Metabolomic Profiling of Guadua Species and Its Correlation with Antioxidant and Cytotoxic Activities

Bamboo plants are widely used in Asian traditional medicine for various health issues and exhibit therapeutic potential. Guadua species are renowned bamboos for their high phenolic compound content, including flavonoids and hydroxycinnamic acid derivatives, and possess noteworthy biological properties. Despite this, there is a notable scarcity of research on the chemical and biological aspects of Latin American bamboo leaf extracts (BLEs), especially concerning the Guadua genus. This study aimed to employ a metabolomics approach to integrate the phytochemical and activity profiles of BLEs to identify potential bioactive markers. We determined the metabolic fingerprints of 30 BLEs through HPTLC, HPLC-DAD, UHPLC-QTOF-MS, and 1H-NMR analyses and screened for antioxidant and cytotoxic activities using ABTS, DPPH, and MTT methods. Ultimately, correlation analyses were performed by using chemometric methods and molecular networking. Our findings present a comprehensive chemical characterization, encompassing 40 flavonoids and 9 cinnamic acid derivatives. Notably, most of these compounds have been reported for the first time within the genus, signifying novel discoveries. Additionally, certain compounds identified in other species of the subfamily Bambusoideae provide valuable comparative insights. These compounds demonstrated a significant correlation with antioxidant potential, with values exceeding 100 and 30 μmol of TE/g of extract for ABTS and DPPH, respectively, in the samples. Extracts from G. incana and G. angustifolia exhibited potent cytotoxic effects with IC50 values of 1.23 and 4.73 μg/mL against HCT-116 colon cancer cells, respectively. Notably, glycosylated flavones showed a strong correlation with cytotoxicity. These new findings significantly contribute to our understanding of the chemical composition and biological properties of these often overlooked bamboo species, providing them with important added value and alternative use.


Table of Contents
1. Figure S1.UV spectra of the major peaks found in the BLEs obtained by HPLC-DAD.S1.Phytochemicals tentatively identified in BLEs using UHPLC-QTOF-MS.

Table
3. Figure S2.Chemical structures of the metabolites were found in BLEs. 4. Figure S3.MS 2 spectrum match feature of GNPS showing the similarity of fragments patterns of the experimental and library data.
5. Figure S4.PCA score plot including Quality Controls (QCs) and all samples from UHPLC-QTOF-MS analysis.6.Table S2.List and information on the environmental variables of collection of the Guadua species used for the study.
7. Table S3.Comparison of retention times of reference standards between HPLC-DAD and UHPLC-QTOF-MS analyses.
11. Table S5.Parameters used in the MZmine 2.53 software for processing the data obtained by UHPLC-QTOF-MS.
12. Table S6.Parameters used in the NMRProcFlow software for processing the data obtained by 1 H-NMR.13. Figure S7.Calibration curves obtained for the determination of antioxidant capacity by ABTS and DPPH.

References
Brief description of contents: (i) UV spectra of the major peaks found in the BLEs obtained by HPLC-DAD; (ii) Phytochemicals tentatively identified in BLEs using UHPLC-QTOF-MS; (iii) Chemical structures of the metabolites were found in BLEs; (iv) MS 2 spectrum match feature of GNPS showing the similarity of fragments patterns of the experimental and library data; (v) PCA score plot including Quality Controls (QCs) and all samples from UHPLC-QTOF-MS analysis; (vi) List and information on the environmental variables of collection of the Guadua species used for the study; (vii) Comparison of retention times of reference standards between HPLC-DAD and UHPLC-QTOF-MS analyses; (viii) 1 H-NMR spectra obtained for the reference standards (Quercetin and Rutin); (ix) Calibration curves obtained for the determination of antioxidant capacity by ABTS and DPPH (x) Parameters used in the GNPS Classical Molecular Networking platform, MZmine version 2.53, and NMRProcFlow software for processing the data obtained by UHPLC-QTOF-MS, and 1 H-NMR.

Figure S1
. UV spectra of the major peaks found in the BLEs obtained by HPLC-DAD.The retention time of each peak and its corresponding spectrum are described in highlighted green (t R /1.00).

Figure S3 .
Figure S3.MS 2 spectrum match feature of GNPS showing the similarity of fragments patterns of the experimental and library data.

Figure S4 .
Figure S4.PCA score plot including Quality Controls (QCs) and all samples from UHPLC-QTOF-MS analysis in both negative and positive ion mode.

Figure S7 .
Figure S7.Calibration curves obtained for the determination of antioxidant capacity by ABTS and DPPH.

Table S2 .
List and information on the environmental variables of collection of the Guadua species used for the study.

Table S3 .
Comparison of retention times of reference standards between HPLC-DAD and UHPLC-QTOF-MS analyses.

Table S4 .
Parameters used in the GNPS Classical Molecular Networking.

Table S6 .
Parameters used in the NMRProcFlow software for processing the data obtained by 1 H-NMR.