Biochemical Systematics and Ecology 96 (2021) 104262
12
precocious; begin in autumn and finishing in early summer with only
one spawning event in late summer. According to Riesgo et al.
(2012), oogenesis was a continuous process in E. sepositus, with only
one spawning event in August–September. These data justify the
presence of important amount of saponins in the female gonads
throughout the maturation. In male individuals, the GSI values
indicate that spermatogenesis was restricted to 5–6 months in springsummer. This result concord with the result obtained by Riesgo et al.
(2012) and can explain the low gonads content of saponins observed
in male individuals in early development stage (autumn and spring);
• The seasonal variation of saponins amount in whole body, gonads
and oral body wall of both sexes was significant, with higher rates of
saponins in spawning season over the others.
4. Conclusion
HR-ESI-MS and HR-ESI-MS/MS were applied to study the diversity
and body distribution of saponins in E. sepositus. This technique was used
to monitor the parent saponins and their mass fragmentation patterns in
the MS and MS
2
experiments.
The comparison has been made in respect to body, sex and seasons
components. Our results reported the presence of twelve different
saponin congeners, which highlights the diversity of saponins identified
in a Mediterranean starfish E. sepositus. With the analytical techniques
used, only 8 structures have been elucidated. The distribution of these
saponins in stomach, pyloric caeca, gonads, oral body wall and aboral
body wall of the starfish E. sepositus showed qualitative and quantitative
differences based on MS analysis and semi-quantitative study. The
saponin mixtures in each organ were rich and complex (5–7 different
saponins) with usually one or two saponins differing from one body
compartment to the other and from sex’s individuals to others. Sepositoside A and luzonicoside A were the most abundant saponins in the five
studied organs.
The obtained data indicate high significant inter-organs variability of
saponin amounts in both sexes at different seasons. It is likely to
conclude that the level of saponins in each organ seems to depend on the
season in accordance with the stage of development.
High content of gonads saponin contributed to the maximal saponin
content of whole body in summer (spawning season).
Saponins congeners were located mainly in gonads and oral body
during spawning period, but were located at early developmental stage
in stomach when starfish are actively feeding. The presence of saponins
in the reproductive and digestive organs in such manner suggests that
these compounds are involved in the reproduction and digestion of food.
At this point of the work, HR-ESI-MS and HR-ESI-MS/MS analyses
seems to be efficient for a direct screening of saponin distribution within
the organ extracts of E. sepositus. The identification of keyfragmentations can be very useful to highlighted isomeric saponins by
running HR-ESI-MS/MS analyses. However, an additional characterization step based on a chromatographic separation prior to the mass
spectrometry analyses is required to distinct isomeric saponins.
In the future, chromatography methodologies will be used to separate and purify different saponins from these saponin extracts in order to
clarify the biological roles of each saponin in E. sepositus.
Author contributions
Fouzia Bachari-Houma, Didier Hauchard, Mohamed Chibane and
Vincent Ferri` eres designed the study. Bouchra Dahmoune, Jean-Paul
Gu´ egan, and Philippe J´ ehan performed the experiments. Farid Dahmoune and Lotfi Mouni participated in study design, data collection and
analysis. Bouchra Dahmoune and Farid Dahmoune wrote the manuscript. Cherifa Aissou-Akrour with all the authors revised the
manuscript.
CRediT authorship contribution statement
Bouchra Dahmoune: Conceptualization, Formal analysis, Data
curation, Writing – original draft, Writing – review & editing, Funding
acquisition, acquisition of data, analysis and/or interpretation of data,
Drafting the manuscript, revising the manuscript critically for important
intellectual content. Fouzia Bachari-Houma: Conceptualization,
Writing – review & editing, Conception and design of study, revising the
manuscript critically for important intellectual content. Mohamed
Chibane: Conceptualization, Writing – review & editing, Conception
and design of study, revising the manuscript critically for important
intellectual content. Philippe J´ ehan: Formal analysis, Data curation,
Writing – original draft, Writing – review & editing, Funding acquisition,
acquisition of data, analysis and/or interpretation of data, revising the
manuscript critically for important intellectual content. Jean-Paul
Guegan: Formal analysis, Data curation, Writing – original draft,
Writing – review & editing, Funding acquisition, acquisition of data,
analysis and/or interpretation of data, revising the manuscript critically
for important intellectual content. Farid Dahmoune: Formal analysis,
Data curation, analysis and/or interpretation of data. Cherifa AissouAkrour: revising the manuscript critically for important intellectual
content. Lotfi Mouni: Formal analysis, Data curation, analysis and/or
interpretation of data. Vincent Ferri` eres: Conceptualization, Writing –
review & editing, Conception and design of study, revising the manuscript critically for important intellectual content. Didier Hauchard:
Conceptualization, Writing – review & editing, Conception and design of
study, revising the manuscript critically for important intellectual
content.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgements
We would like to express our sincerest thanks to the Algerian Ministry of Higher Education and Scientific Research, and Campus France
for their scholarship within the framework of Algerian-French cooperation PROFAS B+.
We thank the staff of the laboratories who participated in the realization of this work particularly Isabelle Soutrel for their valuable technical assistance and support, and for providing facilities to realize this
search.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.bse.2021.104262.
References
Dahmoune, B., Gu´ egan, J.P., Vives, T., Soutrel, I., J´ ehan, P., Loutfi, M., Chibane, M.,
Ferri` eres, V., Bachari, H.F., Hauchard, D., 2019. Saponines macrocycliques d’
Echinaster sepositus Extractions et caract´ erisations par RMN, UPLC/MS, MS/MS.
Personnal communication. Journ´ ees Scientifiques SCF-BPL 1-3 Avril 2019. Castel
Saint-Anne, Tregastel. p.65.
De Simone, F., Dini, A., Finamore, E., Minale, L., Pizza, C., Ruccio, R., Riccio, R.,
Zollo, F., 1981. Starfish saponins. Part 5. Structure of seposituside A, a novel
steroidal Glycoside from the starfish Echinaster sepositus. J. Chem. Soc., Perkin
Trans. I, 1855–1862. https://doi.org/10.1039/P19810001855.
De Simone, F., Dini, A., Minale, L., Ricclo, R., Zollo, F., 1980. The sterols OF the asteroid
ECHINASTER sepositus*. Comp. Biochem. Physiol. 66, 351–357. https://doi.org/
10.1016/0305-0491(80)90216-3.
Demeyer, M., De Winter, J., Caulier, G., Eeckhaut, I., Flammang, P., Gerbaux, P., 2014.
Molecular diversity and body distribution of saponins in the sea star Asterias rubens
by mass spectrometry. Comp. Biochem. Physiol. B Biochem. Mol. Bio 168, 1–11.
https://doi.org/10.1016/j.cbpb.2013.10.004.
B. Dahmoune et al.
12
precocious; begin in autumn and finishing in early summer with only
one spawning event in late summer. According to Riesgo et al.
(2012), oogenesis was a continuous process in E. sepositus, with only
one spawning event in August–September. These data justify the
presence of important amount of saponins in the female gonads
throughout the maturation. In male individuals, the GSI values
indicate that spermatogenesis was restricted to 5–6 months in springsummer. This result concord with the result obtained by Riesgo et al.
(2012) and can explain the low gonads content of saponins observed
in male individuals in early development stage (autumn and spring);
• The seasonal variation of saponins amount in whole body, gonads
and oral body wall of both sexes was significant, with higher rates of
saponins in spawning season over the others.
4. Conclusion
HR-ESI-MS and HR-ESI-MS/MS were applied to study the diversity
and body distribution of saponins in E. sepositus. This technique was used
to monitor the parent saponins and their mass fragmentation patterns in
the MS and MS
2
experiments.
The comparison has been made in respect to body, sex and seasons
components. Our results reported the presence of twelve different
saponin congeners, which highlights the diversity of saponins identified
in a Mediterranean starfish E. sepositus. With the analytical techniques
used, only 8 structures have been elucidated. The distribution of these
saponins in stomach, pyloric caeca, gonads, oral body wall and aboral
body wall of the starfish E. sepositus showed qualitative and quantitative
differences based on MS analysis and semi-quantitative study. The
saponin mixtures in each organ were rich and complex (5–7 different
saponins) with usually one or two saponins differing from one body
compartment to the other and from sex’s individuals to others. Sepositoside A and luzonicoside A were the most abundant saponins in the five
studied organs.
The obtained data indicate high significant inter-organs variability of
saponin amounts in both sexes at different seasons. It is likely to
conclude that the level of saponins in each organ seems to depend on the
season in accordance with the stage of development.
High content of gonads saponin contributed to the maximal saponin
content of whole body in summer (spawning season).
Saponins congeners were located mainly in gonads and oral body
during spawning period, but were located at early developmental stage
in stomach when starfish are actively feeding. The presence of saponins
in the reproductive and digestive organs in such manner suggests that
these compounds are involved in the reproduction and digestion of food.
At this point of the work, HR-ESI-MS and HR-ESI-MS/MS analyses
seems to be efficient for a direct screening of saponin distribution within
the organ extracts of E. sepositus. The identification of keyfragmentations can be very useful to highlighted isomeric saponins by
running HR-ESI-MS/MS analyses. However, an additional characterization step based on a chromatographic separation prior to the mass
spectrometry analyses is required to distinct isomeric saponins.
In the future, chromatography methodologies will be used to separate and purify different saponins from these saponin extracts in order to
clarify the biological roles of each saponin in E. sepositus.
Author contributions
Fouzia Bachari-Houma, Didier Hauchard, Mohamed Chibane and
Vincent Ferri` eres designed the study. Bouchra Dahmoune, Jean-Paul
Gu´ egan, and Philippe J´ ehan performed the experiments. Farid Dahmoune and Lotfi Mouni participated in study design, data collection and
analysis. Bouchra Dahmoune and Farid Dahmoune wrote the manuscript. Cherifa Aissou-Akrour with all the authors revised the
manuscript.
CRediT authorship contribution statement
Bouchra Dahmoune: Conceptualization, Formal analysis, Data
curation, Writing – original draft, Writing – review & editing, Funding
acquisition, acquisition of data, analysis and/or interpretation of data,
Drafting the manuscript, revising the manuscript critically for important
intellectual content. Fouzia Bachari-Houma: Conceptualization,
Writing – review & editing, Conception and design of study, revising the
manuscript critically for important intellectual content. Mohamed
Chibane: Conceptualization, Writing – review & editing, Conception
and design of study, revising the manuscript critically for important
intellectual content. Philippe J´ ehan: Formal analysis, Data curation,
Writing – original draft, Writing – review & editing, Funding acquisition,
acquisition of data, analysis and/or interpretation of data, revising the
manuscript critically for important intellectual content. Jean-Paul
Guegan: Formal analysis, Data curation, Writing – original draft,
Writing – review & editing, Funding acquisition, acquisition of data,
analysis and/or interpretation of data, revising the manuscript critically
for important intellectual content. Farid Dahmoune: Formal analysis,
Data curation, analysis and/or interpretation of data. Cherifa AissouAkrour: revising the manuscript critically for important intellectual
content. Lotfi Mouni: Formal analysis, Data curation, analysis and/or
interpretation of data. Vincent Ferri` eres: Conceptualization, Writing –
review & editing, Conception and design of study, revising the manuscript critically for important intellectual content. Didier Hauchard:
Conceptualization, Writing – review & editing, Conception and design of
study, revising the manuscript critically for important intellectual
content.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgements
We would like to express our sincerest thanks to the Algerian Ministry of Higher Education and Scientific Research, and Campus France
for their scholarship within the framework of Algerian-French cooperation PROFAS B+.
We thank the staff of the laboratories who participated in the realization of this work particularly Isabelle Soutrel for their valuable technical assistance and support, and for providing facilities to realize this
search.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.bse.2021.104262.
References
Dahmoune, B., Gu´ egan, J.P., Vives, T., Soutrel, I., J´ ehan, P., Loutfi, M., Chibane, M.,
Ferri` eres, V., Bachari, H.F., Hauchard, D., 2019. Saponines macrocycliques d’
Echinaster sepositus Extractions et caract´ erisations par RMN, UPLC/MS, MS/MS.
Personnal communication. Journ´ ees Scientifiques SCF-BPL 1-3 Avril 2019. Castel
Saint-Anne, Tregastel. p.65.
De Simone, F., Dini, A., Finamore, E., Minale, L., Pizza, C., Ruccio, R., Riccio, R.,
Zollo, F., 1981. Starfish saponins. Part 5. Structure of seposituside A, a novel
steroidal Glycoside from the starfish Echinaster sepositus. J. Chem. Soc., Perkin
Trans. I, 1855–1862. https://doi.org/10.1039/P19810001855.
De Simone, F., Dini, A., Minale, L., Ricclo, R., Zollo, F., 1980. The sterols OF the asteroid
ECHINASTER sepositus*. Comp. Biochem. Physiol. 66, 351–357. https://doi.org/
10.1016/0305-0491(80)90216-3.
Demeyer, M., De Winter, J., Caulier, G., Eeckhaut, I., Flammang, P., Gerbaux, P., 2014.
Molecular diversity and body distribution of saponins in the sea star Asterias rubens
by mass spectrometry. Comp. Biochem. Physiol. B Biochem. Mol. Bio 168, 1–11.
https://doi.org/10.1016/j.cbpb.2013.10.004.
B. Dahmoune et al.