Quantification of appetite suppressing steroid glycosides from Hoodia gordonii in dried plant material, purified extracts and food products using HPLC-UV and HPLC-MS methods.
High-performance liquid chromatography (HPLC)-UV and HPLC-Mass Spectrometry (MS) methods were developed for the quantitative analysis of the family of Hoodia gordonii steroid glycosides with appetite suppressing properties in dried plant material, in purified and enriched extracts and in various prototype food-products fortified with H. gordonii extracts. For solid materials, e.g. dried plants or for non-fatty foods, extraction of the steroid glycosides is performed using methanol. For products where the steroid glycosides are present in an oil matrix, direct injection of the oil after dilution in tetrahydrofuran is applied. The HPLC separation is performed on an octyl-modified reversed-phase column in the gradient mode with UV detection at lambda = 220 nm. Quantification is performed against an external calibration line prepared using either one of the pure steroid glycosides or geranyl-tiglate. Short- and long-term repeatabilities of the methods are better than 3 and 6%, respectively. Recoveries are better than 85%, even in the analysis of the least abundant steroid glycosides in a complex yoghurt drink. Linearity is better than 3-4 orders of magnitude and the detection limits are below approximately 2 microg g(-1) for the individual steroid glycosides in dried plant material and food products. HPLC-MS is used to confirm that the steroid glycosides contain the characteristic steroid core, the carbohydrate chain and the tigloyl group.
Janssen
Hans-Gerd
HG
Unilever Food and Health Research Institute, Advanced Measurement and Imaging, Unilever Research and Development, P.O. Box 114, 3130 AC Vlaardingen, The Netherlands. hans-gerd.janssen@unilever.com
Swindells
Chris
C
Gunning
Philip
P
Wang
Weijun
W
Grün
Christian
C
Mahabir
Krishna
K
Maharaj
Vinesh J
VJ
Apps
Peter J
PJ
D029062:Apocynaceae / Q000737:chemistry*; D001067:Appetite Depressants / Q000737:chemistry / Q000302:isolation & purification*; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003890:Desiccation; D005504:Food Analysis; D006027:Glycosides / Q000737:chemistry / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010936:Plant Extracts / Q000737:chemistry; D013256:Steroids / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D001067:Appetite Depressants; D006027:Glycosides; D010936:Plant Extracts; D013256:Steroids