Abstract
Tomato steroidal glycoalkaloids (tSGAs) are a class of cholesterol-derived metabolites uniquely produced by the tomato clade. These compounds provide protection against biotic stress due to their fungicidal and insecticidal properties. Although commonly reported as being anti-nutritional, both in vitro as well as pre-clinical animal studies have indicated that some tSGAs may have a beneficial impact on human health. However, the paucity of quantitative extraction and analysis methods presents a major obstacle for determining the biological and nutritional functions of tSGAs. To address this problem, we developed and validated the first comprehensive extraction and ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) quantification method for tSGAs. Our extraction method allows for up to 16 samples to be extracted simultaneously in 20 min with 93.0 ± 6.8 and 100.8 ± 13.1% recovery rates for tomatidine and alpha-tomatine, respectively. Our UHPLC-MS/MS method was able to chromatographically separate analytes derived from 18 tSGA peaks representing 9 different tSGA masses, as well as two internal standards, in 13 min. Tomato steroidal glycoalkaloids that did not have available standards were annotated using high resolution mass spectrometry as well as product ion scans that provided fragmentation data. Lastly, we utilized our method to survey a variety of commonly consumed tomato-based products. Total tSGA concentrations ranged from 0.2 to 3.4 mg/serving and represent some of the first reported tSGA concentrations in tomato-based products. Our validation studies indicate that our method is sensitive, robust, and able to be used for a variety of applications where concentrations of biologically relevant tSGAs need to be quantified.
Highlights
Solanaceous plants produce a spectrum of cholesterol derived compounds called steroidal glycoalkaloids
To address the lack of suitable approaches to extract and quantify Tomato steroidal glycoalkaloids (tSGAs), we developed and validated a high-throughput extraction and ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method suitable for tomato and tomato-based products
Development of High-Throughput Extraction Method Generally, tomato samples are pulverized in a mortar in the presence of liquid nitrogen or homogenized using a blender prior to extracting tSGAs
Summary
Solanaceous plants produce a spectrum of cholesterol derived compounds called steroidal glycoalkaloids. Emerging evidence suggests that some tSGAs may play a role in positive health outcomes associated with tomato consumption (Cayen, 1971; Lee et al, 2004; Choi et al, 2012; Cooperstone et al, 2017). While these compounds continue to be evaluated both in planta and in vivo, there is a lack of quantitative and validated methods to extract and measure tSGAs from tomatoes; a critical need for additional research in this area
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