Natural Product Research

Formerly Natural Product Letters

ISSN: 1478-6419 (Print) 1478-6427 (Online) Journal homepage: https://www.tandfonline.com/loi/gnpl20

The chemical profile and biological activity of

different extracts of Sapindus mukorossi Gaertn.
against Cutibacterium acnes

Min-Ping Wei, Jin-Dan Qiu, Lu Li, Yun-Fei Xie, Ya-Hui Guo, Hang Yu, Yu-Liang
Cheng, He Qian & Wei-Rong Yao

To cite this article: Min-Ping Wei, Jin-Dan Qiu, Lu Li, Yun-Fei Xie, Ya-Hui Guo, Hang Yu, Yu-Liang
Cheng, He Qian & Wei-Rong Yao (2020): The chemical profile and biological activity of different
extracts of Sapindus�mukorossi Gaertn. against Cutibacterium�acnes, Natural Product Research,
DOI: 10.1080/14786419.2020.1715399

To link to this article: https://doi.org/10.1080/14786419.2020.1715399

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Published online: 29 Jan 2020.

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NATURAL PRODUCT RESEARCH
https://doi.org/10.1080/14786419.2020.1715399

SHORT COMMUNICATION

The chemical profile and biological activity of different

extracts of Sapindus mukorossi Gaertn. against
Cutibacterium acnes
Min-Ping Weia,b,c, Jin-Dan Qiua,b,c, Lu Lia,b,c, Yun-Fei Xiea,b,c, Ya-Hui Guoa,b
c
, Hang Yua,b,c, Yu-Liang Chenga,b,c, He Qiana,b,c and Wei-Rong Yaoa,b,c
a
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province,
China; bSchool of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province, China;
c
Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province, Jiangnan
University, Wuxi, Jiangsu Province, China

ABSTRACT ARTICLE HISTORY

This study evaluated the antibacterial activities of different Received 7 September 2019
extracts of Sapindus mukorossi Gaertn. (S. mukorossi) on Accepted 4 January 2020
Cutibacterium acnes (C. acnes). The extract solvent and procedure
were screened, based on the yield of saponins and minimum KEYWORDS
inhibitory concentration (MIC). The results showed that the opti- Sapindus mukorossi Gaertn;
antibacterial fraction;
mized product, fermentation and ethyl acetate extract by adding Cutibacterium acnes;
isoamyl alcohol from water extract of S. mukorossi (SWFEAI), had chemical profile
the highest yield of saponins (7.83 ± 0.26%) and the best antibac-
terial activity (MIC ¼ 0.125 mg/mL) on C. acnes. The destroyed
bacterial cell membrane and wall were observed by transmission
electron microscopy, which then resulted in cell lysis and death.
Furthermore, 20 compounds of SWFEAI were detected, among
which oleanane-type triterpenoid saponins with molecular
weights of 734, 750, 882, 924 and 966 were speculated to contrib-
ute to the antibacterial activities of SWFEAI. The results showed
that SWFEAI could be a natural anti-acne agent.

CONTACT Wei-Rong Yao yaoweirongcn@jiangnan.edu.cn State Key Laboratory of Food Science and
Technology, Jiangnan University, Wuxi 214122, Jiangsu Province, China
Supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2020.1715399.
ß 2020 Informa UK Limited, trading as Taylor & Francis Group
2 M.-P. WEI ET AL.

1. Introduction
Cutibacterium acnes (C. acnes), an anaerobic Gram-positive bacterium, is thought to
play an important role in the pathogenesis of inflamed lesions in acne vulgaris, a com-
mon skin disease. Antibiotics such as benzoyl peroxide, tetracycline, and erythromycin
are typical therapeutic agents for C. acnes-induced inflammatory skin diseases, which
act by inhibiting inflammation or killing the bacteria (Lee et al. 2014). However, these
antibiotics have been known to induce side effects, such as dryness, redness, irritation
of skin and hypopigmentation, so still limit their uses (Sharma et al. 2014). Antibiotics
have been used for more than 40 years against C. acnes (Abdel Fattah and Darwish
2013). In the 1970s, evidence of resistance to topical erythromycin and clindamycin
was reported due to improper application and supervision of antibiotics, and since
then, antibiotic resistance in acne has been increasing worldwide (Dreno 2016).
Therefore, it is necessary to develop therapeutic agents for acne that have no side
effects but exhibit high antibacterial activities.
The genus Sapindus belongs to the family Sapindaceae and has proximately 2000
species, of which Sapindus mukorossi Gaertn. (S. mukorossi) is a deciduous tree that is
largely distributed in upper reaches of Indo-Gangetic plains, Shivaliks and sub-
Himalayan tracks at altitudes between 200 m and 1500 m (Sultana et al. 2012). Its fruit
pericarp is rich in saponins such as Sapindoside A, B, C, L and M (Ling et al. 2019),
which are bioactive compounds with a number of potential applications, ranging from
surfactants to physiologically active agents (Dobhal et al. 2007). For example, triterpen-
oid saponins isolated from the fruits of S. mukorossi showed inhibition activities on
the proliferation of human cancer cells (Zhang et al. 2014) and human lung adenocar-
cinoma cells (Zhang et al. 2016). Besides, the methanol extracts of S. mukorossi stem
bark displayed anti-oxidant activity for DPPH with IC50 value of 162.5 – 530.7 lg/mL
(Shah et al. 2017). Microbiologically, water extract of S. mukorossi pericarp showed
strong inhibitory activity against Candida albicans growth with an MIC of 0.039 mg/mL
(Li et al. 2019), and the crude ethanolic extract of S. mukorossi has previously shown
inhibitory activity against C. acnes growth with an MIC of 40 mg/mL (Huang 2014), but
detailed information about the possible mechanism of action was not reported. The
objective of this study was to screen the active fractions of S. mukorossi against C.
acnes based on the yield of saponins and the minimum inhibitory concentrations
(MIC). Furthermore, the extract with the lowest MIC against C. acnes was analyzed via
high performance liquid chromatography-mass spectrometry (HPLC-MS).

2. Results and discussion

2.1. Antibacterial screening of S. mukorossi fractions
A preliminary antibacterial screening of the various fractions from S. mukorossi was
performed (Figure S1, Supplementary material). The antibacterial activity of each frac-
tion against C. acnes are presented in Figure S2 (Supplementary material). The sapo-
nins yield of water extract of S. mukorossi (SW) was the highest, making up
56.84 ± 5.01% (P < 0.05) of SW, however, this extract showed the lowest antibacterial
activity with an MIC of 4.0 mg/mL. While the fermented water extract, subsequently
 NATURAL PRODUCT RESEARCH 3

extracted using ethyl acetate (SWFEA) had a lower saponin yield of 4.59 ± 0.31%
(P < 0.05) but it exhibited the best the antibacterial effect on C. acnes (ATCC 6919)
with an MIC of 0.5 mg/mL, which was lower than that of erythromycin on C. acnes
with an MIC of 0.25 mg/mL. Based on the yield and antibacterial activity, the SWFEA
extract was selected for the subsequent experiments.

2.2. Optimization of the extraction conditions from SWFEA

The subsequent extraction conditions of SWFEA were optimized by changing several
parameters including the extraction solvent, the ratio of SWFEA to extraction solvent,
shaking speed, number of extractions and extraction duration, as shown in Figure S3
(Supplementary material). To summarize the above single parameter study, a mixture
of 25% isoamyl alcohol in ethyl acetate (% v/v) was used as the extractant, its total
volume to SWFEA was 2-fold, and the SWFEA was extracted three times, with 4 min
for each time with shaking frequency of 70 r/min and an extract called SWFEAI
obtained with a higher yield of saponin content, making up 7.83 ± 0.26% of the extract
and an increase in antibacterial activity with an MIC of 0.125 mg/mL. Compared with
SWFEA, the yield of SWFEAI increased by 3.24 ± 0.15%, and the value of MIC decreased
from 0.50 to 0.125 mg/mL. The antibacterial activity of SWFEAI on C. acnes was better
than that of the total saponins of Polyphylla with an MIC of 5.0 mg/mL, which was
probably because of the different raw material or chemical profile (Wang et al. 2016).

2.3. The effect of SWFEAI on the release of macromolecule substances

As presented in Figure S4A (Supplementary material), the optical density in the control
was almost unchanged as time was prolonged, while the optical density in the sam-
ples with 1MIC and 2MIC SWFEAI increased rapidly and reached the highest level at
8 h. During incubation for 14 h, the optical density of the 2MIC sample was higher
than that of the 1MIC sample, and the optical density at 8 h showed a good dose-
response with an r value of 0.9780 (Figure S4B, Supplementary material). A published
study revealed that Mangifera indica kernel extracts could induce the cytoplasmic leak-
age from P. acnes, which was in agreement with the results of current study
(Poomanee et al. 2018), indicating that SWFEAI could break the integrity of C. acnes
cell membranes, resulting in the release of macromolecule substances, such as DNA
and RNA.

2.4. The effect of SWFEAI on the permeability of cell walls

Compared to the control group, the alkaline phosphatase (AKP) activity of C. acnes
sharply increased after SWFEAI treatment at 1MIC or 2MIC (P < 0.05) (Figure S4C,
Supplementary material), and showed a good dose-response relationship (r ¼ 0.9872).
Zanthoxylum bungeanum has been reported to cause the leakage of AKP and inhibit
the growth of C. acnes, which was consistent with the results obtained in this study
(Wang et al. 2017), so it can be inferred that SWFEAI could destroy cell wall permeabil-
ity, resulting in the leakage of AKP.
4 M.-P. WEI ET AL.

2.5. The effects of SWFEAI on C. acnes cell morphology

The morphological changes in bacteria treated with the SWFEAI extract were observed
using Transmission Electron Microscopy (TEM), as shown in Figure S5 (Supplementary
material). The cells of untreated C. acnes showed normal morphological characters
with distinct cell walls, which were long, spindle-shaped, smooth, and lined with cell
membranes (Figure S5A, Supplementary material). Damage to C. acnes cell walls was
observed for bacterial cells treated with 0.25 mg/mL and 0.50 mg/mL SWFEAI for 24 h
(Figure S5B, C, Supplementary material). The cytoplasmic volume decreased and the
cell membrane invaginated with notable structural disorganization within the cell cyto-
plasm, indicated by the black arrows in Figure S5B, C (Supplementary material). In
addition, the cells treated with 0.25 mg/mL SWFEAI exhibited abnormal changes in cell
content material, however, at 0.50 mg/mL, the cell walls were found to be completely
lysed and cell debris was observed. A previous study reported C. acnes treated with
the purified honeybee venom lost the integrity of surface architecture (Sang et al.
2013), which was in accordance with the results of the current study.

2.6. Detection of chemical profile of SWFEAI via HPLC-MS

The potential chemical profile of SWFEAI was detected using HPLC-MS in negative ion
mode. The chromatogram is shown in Figure S6 (Supplementary material). Combined
with the relevant literature (Dobhal et al. 2007; Ling et al. 2019; Wu et al. 2014), 20
compounds were speculated, among which sesquiterpene glucosides and saponins
were detected with retention time ranging from 8 to 20 min (Table S1, Supplementary
material). Compounds with molecular weights (Mr) of 692, 1002, 1146, 1148, 1190 and
1232 were associated with sesquiterpene glycoside groups, while compounds with
molecular weights (Mr) of 734, 750, 792, 882, 924 and 966 were identified as triterpen-
oid saponins, primarily the oleanane type, excepting the compound with Mr of 792
which is of the Hederagenin type (Wu et al. 2014). Oleanane-type triterpenoid sapo-
nins had been reported to possess wide-spectrum antimicrobial activity, therefore the
observed antibacterial activity against C. acnes is potentially due to the presence of
these compounds within the SWFEAI extract (Tamura et al. 2001). In order to obtain
the specific compounds responsible for the antibacterial activity, further purification
is required.

3. Conclusion
In this study, different extracts of S. mukorossi against C. acnes was first carried out,
among which SWFEA exhibited the best the antibacterial effect on C. acnes with an
MIC of 0.5 mg/mL but with a lower saponin yield of 4.59 ± 0.31%. Therefore, the subse-
quent extraction conditions of SWFEA were optimized, and an extract called SWFEAI
was obtained with a higher yield (7.83 ± 0.26%) and the best antibacterial effect (MIC
¼ 0.125 mg/mL). Changes of permeability and integrity of the cell membrane, damage
of the cell wall, and further leakage out of AKP inside the cell were determined as the
bactericidal mechanisms of SWFEAI against C. acnes. Furthermore, the chemical profile
of SWFEAI demonstrated that oleanane-type triterpenoid saponins contributed to the
 NATURAL PRODUCT RESEARCH 5

antibacterial activities of SWFEAI. Therefore, the results of this study would provide a
theoretical basis for supporting SWFEAI as a candidate for future anti-acne agents with
reasonably high antibacterial.

Disclosure statement
There is no conflict of interest of any authors in relation to the submission.

Funding
The work described in this article was supported by the National First-class Discipline Program
of Food Science and Technology (JUFSTR20180509).

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