Extraction And Identification Of Anthocyanins From Four Flower Organs Of Brassica Napus L.

As the main oilseed crops in China,Brassica napus is a significant source of edible vegetable oil,protein feed,energy crops and industrial raw materials.Flower color is one of the important ornamental traits of B.napus.The study of anthocyanidin is the basis of flower color improvement,and the clarification of the relationship between flower color and anthocyanidin makes the flower breeding work more directional.In this paper,with Brassica napus L.white flower line W01,yellow flower line Zhong Shuang 11,pink flower line P05 and orange flower line R03 as materials,high performance liquid chromatography was applied to study the chromategraphic separa tion conditions and optimization of ultrasound-assisted extraction of anthocyanins in Brassica napus L.floral organs,and the total anthocyanins content was measured.Also,ultra high performance liquid chromatography-tandem mass spectrometry technology(UPLC-HESI-MS)was used to identify and analyze anthocyanins extracted by ultraso nic assisted method.The main results were as follows:1.An HPLC test method was constructed,which can optimize the conditions for the extraction of anthocyanins by ultrasonic assisted extraction.Through singlefactor test and orthogonal experiments,the effect of types of organic solvents,the concentra tion of ethanol,solid-liquid ratio,extraction time and extraction times on anthocyanin extraction from flower organs of B.napus.were analyzed and the optimization of ultrasound-assisted extraction of anthocyanins were determined.2.The total amount of anthocyanins in floral organs of four flower materials at different developmental stages were analyzed by HPLC method using cyanidin-3-Oglucoside as standard.The anthocyanins were not detected in the stalks and sepals of the four flower materials.Whereas anthocyanins were found in the petals,anthers and pistils of pink and orange flowers.3.The components of anthocyanins in stalks,sepals,petals,anthers and pistils of four flower materials were analyzed by UPLC-HESI-MS.The results showed that fifteen types of anthocyanins were identified,including delphinidin-3-caffeoylglucoside-5-glucoside,delphinidin-3,5-glucoside,delphinidin-3-O-?-glucopyranoside,Cyanidin-3-diglucoside-5-glucoside,cyanidin-3,5-O-glucoside,cyanidin-3-O-(2''-O-beta-glucopyranoside-beta-glucopyranoside),cyanidin3-O-glucoside,petunidin-3,5-O-hexanoside,petunidin-3-O-sophoroside,petunidin-3-O-beta-glucopyranoside and petunidin-3-caffeoylglucoside-5-glucoside.Among them,delphinidin-3-caffeoylglucoside-5-glucoside,delphinidin-3,5-glucoside,cyanidin-3,5-O-glucoside and petunidin-3,5-Ohexanoside have one isomer.4.The content and proportion of delphinidin-3-O-beta-pyranoside,cyanidin-3-Oglucoside and petunidin-3-O-beta-glucopyranoside determine the shading of color of orange and pink flowers,while cyanidin-3-O-(2''-O-beta-glucopyranoside-betaglucopyranoside)and petunidin-3-O-sophoroside play a key role in the color of orange and pink flowers.The content and proportion of delphinidin-3,5-glucoside,cyanidin-3,5-O-glucoside and petunidin-3,5-O-hexoside determine the color of orange flower anther;The content and proportion of delphinidin-3,5-glucoside and cyanidin-3-O-(2''-O-beta-glucopyranoside-beta-glucopyranoside)play a decisive role in the color of pink flower anthers.The content and proportion of delphinidin-3-caffeoylglucoside-5-glucoside and its isomers and petunidin-3-caffeoylglucoside-5-glucoside play a decisive role in the color of orange flower pistils.The contents of delphinidin-3-caffeoylglucoside-5-glucoside and petunidin-3-caffeoyl-glucoside-5-glucoside have great influence on the color of pink flower pistils.