Identification of Silymarin Constituents: An Improved HPLC–MS Method

Abstract

A high-performance liquid chromatographic/ tandem mass spectrometric method was developed for the determination of the major bioactive flavonolignans in silymarin, a herbal remedy extracted from the milk thistle Silybum marianum. In this study, eight active components of silymarin with the same elemental composition, including silychristins A and B, silydianin, silybin A and B, isosilybin A and B and an unknown compound were completely separated. Furthermore, three additional components were detected and partly separated; presumably two silybin stereoisomers and one isosilybin stereoisomer. The collision-induced dissociation (CID) MS/MS spectra of these silymarin constituents were studied: the spectral similarity values of the component pairs were determined, and simple criteria were found for distinguishing the components.Keywords High-performance liquid chromatography Á Tandem mass spectrometry Á Milk thistle Á Silymarin Á Silybin Electronic supplementary material The online version of this article (

Silibinin suppresses EMT-driven erlotinib resistance by reversing the high miR-21/low miR-200c signature in vivo

“…The milk thistle extract composition was analyzed and quantified by HPLC–145 DAD-ESI-MS/MS on an Agilent LC 1100 series (Agilent Technologies, Inc., Palo Alto, CA, USA). The retention time, UV spectra, and MS/MS data of the sample peaks were compared to data reported in the literature727374. Quantitation of the main flavonolignan compounds was performed using ChemStation for LC 3D software (Agilent Technologies Life Sciences and Chemical Analysis, Waldbronn, Germany), and the silibinin equivalents were determined with silibinin standards (Sigma–Aldrich).…”

Simple and Rapid HPLC Separation and Quantification of Flavonoid, Flavonolignans, and 2,3-Dehydroflavonolignans in Silymarin

“…This result, in line with previously published reports [2,26], clearly shows that the calibration curves strictly for each individual silymarin component must be measured to precisely determine their content. Generalization of one calibration curve, such as analysis based only on the most common compound silybin [3,27,29], leads to gross inaccuracies in the quantification.…”

“…The flowers, roots [1], and mainly the fruits (achenes) contain a rather unique type of polyphenols, the flavonolignans such as isosilychristin, silychristins A and B, silydianin, silybins A and B, isosilybins A and B (Figure 1). The complex extract containing them and known as silymarin (SM) [2][3][4] is obtained from milk thistle fruits and contains besides the flavonolignans also their biogenetic precursor the flavanonol taxifolin [5]. The corresponding 2,3-dehydroderivatives of flavonolignans such as 2,3-dehydrosilybin and 2,3-dehydrosilychristin ( Figure 1) are tentative silymarin components as well although they used to be considered possible artifacts or oxidative products arising from processing and storage [6].…”

Rapid qualitative analysis of 2 flavonoids, rutin and silybin, in medical pills by direct analysis in real‐time mass spectrometry (DART‐MS) combined with in situ derivatization

“…Because of the extensive medical applications of flavonoids, there is a strong need for the development of highly sensitive and effective analytical methods for the characterization of these compounds. Liquid chromatography (LC) and its coupling with mass spectrometry (LC‐MS) are conventionally used analyze flavonoids and silymarin . However, these techniques are quite time‐consuming and labor intensive.…”

“…Liquid chromatography (LC) and its coupling with mass spectrometry (LC-MS) are conventionally used analyze flavonoids 4 and silymarin. 5 However, these techniques are quite time-consuming and labor intensive. In contrast, ambient mass spectrometric methods, eg, direct analysis in real-time mass spectrometry (DART-MS), enable the examination of solid samples in open air without sample preparation, thereby allowing for rapid analysis.…”