Optimization Of HPLC Method For Determination Of Folic Acid In Foxtail Millet And Evaluation Of Germplasm Resources

Millet is known as “The first of the grain”,it is rich in a variety of nutrients to attract many scholars to explore its nutritional value.Folic acid is one of water-soluble B vitamin,which is widely involved in the transfer of one-carbon unit in cells,and has the function of regulating the development of embryonic nerve cells and lowering blood lipids.However,there are few studies on folic acid in millet seeds at present.In this study,a single-factor test and the response surface optimization test was designed for the extraction of folic acid from foxtail millet by three enzymatic methods and the content of folic acid in foxtail millet was measured by High Performance Liquid Chromatography(HPLC).We used 302 foxtail millet germplasm resources in Jinzhong and Jinnan ecoregion,then content differences were statistically analyzed.The total content of folic acid was revealed,which laid a theoretical foundation for molecular breeding of high folic acid millet germplasm in the future.The main findings were described as follows:(1)Trienzyme method was used to extract folic acid from foxtail millet with single factor test and response surface optimization test.The optimum extraction conditions were as follows: 1 mL of extract,20?L of amylase,150 ?L of protease,25 ?L of rat serum,and 1 mg/L of extract,20 ?L of amylase,150 ?L of protease and 25 ?L of rat serum.(2)Folic acid content was measured in foxtail millet by high performance liquid chromatography(HPLC)for single factor test and response surface optimization test.The optimum conditions were as follows: flow rate 1 mL / min,mobile phase pH 7,wavelength 280 nm,column temperature 30?.(3)The above method was used to extract and determine the folic acid content from 302 foxtail millet planted in two different ecological regions of Jinzhong and Jinnan ecoregion.The variation of folate content in Jinzhong ecoregion was 0.12~2.76 ?g/g,average folate content was 1.28 ?g/g,standard deviation was 0.38,coefficient of variation was 30.0 %.The variation of folate content in Jinnan ecoregion was 0.29~2.53 ?g/g,the average folate content was 1.21 ?g/g,the standard deviation was 0.44,and the coefficient of variation was 37.0 %.The idioplasm of high folic acid millets are ‘Jingu 21',‘cat's shitvalley',‘Dadanpi',‘Nenxuan 13',‘Yugu 1',etc.The idioplasm of low folic acid millets are ‘Jingu 23',‘Fenxuan 3',‘Liquor Valley' and ‘Longgu 4',etc.