Rapid isolation and characterization of crocins, picrocrocin, and crocetin from saffron using centrifugal partition chromatography and LC-MS.
Evangelia KarkoulaApostolis AngelisNikolaos-Stavros KoulakiotisEvangelos GikasMaria HalabalakiAnthony TsarbopoulosAlexios-Leandros SkaltsounisPublished in: Journal of separation science (2018)
This study demonstrates a simple method for one-step isolation of the main secondary metabolites of a hydroalcoholic extract of Crocus sativus stigmas (saffron) using step-gradient centrifugal partition chromatography. The analysis was performed in dual and elution-extrusion mode, using five biphasic systems of the solvents heptane/ethyl acetate/butanol/ethanol/water in ratios of 4:10:0:4:10, 1:13:0:4:10, 1:12:1:4:10, 1:10:3:4:10, and 1:7:6:4:10. Five major crocins, picrocrocin, and crocetin were directly isolated in one step. Scaling up to preparative level, allowed the recovery of significantly high quantities of pure compounds, especially trans-crocin-4, saffron's principal crocin. Comparing dual-mode and elution-extrusion, in dual-mode, the trans-crocin-4 containing fractions were co-eluted with a high amount of free β-d-glucose. In contrast, absence of free β-d-glucose was observed in the corresponding trans-crocin-4 fractions obtained by the second method denoting its superiority against dual-mode. Initiating analysis with the 4th solvent-system afforded selective isolation of trans-crocin-4, with reduction in experimental time and solvent consumption. Structure elucidation was performed by nuclear magnetic resonance spectroscopy, liquid chromatography with mass spectrometry, and high-resolution tandem mass spectrometry. The proposed methodology comprises an integrated approach for the purification and characterization of biologically active saffron components in a fast, selective, and environmentally friendly manner.
Keyphrases
- liquid chromatography
- tandem mass spectrometry
- mass spectrometry
- ultra high performance liquid chromatography
- high performance liquid chromatography
- high resolution
- gas chromatography
- high resolution mass spectrometry
- simultaneous determination
- ionic liquid
- solid phase extraction
- capillary electrophoresis
- magnetic resonance
- blood pressure
- metabolic syndrome
- ms ms
- magnetic resonance imaging
- type diabetes
- high speed
- adipose tissue