食用菌中L-色氨酸HPLC检测方法的建立  

Establishment of HPLC Detection Method for L-tryptophan in Edible Fungi

作　　者：游金坤[1] 孙达锋[1] 代秋琼 曹燕妮 邓雅元[1] 华蓉[1] YOU Jinkun;SUN Dafeng;DAI Qiuqiong;CAO Yanni;DENG Yayuan;HUA Rong(Kunming Edible Fungi Institute of All China Federation of Supply and Marking Cooperatives,Kunming 650221,China;Yunnan Yunjun Technology(Group)Co.,Ltd.,Kunming 650221,China;Yunnan Academy of Edible Fungi Industry Development,Kunming 650221,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]云南云菌科技(集团)有限公司,云南昆明650221 [3]云南省食用菌产业发展研究院,云南昆明650221

出　　处：《中国食用菌》2024年第6期72-77,共6页Edible Fungi of China

基　　金：科技人才与平台计划(院士专家工作站)(202305AF150187);云南省省市一体化专项(202402AN360003)。

摘　　要：建立了高效液相色谱(high performance liquid chromatography,HPLC)测定食用菌中L-色氨酸含量的方法,并进行了方法学验证。采用Acclaim^(TM)120 C18色谱柱(4.6 mm×250 mm,5μm),以0.05 moL·L^(-1)乙酸钠缓冲液(用冰乙酸将pH调至4.0~5.0)和纯甲醇为流动相(体积比为70∶30)等度洗脱,流速为1.0 mL·min^(-1),检测波长为280 nm,柱温为30℃,进样量为20μL。结果显示L-色氨酸在4~100μg·mL^(-1)线性关系良好,R^(2)>0.9955。精密度、重复性、24 h内稳定性的相对标准偏差均小于15%(n=6),加标回收率为87.5%(相对标准偏差为0.63%)。说明建立的高效液相色谱检测方法简单、快速、灵敏度高、特异性强、分离度好,适用于食用菌中L-色氨酸含量的测定。Methodological validation was conducted on the established high performance liquid chromatography(HPLC) method for determining L-tryptophan content in edible fungi.Using an Acclaim~(TM)120 C18 chromatography column(4.6 mm × 250 mm,5 μm),isometric elution was performed with 0.05 mol·L~(-1) sodium acetate buffer(pH adjusted to 4.0-5.0 with glacial acetic acid)and pure methanol as mobile phases(the volume ratio is 70 ∶ 30),with a flow rate of 1.0 mL·min~(-1),detection wavelength of 280 nm,column temperature of 30 ℃,and injection volume of 20 μL.The results showed that L-tryptophan showed a good linear relationship in the range of 4-100 μg·mL~(-1),and the R~2>0.995 5.The relative standard deviation of precision,repeatability and stability within 24 hours is less than 15%(n=6),and the spiked recovery rate was 87.5%(relative standard deviation was0.63%).The established HPLC detection method is simple,fast,highly sensitive,specific,and has good separation,it is suitable for the determination of L-tryptophan content in edible fungi.

关 键 词：食用菌 L-色氨酸 高效液相色谱 

分 类 号：S646.9[农业科学—蔬菜学]