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\DOI{10.5802/crchim.399}
\datereceived{2024-10-29}
\daterevised{2025-01-26}
\datererevised{2025-03-04}
\dateaccepted{2025-04-09}
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\dateposted{2025-05-06}
\begin{document}

\begin{noXML}

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\title{Use of \textit{Carapa guianensis} Aubl. agro-industrial waste as
an alternative for obtaining bioproducts}

\alttitle{Extraction de bioproduits \`{a} partir de d\'{e}chets
agro-industriels de \textit{Carapa guianensis}}

\author{\firstname{Kyouk Isabel} \lastname{Portilho dos Santos}\CDRorcid{0000-0003-2843-791X}\IsCorresp}
\address{Programa de P\'{o}s Gradua\c{c}\~{a}o em Biodiversidade e
Biotecnologia da Rede BIONORTE, Universidade Federal do Par\'{a}
(PPG-BIONORTE/ICB/UFPA), Rua Augusto Correa, 01, CEP 66075-110,
Bel\'{e}m, PA, Brazil}
\email[K. I. Portilho dos Santos]{kyoukportilho@yahoo.com.br}

\author{\firstname{Renan} \lastname{Campos e Silva}\CDRorcid{0000-0002-0846-9140}}
\address{Laborat\'{o}rio Central de Extra\c{c}\~{a}o, Programa
de P\'{o}s-Gradua\c{c}\~{a}o em Qu\'{i}mica, Universidade Federal do
Par\'{a} (CE/PPGQ/ICEN/UFPA), Rua Augusto Correa, 01, CEP 66075-110,
Bel\'{e}m, PA, Brazil}

\author{\firstname{Anderson}\nobreakauthor\lastname{de\nobreakauthor Santana\nobreakauthor Botelho}\CDRorcid{0000-0002-3143-4608}}
\addressSameAs{2}{Laborat\'{o}rio Central de Extra\c{c}\~{a}o, Programa
de P\'{o}s-Gradua\c{c}\~{a}o em Qu\'{i}mica, Universidade Federal do
Par\'{a} (CE/PPGQ/ICEN/UFPA), Rua Augusto Correa, 01, CEP 66075-110,
Bel\'{e}m, PA, Brazil}

\author{\firstname{Liderl\^{a}nio}\nobreakauthor\lastname{de\nobreakauthor Almeida\nobreakauthor Ara\'{u}jo}\CDRorcid{0000-0002-4081-4354}}
\address{Universidade do Estado do Par\'{a}, Camet\'{a} 1872, Brazil}

\author{\firstname{Ivete}\nobreakauthor\lastname{Furtado\nobreakauthor Ribeiro\nobreakauthor Caldas}\CDRorcid{0000-0002-2095-101X}}
\addressSameAs{3}{Universidade do Estado do Par\'{a}, Camet\'{a} 1872,
Brazil}

\author{\firstname{Wandson Braamcamp} \lastname{de Souza Pinheiro}\CDRorcid{0000-0003-2977-8863}}
\addressSameAs{1}{Programa de P\'{o}s Gradua\c{c}\~{a}o em Biodiversidade e
Biotecnologia da Rede BIONORTE, Universidade Federal do Par\'{a}
(PPG-BIONORTE/ICB/UFPA), Rua Augusto Correa, 01, CEP 66075-110,
Bel\'{e}m, PA, Brazil}
\addressSameAs{2}{Laborat\'{o}rio Central de Extra\c{c}\~{a}o, Programa de
P\'{o}s-Gradua\c{c}\~{a}o em Qu\'{i}mica, Universidade Federal do
Par\'{a} (CE/PPGQ/ICEN/UFPA), Rua Augusto Correa, 01, CEP 66075-110,
Bel\'{e}m, PA, Brazil}

\keywords{\kwd{Agro-industrial waste}\kwd{\textit{Carapa
guianensis}}\kwd{Bioactive compounds}}

\altkeywords{\kwd{D\'{e}chets agro-industriels}\kwd{\textit{Carapa
guianensis}}\kwd{Compos\'{e}s bioactifs}} 

\begin{abstract}
The processing of \textit{Carapa guianensis} seeds to obtain oil on an
industrial scale generates a significant amount of residue,
approximately 66\% m/m, defined as andiroba agro-industrial residue, a
potential source of biomolecules. The chemical profile of the extracts
from this matrix and the fingerprinting by High Performance Thin Layer
Chromatography (HPTLC), as well as the spectroscopic analysis of the
NMR data (\textsuperscript{1}H, \textsuperscript{13}C) and comparison with data obtained in the
literature, allowed the identification of two new triterpene saponins
in the \textit{Carapa} genus (oleanolic acid
3-O-[\tralicstex{Î±}{${\upalpha}$}-L-ramnopyranosyl-(1\tralicstex{â†’}{${\rightarrow}$}2)-\tralicstex{Î±}{${\upalpha}$}-L-arabinopyranosyl]
and hederagenin
3-O-[\tralicstex{Î±}{${\upalpha}$}-L-ramnopyranosyl-(1\tralicstex{â†’}{${\rightarrow}$}2)-\tralicstex{Î±}{${\upalpha}$}-L-arabinopyranosyl]
(Pulsatilla A)), along with three limonoids reported in the species. In
this way, the research showed that even after obtaining andiroba oil,
the agro-industrial residue generated still has a wealth of
phytochemicals, biomass that can be used as an alternative in the
search for bioproducts. 
\end{abstract}

\begin{altabstract}
Le traitement des graines de \textit{Carapa guianensis} pour obtenir de
l'huile \`{a} \'{e}chelle industrielle g\'{e}n\`{e}re une quantit\'{e}
importante de r\'{e}sidus (ratio massique d'environ 66 \%). Ces
r\'{e}sidus peuvent repr\'{e}senter une source potentielle de
biomol\'{e}cules. La caract\'{e}risation des extraits correspondants
par chromatographie sur couche mince \`{a} haute performance et par
spectroscopie RMN (\textsuperscript{1}H, \textsuperscript{13}C) coupl\'{e}e \`{a} une synth\`{e}se
bibliographique, ont permis d'identifier deux nouvelles saponines
triterp\'{e}niques (acide ol\'{e}anolique
3-O-[\tralicstex{Î±}{${\upalpha}$}-L-ramnopyranosyl-(1\tralicstex{â†’}{${\rightarrow}$}2)-\tralicstex{Î±}{${\upalpha}$}-L-arabinopyranosyl] 
et h\'{e}d\'{e}rag\'{e}nine 
3-O-[\tralicstex{Î±}{${\upalpha}$}-L-ramnopyranosyl-(1\tralicstex{â†’}{${\rightarrow}$}2)-\tralicstex{Î±}{${\upalpha}$}-L-arabinopyranosyl] 
(Pulsatilla A)), ainsi que trois limono\"{i}des rapport\'{e}s dans
l'esp\`{e}ce. Ce travail de recherche a d\'{e}montr\'{e} que le
r\'{e}sidu agro-industriel du proc\'{e}d\'{e} d'extraction d'huile
\`{a} partir des graines de \textit{Carapa guianensis} est riche en
compos\'{e}s phytochimiques, qui peuvent \^{e}tre utilis\'{e}s pour
l'extraction de bioproduits.
\end{altabstract}

\shortrunauthors

\maketitle

%{\vspace*{-6pt}}

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\section{Introduction} \label{sec1}

Triterpenoids are molecules that can undergo a variety of structural
modifications. Among these compounds, we highlight pentacyclic
triterpenoids such as lupeol, ${\upalpha}$-amyrin and
${\upbeta}$-amyrin, which are often found in the form of glycosylated
structures called triterpene saponins~\cite{1,2}. They are glycosides
that have antiviral, antitumor and hepatoprotective
properties~\cite{3,4}. In addition to the triterpenoids,
tetranortriterpenes (limonoids) are formed from euphol/tirucallol as a
result of the cyclization of the side chain with the loss of four
carbon atoms giving rise to the formation of a furan ring~\cite{5}.
Chemotaxonomic markers of the Meliaceae family, these compounds make
the species \textit{Carapa guianensis} Aubl.\ an important raw material
for traditional Amazonian medicine. Of the plant's parts, the seed
stands out, from which an oil popularly known as andiroba oil is
extracted. This oil is widely used in the treatment of numerous
pathologies, as it has anti-inflammatory, analgesic, antiarthritic,
antitumor, larvicidal and antimicrobial properties~\cite{6,7,8}. Among
the numerous phytochemical compounds extracted from various parts of
plants in the Meliaceae family, limonoids stand out as the most
significant. These compounds are noted for their diverse biological
activities, including insecticidal, antifungal, antimalarial,
antibacterial, antiviral, and anticancer properties~\cite{9,10}.

On an industrial scale, andiroba oil is obtained by cold-pressing the
seeds (kernels and husks) without adding organic solvents, which
consists in a continuous and clean extraction, free of any chemical
additives, generating a solid by-product~\cite{11}. The quality of the
by-product is a reflection of the process used to obtain the oil, as
pressing the seeds allows for greater efficiency in the extraction of
lipid metabolites. However, the downside is the significantly low yield
of vegetable oil extracted~\cite{12}, which results in a greater need
for raw materials to obtain a considerable amount of product,
generating an accumulation of agro-industrial waste. In this case, cold
pressing extracts 34\% m/m of extra virgin oil and leaves a significant
percentage of residue, approximately 66\% m/m, of which only a small
part is used in the production of repellent candles~\cite{11,13}. The
rest is discarded, burned, used in compost, or occasionally used as a
complement to pig feed~\cite{14}.

This agro-industrial residue, when combined with well-controlled
production processes and appropriate pretreatment, can be used as a
primary material to obtain substances and sub-extracts of economic and
pharmacological interest, since even after the targeted processing, the
resulting residue can still contain a number of important 
compounds~\cite{14,15,16}. In general, the use of agro-industrial waste
has become a useful alternative to issues of sustainability and
scarcity of resources, as well as being a source of bioproducts.
Indeed, they are derived from peels and seeds, which can still contain
fibers and bioactive compounds after industrial processing, thus
presenting numerous technological applications, such as in the
production of pharmaceuticals and cosmetics~\cite{17,18}. Furthermore,
the use of by-products is proving to be a tool for enhancing the value
of the production chain.

Therefore, given the economic importance of the \textit{Carapa
guianensis} species and the fact that industrial processing to obtain
oil from its seeds generates a significant percentage of
agro-industrial residue, the aim of this research was to investigate
the chemical composition of the agro-industrial residue from 
\textit{C.~guianensis} seeds in order to identify the presence of
substances of economic and pharmacological interest that add value to
the residue.{\vspace*{-6pt}}

\section{Material and methods}\label{sec2}
{\vspace*{-3pt}}
\subsection{Chemical products and materials}\label{ssec21}

All the chemical reagents and solvents used for extractions,
chromatographic and spectroscopic analyses were of analytical grade,
from TEDIA$^{\text{\textregistered}}$\ COMPANY (Fairfield, USA). Column
chromatography (CC) was performed using normal phase silica gel
(60--200~${\upmu}$m) (SiliCycle Inc., Quebec, Canada). In addition,
thin layer chromatography (TLC) was carried out on SiliaPlate TLC
chromatography plates with aluminum and glass supports, silica, 
200~${\upmu}$m, 20 ${\times}$ 20~cm, F 254 (SiliCycle Inc., Quebec,
Canada). The structural elucidation of the isolated substances was
carried out using one-dimensional nuclear magnetic resonance (NMR). The
NMR spectra were obtained on a VARIAN$^{\text{\textregistered}}$\ Mercury Plus
spectrometer operating at 300~MHz for $^{1}$H and 75~MHz for $^{13}$C.
The chemical shifts were recorded with dimensionless ${\updelta}$
values (ppm), with the signal from the deuterated solvent as the
internal reference. The solvents were CDCl\tsub{3} and
C\tsub{5}D\tsub{5}N from CAMBRIDGE ISOTOPE
LABORATORIES$^{\text{\textregistered}}$.

{\vspace*{-6pt}}

\subsection{Obtaining and processing the agro-industrial residue
\textit{Carapa guianensis}}\label{ssec22}

The extracted cake from the mechanical pressing of
\textit{C.~guianensis} seeds was purchased from the local industry in
the municipality of Ananindeua, in the state of Par\'{a}, in the
northern region of Brazil (SisGen registration number A3B8C54). This
cake was dehydrated in an oven at a temperature ($T$) of 
45~\textdegree C and humidity ($U$\%) of 20\% until it reached a
constant mass. The dehydrated material was crushed and sieved through a
40 mesh, and the resulting sample was vacuum-packed and stored at
25~\textdegree C.

{\vspace*{-6pt}}

\subsection{Preparation of the extracts}\label{ssec23}

The phytochemical analysis of the \textit{C.~guianensis}
agro-industrial residue was based on preliminary tests in triplicates,
assessing the extraction method (cold and hot) and the extraction
efficiency of the organic solvents used (hexane, ethyl acetate, and
ethanol). In both extractions, a sample mass/solvent volume ratio of
1:10 m/v was adopted. The cold extractions using a single solvent and
successive extractions (increasing polarity gradient) lasted 24~h at
room temperature each. Each extraction was carried out in triplicate in
a single batch, with the solvents renewed twice. The hot Soxhlet
extractions lasted three hours for each solvent. All extract solutions
obtained were concentrated to solids in a Syncore rotary evaporator
(B\"{U}CHI - Flawil, Switzerland).{\vspace*{-6pt}}

\subsection{Characterization of the extracts by HPTLC}\label{ssec24}

The chemical profile of the extracts was obtained by HPTLC (High
Performance Thin-Layer
Chromatography - CAMAG$^{\text{\textregistered}}$). The samples were
deposited in spot form, 10~${\upmu}$g/spot for the extracts and 
1~${\upmu}$g/spot for the limonoid standard (7-desacetoxy-7-oxogedunin)
on silica gel F-254 60~\AA{} chromatoplates with glass support
(SILICYCLE$^{\text{\textregistered}}$) in an ATS4 (Automatic TLC
Sample4) module. The chromatoplates were then eluted in the AMD2
(Automated Multiple Development) module in gradient mode, following the
12-step adjustment with decreasing polarity and a chromatographic path
of 80~mm, using a mixture of organic solvents as the mobile phase:
hexane/ethyl acetate/methanol (1\% formic acid), ranging linearly from
70/15/15\% to 100/0/0\%. It was then developed using a selective
reagent for terpenes and limonoids (VSA) and scanned using the TLC
Visualizer photodocumenter in white light. The data was stored and
processed in the WinCats - Planar Chromatography Manager software,
version 1.4.6 (CAMAG, Muttenz, Switzerland).{\vspace*{-6pt}}

\subsection{Fractionation and isolation}\label{ssec25}

The fractionation of 50~g of the extract obtained from the hot
ethanolic extraction of the agro-industrial residue of
\textit{C.~guianensis} (E9) was performed by column chromatography
(CC), using silica Flash G60 (60--200~${\upmu}$m)
(SILICYCLE$^{\text{\textregistered}}$) as the stationary phase and, as
a mobile phase, a gradient of increasing polarity with seven
hexane/ethyl acetate/methanol systems: (1) 90:0.5:0.5, (2) 80:10:10,
(3) 70:15:15, (4)~60:20:20, (5) 50:25:25, (6) 00:50:50, and (7)
00:00:100. The fractions obtained were analyzed by HPTLC and combined
based on similarities into 4 fractions: F1~(2.0 g), F2 (2.77 g), F3
(1.62 g), and F4 (3.65 g).

Fractions F1 to F3 were subjected to silica gel column chromatography
using mixtures of hexane, ethyl acetate, and methanol as eluents, in
ascending order of polarity. The fractions were monitored by TLC and
revealed with VSA and visible light. Fraction F4 was also subjected to
silica gel column chromatography, but using dichloromethane (100\%) as
the mobile phase. The resulting chromatographic fractions were analyzed
by TLC and combined into fractions 55 to 80, which were purified by
preparative thin layer chromatography. Two 50~mg injections of samples
solubilized in methanol were deposited as a 180~mm ${\times}$ 3~mm band
on preparative TLC and eluted in a 15\% dichloromethane/methanol
system, following a 150~mm chromatographic path. The chromatographic
bands were demarcated and removed, each sample was solubilized in
methanol and filtered. All the substances were identified by 1D
($^{1}$H and $^{13}$C) and 2D ($^{1}$H--$^{13}$C HETCOR, HSQC, and
HMBC) NMR spectroscopy , and the data compared with literature:

Gedunin (S1): From F1, fractions 56 to 72, a white amorphous solid
(15~mg); $^{1}$H NMR (CDCl\tsub{3}) and $^{13}$C NMR (CDCl\tsub{3})
(see Tables~\ref{tab2} and \ref{tab3}; Section S1, Supplementary
Material).

6${\upalpha}${-Acetoxygedunin (S2): From F2, fractions 33 to 36 (white
crystals) (18~mg);} $^{1}${H NMR (CDCl}\tsub{3}{) and} $^{13}$C NMR
(CDCl\tsub{3}) (see Tables~\ref{tab2} and~\ref{tab3}; Section S2,
Supplementary Material).

7-Desacetoxy-7-oxogedunin (S3): From F3, fractions 38 to 52 (white
crystals) (15~mg), $^{1}$H NMR (CDCl\tsub{3}) and $^{13}$C NMR
(CDCl\tsub{3}) (see Tables~\ref{tab2} and~\ref{tab3}; Section S3,
Supplementary Material).

Oleanolic acid
3-O-[${\upalpha}$-L-ramnopyranosyl-(1${\rightarrow}$2)-${\upalpha}$-L-arabinopyranosyl] (S4): 
From F4, a white amorphous solid (38~mg); $^{1}$H NMR
(C\tsub{5}D\tsub{5}N) and $^{13}$C NMR (C\tsub{5}D\tsub{5}N) (see
Tables~\ref{tab4} and~\ref{tab5}; Section S4, Supplementary Material).

Hederagenin
3-O-[${\upalpha}$-L-ramnopyranosyl-(1${\rightarrow}$2)-${\upalpha}$-L-arabinopyranosyl] 
(Pulsatilla~A) (S5): From F4, a white amorphous solid (15~mg); $^{1}$H
NMR (C\tsub{5}D\tsub{5}N) and $^{13}$C NMR (C\tsub{5}D\tsub{5}N) (see
Tables~\ref{tab4} and~\ref{tab5}; Section S5, Supplementary Material).
{\vspace*{-5pt}}

\subsection{Fingerprint of hot ethanolic extract (E9) and commercial
\textit{C.~guianensis} oil}\label{ssec26}

The fingerprint of E9 and commercial andiroba oil was evaluated
according to Section~\ref{ssec24}, this time by injecting the masses of
the substances isolated and identified in this work (S1, S2, S3, S4,
and S5).

\section{Results and discussion}\label{sec3}
\subsection{Extractor efficiency}\label{ssec31}

Regardless of the solvent used, hot extraction gave higher mass
percentage yields than cold extraction, with a percentage increase in
yield ranging from 38.96 to 53.18\%. When comparing the extracting
capacity of each solvent, ethanol gave the highest mass yield
(26.96\%).

When the extraction was carried out with solvents in succession in
increasing order of polarity, hexane was the one with the highest mass
yield (12.14\%). The cold extraction method with successive solvents
could therefore be more suitable to extract low polarity components
with greater affinity for hexane.

Thus, this work uses a chemical investigation approach to investigate
the agro-industrial residue of \textit{C.~guianensis}. The choice of
the extracting solvents was according to their selectivity with low,
medium, and high polarity molecules, which allowed phytochemical
screening. The ethanolic extract was chosen for the other studies due
to its higher degradation rate. Above all, the preference for ethanol
arises from its prevalence in biological studies using plant extracts,
due to its low toxicity and low persistence in Nature. In addition, its
low cost and large-scale production from renewable sources make it one
of the safest solvents in the recovery of natural\unskip\break assets~\cite{19}.

At the same time, green chemistry approaches have been developed to
mitigate the undesirable effects of traditional extraction protocols,
aiming to replace organic solvents with environmentally friendly
alternatives. In this scenario, natural deep eutectic solvents (NADESs)
have emerged as a pioneering proposal for the recovery of
phytochemicals, as they offer advantages such as low toxicity,
biodegradability, solute stabilization, sustainability, simple
preparation, and tunable viscosity and density~\cite{20}. However, it
is important to emphasize that for efficient extraction, some
parameters must be considered, such as the physical properties of the
solvent. For example, NADESs present hydrogen bond networks between the
constituents, making them viscous, which compromises the extraction
efficiency and, consequently, the affinity capacity of the target
compounds with their phase. Therefore, there is a need to adjust this
property~\cite{21,22}. This implies additional investigations on the
use of NADESs in the activities of isolation, separation, and recovery
of bioactive compounds present in plant matrices, in addition to the
elaboration and evaluation of extraction protocols using NADESs, the
stability of biomolecules, production costs and their effects on human
life and health, before their use in the pharmaceutical
industry~\cite{21}.

\subsection{Chemical profile by high performance thin layer
chromatography (HPTLC)}\label{ssec32}

Chemical profile by HPTLC made it possible to evaluate the metabolic
complexity of the compounds present in the extracts of the
agro-industrial residue of \textit{C.~guianensis}. Following the
methods described by~\cite{15}, the use of vanillin--sulphuric acid
solution (VSA) as a chemical developer allows the detection of fatty
acids (blue), as well as terpenes, limonoids (chemotaxonomic marker)
and steroids (purple). It was therefore possible to determine the
occurrence of substances from these classes in all the extracts,
because bands were formed with blue and/or purple tones after the plate
was revealed with VSA. The Rf comparison with the standard
7-desacetoxy-7-oxogedunin (track 1 in Figure~\ref{fig1} Rf 0.53) made
it possible to observe the presence of the limonoid
7-desacetoxy-7-oxogedunin in some extracts (0.53), based on the
proportion of mass and intensity of coloration. It is notably observed
in the hot ethanolic extract (tracks 18--19 in Figure~\ref{fig1}), in
addition to the diversity of compounds displayed through
chromatographic bands throughout the chromatoplate, thus proving that
this biomass still has a wealth of biomolecules even after extracting
andiroba oil.

\begin{figure*}
\includegraphics{fig01}
{\vspace*{2pt}}
\caption{\label{fig1}Result of the analysis of the chemical profile,
via HPTLC, of the extracts obtained cold and hot, with the solvents
Hex, AcOEt and EtOH (each in duplicate, see Table~\ref{tab1}) and the
limonoid standard 7-desacetoxy-7-oxogedunin (track 1). Elution system,
gradient mode: Hexane/AcOEt/MeOH (1\% HCOOH). VSA derivatizer.}
{\vspace*{2pt}}
\end{figure*}

Table~\ref{tab1} identifies, by means of acronyms, all the extracts
obtained from the andiroba agro-industrial residue, in the cold and hot
method, with the solvents: hexane, ethyl acetate and ethanol, as well
as the standard.

%tab1
\begin{table*}
\caption{\label{tab1}Names of cold and hot extracts and the limonoid
standard}
\begin{tabular}{ccccccccc}
\thead
\multicolumn{2}{c}{\morerows{1}{\raisebox{-2pc}{Standard}}}
&amp; \morerows{1}{\raisebox{-1pc}{\parbox[t]{5pc}{\centering Organic solvents}}}
&amp; \multicolumn{4}{c}{Cold extraction} &amp; \multicolumn{2}{c}{Hot
extraction} \\ \cline{4-7}\cline{8-9}
&amp; &amp;  &amp; 
\multicolumn{2}{c}{\parbox[t]{5pc}{\centering Single solvent maceration}} &amp; 
\multicolumn{2}{c}{\parbox[t]{5pc}{\centering Successive maceration}}\raisebox{-15pt}{} &amp; &amp; \\
\cline{1-2}\cline{4-5}\cline{6-7}
7-desacetoxy-7-oxogedunin &amp; Track &amp; &amp; Extract &amp; Track &amp; Extract &amp; Track &amp; Extract &amp; Track \\
\endthead
P &amp; 1 &amp; Hexane &amp; E1 &amp; 2--3 &amp; E2 &amp; 4--5 &amp; E3 &amp; 6--7 \\ 
&amp; &amp; Ethyl acetate &amp; E4 &amp; 8--9 &amp; E5 &amp; 10--11 &amp; E6 &amp; 12--13 \\ 
&amp; &amp; Ethanol &amp; E7 &amp; 14--15 &amp; E8 &amp; 16--17 &amp; E9 &amp; 18--19
\botline
\end{tabular}
\end{table*}

\subsection{Structural identification}\label{ssec33}

Tetranortriterpenes identified in the study, gedunin (S1),
6${\upalpha}$-acetoxygedunin (S2), and 7-desacetoxy-7-oxogedunin (S3)
(Figure~\ref{fig2}), are already known in the species, and the study
only compared the $^{13}$C (CDCl\tsub{3}, 75~MHz) and $^{1}$H
(CDCl\tsub{3}, 300~MHz) NMR signals with data from the literature, in
addition to two-dimensional correlation maps 2D $^{1}$H $^{13}$C
(HETCOR) and $^{1}$H $^{13}$C (HMBC).

\begin{figure*}
\includegraphics{fig02}
\caption{\label{fig2}Limonoids identified in the agro-industrial waste
of \textit{C.~guianensis}.}
\end{figure*}

Gedunin (S1): characteristic signals of hydrogens and aromatic carbons
at $\updelta_{\mathrm{H}}$ 7.41 ($dl$, $J$ 1.5~Hz, 2H) and 6.33 ($t$,
$J$ 1.2~Hz, 1H); $\updelta_{\mathrm{C}}$ 120.4, 143.1, 109.8, and
141.2, respectively, indicating the presence of a furan ring in the
structure (Tables~\ref{tab2} and~\ref{tab3}; Section S1, Figures
S1A--S1C,  Supplementary Material). 

%tab2
\begin{table*}
\caption{\label{tab2}$^{1}$H NMR spectroscopic data for S1--S3\vspace*{-4pt}}
\fontsize{9}{10.1}\selectfont
\tabcolsep2pt
\begin{tabular}{ccccccc}
\thead
\xmorerows{1}{$^{1}$H} &amp; 
S1 &amp; 1$^{\mathrm{a}}$ &amp;
S2 &amp; 2$^{\mathrm{b}}$ &amp;
S3 &amp; 3$^{\mathrm{b}}$ \\\cline{2-3}\cline{4-5}\cline{6-7}
&amp; \multicolumn{2}{c}{$\updelta_{\mathrm{H}}$} 
&amp; \multicolumn{2}{c}{$\updelta_{\mathrm{H}}$}
&amp; \multicolumn{2}{c}{$\updelta_{\mathrm{H}}$} \\
\endthead
1 &amp; 7.09 ($d$ 10.2~Hz) &amp; 7.07 ($d$ 10.2~Hz) &amp; 7.07 ($d$ 10.2~Hz) &amp; 7.07 ($d$ 10.0~Hz) &amp; 7.09 ($d$ 10.2~Hz) &amp; 7.09 ($d$ 10.0~Hz) \\ 
2 &amp; 5.87 ($d$ 10.2~Hz) &amp; 5.84 ($d$ 10.2~Hz) &amp; 5.94 ($d$ 10.0~Hz) &amp; 5.93 ($d$ 10.0~Hz) &amp; 5.91 ($d$ 10.2~Hz) &amp; 5.94 ($d$ 10.0~Hz) \\ 
3 &amp; - &amp; - &amp; - &amp; - &amp; - &amp; - \\ 
4 &amp; - &amp; - &amp; - &amp; - &amp; - &amp; - \\ 
5 &amp; 
\parbox[t]{5pc}{\centering 2.16 ($dd$ 12.9 and 2.4~Hz)} &amp; 
\parbox[t]{5pc}{\centering 2.12 ($dd$ 12.9 and 2.4~Hz)} &amp; 
\parbox[t]{5pc}{\centering 2.52 ($d$ 12.6~Hz)} &amp; 
\parbox[t]{5pc}{\centering 2.52 ($d$ 13.0~Hz)} &amp; 
\parbox[t]{5pc}{\centering 2.16 ($dd$ 14.4 and 3.0~Hz)} &amp; 
\parbox[t]{5pc}{\centering 2.17 ($dd$ 14.0 and 3.0~Hz)} \\ 
6 &amp; 1.80--1.92 ($m$) &amp; 
\parbox[t]{5pc}{\centering 1.79 ($t$ 12.0~Hz) 1.92 ($d$ 12.0~Hz)} &amp; 
\parbox[t]{5pc}{\centering 5.27 ($dd$ 12.6 and 2.4~Hz)} &amp; 
\parbox[t]{5pc}{\centering 5.27 ($dd$ 13.0 and 2.5~Hz)} &amp; 
\parbox[t]{6pc}{\centering ${\upalpha}$ 2.39 ($dd$ 13.8 and 3.3~Hz) ${\upbeta}$ 2.92 ($t$~14.4~Hz)} &amp; 
\parbox[t]{6pc}{\centering ${\upalpha}$ 2.41 ($dd$ 14.0 and 3.0~Hz) ${\upbeta}$ 2.92 ($t$~14.0~Hz)} \\ 
7 &amp; 4.55 ($sl$) &amp; 4.52 ($sl$) &amp; 4.88 ($d$, 2.1~Hz) &amp; 4.89 ($d$, 2.5~Hz) &amp; - &amp; - \\ 
8 &amp; - &amp; - &amp; - &amp; - &amp; - &amp; - \\ 
9 &amp; 
\parbox[t]{5pc}{\centering 2.48 ($dd$ 12.3 and 6.3~Hz)} &amp; 
\parbox[t]{5pc}{\centering 2.46 ($dd$ 12.7 and 6.2~Hz)} &amp; 
\parbox[t]{5pc}{\centering 2.52 ($d$ 12.6~Hz)} &amp; 
\parbox[t]{5pc}{\centering 2.53 ($d$ 13.0~Hz)} &amp; 
\parbox[t]{5pc}{\centering 2.19 ($dd$ 10.5 and 1.8~Hz)} &amp; 
\parbox[t]{5pc}{\centering 2.21 ($dd$ 10.5 and 1.5~Hz)} \\ 
10 &amp; - &amp; - &amp; - &amp; - &amp; - &amp; - \\ 
11 &amp; 1.80--2.03 ($m$) &amp; 1.81--2.00 ($m$) &amp; 1.88--1.99 ($m$) &amp; 1.88--1.99 ($m$) &amp; 1.80--1.99 ($m$) &amp; 1.80--1.99 ($m$) \\ 
12 &amp; 
1.60--1.71 ($m$) &amp; 
\parbox[t]{5pc}{\centering 1.56 ($dd$, 11.0 and 12.0~Hz) 1.70 ($m$)} &amp; 
1.59--1.75 ($m$) &amp; 
1.59--1.74 ($m$) &amp; 
1.48--1.84 ($m$) &amp; 
1.48--1.84 ($m$) \\ 
13 &amp; - &amp; - &amp; - &amp; - &amp; - &amp; - \\ 
14 &amp; - &amp; - &amp; - &amp; - &amp; - &amp; - \\ 
15 &amp; 3.53 ($s$) &amp; 3.50 ($s$) &amp; 3.61 ($s$) &amp; 3.61 ($s$) &amp; 3.85 ($s$) &amp; 3.86 ($s$) \\ 
16 &amp; - &amp; - &amp; - &amp; - &amp; - &amp; - \\ 
17 &amp; 5.61 ($s$) &amp; 5.59 ($s$) &amp; 5.61 ($s$) &amp; 5.61 ($s$) &amp; 5.45 ($s$) &amp; 5.46 ($s$) \\ 
Me-18 &amp; 1.22 ($s$) &amp; 1.22 ($s$) &amp; 1.25 ($s$) &amp; 1.24 ($s$) &amp; 1.12 ($s$) &amp; 1.13 ($s$) \\ 
Me-19 &amp; 1.20 ($s$) &amp; 1.19 ($s$) &amp; 1.21 ($s$) &amp; 1.20 ($s$) &amp; 1.34 ($s$) &amp; 1.35 ($s$) \\ 
20 &amp; - &amp; - &amp; - &amp; - &amp; - &amp; - \\ 
21 &amp; 7.41 ($dl$ 1.5~Hz) &amp; 7.39 ($dl$ 1.3~Hz) &amp; 7.41 ($dl$ 1.0~Hz) &amp; 7.41 ($dl$ 1.0~Hz) &amp; 7.40 ($dl$ 1.0~Hz) &amp; 7.41 ($dl$ 1.0~Hz) \\ 
22 &amp; 6.33 ($t$ 1.2~Hz)  &amp; 6.31 ($t$ 1.3~Hz)  &amp; 6.33 ($t$ 1.2~Hz)  &amp; 6.33 ($t$ 1.5~Hz)  &amp; 6.35 ($dl$ 1.2~Hz) &amp; 6.36 ($dl$ 1.5~Hz) \\ 
23 &amp; 7.41 ($dl$ 1.5~Hz) &amp; 7.39 ($dl$ 1.3~Hz) &amp; 7.41 ($dl$ 1.0~Hz) &amp; 7.41 ($dl$ 1.0~Hz) &amp; 7.38 ($t$ 1.8~Hz)  &amp; 7.39 ($t$ 1.5~Hz) \\ 
Me-28 &amp; 1.06 ($s$) &amp; 1.03 ($s$) &amp; 1.26 ($s$) &amp; 1.25 ($s$) &amp; 1.14 ($s$) &amp; 1.15 ($s$) \\ 
Me-29 &amp; 1.07 ($s$) &amp; 1.04 ($s$) &amp; 1.16 ($s$) &amp; 1.16 ($s$) &amp; 1.12 ($s$) &amp; 1.12 ($s$) \\ 
Me-30 &amp; 1.15 ($s$) &amp; 1.12 ($s$) &amp; 1.24 ($s$) &amp; 1.27 ($s$) &amp; 1.20 ($s$) &amp; 1.21 ($s$) \\ 
6-OCO\underline{CH3} &amp; - &amp; - &amp; 2.03 ($s$) &amp; 2.03 ($s$) &amp; - &amp; - \\ 
7-OCO\underline{CH3} &amp; 2.03 ($s$) &amp; 2.07 (s) &amp; 2.15 ($s$) &amp; 2.15 ($s$) &amp; - &amp; -
\botline
\end{tabular}
\tabnote{Measured in CDCl\tsub{3}. Chemical shifts (${\updelta}$) are in
ppm from a tetramethylsilane reference. $^{\mathrm{a}}$Khalid; Duddeck;
Gonzalez-Sierra~\cite{23}; $^{\mathrm{b}}$Silva; Nunomura~\cite{24}.}
{\vspace*{-5pt}}
\end{table*}

%tab3
\begin{table*}
\caption{\label{tab3}$^{13}$C NMR spectroscopic data for S1--S3{\vspace*{3pt}}}
\fontsize{10}{12.3}\selectfont\tabcolsep9pt
\begin{tabular}{ccccccc}
\thead
\xmorerows{1}{$^{13}$C} &amp; 
S1 &amp; 1$^{\mathrm{a}}$ &amp;
S2 &amp; 2$^{\mathrm{b}}$ &amp;
S3 &amp; 3$^{\mathrm{b}}$ \\ \cline{2-7}
&amp; \multicolumn{2}{c}{$\updelta_{\mathrm{C}}$} 
&amp; \multicolumn{2}{c}{$\updelta_{\mathrm{C}}$}
&amp; \multicolumn{2}{c}{$\updelta_{\mathrm{C}}$} \\
\endthead
\01   &amp; 157.0 &amp; 157.0 &amp; 156.2 &amp; 156.0 &amp; 155.9 &amp; 156.3 \\ 
\02   &amp; 126.0 &amp; 125.9 &amp; 126.6 &amp; 126.7 &amp; 126.3 &amp; 126.8 \\ 
\03   &amp; 204.0 &amp; 204.0 &amp; 204.1 &amp; 204.2 &amp; 203.2 &amp; 203.4 \\ 
\04   &amp; \044.0  &amp; \044.0 &amp; \044.9 &amp; \045.2 &amp; \045.1 &amp; \045.4 \\ 
\05   &amp; \046.0  &amp; \046.0 &amp; \038.4 &amp; \038.6 &amp; \054.5 &amp; \054.8 \\ 
\06   &amp; \023.2  &amp; \023.2 &amp; \069.6 &amp; \069.8 &amp; \036.6 &amp; \036.9 \\ 
\07   &amp; \073.2  &amp; \073.2 &amp; \072.6 &amp; \072.8 &amp; 208.2  &amp; 208.3 \\ 
\08   &amp; \042.6  &amp; \042.6 &amp; \043.0 &amp; \043.8 &amp; \053.5 &amp; \053.6 \\ 
\09   &amp; \039.5  &amp; \039.5 &amp; \047.8 &amp; \047.9 &amp; \047.5 &amp; \047.8 \\ 
10    &amp; \040.0  &amp; \040.0 &amp; \040.6 &amp; \040.8 &amp; \039.5 &amp; \039.8 \\ 
11    &amp; \014.9  &amp; \014.9 &amp; \015.0 &amp; \015.2 &amp; \017.3 &amp; \017.6 \\ 
12    &amp; \026.0  &amp; \025.9 &amp; \025.8 &amp; \026.1 &amp; \032.1 &amp; \032.4 \\ 
13    &amp; \038.7  &amp; \038.7 &amp; \038.7 &amp; \038.9 &amp; \037.7 &amp; \037.9 \\ 
14    &amp; \069.8  &amp; \069.7 &amp; \069.5 &amp; \069.7 &amp; \065.5 &amp; \065.8 \\ 
15    &amp; \056.8  &amp; \056.8 &amp; \056.2 &amp; \056.4 &amp; \053.4 &amp; \053.8 \\ 
16    &amp; 167.5   &amp; 167.4  &amp; 167.1  &amp; 167.3  &amp; 166.8  &amp; 167.0 \\ 
17    &amp; \078.3  &amp; \078.2 &amp; \078.1 &amp; \078.3 &amp; \077.9 &amp; \078.2 \\ 
Me-18 &amp; \017.7  &amp; \017.7 &amp; \017.9 &amp; \018.1 &amp; \020.6 &amp; \020.8 \\ 
Me-19 &amp; \019.7  &amp; \019.7 &amp; \021.4 &amp; \021.6 &amp; \019.8 &amp; \020.0 \\ 
20    &amp; 120.4   &amp; 120.4  &amp; 120.3  &amp; 120.4  &amp; 120.1  &amp; 120.5 \\ 
21    &amp; 143.1   &amp; 143.1  &amp; 141.2  &amp; 141.2  &amp; 141.0  &amp; 141.0 \\ 
22    &amp; 109.8   &amp; 109.8  &amp; 109.8  &amp; 110.0  &amp; 109.7  &amp; 109.9 \\ 
23    &amp; 141.2   &amp; 141.2  &amp; 143.1  &amp; 143.3  &amp; 143.1  &amp; 143.3 \\ 
Me-28 &amp; \027.1  &amp; \027.1 &amp; \031.6 &amp; \031.6 &amp; \026.9 &amp; \027.2 \\ 
Me-29 &amp; \021.2  &amp; \021.2 &amp; \020.2 &amp; \020.5 &amp; \020.9 &amp; \021.2 \\ 
Me-30 &amp; \018.3  &amp; \018.3 &amp; \018.0 &amp; \018.3 &amp; \017.1 &amp; \017.4 \\ 
6-OCO\underline{CH3} &amp; - &amp; - &amp; \021.2 &amp; \021.4 &amp; - &amp; - \\ 
6-O\underline{C}OCH3 &amp; - &amp; - &amp; 170.1 &amp; 170.3 &amp; - &amp; - \\ 
7-OCO\underline{CH3} &amp; \021.0 &amp; \021.0 &amp; \020.9 &amp; \021.1 &amp; - &amp; - \\ 
7-OCOCH3 &amp; 169.9 &amp; 169.9 &amp; 170.0 &amp; 170.2 &amp; - &amp; -
\botline
\end{tabular}
\tabnote{Measured in CDCl\tsub{3}. Chemical shifts (${\updelta}$) are in
ppm from a tetramethylsilane reference. $^{\mathrm{a}}$Khalid; Duddeck;
Gonzalez-Sierra~\cite{23}; $^{\mathrm{b}}$Silva; Nunomura~\cite{24}.}
{\vspace*{3pt}}
\end{table*}

6${\upalpha}$-Acetoxygedunin (S2): typical signals from hydrogens and
aromatic carbons at $\updelta_{\mathrm{H}}$ 7.41 ($dl$, $J$ 1.0~Hz, 1H)
and 6.33 ($t$, $J$ 1.2~Hz, 1H); at $\updelta_{\mathrm{C}}$ 120.3,
141.2, 109.8, and 143.1, respectively, indicating the presence of a
furan ring in the structure (Tables~\ref{tab2}  and~\ref{tab3}; Section
S2, Figures S2A--S2C, Supplementary Material).

7-Deacetoxy-7-oxogedunin (S3): typical signals from hydrogens and
aromatic carbons at $\updelta_{\mathrm{H}}$ 7.40 ($dl$, $J$ 1.0~Hz,
1H), 6.35 ($dl$, $J$ 1.2~Hz, 1H), and 7.38 ($t$, 1.8~Hz, 1H); at
$\updelta_{\mathrm{C}}$ 120.1, 141.0, 109.7, and 143.1, respectively,
indicating the presence of a furan ring in the structure
(Tables~\ref{tab1} and~\ref{tab2}; Section S3, Figures S3A--S3C, 
Supplementary Material).

The findings were compared with those of~\cite{23,24}
(Tables~\ref{tab2} and~\ref{tab3}). These compounds are noted for their
diverse biological activities, including insecticidal, antifungal,
antimalarial, antibacterial, antiviral, and anticancer properties. Such
a variety illustrates their broad applicability, spanning the
agricultural and pharmaceutical sectors. For example, their phytotoxic
properties make them potential bio-inputs for the formulation of new
herbicides in the control of parasitic plants. In clinical trials, the
three compounds showed protective effects against liver damage induced
by D-GalN/LPS in mice, which shows that they are promising for making
hepatoprotective formulations~\cite{9,10}.

The research highlights the unprecedented identification of these
substances in the \textit{Carapa} gender, although the triterpene
saponins identified in this study have already been reported in the
literature (Figure~\ref{fig3}). Therefore, the study only compared the
$^{13}$C (C\tsub{5}D\tsub{5}N, 75~MHz) and $^{1}$H
(C\tsub{5}D\tsub{5}N, 300~MHz) NMR signals with the data
from~\cite{25,26}, (2006) (Tables~\ref{tab4} and~\ref{tab5}) in
addition to two-dimensional correlation maps 2D $^{1}$H $^{13}$C
(HETCOR and HSQC) and $^{1}$H $^{13}$C (HMBC).

\begin{figure*}
\includegraphics{fig03}
\caption{\label{fig3}Triterpene saponins identified in the
agro-industrial residue of \textit{C.~guianensis}.}
\end{figure*}

%tab4
\begin{table*}[p!]
\caption{\label{tab4}$^{1}$H NMR spectroscopic data for S4 and S5\vspace*{-4pt}}
\fontsize{8.9}{9.5}\selectfont
\tabcolsep4.2pt
\begin{tabular}{ccccc}
\thead
\xmorerows{1}{$^{1}$H} &amp; S4 &amp; 4$^{\mathrm{a}}$ &amp; S5 &amp; 5$^{\mathrm{b}}$ \\ \cline{2-5}
&amp; \multicolumn{2}{c}{$\updelta_{\mathrm{H}}$}
&amp; \multicolumn{2}{c}{$\updelta_{\mathrm{H}}$} \\ 
\endthead
1 &amp; 1.42--1.54 ($m$) &amp; - &amp; - &amp; - \\ 
2 &amp; 2.02--2.07 ($m$) &amp; - &amp; 1.20--1.28 ($m$) &amp; - \\ 
3 &amp; 3.26 ($dd$ 12.0 and 3.6~Hz)
&amp; - &amp; 4.28 ($dd$ 9.3 and 5.1~Hz) &amp; - \\ 
4 &amp; - &amp; - &amp; - &amp; - \\ 
5 &amp; 0.80 ($dd$ 11.4 and 5.1~Hz) &amp; - &amp; 1.68--1.76 ($m$) &amp; - \\ 
  &amp; 1.54--1.75 ($m$) &amp; - &amp; - &amp; - \\ 
6 &amp; - &amp; - &amp; - &amp; - \\ 
7 &amp; 1.76--2.04 ($m$) &amp; - &amp; - &amp; - \\ 
8 &amp; - &amp; - &amp; - &amp; - \\ 
9 &amp; 1.54--1.76 ($m$) &amp; - &amp; 1.77--1.81 ($m$) &amp; - \\ 
10 &amp; - &amp; - &amp; - &amp; - \\ 
11 &amp; 1.82--1.94 ($m$) &amp; - &amp; 1.89--1.94 ($m$) &amp; - \\ 
12 &amp; 5.48 ($sl$) &amp; - &amp; 5.46 ($sl$) &amp; - \\ 
13 &amp; - &amp; - &amp; - &amp; - \\ 
14 &amp; - &amp; - &amp; - &amp; - \\ 
15 &amp; 2.11--2.20 ($m$) &amp; - &amp; - &amp; - \\ 
16 &amp; 1.82--1.94 ($m$) &amp; - &amp; 1.89--1.94 ($m$) &amp; - \\ 
17 &amp; - &amp; - &amp; - &amp; - \\ 
18 &amp; 3.30 ($dd$ 12.0 and 3.6~Hz) &amp; - &amp; 3.23--3.32 ($m$) &amp; - \\ 
19 &amp; 1.82 ($s$) &amp; - &amp; - &amp; - \\ 
20 &amp; - &amp; - &amp; - &amp; - \\ 
21 &amp; 1.24--1.52 ($m$) &amp; - &amp; - &amp; - \\ 
22 &amp; 1.76--2.04 ($m$) &amp; - &amp; - &amp; - \\ 
23 &amp; - &amp; - &amp; 3.74 ($t$ 9.6~Hz) &amp; - \\
Me-23 &amp; 1.19 ($s$) &amp; - &amp; -        &amp; - \\ 
Me-24 &amp; 1.08 ($s$) &amp; - &amp; 1.05 ($s$) &amp; - \\ 
Me-25 &amp; 0.85 ($s$) &amp; - &amp; 0.92 ($s$) &amp; - \\ 
Me-26 &amp; 1.00 ($s$) &amp; - &amp; 1.00 ($s$) &amp; - \\ 
Me-27 &amp; 1.31 ($s$) &amp; - &amp; 1.23 ($s$) &amp; - \\ 
28 &amp; - &amp; - &amp; - &amp; - \\ 
Me-29 &amp; 0.97 ($s$) &amp; - &amp; 0.92 ($s$) &amp; - \\ 
Me-30 &amp; 1.02 ($s$) &amp; - &amp; 1.01 ($s$) &amp; - \\ 
1$'$ (Ara)  &amp; 4.91 ($d$ 5.7~Hz) &amp; 4.92 ($d$ 5.3~Hz) &amp; 5.13 ($d$ 6.3~Hz) &amp; 5.12 ($d$ 6.2~Hz) \\ 
2$'$ (Ara)  &amp; 4.54--4.70 ($m$)  &amp; - &amp; 4.58 ($dd$ 6.3 and 7.2~Hz) &amp; - \\ 
3$'$ (Ara)  &amp; 4.26--4.38 ($m$)  &amp; - &amp; 4.12--4.16 ($m$) &amp;  \\ 
4$'$ (Ara)  &amp; 4.26--4.38 ($m$) and 4.65 ($dd$ 3.6 and 9.9~Hz) &amp; - &amp; 4.19--4.22 ($m$) &amp; - \\ 
5$'$ (Ara)  &amp; 3.85 ($dl$ 9.9~Hz) and 4.26--4.38 ($m$) &amp; - &amp; 3.74 ($t$ 96~Hz) 4.20--4.34 ($m$) &amp; - \\ 
1$''$ (Ram) &amp; 6.16 ($sl$) &amp; 6.10 ($sl$) &amp; 6.23 ($sl$) &amp; 6.22 ($sl$) \\ 
2$''$ (Ram) &amp; 4.76 ($sl$) &amp; - &amp; 4.73--4.76 ($m$) &amp; - \\ 
3$''$ (Ram) &amp; 4.54--4.70 ($m$) &amp; - &amp; 4.65 ($dd$ 9.4 and 3.3~Hz) &amp; - \\ 
4$''$ (Ram) &amp; 4.26--4.38 ($m$) &amp; - &amp; 4.31 ($t$ 9.4~Hz) &amp; - \\ 
5$''$ (Ram) &amp; 4.54--4.70 ($m$) &amp; - &amp; 4.68--4.74 ($m$) &amp; - \\ 
Me-6$''$ (Ram) &amp; 1.64 ($d$ 6.3~Hz) &amp; - &amp; 1.64 ($d$ 6.0~Hz) &amp; - 
\botline
\end{tabular}
\tabnote{\fontsize{8.5}{9.5}\selectfont Measured in C\tsub{5}D\tsub{5}N.
Chemical shifts (${\updelta}$) are in ppm from a tetramethylsilane
reference.  $^{\mathrm{a}}$Nakanishi et~al.~\cite{25};
$^{\mathrm{b}}$Panov et~al.~\cite{26}.}
\end{table*}

%tab5
\begin{table}
\caption{\label{tab5}$^{13}$C NMR spectroscopic data for S4 and S5\vspace*{-4pt}}
\fontsize{8.8}{9.3}\selectfont
\begin{tabular}{ccccc}
\thead
\xmorerows{1}{$^{13}$C} &amp; S4 &amp; 4$^{\mathrm{a}}$ &amp; S5 &amp; 5$^{\mathrm{b}}$ \\ \cline{2-5}
 &amp; \multicolumn{2}{c}{$\updelta_{\mathrm{C}}$} 
 &amp; \multicolumn{2}{c}{$\updelta_{\mathrm{C}}$}  \\
\endthead
\01&amp; 39.0 &amp; 38.9 &amp; 39.0 &amp; 39.0 \\ 
\02&amp; 26.7 &amp; 26.5 &amp; 26.2 &amp; 26.2 \\ 
\03&amp; 88.9 &amp; 88.8 &amp; 81.1 &amp; 81.1 \\ 
\04&amp; 39.7 &amp; 39.5 &amp; 43.5 &amp; 43.6 \\ 
\05&amp; 56.0 &amp; 55.9 &amp; 47.7 &amp; 47.7 \\ 
\06&amp; 18.7 &amp; 18.5 &amp; 18.2 &amp; 18.2 \\ 
\07&amp; 33.3 &amp; 33.3 &amp; 32.9 &amp; 32.8 \\ 
\08&amp; 39.9 &amp; 39.8 &amp; 39.8 &amp; 39.8 \\ 
\09&amp; 48.2 &amp; 48.1 &amp; 48.2 &amp; 48.2 \\ 
10 &amp; 37.2 &amp; 37.1 &amp; 36.9 &amp; 36.9 \\ 
11 &amp; 24.0 &amp; 23.8 &amp; 23.9 &amp; 23.8 \\ 
12 &amp; 122.6\0&amp; 122.4\0&amp; 123.0\0&amp; 122.6\0\\ 
13 &amp; 145.0\0&amp; 144.9\0&amp; 145.2\0&amp; 144.8\0\\ 
14 &amp; 42.3 &amp; 42.2 &amp; 42.1 &amp; 42.2 \\ 
15 &amp; 28.5 &amp; 28.4 &amp; 28.4 &amp; 28.3 \\ 
16 &amp; 23.9 &amp; 23.7 &amp; 23.9 &amp; 23.7 \\ 
17 &amp; 46.6 &amp; 46.7 &amp; 46.6 &amp; 46.6 \\ 
18 &amp; 42.1 &amp; 42.0 &amp; 42.2 &amp; 42.0 \\ 
19 &amp; 46.9 &amp; 46.6 &amp; 46.8 &amp; 46.4 \\ 
20 &amp; 31.1 &amp; 31.0 &amp; 31.0 &amp; 30.9 \\ 
21 &amp; 34.4 &amp; 34.3 &amp; 34.3 &amp; 34.2 \\ 
22 &amp; 33.4 &amp; 33.3 &amp; 32.9 &amp; 33.2 \\ 
23 &amp; -  &amp; -  &amp; 64.0 &amp; 64.0 \\ 
Me-23 &amp; 28.2 &amp; 28.1 &amp; - &amp; - \\ 
Me-24 &amp; 17.2 &amp; 17.0 &amp; 14.0 &amp; 14.0 \\ 
Me-25 &amp; 15.7 &amp; 15.5 &amp; 16.1 &amp; 16.1 \\ 
Me-26 &amp; 17.5 &amp; 17.4 &amp; 17.6 &amp; 17.5 \\ 
Me-27 &amp; 26.3 &amp; 26.2 &amp; 26.2 &amp; 26.2 \\ 
28    &amp; 180.6\0&amp; 180.6\0&amp; 181.2\0&amp; 180.2\0 \\ 
Me-29 &amp; 33.5 &amp; 33.3 &amp; 33.3 &amp; 33.2 \\ 
Me-30 &amp; 24.0 &amp; 23.8 &amp; 23.9 &amp; 23.8 \\ 
1$'$ (Ara)     &amp; 105.0\0&amp; 104.6\0&amp; 104.4\0&amp; 104.3\0 \\ 
2$'$ (Ara)     &amp; 76.0 &amp; 76.0 &amp; 75.9 &amp; 75.9 \\ 
3$'$ (Ara)     &amp; 74.2 &amp; 74.0 &amp; 74.6 &amp; 74.5 \\ 
4$'$ (Ara)     &amp; 68.9 &amp; 68.6 &amp; 69.3 &amp; 69.3 \\ 
5$'$ (Ara)     &amp; 64.9 &amp; 64.5 &amp; 65.6 &amp; 65.5 \\ 
1$''$ (Ram)    &amp; 101.9\0&amp; 101.7\0&amp; 101.7\0&amp; 101.7\0 \\ 
2$''$ (Ram)    &amp; 72.5 &amp; 72.4 &amp; 72.4 &amp; 72.3 \\ 
3$''$ (Ram)    &amp; 72.7 &amp; 72.6 &amp; 72.6 &amp; 72.5 \\ 
4$''$ (Ram)    &amp; 74.0 &amp; 73.7 &amp; 74.1 &amp; 74.2 \\ 
5$''$ (Ram)    &amp; 70.0 &amp; 69.9 &amp; 69.8 &amp; 69.8 \\ 
Me-6$''$ (Ram) &amp; 18.8 &amp; 18.5 &amp; 18.6 &amp; 18.6 
\botline
\end{tabular}
\tabnote{Measured in C\tsub{5}D\tsub{5}N. Chemical shifts
(${\updelta}$) are in ppm from a tetramethylsilane reference.
$^{\mathrm{a}}$Nakanishi et~al.~\cite{25}; $^{\mathrm{b}}$Panov 
et~al.~\cite{26}.}
\end{table}

Oleanolic acid
3-O-[${\upalpha}$-L-ramnopyranosyl-(1${\rightarrow}$2)-${\upalpha}$-L-arabinopyranosyl]
(S4): typical hydrogen signals from glycidic units such as arabinose
were observed at $\updelta_{\mathrm{H}}$ 4.91 ($d$, $J$ 5.7~Hz)
(anomeric hydrogen H-1$'$), as well as raminose at
$\updelta_{\mathrm{H}}$ 6.16 ({$sl$}) (anomeric hydrogen H-1$''$), in
addition to the characteristic signals of anomeric carbons for two
sugar units of the arabinose type at $\updelta_{\mathrm{C}}$ 105.1 and
raminose at $\updelta_{\mathrm{C}}$ 101.9. (Tables~\ref{tab4}
and~\ref{tab5}; Section S4, Figures S4A--S4C, Supplementary Material). 

Hederagenin 3-O-[${\upalpha}$-L-rhamnopyranosyl-(1${\rightarrow}$2)-${\upalpha}$-L-arabinopyranosyl]
(Pulsatilla~A) (S5): typical hydrogen signals from glycidic units such
as arabinose were observed at $\updelta_{\mathrm{H}}$ 5.13 ($d$, $J$
6.3~Hz) (anomeric hydrogen H-1$'$) and from raminose at
$\updelta_{\mathrm{H}}$ 6.23 ({$sl$}) (anomeric hydrogen H-1$''$), in
addition to signals characteristic of anomeric carbons for two sugar
units of the arabinose type at $\updelta_{\mathrm{C}}$ 104.4 and
raminose at $\updelta_{\mathrm{C}}$ 101.7, and an oxymethylene carbon
signal at $\updelta_{\mathrm{C}}$ 64.0 (Tables~\ref{tab4} and
\ref{tab5}; Section S5, Figures S5A--S5C, Supplementary Material).

In summary, this phytochemical study provides new information on
\textit{C.~guianensis} and the \textit{Carapa} gender, revealing
greater chemical diversity in the species. Triterpene saponins have
previously been detected in \textit{C.~procera} and
\textit{C.~guianensis} through preliminary tests~\cite{27,28}, but the
present study provides the first structural identification of these
metabolites in \textit{C.~guianensis}. The glycosides identified in the
agro-industrial residues of \textit{C.~guianensis} have important
biological characteristics such as: antiviral, antitumor and
hepatoprotective~\cite{29,30,31,32,33}. Saponins with the aglycones
hederagenin or oleanolic acid in their structures, such as those
identified in this study, have antifungal activities due to their
ability to bind to sterols in the fungal membrane, causing pore
formation and loss of membrane integrity~\cite{34}. These promising
biomolecules could be of interest to the pharmaceutical and cosmetics
industries. The results of this study provide new perspectives for the
reuse of \textit{C.~guianensis} industrial waste as a source of
bioactive substances of industrial interest.

\subsection{HPTLC fingerprint}\label{ssec34}

The fingerprinting of the hot ethanolic extract (E9) and the commercial
oil of \textit{C.~guianensis} (OA), using the substances isolated in
this research (S1, S2, S3, S4 and S5) in duplicate via HPTLC, enabled a
certain similarity to be observed between the chemical composition of
OA and E9. This finding is supported by the observation of
chromatographic bands {suggestive} of limonoids when derivatized with
VSA, as observed for limonoids S1, S2, and S3 isolated from andiroba
agro-industrial residue. This observation can be explained by the fact
that the process of obtaining andiroba oil from the seeds of the
species does not deplete the limonoid content, since substances of this
class are still present in the residue. It was also possible to observe
the presence of substances other than limonoids, which are absent in
andiroba oil, such as triterpene saponins (S4~and~S5), revealing the
chemical potential of the agro-industrial residue of \textit{Carapa
guianensis} Aubl.\ (Figure~\ref{fig4}). \looseness=-1

\begin{figure*}
\includegraphics{fig04}
\caption{\label{fig4}Fingerprint---andiroba oil (OA), hot ethanolic
extract (E9) and isolated substances (S1, S2, S3, S4 and S5) in
duplicate via HPTLC. Elution system, gradient mode: Hexane/AcOEt/MeOH
(1\% HCOOH). VSA derivatizer.}
\end{figure*}

The results obtained in this study are relevant and promising, as they
show that even after obtaining andiroba oil, the agro-industrial
residue of \textit{C.~guianensis} still has a wealth of phytochemicals.
This contributes to the phytochemical study of the species, as well as
identifying the presence of \mbox{substances} of economic and pharmacological
interest. This biomass can be used as an alternative in the search for
\mbox{bioproducts}, thus adding value to the{\unskip\break} residue.

\section{Conclusion}\label{sec4}

Bioprospecting of the agro-industrial residue of \textit{C.~guianensis}
indicated that the most efficient extraction method was a warm
extraction (Soxhlet---EtOH), with the highest percentage yield of
26.96\% and the greatest diversity of secondary metabolites. NMR
analysis and comparison with data from the \mbox{literature} allowed us to
identify 5 compounds present in the agro-industrial residue of
\textit{C.~guianensis}, 3 of which are limonoids (S1, S2 and
S3---gedunin, 6${\upalpha}$-acetoxygedunin and
7-desacetoxy-7-oxogedunin, respectively), important bioactive compounds
that have actions: antinociceptive, anti-inflammatory, antitumor,
hepatoprotective, etc. In addition, we discovered classes of substances
unknown in the \textit{Carapa} genus until this study, such as the
triterpene saponins (S4:
3-O-[${\upalpha}$-L-arabinopyranoside-${\upalpha}$-L-ramnopyranoside-(1${\rightarrow}$2)] 
oleanolic acid and S5: 
3-O-[${\upalpha}$-L-arabinopyranoside-${\upalpha}$-L-ramnopyranoside-(1${\rightarrow}$2)] 
hederagenin (Pulsatilla~A)), glycosides with antiviral, antitumor, and
hepatoprotective properties. The HPTLC fingerprint of the hot ethanolic
extract (E9) and the commercial oil of \textit{C.~guianensis}, using
the substances isolated in this research (S1, S2, S3, S4 and S5),
although qualitative, shows that E9 has lower concentrations of
limonoid components than andiroba oil. However, it is worth noting that
there are chemical substances present in the extract studied that are
absent in andiroba oil, such as substances S4 and S5. In this way, the
research showed that even after obtaining andiroba oil, the
agro-industrial residue generated still has a wealth of phytochemicals,
biomass that can be used as an alternative in the search for
bioproducts.

\section*{Declaration of interests}

The authors do not work for, advise, own shares in, or receive funds
from any organization that could benefit from this article, and have
declared no affiliations other than their research organizations.

\section*{Supplementary data}

Supporting information for this article is available on the journal's
website under \printDOI\ or from the author.  

\CDRsupplementaryTwotypes{supplementary-material}{\cdrattach{crchim-399-suppl.pdf}}

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\refinput{crchim20240890-reference.tex}

\end{document}
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