UPLC法测定不同加工方式梅花鹿鹿茸中的核苷类成分  

Simultaneous determination of nucleosides and nucleobases in Cervi Cornu Pantotrichum of sika deer(Cervus nippon) with different processing methods by UPLC

作　　者：孙印石[1,2] 王燕华[2] 王玉方[1] 陈宝[2] 陈丽雪 

机构地区：[1]中国农业科学院特产研究所,吉林长春130112 [2]吉林农业大学中药材学院,吉林长春130118

出　　处：《中草药》2018年第4期840-846,共7页Chinese Traditional and Herbal Drugs

基　　金：吉林省科技发展计划项目(20170201034YY;20170309002YY;20170311027YY);中国农业科学院科技创新工程项目(CAAS-ASTIP-2016-ISAPS)

摘　　要：目的建立UPLC法同时测定梅花鹿鹿茸(花鹿茸)中胞嘧啶、尿嘧啶、腺嘌呤、鸟嘌呤、次黄嘌呤、黄嘌呤、尿苷、胸腺嘧啶、肌苷、鸟苷、腺苷、2′-脱氧鸟苷、β-胸苷13种核苷类成分含量的方法,研究不同加工方式的花鹿茸核苷类成分含量的差异及其在不同部位中的分布差异。方法水超声辅助提取花鹿茸样品中的核苷类成分,色谱柱Acquity UPLC?HSS T3(100 mm×2.1 mm,1.8μm),乙腈(A)-0.006%甲酸水溶液(B)梯度洗脱,体积流量0.3 mL/min;柱温30℃;进样量3μL;检测波长260 nm。结果 13种成分基本达到基线分离,线性范围内均具有良好的线性关系(r>0.999 6);排血茸和带血茸的蜡片、粉片、纱片部位核苷类成分的总量分别是4.47、3.95、2.68 g/kg和4.14、3.44、2.51 g/kg,煮炸茸和冻干茸3个部位核苷类成分的量分别为4.60、2.95、2.74 g/kg和5.06、4.24、2.31 g/kg。结论就核苷类成分总量而言,排血茸蜡片、粉片、纱片部位高于带血茸,冻干茸蜡片、粉片部位高于煮炸茸,纱片部位低于煮炸茸。Objective To establish the method of simultaneous determination of the content of 13 nucleosides and nucleobases, including cytosine, uracil, adenine, guanine, 6-hydroxypurine, 2,6-dihydroxypurine, uridine, thymine, inosine, guanosine, adenosine, 2＇-deoxyguanosine （2＇-dG）, beta-thymidine, in Cervi Cornu Pantotrichum （CCP） of sika deer （Cervus nippon） by UPLC, and compare the distribution differences of nucleosides and nucleobases in different zones of the CCP with different processing methods. Methods The nucleosides and nucleobases in CCP were extracted by water with assistance of ultrasound. Acquity UPLC HSS T3 column （100 mm×2.1 mm, 1.8 μm） was used as chromatographic column to separate the nucleosides and nucleobases. Thirteen target compounds were eluted with acetonitrile 100% （eluent A） and water plus 0.006% formic acid （eluent B） at a flow rate of 0.3 mL/min. The column temperature was 30℃, and the injection volume was 3 μL and the detection wavelength was 260 nm. Results A total of 13 nucleosides and nucleobases basically reached the baseline separation with a good linearity within linear range （r 〉 0.999 6）. The nucleosides and nucleobases content in wax slices, powder slices, gauze slices of CCP without and with blood were respectively 4.47, 3.95, 2.68 g/kg and 4.14, 3.44, 2.51 g/kg. And those three parts of CCP with boiling and freeze-drying processing were respectively 4.60, 2.95, 2.74 g/kg and 5.06, 4.24, 2.31 g/kg. Conclusion As far as the total content of nucleosides and nucleobases were concerned, the wax slices, powder slices and gauze slices of CCP without blood were all higher than those of CCP with blood. The wax slices, powder slices of CCP with freeze-drying processing were more than those of CCP with boiling processing, while the gauze slices of which were less than those of CCP with boiling processing.

关 键 词：核苷 核酸碱基 胞嘧啶 尿嘧啶 腺嘌呤 鸟嘌呤 次黄嘌呤 黄嘌呤 尿苷 胸腺嘧啶 肌苷 鸟苷 腺苷 2′-脱氧鸟苷 β-胸苷 梅花鹿 鹿茸 排血茸 带血茸 煮炸茸 冻干茸 蜡片 粉片 纱片 UPLC 分布差异 

分 类 号：R286.02[医药卫生—中药学]