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侯思宇,宋 敏,闫陆飞,孙朝霞,韩渊怀,李红英.HPLC法测定谷子籽粒叶酸含量及种质资源评价[J].土壤,2018,50(6):1235-1240. HOU Siyu,Song Min,YAN Lufei,SUN Zhaoxia,HAN Yuanhuai,LI Hongying.Evaluation and Detection of Folic Acid Content of Seeds Among Foxtail Millet Accessions by HPLC Method[J].Soils,2018,50(6):1235-1240 本文二维码信息
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HPLC法测定谷子籽粒叶酸含量及种质资源评价
Evaluation and Detection of Folic Acid Content of Seeds Among Foxtail Millet Accessions by HPLC Method
投稿时间:2018-07-15  修订日期:2018-08-10
DOI:10.13758/j.cnki.tr.2018.06.028
中文关键词:  谷子  高效液相色谱法  叶酸  种质资源
Key Words:Foxtail millet  HPLC  Folic acid  Germplasm resource
基金项目:山西省农谷建设科研专项项目(SXNGJSKYZX201702)资助。
作者单位E-mail
侯思宇 山西农业大学农学院 18635068055@163.com 
宋 敏 山西农业大学农学院  
闫陆飞 山西农业大学农学院  
孙朝霞 山西农业大学农学院  
韩渊怀 山西农业大学农学院 swgctd@163.com 
李红英 山西农业大学农学院  
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中文摘要:
      谷子籽粒中富含叶酸,作为一种功能性食品深受北方人民的喜爱,而作物籽粒,如小米叶酸测定方法及含量分析存在较大差异,为小米品质鉴定及高叶酸谷子种质筛选造成了一定困难。本研究通过“三酶法”提取小米中的叶酸,建立了一种基于高效液相色谱技术(HPLC)高效化、规模化且稳定适用于测定谷子籽粒叶酸含量的方法,对2016 年和2017年种植的45个谷子种质进行叶酸含量测定和分类评价。结果表明:叶酸标准品梯度曲线呈线性方程相关,相关系数为0.99;小米样品叶酸测定过程中精密度、稳定性和重复性的相对标准偏差(RSD)分别为0.81%、1.19% 和1.65%,各项指标符合精密测定标准,准确度高,重复性好;45个谷子种质籽粒叶酸含量分布范围为0.26 ~ 2.56 μg/g(2016年度)和0.53 ~ 2.86 μg/g(2017年度),含量均值分别为1.19和1.84 μg/g,标准差0.47和0.51 μg/g,变异系数39.5% 和27.7%;对上述种质进行正态分布曲线分析,发现叶酸含量整体呈现偏度分布;从中筛选出9个高叶酸(含量≥1.91 μg/g)和3个低叶酸(含量≤1.15 μg/g)的谷子种质。上述结论为小米叶酸含量提供了测定标准和评价体系,也为高叶酸谷子种质创新与利用奠定了基础。
Abstract:
      Foxtail millet is a kind of functional food rich in folic acid, and it is popular in the northern China. However, there are significant differences in folic acid contents in crop grains, such as the determination method and content analysis of folate in Foxtail millet, which make it difficult to identify the quality of millet and select the germplasm with high folate content. In this study, ‘triple-enzymes method’ was used to isolate folic acid from foxtail millet grains and construct a large-scale detection method by HPLC for folic acid content. Meantime, the folic acid contents were assessed among 45 accessions of foxtail millet in 2016 and 2017. The results showed that: based on HPLC, a folic acid content determination method for foxtail millet was successfully established. Folic acid standard curve showed the linear correlation between folic acid standard and peak area value, the correlation coefficient was 0.99. The relative standard deviation (RSD) was used to evaluate the precision, stability and repeatability in the detection process of folic acid content, and they were 0.81%, 1.19% and 1.65%, respectively. The range of grain folic acid content of 45 millet varieties were 0.26 ~ 2.56 μg/g in 2016 and 0.53 ~ 2.86 μg/g in 2017, with average values of 1.19 and 1.84 μg/g, standard deviation of 0.47 and 0.51 μg/g, and variation coefficient of 39.5% and 27.7%, respectively. The gaussian distribution had a little skewness. Finally, 9 high folic acid (folate content≥1.91 μg/g) and 3 low folic acid (folate content≤1.15 μg/g) accessions were screened from 45 millet accessions, respectively. This study not only provided the standard determination and evaluation system of folate in foxtail millet, and also laied a foundation for the innovation and utilization of high folate germplasm of foxtail millet.
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