Method for measuring glycoside in tobacco through enzyme hydrolysis-gas chromatography-mass spectrum combination
A technology of gas chromatography and enzymatic hydrolysis, which is applied in the field of enzymatic hydrolysis-gas chromatography-mass spectrometry for the determination of glycosides in tobacco, can solve the problems of less qualitative confirmation information and poor separation ability, and achieve high accuracy, improve precision, and optimize Effect of enzymatic hydrolysis conditions
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Embodiment 1
[0029] The method for the determination of glycosides in tobacco by enzymatic hydrolysis-gas chromatography-mass spectrometry of the present invention comprises the following steps:
[0030] (1) Extraction: Accurately weigh 4.0000 g of tobacco powder into a stoppered Erlenmeyer flask, add 250 μg of phenyl-β-D-glucopyranoside internal standard 1, and add 40 mL of methanol for ultrasonic extraction for 30 min (100 Hz at room temperature ), let stand for 15 minutes after the ultrasonication, and filter the supernatant, add 40 mL of methanol to the residue for ultrasonic extraction for 30 minutes (100 Hz at room temperature), combine the filtrates after the ultrasonication, and concentrate the methanol to 2 mL in a rotary concentrator;
[0031] (2) Purification: After dissolving and mixing with 25 mL of deionized water, transfer it to the XAD-2 macroporous resin glass column. The adsorbed glycosides are first rinsed with 50 mL of water to remove water-soluble sugars, amino acids an...
Embodiment 2
[0040] Tobacco glycosides were determined by enzymatic hydrolysis-gas chromatography-mass spectrometry for flue-cured tobacco, sun-cured tobacco and oriental tobacco by using the method of Example 1 of the present invention. The varieties of the three types of tobacco are K326, Kenner and Basma, and the parts are all in the middle leaves. All the tobacco leaves have not been rebaked and alcoholized. The chromatogram and glycoside content of relative quantitative determination are as Figure 2-5 and shown in Table 2.
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