GC-MS (gas chromatography-mass spectrometer) targeted tobacco sample sterol extraction method

Abstract

The invention discloses a GC-MS (gas chromatography-mass spectrometer) targeted tobacco sample sterol extraction method. The method comprises the following process steps: (1) freeze-drying a tobacco sample, and grinding the tobacco sample through a grinder; (2) adding 1.5ml of normal hexane in 50mg of ground tobacco sample, and performing ultrasonic extraction; (3) extracting the sample, performing high-speed centrifugation at room temperature, transferring a supernatant into a chromatographic bottle, and blow-drying with nitrogen to obtain a solution I; (4) adding 1.2ml of 80% methanol and 20% KOH solution into a precipitate; (5) performing high-speed centrifugation, transferring a supernatant into the chromatographic bottle, adding 1.0ml of normal hexane, and performing vibration and standing; (6) extracting a supernatant, adding 1.0ml of saturated NaCl, taking a supernatant as a solution II, combining the solution I and the solution II, blow-drying with nitrogen, and adding 0.5ml of chloroform for dissolution to obtain a tobacco sample sterol sampling analysis solution specially suitable for GC-MS measurement. The method has the advantages that the extraction method is optimally suitable for tobacco leaf sterol; the use amount of the sample is small and is only 50mg; few types of organic solvents are used, the use amount of the organic solvents is low, and the organic solvents are relatively cheap and small in toxicity; the extraction time is short, and the sample reproducibility is good.

Description

[0001] technical field [0002] The invention relates to a method for extracting sterols from fresh tobacco leaf samples for tobacco metabolomics, specifically a GC-MS targeted tobacco sample sterol extraction method. The invention uses GC-MS as an analysis platform and adopts SIM ion mode to analyze suitable tobacco Optimal extraction method of leaf sterols. Background technique [0003] Phytosterols are a class of steroidal compounds that widely exist in nature. They belong to saturated or unsaturated secondary alcohols. In the past, there were many studies on animal sterols, but phytosterols were neglected. Tobacco contains a variety of sterol compounds, mainly cholesterol, stigmasterol, campesterol and β-sitosterol, which are one of the main components of tobacco leaf oil. However, when tobacco leaves are smoked, the parent polycyclic structure of sterols can also form fused-ring aromatic hydrocarbons, such as stigmasterol pyrolyzed at 750°C to generate benzo[α]pyrene (...

AI Technical Summary

Problems solved by technology

At present, there is no general extraction method for sterols from fresh tobacco leaves. Only based on the extraction methods of other plants and roasted tobacco leaves, combined with the detection method (GC-MS), a general extraction method for sterols from fresh tobacco leaves can be established.

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