食用菌中6种核苷酸的UHPLC检测方法的建立  

Establishment of UHPLC Detection Method for 6 Nucleotides in Edible Fungi

作　　者：代秋琼 孙达锋[1,2] 华蓉 陈勇 曹燕妮[2] 邓雅元 吴素蕊[2] 游金坤 DAI Qiu-qiong;SUN Da-feng;HUA Rong;CHEN Yong;CAO Yan-ni;DENG Ya-yuan;WU Su-rui;YOU Jin-kun(Kunming Edible Fungi Institute of All China Federation of Supply and Marking Cooperatives,Kunming 650221,China;Yunnan Edible Fungi Industry Development Research Institute,Kunming 650221,China;Yunnan Yunjun Technology(Group)Co.,Ltd.,Kunming 650221,China;National Engineering Research Centre for Biochemistry,Nanjing University of Technology,Nanjing 210009,China)

机构地区：[1]中华全国供销合作总社昆明食用菌研究所,云南昆明650221 [2]云南省食用菌产业发展研究院,云南昆明650221 [3]云南云菌科技(集团)有限公司,云南昆明650221 [4]南京工业大学国家生化工程研究中心,江苏南京210009

出　　处：《中国食用菌》2023年第5期58-61,69,共5页Edible Fungi of China

基　　金：科技人才与平台计划(院士专家工作站)(202305AF150187)。

摘　　要：对建立的UHPLC测定食用菌中尿嘧啶核苷酸、胞嘧啶核苷酸、鸟嘌呤核苷酸、肌苷5’-核苷酸、5’-磷酸腺苷、黄嘌呤核苷酸含量的方法进行方法学验证。采用YMC-Pack QDS-AQ色谱柱(250 mm×4.6 mm,5μm),以甲醇和10 mmol·L^(-1)乙酸铵(乙酸调pH至4.0)为流动相进行梯度洗脱,流速为0.8 mL·min^(-1),检测波长为260 nm,柱温为30℃,进样量为10μL。结果显示建立的UHPLC检测方法下6种核苷酸在4~100μg·mL^(-1)范围内线性关系良好,R^(2)>0.9997。精密度、重复性、24 h内稳定性RSD(n=6)均小于15%,加标回收率为90.61%~117.47%(RSD为2.12%~7.72%)。建立的食用菌中6种核苷酸UHPLC检测方法简单、快速、灵敏度高、特异性强、分离度好,适用于食用菌中6种核苷酸的含量测定。To establish a UHPLC method for the determination of uracil nucleotide,cytosine nucleotide,gunine nucleotide,ino-sine 5’-nucleotide,5’-adenosine phosphate and xanthine nucleotide in edible fungi,and complete method verification.The chromatographic conditions were YMC-Pack QDS-AQ column(250 mm×4.6 mm,5μm),gradient elution was performed with methanol and 10 mmol·L^(-1) ammonium acetate(adjust pH to 4.0 with acetic acid)as flowing phase,flow rate 0.8 mL·min^(-1),detection wavelength 260 nm,column temperature 30℃and injection volume 10μL.The results showed that the linear rela-tionship of the 6 nucleotides in the range of 4-100μg·mL^(-1) is good under the established UHPLC detection method,R^(2)>0.9997.The RSD(n=6)of precision,repetitiveness and stability within 24 hours were all less than 15%,the bidding recovery rateswere 90.61%-117.47%(RSD were 2.12%-7.72%).The established UHPLC detection method of 6 nucleotides in edible fungi is sim-ple,fast,high sensitivity,strong specificity,good separation,which is suitable for the measurement of 6 nucleotides in edible fungus.

关 键 词：食用菌 核苷酸 含量测定 超高效液相色谱 

分 类 号：S646.9[农业科学—蔬菜学]