澳洲坚果各部位蜀黍苷含量的测定
Determination of Dhurrin Content in Different Parts of Macadamia
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摘要: 为检测澳洲坚果果仁、花、叶、壳和青皮各部位的蜀黍苷含量,并为澳洲坚果各部位的开发利用提供依据,以苦杏仁苷为内标物质,采用高效液相色谱法,Diamonsil C18(150 mm×4.6 mm,5 μm)色谱柱,柱温35℃,自动进样器进样量5 μL,总流速1.0 mL/min,乙腈(A)-0.1%甲酸水溶液(B)为流动相,使用蒸发光散射检测器(ELSD)进行检测。结果表明:蜀黍苷和苦杏仁苷标准品的浓度比(X)与对应的峰面积比(Y)所作标准曲线方程为Y=0.465389X2+0.349685X-0.000520968,决定系数R2=0.9977,RSD为2.60%。方法的平均回收率为97.93%±2.35%,RSD为2.39%。对澳洲坚果各部位蜀黍苷含量进行测定发现,澳洲坚果花中蜀黍苷含量最高,达(49.92±0.96)mg/g,叶次之,果仁中蜀黍苷含量最低,仅为(0.64±0.01)mg/g。且不同品种青皮中蜀黍苷含量亦不同。HPLC-ELSD内标法测定蜀黍苷快速简便,准确度高,重复性好,成本较低,测定结果可靠。澳洲坚果花、叶中蜀黍苷含量较高,使用时应适当考虑脱除其中的蜀黍苷。澳洲坚果仁中仅含微量蜀黍苷,在安全使用范围之内。Abstract: HPLC-ELSD internal standard method was explored for the determination of dhurrin content in macadamia nuts,flowers,leaves,shells and husks to lay the foundation for the further application of macadamia. Amygdalin was chosen as the internal standard object in the experiment. Analysis was carried out using HPLC,equipped with an Diamonsil C18 column(150 mm×4.6 mm,5 μm)thermostated at 35℃. Chromatography was performed with the injection volume of 5 μL and total flow rate of 1.0 mL/min,by eluting with a gradient of acetonitrile(A)and 0.1% formic acid water solution(B). ELSD detector was used for dhurrin detection. Results showed that the standard curve for the concentration ratio(X)of dhurrin and amygdalin and their corresponding peak area ratio(Y)was Y=0.465389X2+0.349685X-0.000520968,with a fit of R2=0.9977,RSD was 2.60%.Recovery rate of this method was 97.93±2.35%,RSD=2.39%. Dhurrin content in macadamia flowers was the highest(49.92±0.96) mg/g,followed by macadamia leaves,and the lowest dhurrin content was found in macadmaia nuts(0.64±0.01) mg/g.Besides,dhurrin contents in different cultivars of macadamia husks was also varied. HPLC-ELSD internal standard method is a rapid,simple,accurate and reproducible method for dhurrin determination,with relatively low cost and reliable quantitative results. Appropriate consideration should be given to the removal of dhurrin in macadamia flowers and leaves before using due to their relatively high level of dhurrin content. While macadamia nuts contain only trace amounts of dhurrin,which is safe to use.