Protocol detail

Quantitative GC-MS Analysis of Plant Sterols

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Sterols were silylated with N,O-bis[trimethylsilyltrifluoro-acetamide] (BSTFA), trimethylchlorosilane (TMCS), and pyridine at 70 °C for 1 h. Trimethylsilyl (TMS) derivatives of sterols were analyzed with GC-MS (Shimadzu) equipped with a Phenomenex (USA) ZB-5 Guardian column (30 m × 0.25 mm × 0.25 μm). Sterols were identified using characteristic ions^{78 (link)} and quantified using authentic standard solutions of plant sterol mixture from Larodan (including 53% β-sitosterol, 7% stigmasterol, 26% campesterol, 13% brassicasterol), and cholesterol, desmosterol, ergosterol, and fucosterol from Sigma-Aldrich. The recovery percentage of the sterol samples was calculated using 5-α-cholestane (Sigma-Aldrich) as an internal standard.

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Taipale S.J., Peltomaa E., Kukkonen J.V., Kainz M.J., Kautonen P, & Tiirola M. (2019). Tracing the fate of microplastic carbon in the aquatic food web by compound-specific isotope analysis. Scientific Reports, 9, 19894.

Publication 2019

GC-MS ZB-5 Guardian column β-sitosterol desmosterol ergosterol fucosterol 5-α-cholestane

Acetamide Brassicasterol Bstfa Campesterol Cholestane Cholesterol Derivatives Desmosterol Ergosterol Fucosterol Gc ms Guardian Plant sterol Pyridine Sitosterol Sterol Stigmasterol Trimethylchlorosilane

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Variable analysis

independent variables

Silylation reagents used: N,O-bis[trimethylsilyltrifluoro-acetamide] (BSTFA), trimethylchlorosilane (TMCS), and pyridine
Silylation temperature: 70 °C
Silylation time: 1 h

dependent variables

Sterol composition and concentrations as determined by GC-MS analysis

control variables

GC-MS instrument: Shimadzu
GC column: Phenomenex ZB-5 Guardian column (30 m × 0.25 mm × 0.25 μm)
Authentic standard solutions used for sterol identification and quantification: plant sterol mixture from Larodan (including 53% β-sitosterol, 7% stigmasterol, 26% campesterol, 13% brassicasterol), and cholesterol, desmosterol, ergosterol, and fucosterol from Sigma-Aldrich
Internal standard used for recovery calculation: 5-α-cholestane (Sigma-Aldrich)

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