Development of Practical HPLC Methods for the Separation and Determination of Eggplant Steroidal Glycoalkaloids and their Aglycones

Abstract

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.

Simple and Reliable Methods for the Determination of Three Steroidal Glycosides in the Eight Species of Solanum by Reversed‐phase HPLC Coupled with Diode Array Detection

“…Although there are several analytical methods for the determination of steroidal glycoalkaloids and their aglycones (Laurila et al, 1999;Eanes et al, 2008;Paul et al, 2008;Skarkova et al, 2008;Shanker et al, 2011), but HPLC-photodiode array (PDA) is the most widely used and most versatile method, because it is rapid, accurate, relatively easy-to-use and reproducible. Furthermore, individual steroidal glycosides and their aglycones can be determined without any derivatisation and HPLC equipment is widely availabile around the world.…”

“…Similarly several indiosides from S. violaceum have shown significant proliferation inhibition against Bel-7402 cells (Ma et al, 2006). Although there are several analytical methods for the determination of steroidal glycoalkaloids and their aglycones, such as GC-MS (Laurila et al, 1999), LC-MS (Eanes et al, 2008), MALDI-TOF/MS (Eanes et al, 2008), HPLC-UV and HPTLC (Eanes et al, 2008;Paul et al, 2008;Skarkova et al, 2008;Shanker et al, 2011), but there is no report on the qualitative and quantitative estimation of the three markers, ISD, SM and SN in S. violaceum or in any other species of the genus Solanum.…”

Development of an Efficient Extraction Method for Separating Solasodine, an Steroidal Alkaloid, From an Oily Matrix: A Comparison Between LLE and SPE

“…Different analytical techniques such as high performance thin layer chromatography (HPTLC) [8], gas chromatography (GC) [9], high performance liquid chromatography (HPLC) [10] and potentiometry [11,12] have been reported for the determination of steroidal alkaloids in herbal drugs. Most of these compounds have a low volatility and are susceptible to elevated temperatures, so GC is not usually the method of choice for analysis of steroidal alkaloids [13], whereas, HPLC has been used for the determination of a variety of steroids and alkaloids [10].…”

Variations in the cytotoxic glycoalkaloids solamargine and solasonine in different parts of the Solanum incanum plant during its growth and development in Oman

“…A useful analysis method consists of three major steps: extraction of the alkaloids with aqueous or non-aqueous solvents, pre-purification or removal of interfering substances (impurities), and chromatographic procedures [4,10]. Different methodologies have been proposed to analyse Solanaceae glycoalkaloids in foodstuffs and plant materials, including colorimetric, chromatographic, or immunoassay methods [9,19]. The chromatographic methods, such as high-performance thin-layer chromatography (HPTLC), high-performance liquid chromatography (HPLC), and gas chromatography (GC), can be used to determine the levels of solasodine and its glycosides from their elaborating plant [3,10,11,[15][16][17]19,20].…”

“…Different methodologies have been proposed to analyse Solanaceae glycoalkaloids in foodstuffs and plant materials, including colorimetric, chromatographic, or immunoassay methods [9,19]. The chromatographic methods, such as high-performance thin-layer chromatography (HPTLC), high-performance liquid chromatography (HPLC), and gas chromatography (GC), can be used to determine the levels of solasodine and its glycosides from their elaborating plant [3,10,11,[15][16][17]19,20]. Among the various analytical techniques, high-performance thin-layer chromatography (HPTLC), in particular, appears to be suitable for phytomolecules of varying natures and provides a rational approach in the authentication and quality assessment of crude medicinal herbs and their formulations [18].…”