Comparison of different tandem mass spectrometric techniques (ESI-IT, ESI- and IP-MALDI-QRTOF and vMALDI-TOF/RTOF) for the analysis of crocins and picrocrocin from the stigmas of Crocus sativus L.
BACKGROUND
The expensive spice saffron originating from the stigmas of Crocus sativus L. and also applied in traditional Chinese medicine (TCM) constitutes a complex mixture of glycoconjugates varying not only in the aglycon structure, but also in glycosylation pattern. Therefore, various tandem mass spectrometric techniques were evaluated for their usefulness in structural elucidation.


METHODS
Three selected constituents of the stigmas of Crocus sativus L., trans- and cis-crocin-4 as well as picrocrocin, were isolated and purified by HPLC and finally analyzed by ESI-MS (ion trap, QqRTOF), IP-MALDI-MS (QqRTOF) and vMALDI-MS (TOF/RTOF) in combination with tandem mass spectrometry in collision energy regimes ranging from a few eV (LE) to 20 keV (HE) collisions for the first time. These data aid in structurally elucidating minor, unknown glycoconjugates originating from this plant-derived spice.


RESULTS
LE-CID of isomeric crocins on either an ion trap with ESI or a QqRTOF-instrument with ESI or IP-MALDI as desorption/ionization technique only yielded a limited number of structurally diagnostic sodiated product ions related to the carbohydrate moiety as well as to the intact aglycon in contrast to true HE-CID. The low MW constituent picrocrocin did not yield useful LE-CID spectra, but showed a high number of structurally diagnostic product ions by HE-CID utilizing a vMALDI TOF/RTOF-instrument.


CONCLUSIONS
The highest number of structurally diagnostic product ions allowing also determination of the carbohydrate linkage of the gentiobiose-moiety of isomeric crocins ((0,4)A(2), (3,5)A(2) product ions indicating a 1→6 carbohydrate linkage) was only achievable by HE-CID. Fragmentation of the aglycon was not observed by any collision energy regime applied.
Laboratory of Instrumental Analysis, Department of Pharmacy, University of Patras, Patras, Greece.
Nikolaos Stavros
NS
Koulakiotis
Ernst
E
Pittenauer
Maria
M
Halabalaki
Anthony
A
Tsarbopoulos
Günter
G
Allmaier
D002338:Carotenoids / Q000032:analysis* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D027622:Crocus / Q000737:chemistry*; D053138:Cyclohexenes / Q000032:analysis* / Q000302:isolation & purification; D005960:Glucosides / Q000032:analysis* / Q000302:isolation & purification; D010936:Plant Extracts / Q000032:analysis / Q000302:isolation & purification; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013729:Terpenes / Q000032:analysis* / Q000302:isolation & purification
D053138:Cyclohexenes; D005960:Glucosides; D010936:Plant Extracts; D013729:Terpenes; D002338:Carotenoids; C029036:crocin; C087962:picrocrocin