威廉环毛蚓中8种核苷提取工艺优化及含量测定  

Extraction process optimization and content determination of eight nucleosides from Pheretima guillelmi

作　　者：宇泉霖 吴雪纯 邱旖 宋嘉佳 蒋巧萍 孙常胜 吴婧楠 蔡程科[1] 王洪飞 YU Quan-lin;WU Xue-chun;QIU Yi;SONG Jia-jia;JIANG Qiao-ping;SUN Chang-sheng;WU Jing-nan;CAI Cheng-ke;WANG Hong-fei(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102400,China;Beijing Quantum Hi-tech Pharmaceutical Technical Co.,Ltd.,Beijing 102200,China)

机构地区：[1]北京中医药大学中药学院,北京102400 [2]北京量子高科制药有限公司,北京102200

出　　处：《中成药》2024年第8期2526-2530,共5页Chinese Traditional Patent Medicine

基　　金：国家科技重大专项研发项目(2011ZX09201-201);北京市科技计划专项研发项目(Z16010101390)。

摘　　要：目的优化威廉环毛蚓中尿嘧啶、次黄嘌呤、黄嘌呤、尿苷、胸腺嘧啶、肌苷、鸟苷、2′-脱氧鸟苷提取工艺,并测定其含量。方法以料液比、超声时间、超声温度为影响因素,次黄嘌呤、总核苷含量为评价指标,正交试验优化提取工艺。HPLC法测定各核苷含量,分析采用Agilent C_(18)色谱柱(4.6 mm×250 mm,5μm);流动相甲醇-水,梯度洗脱;体积流量1 mL/min;柱温30℃;检测波长260 nm。结果最佳条件为料液比1∶250,超声时间30 min,超声温度60℃。8种核苷在各自范围内线性关系良好(R^(2)>0.9990),平均加样回收率99.11%~103.27%,RSD 0.85%~2.89%。结论该方法稳定可靠,可用于威廉环毛蚓中核苷的提取与含量测定。AIM To optimize the extraction process for uracil,hypoxanthine,xanthine,uridine,thymine,inosine,guanosine and 2′-deoxyguanosine from Pheretima guillelmi(Michaelsen),and to determine their contents.METHODS With solid-liquid ratio,ultrasonic time and ultrasonic temperature as influencing factors,contents of hypoxanthine and total nucleosides as evaluation indices,the extraction process was optimized by orthogonal test.HPLC was adopted in the content determination of varioud nucleosides,the analysis was performed on a 30℃thermostatic Agilent C_(18)column(4.6 mm×250 mm,5μm),with the mobile phase comprising of methanol-water flowing at 1 mL/min in a gradient elution manner,and the detection wavelength was set at 260 nm.RESULTS The optimal conditions were determined to be 1∶250 for solid-liquid ratio,60 min for ultrasonic time,and 60℃for ultrasonic temperature.Eight nucleosides showed good linear relationships within their own ranges(R^(2)>0.9990),whose average recoveries were 99.11%-103.27%with the RSDs of 0.85%-2.89%.CONCLUSION This stable and reliable method can be used for the extraction and content determination of nucleosides from P.guillelmi.

关 键 词：威廉环毛蚓 核苷 提取工艺 含量测定 正交试验 HPLC 

分 类 号：R284.1[医药卫生—中药学] R284.2[医药卫生—中医学]