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建立了一种用于烟草中游离氨基酸测定的反相高效液相色谱法。实验采用超声波水解、邻苯二甲醛/3-巯基丙酸作为衍生剂进行柱前衍生。色谱柱为依利特C18柱(4.6mmi.d.×250mm,5μm),流动相A为18mmol/L的醋酸钠溶液(PH7.2)含体积分数为0.002%的三乙胺和0.3%的四氢呋喃,流动相B组成为:100mmol/L的醋酸钠溶液(PH7.2)-乙腈-甲醇(体积比为1:2:2),流速为1.0ml/min,柱温为40℃。荧光检测器,激发波长350nm,发射波长450nm。方法的回收率为95.3%￣100.7%,RSD为2.32%￣9.24%(n=6)。该方法简便、准确、重现性好。测定不同肥料配比生产的白肋烟烟叶中17种游离氨基酸的含量。结果表明,不论有机肥与无机肥怎样配比,天冬氨酸的含量与各氨基酸相比都是最高的;随着饼肥的加入,大部分氨基酸的含量是先增加后逐渐降低的趋势,15%饼肥+85%无机肥与30%饼肥+70%无机肥配比时,大部分游离氨基酸的含量接近,30%饼肥+70%无机肥配比时的游离氨基酸总量最高,综合考虑,30%饼肥+70%无机肥配比时的烟叶质量最好。
An RP-HPLC method for the determination of free amino acids in tobacco was established. Experiments using ultrasonic hydrolysis, o-phthalaldehyde / 3-mercaptopropionic acid derivative as a pre-column derivatization. The column was Elitel C18 column (4.6mmi.d. × 250mm, 5μm), the mobile phase A was 18mmol / L sodium acetate solution (pH7.2) containing 0.002% of triethylamine and 0.3% of tetrahydrofuran The mobile phase B consisted of 100 mmol / L sodium acetate solution (pH7.2) -acetonitrile-methanol (1: 2: 2 by volume) at a flow rate of 1.0 ml / min and a column temperature of 40 ° C. Fluorescence detector, excitation wavelength 350nm, emission wavelength 450nm. The recoveries were 95.3% ~ 100.7% and RSDs were 2.32% ~ 9.24% (n = 6). The method is simple, accurate and reproducible. The content of 17 kinds of free amino acids in burley tobacco produced by different fertilizer ratio was determined. The results showed that the content of aspartic acid was the highest among all the amino acids regardless of the ratio of organic fertilizer and inorganic fertilizer. With the addition of cake fertilizer, most of the amino acid content increased first and then decreased gradually. 15 The content of most free amino acids was close to the ratio of 30% cake fertilizer + 85% inorganic fertilizer and 30% cake fertilizer + 70% inorganic fertilizer, and the total amount of free amino acids was the highest when 30% cake fertilizer + 70% inorganic fertilizer was combined. % Of cake fertilizer + 70% inorganic fertilizer when the best quality tobacco.