Continuous vs. segmented second-dimension system gradients for comprehensive two-dimensional liquid chromatography of sugarcane (Saccharum spp.)
- 388 Downloads
A comprehensive two-dimensional liquid chromatography system in combination with photodiode array and mass spectrometry detection was developed for analysis of polyphenols in sugarcane (Saccharum spp.) leaf extracts. To achieve this, a micro cyano column and a partially porous octodecylsilica column were used in the first and the second dimension, respectively. The choice of the cyano column over other reversed-phase columns tested for the first-dimension separation was due to its lower correlation selectivity with respect to the octodecylsilica column, which was used for the second-dimension separation. Even when reversed-phase mode was used in both dimensions, a satisfactory degree of orthogonality was achieved by use of different gradient elution modes in the second dimension. By means of the setup investigated, 38 polyphenolic compounds were detected, and among them 24 were positively identified by means of complementary data from photodiode array and mass spectrometry detection and an in-house database. This is the first time such a powerful analytical technique has been used for polyphenolic characterization of sugarcane extracts.
KeywordsPolyphenols Comprehensive two-dimensional liquid chromatography Mass spectrometry Cyano column Sugarcane
The authors gratefully acknowledge Shimadzu and Sigma-Aldrich/Supelco for their continuous support. G.M.L. thanks the Conselho Nacional de Desenvolvimento Científico e Tecnológico for a scholarship (process number BEX 5605/13-0).
- 1.CONAB Companhia Nacional de Abastecimento (2013). Conab - Companhia Nacional de Abastecimento. http://www.conab.gov.br/. Accessed 29 Nov 2013
- 2.Li X, Shun Y, Bin T, Xue L, Chunmei J, Hang S (2010) Determination and comparison of flavonoids and anthocyanins in Chinese sugarcane tips, stems, roots and leaves. J Sep Sci 33:1216–1223Google Scholar
- 3.Colombo R, Yariwake JH, Queiroz EF, Ndjoko K, Hostettmann K (2005) On-line identification of sugarcane (Saccharum officinarum L.) methoxyflavones by liquid chromatography–UV detection using post-column derivatization and liquid chromatography–mass spectrometry. J Chromatogr A 1082:51–59CrossRefGoogle Scholar
- 6.Cavalheiro AJ, Coutinho ID, Leme GM, Silva AA, Silva APD (2012) In: Stradiotto NR, Lemos EG (eds) Bioenergia: pesquisa, desenvolvimento e inovação. São Paulo, Cultura AcadêmicaGoogle Scholar
- 19.Montero L, Herrero M, Prodanov M, Ibanàñez E, Cifuentes A (2013) Characterization of grape seed procyanidins by comprehensive two-dimensional hydrophilic interaction × reversed phase liquid chromatography coupled to diode array detection and tandem mass spectrometry. Anal Bioanal Chem 405:4627–4638CrossRefGoogle Scholar
- 33.Scoparo CT, Souza LM, Dartora N, Sassaki GL, Gorin PAJ, Iacomini M (2012) Analysis of Camellia sinensis green and black teas via ultra high performance liquid chromatography assisted by liquid-liquid partition and two-dimensional liquid chromatography (size exclusion × reversed phase). J Chromatogr A 1222:29–37CrossRefGoogle Scholar
- 39.Snyder LR, Kirkland JJ, Loyd JW (2010) Introduction to modern liquid chromatography. Wiley, TotowaGoogle Scholar