A practical set of HPLC methods was developed for the separation and determination of the eggplant steroidal glycoalkaloids, solanine, chaconine, solasonine, solamargine, and their aglycones, solasodine and solanidine. A gradient method was initially developed, but proved to be neither robust nor practical. Three separate isocratic methods using acetonitrile and ammonium dihydrogen phosphate were developed and shown to be more repeatable, less subject to fluctuations in mobile phase composition, and less time consuming. The effect of adjusting buffer pH, column temperature, and buffer type (triethylammonium phosphate vs. ammonium dihydrogen phosphate) were evaluated. It was also discovered that, by addition of 10% methanol to the acetonitrile portion of the mobile phase, more control over the separations was possible. The use of methanol as a mobile phase entrainer greatly improved separations in some cases and its effectiveness was also dependent upon column temperature. Assessments of the method recovery, limit of detection, and limit of quantitation were made using extracts from S. melongena and S. linnaeanum.
Solid-phase microextraction (SPME), followed by on-fiber derivatization was investigated for the analysis of the steroidal glycoalkaloid aglycones, solasodine and solanidine. The aglycones were first extracted by direct immersion of the SPME fiber in the sample medium and then derivatized on the fiber in a separate step using 1-(trimethylsilyl)imidazole (TMSI). The derivatized compounds were then desorbed from the SPME fiber and detected by GC-MS. Polydimethylsiloxane/Divinylbenzene (PDMS-DVB), Carboxen/Polydimethylsiloxane (CAR-PDMS), and Carbowax/Divinylbenzene (CW-DVB) fibers were employed with the CW-DVB fibers being the most successful, as expected. Closed-end capillary tubes were used to hold the extraction media. Both aglycones were successfully extracted, derivatized, and detected by GC-MS. Solasodine always required derivatization, but solanidine did not.The same method was successfully applied to cholesterol so that it could be used as an internal standard. Also, using the closed-end capillary tubes, a two-phase extraction system was also investigated, whereby the fiber was only exposed to the phase in which it was presumed to be less damaged. However, in all cases, fiber degradation was significant, preventing the use of extended extraction times and limiting reuse of the fibers. However, the results represent a first look into the feasibility of the method. With the development of more suitable SPME phases, this method could potentially provide a complementary route for routine determinations of glycoalkaloids for both research and food quality control.
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