FT-NIR spectroscopy and RP-HPLC combined with multivariate analysis reveals differences in plant cell suspension cultures of Thevetia peruviana treated with salicylic acid and methyl jasmonate

Plant cell suspension culture of T. peruviana is a feasible biotechnological platform for the production of secondary metabolites with anti-proliferative/cytotoxic activity, as phenolic compounds (PC); however, different in in vitro growth conditions may affect the production, demanding strategies to increase the metabolite biosynthesis, as well as the development of sensitive and rapid analytical methods for metabolite monitoring. The Fourier transform near-infrared (FT-NIR) spectroscopy and Reversed-phase high-performance liquid chromatography (RP-HPLC) combined with Multivariate analysis (MVA) were used to detect significant differences in the PC production in cultures treated with two elicitors. The results suggest that the FT-NIR-MVA is useful for discriminating samples according to the treatment, showed significant influence of the PC signal. RP-HPLC-MVA showed that the elicitor effect occurs at 72 h post-elicitation. Detection of dihydroquercetin (maximum concentration = 12.59 mg/L), a flavonoid with anti-cancer properties, is highlighted. Future studies will be aimed at scaling this culture to increase the productivity of dihydroquercetin.

Keywords: 2,4-D, 2,4-dichlorophenoxy acetic acid; CGA, chlorogenic acid; COW, Correlation Optimized Warping; DHQ, dihydroquercetin; DV1, first derivatives; DV2, second derivatives; DW, dry weight; FT-NIR; FT-NIR, fourier transform near-infrared spectroscopy; FW, fresh weight; GAE, gallic acid equivalents; KT, Kinetin; MVA, multivariate analysis; MeJA, Methyl jasmonate; Multivariate analysis; OPLS-DA, orthogonal partial least square-discriminant analysis; PC, phenolic compounds; PCA, principal component analysis; PLS, partial least square-discriminant analysis; Plant cell culture; RP-HPLC; RP-HPLC, reversed phase-high performance liquid chromatography; SA, salicylic acid; SG, Savitzky Golay; SH, Schenk and Hildebrandt; SNV, Standard Normal Variate; Thevetia peruviana.