Abstract
Silybin is a flavonolignan extracted from Silybum marianum with chemopreventive activity against various cancers, including breast. This study was designed to develop an HPLC-MS/MS method for the determination of silybin in human plasma, urine and breast tissue in early breast cancer patients undergoing Siliphos® supplementation, an oral silybin-phosphatidylcholine complex. The determination of silybin was carried out by liquid–liquid extraction with methyl-tert-butyl ether (MTBE); total silybin concentration was determined by treating the samples with β–glucuronidase, while for the determination of free silybin, the hydrolytic step was omitted. Naringenin and naproxen were selected as internal standards. The detection of the analyte was carried out by mass spectrometry and by chromatography. The HPLC-MS/MS method was evaluated in terms of selectivity, linearity, limit of quantification, precision and accuracy, and carryover. The method proved to be selective, linear, precise and accurate for the determination of silybin. To the best of our knowledge, this presents the first analytical method with the capacity to quantify the major bioactive components of milk thistle in three different biological matrices with a lower limit of quantification of 0.5 ng/mL for plasma. Silybin phosphatidylcholine, taken orally, can deliver high blood concentrations of silybin, which selectively accumulates in breast tumor tissue.
Highlights
The incidence of cancer and other degenerative disorders continues to rise in an increasingly aging population
At first apigenin was selected as internal standard, since at the chromatographic conditions established, naringenin was eluted at the same retention time of silybin
The experimental results demonstrated that naringenin behavior was more similar to silybin than apigenin, and the former was chosen as internal standard
Summary
The incidence of cancer and other degenerative disorders continues to rise in an increasingly aging population. Dietary phytochemicals are considered to be a major source of novel and safe potential cancer chemopreventive agents, mostly due to their long treatment regimen [1,2]. Most of the antitumor activity of silybin has been discovered through in vitro studies in tumor cell lines [7,8]. Fewer studies have investigated the antitumor effect exerted by in vivo supplementation with silybin on mice previously treated with carcinogens or on nude mice bearing human xenografts [9,10,11,12,13]. Some investigations have reported the effect of silymarin in the prevention and treatment of liver diseases and primary liver cancer in in vitro and in vivo models [4]. Recent data in animals reported the role of silybin in the chemoprevention of several cancer types, including breast [14,15]
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