摘要
以‘草莓果冻’海棠为试材,采用高效液相色谱结合固相萃取法建立了一种用于海棠果实中脱落酸(ABA)含量检测的方法。通过优化前处理及色谱条件,最终确定取样量为3g,用80%预冷甲醇浸提12h,经不溶性PVPP吸附酚类等杂质,石油醚脱色,乙酸乙酯萃取,C18固相萃取小柱进一步纯化,再经甲醇∶1.0%乙酸溶液(45∶55,v/v),流速为1.0mL·min^(-1)进行洗脱,C18色谱柱分离,进样体积20μL,检测波长为254nm,柱温45℃。结果表明:该方法在0.01~0.50μg·mL^(-1)范围内线性关系良好,R^2=0.999 8,3个浓度水平下平均回收率均大于70%。采用该方法对‘草莓果冻’海棠半同胞子代中14个单株落果初期果实进行了测定,发现果实中脱落酸含量最低为0.026 9μg·g^(-1),最高含量为0.357 9μg·g^(-1),差别较大,该浓度与已有报道中的测定结果处于同一数量级水平。该方法的建立对于海棠的选育和生长发育进一步研究具有重要意义。
＇Strawberry jelly＇crabapple（Malus sp.）was used as test material,a reliable method for determination of abscisic acid（ABA）content in crabapple was established using the high performance liquid chromatography（HPLC）combined with solid phase extraction（SPE）.By optimizing the sample pre-treatment and the chromatographic conditions,the final experimental process as follows,sample weight for 3 g,with 80% precooling methanol extracting for 12 hours,using the insoluble PVPP for adsorbing the phenolic compounds,then the extracts was decolorize by petroleum ether,and ABA was extracted by ethyl acetate,then further purification using C18 cartridge.The method was performed with a reverse-phase C18 column and using the mixture of methanol and water（containing 1.0%acetic acid）with the volume ratio of 45∶55 as mobile phase;the sample injection volume was 20μL and the flow rate was 1.0 mL· min^-1;the detection wavelength was 254 nm and the column temperature was 45 ℃.The results showed that the ABA were separated well by this method,which calibration curves had good linearity over the range from 0.01 to 0.50μg·mL^-1 with a correlation coefficient of 0.999 8,the average recovery rate for ABA was larger than 70% at three concentration levels.The abscisic acid content was lowest as 0.026 9μg·g^-1,and the highest as 0.357 9μg·g^-1 with great difference,at the concentration level of previous reported.This method showed great significance for the further research of crabapples breeding and growth.
出处
《北方园艺》
CAS
北大核心
2018年第4期138-144,共7页
Northern Horticulture
基金
河北科技师范学院应用开发研究基金资助项目(2017-3)
国家林业948资助项目(2011-4-26)
秦皇岛市农业科学研究院计划资助项目(2015-21)
关键词
海棠
高效液相色谱(HPLC)
脱落酸(ABA)
含量
crabapple （Malus sp. ）
high performance liquid chromatography （HPLC）
abscisic acid（ABA）
content
