, 40 μL of Folin–Ciocalteu reagent (Sigma-Aldrich, St. Louis, MO, USA) was added to wells containing the samples. After adding 100 μL of 7.5% sodium carbonate (Na2CO3), the samples were left to react for 30 min at 37 °C in the dark. Subsequently, the samples’ absorbance was measured at 760 nm using a microplate reader (Epoch; BioTek, Winooski, VT, USA). The TPC was then constructed using a standard curve built with various tannic acid concentrations (200–12.5 µg/mL).

2.5. Total Flavonoid Content Assay

The TFC of the extracts was determined using a modified method adapted from a previous study [16]. First, a stock solution (2.5 mg/mL) of each sample was serially diluted to achieve three concentrations (2.5–0.63 mg/mL). Subsequently, 100 μL of each diluted sample was transferred into the wells of a 96-well microtiter plate. An equal volume (100 μL) of 2% aluminum chloride hexahydrate (AlCl3·6H2O) solution was added to the wells. The plate was incubated at room temperature for 10 min and the absorbance was recorded at 430 nm using a microplate reader (Epoch; BioTek, Winooski, VT, USA). The TFC was then calculated using a standard curve generated using various quercetin concentrations (200–12.5 µg/mL).

2.6. Phytochemical Detection by Ultra-Performance Liquid Chromatography (UPLC), Quadrupole Time-of-Flight Mass Spectrometry (QTOF-MS), and Electrospray Ionization (ESI) (UPLC-QTOF-ESI-MS/MS)

The analysis was conducted utilizing an LC system with Thermo Vanquish UHPLC equipment (Thermo Fisher Scientific, Waltham, MA, USA) fitted with a Waters Cortex T3 column (150 mm × 2.1 mm, particle size 1.6 μm). The column temperature was maintained at 45 °C. The mobile phase consisted of acetonitrile (eluent B: 0.1% HCOOH) and water (eluent A: 0.1% HCOOH). A flow rate of 0.25 mL/min and a gradient elution program (97% A from 0 to 0.5 min, 85% A at 15 min, 0% A from 50 to 55 min, and 3% B from 55.1 to 60 min) were employed. For spectrometric analysis, a Triple TOF 5600+ System (AB SCIEX, Marlborough, MA, USA) equipped with a heated electrospray ion source was used. The mass spectrometer was run in both positive and negative ion modes to produce survey full-scan MS spectra (m/z 100–1500) using a quadrupole system at a resolution of 70,000. The spray voltage was set to 3.5 kV in positive ion mode. The 13 most energetic precursor ions were chosen for MS2 fragmentation at a resolution of 17,500. The MS specifications also included a capillary temperature of 320 °C, a sheath gas of 50 AU, a sweep gas of 0 AU, and an auxiliary gas of 10 AU.

2.7. Quantification of Flavonoid Glycosides by HPLC

The HPLC analysis was conducted using a Waters Alliance e2695 separation module (Milford, MA, USA) coupled with a Waters 2998 PDA detector, following a previously described method with modifications [17]. The separation was performed on an INNO C18 column (25 cm × 4.6 mm, 5 µm). The mobile phase of the gradient elution system consisted of 0.1% TFA in water (A) and ACN (B), with the following gradient elution system: 95% A from 0 to 10 min, 0% A at 40 min, and 95% B from 50 to 60 min. The sample injection volume was 10 µL, with a flow rate of 1.0 mL/min. Detection was performed at 254 nm.

2.8. Calibration Curves

Standard stock solutions were prepared by dissolving the reference compounds in methanol (MeOH) to generate a concentration of 1 mg/mL. Working solutions used to construct the calibration curve were prepared by serially diluting the selected stock solutions to the desired concentrations (0.500–0.03125 mg/mL). The samples were dissolved in the same solvent and each extract had a final concentration of 15 mg/mL. The standard and sample solutions were filtered using a 0.45 µm polyvinylidene fluoride (PVDF) filter before use. Calibration functions for the standard compounds were calculated based on peak areas (Y), concentrations (X, μg/10 μL), and mean values ± standard deviation (SD) (n = 3).

2.9. Statistical Analysis

The results are expressed as mean ± SD, and all analyses were conducted in triplicate. Data were normalized using one-way analysis of variance (ANOVA) and Tukey’s post hoc test. All statistical tests were performed using GraphPad Prism 8.0.2 software (GraphPad Software, Boston, MA, USA). p-values < 0.05 were considered statistically significant.

3. Results and Discussion

3.1. Total Polyphenol and Total Flavonoid (TPC and TFC) Content

The TPC of S. mukorossi extracts SLE, SST, SFR, SPF, SSE, DLE, DST, DFR, DPF, and DSE were evaluated. The results varied significantly (Figure 3). Generally, SLE and DLE exhibited the highest TPC among all the extracts with values of 2.70 mg tannic acid equivalent (TAE)/g extract and 2.90 mg TAE/g extract, respectively. This was followed by SST (2.38 mg TAE/g extract), DFR (2.11 mg TAE/g extract), and DST (1.66 mg TAE/g extract). In comparison, the remaining samples exhibited significantly lower TPCs with DPE, SSE, SPE, DSE, and SFR showing values of 0.43 mg TAE/g extract, 0.35 mg TAE/g extract, 0.30 mg TAE/g extract, 0.06 mg TAE/g extract, and 0.04 mg TAE/g extract, respectively, indicating minimal polyphenolic contributions to the overall phytochemical profile.
Similarly, the total flavonoid content (TFC) of the tested extracts followed a similar trend to TPC with SLE and DLE exhibiting the highest TFC values at 40.71 mg quercetin equivalent (QE)/g extract and 41.07 mg QE/g extract, respectively. This was followed by SST (28.14 mg QE/g extract), DST (18.08 mg QE/g extract), DFR (5.33 mg QE/g extract), SPE (2.99 mg QE/g extract), DPE (2.10 mg QE/g extract), DSE (2.09 mg QE/g extract), SSE (1.74 mg QE/g extract), and SFR (0.80 mg QE/g extract). A previous study by Singh and Kumari (2015) [18] examined the TPC and TFC of S. mukorossi leaves and fruits extracted with different solvents (ethanol, methanol, and acetone). The results showed that the TPC of the leaves was highest when extracted with water compared to methanol and ethanol, whereas the TFC of the leaves would be highest if extracted with ethanol. For the fruits, TPC was highest when extracted with methanol, whereas TFC was highest in the ethanolic extracts. Conversely, a study by the same authors that analyzed the callus of the plant reported that ethanol extraction yielded high TPC, whereas methanol extraction gave the highest TFC compared to ethanol and water. Another study evaluated the TPC and TFC of S. mukorossi partitioned using solvents of varying polarities [19]. Compared to the results of the present study, their TPC values were comparable to those of TFC. Differences in methods and samples may explain the variation in results between these studies and the present work. Furthermore, the present study used only ethanol as the extraction solvent, which is widely used and generally recognized as safe (GRAS) [20].

3.2. UPLC-QTOF-ESI-MS/MS Profiling

To characterize the flavonoid glycosides in S. mukorossi extracts, a representative sample (SLE) was subjected to ultra-performance liquid chromatography-quadrupole time-of-flight-electrospray ionization-mass spectrometry/mass spectrometry (UPLC-QTOF-ESI-MS/MS), resulting in the emergence of three prominent peaks. The base peak chromatogram and the MS2 spectra of these peaks in SLE are shown in Figure 4.
A UPLC-QTOF-ESI-MS/MS system was employed for these studies. This versatile and highly sensitive technique enabled the detection of the 13 most energetic peaks, which were identified by comparing their molecular weights and fragmentation patterns with spectral libraries. All phytochemicals were flavonoid glycosides (Table 1).
Peak 1 (molecular weight = 772.2) eluted at 13.60 min and was identified as quercetin 3-rutinoside-7-glucoside. The fragmentation pattern exhibited classic losses of rhamnose and glucose, indicating the presence of two sugar moieties linked to the quercetin backbone. Similarly, Peak 2, with a retention time of 15.71 min and a molecular weight of 594.2, was identified as vicenin-2, a C-glycosyl flavonoid [21]. Its MS2 spectrum supported its identity as a di-C-glycosyl apigenin derivative by exhibiting neutral losses of glucose [22]. Peak 3 was identified as schaftoside based on its molecular weight of 564.1 and fragment ions at m/z 443.1 and m/z 353.1, suggesting consecutive sugar moiety losses, consistent with its glycosylated structure [23].
Peak 4 with a molecular weight of 610.2 eluted at 19.67 min and was identified as rutin (quercetin-3-rutinoside) [24]. Its fragmentation pattern showed distinct losses of rhamnose and glucose, supporting its identity. Peak 5, observed at 20.06 min with a molecular weight of 464.1, was identified as hyperoside (quercetin-3-galactoside) based on the loss of galactose in its fragmentation spectrum [25]. Peak 6, with a molecular weight of 594.2 was identified as nicotiflorin (kaempferol-3-rutinoside) and eluted at 21.04 min [26]. Its MS2 spectrum displayed fragment ions, indicating the loss of rutinose. Peak 7 with a molecular weight of 624.2, was tentatively identified as narcissin (isorhamnetin-3-rutinoside) due to the loss of rhamnose during fragmentation, further confirming its structure [27].
A previous study used LC-MS/MS to rapidly identify the phytochemical constituents of the fruits of S. mukorossi [28], detecting 31 compounds, of which 16 were acyclic sesquiterpene oligoglycosides and 15 were triterpenoid saponins. These results were not unexpected, as the fruits of this plant are the ones most utilized because of their rich saponin content [29].

3.3. HPLC Quantitative Analysis

To quantify the prominent compounds in the extracts, high-performance liquid chromatography was employed. The analysis quantified three major flavonoid glycosides, rutin (1), nicotiflorin (2), and narcissin (3) from the extracts of S. mukorossi collected from the Suwon and Daegu regions. With the methods employed, the standard compounds demonstrated good retention times and linearity (Figure 5, Table 2).
The results revealed variations in their contents across different plant parts (Figure 6). In Suwon samples, the highest rutin (1) content (54.37 mg/g) was observed in the leaf extract (SLE), contributing to a total content of 78.31 mg/g (Table 3). Nicotiflorin (2) and narcissin (3) were also abundant in SLE at 14.25 mg/g and 5.19 mg/g, respectively.
Conversely, SST exhibited significantly lower levels of all three compounds, with a total content of 15.29 mg/g. In other plant parts, including the SFR, SPF, and SSE, most compounds were either not detected (ND) or present in trace amounts (tr), with the total content in SPF being only 1.89 mg/g. These results highlight that the leaves were the richest source of flavonoid glycosides among the Suwon samples.
Similarly, Daegu samples exhibited a similar pattern, where DLE contained the highest rutin (1) content (70.21 mg/g), significantly surpassing other plant parts and contributing to a total content of 85.44 mg/g (Figure 7).
Nicotiflorin (2) and narcissin (3) were also prominent in DLE at 11.21 mg/g and 4.02 mg/g, respectively. The stem (DST) and fruit receptacle (DFR) had lower flavonoid glycoside contents, with total contents of 7.06 mg/g and 5.35 mg/g, respectively.
As in Suwon, most compounds were not detected or were present in trace amounts in the flesh (DPF) and seeds (DSE) from the Daegu samples. The significantly higher rutin (1) content in Daegu leaf extracts than that in Suwon leaf extracts (70.21 mg/g vs. 54.37 mg/g) highlights potential regional differences in secondary metabolite biosynthesis, possibly influenced by climatic and soil conditions [30]. The variations in flavonoid glycoside content among different plant parts and regions suggest that environmental factors and plant morphology significantly influence phytochemical accumulation [31]. Leaves consistently exhibit the highest flavonoid glycoside content, making them a promising source of bioactive compounds. Generally, leaves contain a higher flavonoid glycoside content compared to other plant tissues because they are more directly exposed to harmful UV rays [32]. On the other hand, the rest of the plant parts that are not rich in flavonoid glycosides may be abundant in other phytochemicals; for example, the fruit pericarp is known to have the most saponins [33]. These results underscore the importance of selecting the appropriate plant parts to maximize the yield of bioactive compounds for pharmaceutical and nutraceutical applications.
This study is the first to report on the flavonoid glycosides present in the leaves of S. mukorossi using the UPLC-QTOF-ESI-MS/MS method and the quantification of their prominent peaks by HPLC. Successful quantification of rutin (1), nicotiflorin (2), and narcissin (3) provides valuable insights into the phytochemical profile of S. mukorossi, emphasizing its potential as a source of health-beneficial flavonoid glycosides. Specifically, these compounds were abundant in the leaves of both the Suwon and Daegu samples, indicating that they have the potential to be a source of anticancer [34], anti-inflammatory [35], and insecticide [36] agents.
The findings of this study have the potential to advance the utilization of S. mukorossi beyond its traditional applications and contribute to the development of sustainable bio-based industries. By identifying and quantifying the key flavonoid glycosides, this research not only provides a foundation for future pharmacological studies but also opens avenues for eco-friendly product innovations in health and personal care.

4. Conclusions

This study elucidates the phytochemical composition of S. mukorossi from two distinct locations in Korea, highlighting its diverse bioactive component profiles. The high total polyphenol and flavonoid contents of the leaf extracts from Suwon and Daegu suggest the potential of these plant parts as sources of bioactive compounds, particularly rutin (1), the most prevalent flavonoid glycoside. The presence of this phytochemical, along with nicotiflorin (2) and narcissin (3), underscores the therapeutic potential of S. mukorossi, particularly its antioxidant and anti-inflammatory properties. The successful identification of flavonoid glycosides and saponins using UPLC-QTOF-ESI-MS/MS and HPLC reinforces the potential of S. mukorossi as a source of compounds with pharmacological, nutraceutical, and cosmetic applications. The diverse phytochemical distribution among the plant parts warrants further research to identify the most suitable parts for extracting these chemicals. The findings of this study open avenues for further exploration of the health benefits of S. mukorossi, particularly in the development of sustainable plant-based products for medicinal and industrial purposes. These results address the gaps in the literature regarding the flavonoid profile of S. mukorossi and contribute to its already well-established medicinal and industrial uses.

Author Contributions

HPLC/PDA, TPC, and TCC analyses, N.P.U.; UPLC-QTOF-ESI-MS/MS analysis, H.-D.L.; Resources, data curation, and funding, J.K. and K.C.; Supervision and editing: S.L. All authors have read and agreed to the published version of the manuscript.

Funding

This study was supported by a grant (Project No. RS-2024-00404250) obtained from the Korea Forest Service (Daejeon, Republic of Korea).

Data Availability Statement

All data are available upon request.

Acknowledgments

We express our gratitude to Hyo-Joo Jun for participating in the Practical and Research Engagement (PRE) Program (2024) of Chung-Ang University, Anseong, Republic of Korea.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Chen, C.; Nien, C.; Chen, L.; Huang, K.; Chang, W.; Huang, H. Effects of Sapindus mukorossi seed oil on skin wound healing: In vivo and in vitro testing. Int. J. Mol. Sci. 2019, 20, 2579. [Google Scholar] [CrossRef] [PubMed]
  2. Agnihotri, S.; Dobhal, P.; Ashfaqullah, S.; Chauhan, H.K.; Tamta, S. Review of the botany, traditional uses, pharmacology, threats and conservation of Zanthoxylum armatum (Rutaceae). S. Afr. J. Bot. 2022, 150, 920–927. [Google Scholar] [CrossRef]
  3. Vlčko, T.; Rathod, N.B.; Kulawik, P.; Ozogul, Y.; Ozogul, F. The impact of aromatic plant-derived bioactive compounds on seafood quality and safety. Adv. Food Nutr. Res. 2022, 99, 275–339. [Google Scholar]
  4. Xu, Y.; Gao, Y.; Chen, Z.; Liu, J.; Wang, X.; Jia, L. Metabolomics analysis of the soapberry (Sapindus mukorossi Gaertn.) pericarp during fruit development and ripening based on UHPLC-HRMS. Sci. Rep. 2021, 11, 11657. [Google Scholar] [CrossRef]
  5. Zhao, Z.; Zhang, A.; Song, L.; He, C.; He, H. Evaluation of Sapindus mukorossi Gaertn flower water extract on in vitro anti-acne activity. Curr. Issues Mol. Biol. 2025, 47, 316. [Google Scholar] [CrossRef]
  6. Upadhyay, A.; Singh, D. Pharmacological effects of Sapindus mukorossi. Rev. Inst. Med. Trop. Sao Paulo 2012, 54, 273–280. [Google Scholar] [CrossRef]
  7. Timilsena, Y.P.; Phosanam, A.; Stockmann, R. Perspectives on saponins: Food functionality and applications. Int. J. Mol. Sci. 2023, 24, 13538. [Google Scholar] [CrossRef]
  8. Zhou, M.; Fan, J.; Gao, Y.; Zheng, C.; Xu, Y.; Jia, L.; An, X.; Chen, Z. Identification and analysis of UGT genes associated with triterpenoid saponin in soapberry (Sapindus mukorossi Gaertn.). BMC Plant Biol. 2024, 24, 588. [Google Scholar] [CrossRef]
  9. Meshram, P.D.; Shingade, S.; Madankar, C.S. Comparative study of saponin for surfactant properties and potential application in personal care products. Mater. Today Proc. 2021, 45, 5010–5013. [Google Scholar] [CrossRef]
  10. Roy, A.; Khan, A.; Ahmad, I.; Alghamdi, S.; Rajab, B.S.; Babalghith, A.O.; Alshahrani, M.Y.; Islam, S.; Islam, M.R. Flavonoids a bioactive compound from medicinal plants and its therapeutic applications. BioMed Res. Int. 2022, 2022, 83. [Google Scholar] [CrossRef]
  11. Xu, Y.; Zhou, S.; Chen, Z.; Zhao, G.; Liu, J.; Wang, L.; Wang, X.; Jia, L.; Zhang, D. Contents of the total saponins and total flavonoids in different organs of Sapindus mukorossi. J. Nanjing For. Univ. (Nat. Sci. Ed.) 2021, 45, 83. [Google Scholar]
  12. Sochacki, M.; Vogt, O. Triterpenoid saponins from washnut (Sapindus mukorossi Gaertn.)—A source of natural surfactants and other active components. Plants 2022, 11, 2355. [Google Scholar] [CrossRef] [PubMed]
  13. da Silva, F.M.A.; Da Silva, K.P.A.; De Oliveira, L.P.M.; Costa, E.V.; Koolen, H.H.; Pinheiro, M.L.B.; De Souza, A.Q.L.; De Souza, A.D.L. Flavonoid glycosides and their putative human metabolites as potential inhibitors of the SARS-CoV-2 main protease (Mpro) and RNA-dependent RNA polymerase (RdRp). Mem. Inst. Oswaldo Cruz 2020, 115, e200207. [Google Scholar] [CrossRef] [PubMed]
  14. Rani, I.; Kalsi, A.; Kaur, G.; Sharma, P.; Gupta, S.; Gautam, R.K.; Chopra, H.; Bibi, S.; Ahmad, S.U.; Singh, I.; et al. Modern drug discovery applications for the identification of novel candidates for COVID-19 infections. Ann. Med. Surg. 2022, 80, 104294. [Google Scholar] [CrossRef]
  15. Chehlh, T.C.; Rincon-Cervera, M.A.; Gomez-Mercado, F.; Lopez-Ruiz, R.; Gallon-Bedoya, M.; Ezzaitouni, M.; Guil-Guerrero, J.L. Wild asparagus shoots constitute a healthy source of bioactive compounds. Molecules 2023, 28, 5786. [Google Scholar] [CrossRef]
  16. Lee, C.; Cho, H.; Shim, J.; Tran, G.H.; Lee, H.; Ahn, K.H.; Yoo, E.; Chung, M.J.; Lee, S. Characteristics of phenolic compounds in Peucedanum japonicum according to various stem and seed colors. Molecules 2023, 28, 6266. [Google Scholar] [CrossRef]
  17. Ma, C. Simultaneous quantification of eight compounds of Lonicera japonica by HPLC-DAD. Nat. Prod. Sci. 2024, 30, 52–58. [Google Scholar] [CrossRef]
  18. Singh, R.; Kumari, N. Comparative determination of phytochemicals and antioxidant activity from leaf and fruit of Sapindus mukorossi Gaertn—A valuable medicinal tree. Ind. Crops Prod. 2015, 73, 1–8. [Google Scholar] [CrossRef]
  19. Shah, M.; Parveen, Z.; Khan, M.R. Evaluation of antioxidant, anti-inflammatory, analgesic and antipyretic activities of the stem bark of Sapindus mukorossi. BMC Complement. Altern. Med. 2017, 17, 526. [Google Scholar] [CrossRef]
  20. Jiménez-Moreno, N.; Volpe, F.; Moler, J.A.; Esparza, I.; Ancín-Azpilicueta, C. Impact of extraction conditions on the phenolic composition and antioxidant capacity of grape stem extracts. Antioxidants 2019, 8, 597. [Google Scholar] [CrossRef]
  21. Silva, D.B.; Turatti, I.C.C.; Gouveia, D.R.; Ernst, M.; Teixeira, S.P.; Lopes, N.P. Mass spectrometry of flavonoid vicenin-2, based sunlight barriers in Lychnophora species. Sci. Rep. 2014, 4, 4309. [Google Scholar] [CrossRef] [PubMed]
  22. Islam, M.N.; Ishita, I.J.; Jung, H.A.; Choi, J.S. Vicenin 2 isolated from Artemisia capillaris exhibited potent anti-glycation properties. Food Chem. Toxicol. 2014, 69, 55–62. [Google Scholar] [CrossRef] [PubMed]
  23. Liu, R.; Meng, C.; Zhang, Z.; Ma, H.; Lv, T.; Xie, S.; Liu, Y.; Wang, C. Comparative metabolism of schaftoside in healthy and calcium oxalate kidney stone rats by UHPLC-Q-TOF-MS/MS method. Anal. Biochem. 2020, 597, 113673. [Google Scholar] [CrossRef]
  24. Kim, Y.; Pyeon, J.; Lee, J.; Kim, E.; La, I.; Lee, O.; Kim, K.; Sung, J.; Kim, Y. Chemical fingerprint analysis of fermented Morinda citrifolia L. (noni) juice by UHPLC Q-TOF/MS combined with chemometric analysis. Appl. Biol. Chem. 2024, 67, 59. [Google Scholar] [CrossRef]
  25. Fan, S.; Ma, J.; Yuan, X.; Wang, X.; Wang, Y.; Zhang, Y. Determination of icariside, hyperoside and psoralen in food by liquid chromatography-tandem mass spectrometry. J. Future Foods 2023, 3, 263–272. [Google Scholar] [CrossRef]
  26. Teoh, W.Y.; Yong, Y.S.; Razali, F.N.; Stephenie, S.; Shah, M.D.; Tan, J.K.; Gnanaraj, C.; Esa, N.M. LC-MS/MS and GC-MS analysis for the identification of bioactive metabolites responsible for the antioxidant and antibacterial activities of Lygodium microphyllum (Cav.) R. Br. Separations 2023, 10, 215. [Google Scholar] [CrossRef]
  27. Vijayan, K.P.R.; Raghu, A.V. Tentative characterization of phenolic compounds in three species of the genus Embelia by liquid chromatography coupled with mass spectrometry analysis. Spectrosc. Lett. 2019, 52, 653–670. [Google Scholar] [CrossRef]
  28. Ling, Y.; Zhang, Q.; Zhong, W.; Chen, M.; Gong, H.; He, S.; Liang, R.; Lv, J.; Song, L. Rapid identification and analysis of the major chemical constituents from the fruits of Sapindus mukorossi by HPLC-ESI-QTOF-MS/MS. Nat. Prod. Res. 2019, 34, 2144–2150. [Google Scholar] [CrossRef]
  29. Sun, C.; Wang, J.; Duan, J.; Zhao, G.; Weng, X.; Jia, L. Association of fruit and seed traits of Sapindus mukorossi germplasm with environmental factors in southern China. Forests 2017, 8, 491. [Google Scholar] [CrossRef]
  30. Aboud, N.M.A. Unlocking the genetic potential: Strategies for enhancing secondary metabolite biosynthesis in plants. J. Saudi Soc. Agric. Sci. 2024, 23, 542–554. [Google Scholar] [CrossRef]
  31. Deng, B.; Cao, Y.; Fang, S.; Shang, X.; Yang, W.; Qian, C. Variation and stability of growth and leaf flavonoid content in Cyclocarya paliurus across environments. Ind. Crops Prod. 2015, 76, 386–393. [Google Scholar] [CrossRef]
  32. Bai, Y.; Gu, Y.; Liu, S.; Jiang, L.; Han, M.; Geng, D. Flavonoids metabolism and physiological response to ultraviolet treatments in Tetrastigma hemsleyanum Diels et Gilg. Front. Plant Sci. 2022, 13, 926197. [Google Scholar] [CrossRef] [PubMed]
  33. Wisetkomolmat, J.; Suppakittpaisarn, P.; Sommano, S.R. Detergent plants of northern Thailand: Potential sources of natural saponins. Resources 2019, 8, 10. [Google Scholar] [CrossRef]
  34. Al-Rajhi, A.M.H.; Ghany, T.M.A. In vitro repress of breast cancer by bio-product of edible Pleurotus ostreatus loaded with chitosan nanoparticles. Appl. Biol. Chem. 2023, 66, 33. [Google Scholar] [CrossRef]
  35. Yu, S.; Guo, Q.; Jia, T.; Zhang, X.; Guo, D.; Jia, Y.; Li, J.; Sun, J. Mechanism of action of nicotiflorin from Tricyrtis maculata in the treatment of acute myocardial infarction: From network pharmacology to experimental pharmacology. Drug Des. Dev. Ther. 2021, 15, 2179–2191. [Google Scholar] [CrossRef]
  36. El-Alfy, M.S.; Mostafa, M.E.; Dawidar, A.M.; Abdel-Mogib, M. Phytochemical composition and green insecticides from Citrus aurantifolia fruit peels against whitefly, Bemisia tabaci. Appl. Biol. Chem. 2024, 67, 85. [Google Scholar] [CrossRef]
Horticulturae 11 00682 g001
Figure 1. The collection sites of the whole S. mukorossi plant (A) and its fruit (B) that were used in the study.
Figure 1. The collection sites of the whole S. mukorossi plant (A) and its fruit (B) that were used in the study.
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Figure 2. Chemical structures of the standard compounds rutin (1), nicotiflorin (2), and narcissin (3).
Figure 2. Chemical structures of the standard compounds rutin (1), nicotiflorin (2), and narcissin (3).
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Figure 3. Total polyphenol and total flavonoid content (TPC and TFC) of the different S. mukorossi extracts. TAE, tannic acid equivalent; QE, quercetin equivalent. The results are expressed as mean ± SD. Different letters in each column indicate significant differences at p < 0.05.
Figure 3. Total polyphenol and total flavonoid content (TPC and TFC) of the different S. mukorossi extracts. TAE, tannic acid equivalent; QE, quercetin equivalent. The results are expressed as mean ± SD. Different letters in each column indicate significant differences at p < 0.05.
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Figure 4. Base peak chromatogram (a) of SLE analyzed in negative ionization mode and the MS2 spectra of rutin (b), nicotiflorin (c), and narcissin (d).
Figure 4. Base peak chromatogram (a) of SLE analyzed in negative ionization mode and the MS2 spectra of rutin (b), nicotiflorin (c), and narcissin (d).
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Figure 5. High-performance liquid chromatography (HPLC) chromatogram of the standard compounds: rutin (1), nicotiflorin (2), and narcissin (3).
Figure 5. High-performance liquid chromatography (HPLC) chromatogram of the standard compounds: rutin (1), nicotiflorin (2), and narcissin (3).
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Figure 6. HPLC chromatograms of SLE (a), SST (b), and SPF (c) showing peaks corresponding to rutin (1), nicotiflorin (2), and narcissin (3).
Figure 6. HPLC chromatograms of SLE (a), SST (b), and SPF (c) showing peaks corresponding to rutin (1), nicotiflorin (2), and narcissin (3).
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Figure 7. HPLC chromatograms of DLE (a), DST (b), and DFR (c) showing peaks corresponding to rutin (1), nicotiflorin (2), and narcissin (3).
Figure 7. HPLC chromatograms of DLE (a), DST (b), and DFR (c) showing peaks corresponding to rutin (1), nicotiflorin (2), and narcissin (3).
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Table 1. Proposed structures by UPLC-QTOF-ESI-MS/MS analysis (negative mode).
Table 1. Proposed structures by UPLC-QTOF-ESI-MS/MS analysis (negative mode).
a tRMolecular FormulaMolecular WeightTentative Identification
13.60C33H40O21772.2Quercetin 3-rutinoside-7-glucoside
15.71C27H30O15594.2Vicenin-2
18.00C26H28O14564.1Schaftoside
19.67C27H30O16610.2Rutin
20.06C21H20O12464.1Hyperoside
21.04C27H30O15594.2Nicotiflorin
21.40C28H32O16624.2Narcissin
a Retention time.
Table 2. Calibration data for rutin (1), nicotiflorin (2), and narcissin (3).
Table 2. Calibration data for rutin (1), nicotiflorin (2), and narcissin (3).
Compounda tRRegression Equationb R2
120.77y = 9846.2x + 95,2800.9991
221.51y = 10,370x + 72,1850.9997
321.63y = 16,616x +323,2070.9983
a Retention time. b Coefficient of determination.
Table 3. HPLC quantification of rutin (1), nicotiflorin (2), and narcissin (3) in each S. mukorossi extract.
Table 3. HPLC quantification of rutin (1), nicotiflorin (2), and narcissin (3) in each S. mukorossi extract.
SampleContent (mg/g)
123Total
SLE54.37 ± 0.63 b14.25 ± 0.38 a5.19 ± 0.25 a78.31
SST11.59 ± 0.10 c0.92 ± 0.04 c2.78 ± 0.04 c15.29
SFR1 NDNDNDND
SPF1.89 ± 0.07 ftrtr1.89
SSE2 trtrtrtr
DLE70.21 ± 0.23 a11.21 ± 0.11 b4.02 ± 0.05 b85.44
DST5.89 ± 0.65 d0.41 ± 0.04 d0.76 ± 0.08 d7.06
DFR4.71 ± 0.20 e0.02 ± 0.12 d0.62 ± 0.17 d5.35
DPFtrtrtrtr
DSENDNDNDND
1 Not detected. 2 Trace. Different lower-case letters in each column indicate statistically significant differences at p < 0.05.
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Uy, N.P.; Lee, H.-D.; Ku, J.; Choi, K.; Lee, S. UPLC-QTOF-ESI-MS/MS-Based Comparative Study of Phytochemicals in Sapindus mukorossi. Horticulturae 2025, 11, 682. https://doi.org/10.3390/horticulturae11060682

AMA Style

Uy NP, Lee H-D, Ku J, Choi K, Lee S. UPLC-QTOF-ESI-MS/MS-Based Comparative Study of Phytochemicals in Sapindus mukorossi. Horticulturae. 2025; 11(6):682. https://doi.org/10.3390/horticulturae11060682

Chicago/Turabian Style

Uy, Neil Patrick, Hak-Dong Lee, Jajung Ku, Kyung Choi, and Sanghyun Lee. 2025. "UPLC-QTOF-ESI-MS/MS-Based Comparative Study of Phytochemicals in Sapindus mukorossi" Horticulturae 11, no. 6: 682. https://doi.org/10.3390/horticulturae11060682

APA Style

Uy, N. P., Lee, H.-D., Ku, J., Choi, K., & Lee, S. (2025). UPLC-QTOF-ESI-MS/MS-Based Comparative Study of Phytochemicals in Sapindus mukorossi. Horticulturae, 11(6), 682. https://doi.org/10.3390/horticulturae11060682

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