Pretreatment method for simultaneous determination of free-state and binding-state sterols in tobaccos
A free state and combined state technology, applied in the direction of measuring devices, material separation, instruments, etc., can solve problems such as unsatisfactory, complex processing, and high operational safety requirements, and achieve the effects of improving accuracy, reducing experimental errors, and simplifying experimental operations.
Inactive Publication Date: 2019-05-10
CHINA TOBACCO YUNNAN IND
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Problems solved by technology
Stedman et al. used the gravimetric method to determine the free and bound sterols in tobacco. This method has low requirements for instruments, but has high requirements for operational safety, long analysis time, and many interference factors. It can only give the total sterol content, not separately. Content of various sterols
Using liquid chromatography to measure sterols in plants, without derivatization treatment, only free sterols can be measured; using gas chromatography to measure sterols in plants requires derivatization, and the pretreatment is complicated
Therefore, none of them can meet the purpose of simultaneously and efficiently measuring the content of free and bound sterols in tobacco.
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Embodiment 1
[0020] A high-efficiency pretreatment method for the simultaneous determination of free and bound sterols in tobacco, which is carried out according to the following steps: Step 1: Pulverize the tobacco sample to be tested, pass through a 40-mesh sieve, and weigh 0.25 g of the sample in an outer tube sample bottle , add 5.0 mL of 2.0 mol / L hydrochloric acid-ethanol solution and 200 μL of 6-ketocholestanol internal standard solution with a concentration of 0.5 mg / mL; Step 2: Ultrasonic hydrolysis in a constant temperature ultrasonic water bath at 60 °C for 1 h;
[0021] Step 3: Cool for 5 minutes after hydrolysis, open the sealing cap of the outer tube, add 10 mL of 4moL / L potassium hydroxide-ethanol solution to the extract after acid hydrolysis, tighten the sealing cap again, and place in a constant temperature ultrasonic water bath at 60 ℃ Ultrasonic saponification for 40 min;
[0022] Step 4: After cooling for 5 minutes, add 10 mL of isooctane and 10 mL of saturated potassiu...
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Abstract
The invention relates to an efficient pretreatment method for simultaneous determination of free-state and binding-state sterols in tobaccos. 2.0 mol/L of a hydrochloric acid-ethanol solution is usedto finish hydrolysis in a 60 DEG C constant-temperature ultrasonic water bath for 1 hour; 4 mol/L of a potassium hydroxide-ethanol solution is used to finish saponification in the 60 DEG C constant-temperature ultrasonic water bath for 40 minutes; isooctane and a saturated potassium chloride solution is oscillated for 5 minutes, and standing is performed for 10 minutes to finish extraction of total sterol; and drying is finished through anhydrous sodium sulfate between sieve plates, filtering of a sample is finished by the sieve plates, then the total sterol is exported from a sample exportingtube in the isooctane, and liquid chromatography-tandem mass spectrometry can be directly adopted for detection, thereby finishing the determination of the sterols in the tobaccos. The hydrolysis, saponification, extraction, drying and filtering operations of the pretreatment of the sterols in the tobaccos are finished in a pretreatment device; and the to-be-analyzed sample can be obtained without sample transfer in the whole sample pretreatment process, so that the experiment operation is greatly simplified and the pretreatment efficiency is improved.
Description
technical field [0001] The invention relates to a high-efficiency pretreatment method for simultaneous determination of free and bound sterols in tobacco, belonging to the technical field of tobacco chemical component determination. Background technique [0002] The sterol compounds in tobacco mainly include stigmasterol, β-sitosterol, campesterol and cholesterol, and mildewed tobacco leaves also contain ergosterol. Among them, the content of stigmasterol is the highest and the content of cholesterol is the lowest. Sterols exist in tobacco in free and bound states, including sterol esters with fatty acids, sterol glycosides and acyl sterol glycosides with sugars. [0003] Sterols in tobacco can regulate the growth and development of tobacco, respond to adversity stress, and are also one of the main components of tobacco leaf oil. However, sterols after combustion have a greater impact on the safety and quality of tobacco. Content is of great significance for tobacco qualit...
Claims
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IPC IPC(8): G01N30/02G01N30/06
Inventor 曾婉俐陈章玉李雪梅向海英李晶高茜杨光宇
Owner CHINA TOBACCO YUNNAN IND
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