Development of a gas chromatography method for quantification of triterpenes in the commercial oleoresins from Protium species

Abstract

Crude non-volatile oleoresin from Burseraceae species (breu), a non-timber forest product of the Amazon, still lacks methods for its quality control even though its pharmacological activities have been associated to the major triterpenes α-amyrin (αAA), β-amyrin, α-amyrenone, β-amyrenone and lupeol (LU). Its chemical characterization has usually been determined by gas chromatography (GC) using columns with phases of low polarity, which are ineffective in the separation of αAA and LU. The present study aimed to develop a GC method to characterize and quantify triterpenes in commercial breu with good selectivity and resolution. Columns with phases of different polarity (DB-35, DB-1701, DB-17HT and DB-Innowax) were tested, and DB-17HT separated αAA and LU without signal suppression or high retention of other constituents. This column allowed the quantification of breu triterpenes using flame ionization detection and cholesterol as a reference standard. Good similarity between the calibration curves of cholesterol and αAA was verified. Internal and external standardizations led to different (p < 0.05) quantifications, but the latter showed less deviation (RSD ≤ 3%). This work provides a reproducible GC method for the quality control of commercial breu, clearly evidencing the presence of lupeol and allowing the accurate quantification of major triterpenes.

Quali–Quantitative Characterization of Volatile and Non-Volatile Compounds in Protium heptaphyllum (Aubl.) Marchand Resin by GC–MS Validated Method, GC–FID and HPLC–HRMS2

“…In our work, the ursanic and oleanic triterpenes ratio is 1.3 in all analyzed samples (PHR, ATCE and AMCE). The observed value is slightly higher than Silva et al (1.1) [ 11 ] and lower than Siani et al (3.0) [ 8 ] and Neves et al (2.0) [ 12 ]. Different ursanic–oleanic abundance ratios found by Neves et al in their GC–MS quantitation experiment could be also explained by using α-amyrin as a unique calibration standard also applied to β-amyrin quantitation.…”

“…Phytochemical investigations of Protium heptaphyllum have been usually conducted on oleum resin essential oils from different Protium species, mainly by GC–FID and GC–MS techniques [ 1 , 9 , 10 ]. In other studies, chemical derivatization and GC–MS analyses were performed in order to characterize and quantify α and β-amyrins and triterpenes in oleum resin extracts of Protium species by the use of toxic organic solvents, such as hexane, chloroform and dichloromethane [ 8 , 11 , 12 ]. Alternatively, 1 H- and 13 C-NMR were performed in order to investigate the triterpenes present in the nonvolatile fraction [ 13 ], though both NMR techniques suffer low feasibility due to high analytical costs and complex sample preparation.…”

Óleo essencial dos pecíolos de Protium ovatum Engl. (Burseraceae) apresenta atividade bioativa e antifúngica?

“…São descritos também, diversas ações sobre modelos biológicos para os óleos essenciais extraídos da resina de P. giganteum, P. aracouchine, P. grandifolium, P. hebetatum, P. paniculatum, P. heptaphyllum e P. subserratum conforme aborda nos estudos de Díaz et al (2011), Freitas et al (2011), Merali et al (2018, Piva et al (2019), Nogueira et al (2019), Figueiredo et al (2019, Kozin et al (2019), Silva et al (2020), Barata et al (2020) e Mannino et al (2021). Os efeitos positivos sobre os mecanismos biológicos não se limitam apenas aos óleos essenciais e óleos-resinas, os extratos vegetais extraídos a partir dos principais solventes etanol, água ou metanol demonstram incríveis atividades biológicas, que estão envolvidas pela presença de fitomoléculas distribuídas entre diversos grupos de cumarinas, esteroides, flavonoides, lignanas dentre outros derivados produzidos a partir do metabolismo especial desses vegetais (Ahujaa et al, 2019;Neves et al, 2020;Barata et al, 2020;Alves et al, 2020;Amparo et al, 2022).…”