TGDATCM-00122021-甘草煮散饮片

ICS11.120.99CCSC273 T/GDATCM0012－2021（完成时间：20219月2021-12-242022-03-24发布T/GDATCMT/GDATCM目前 引 范 附录 附录 T/GDATCMT/GDATCM前T/GDATCMT/GDATCM引甘草煮散饮片来源于豆科植物甘草GlycyrrhizauralensisFisch.干燥根和根茎炮制成饮片后的加工T/GDATCMT/GDATCM【制法】取甘草（厚片），制成粒度为0.8～10.0mm【鉴别】（1）本品粉末淡棕黄色。纤维成束，直径8～14μm，壁厚，微木化，周围薄壁细胞含T/GDATCMT/GDATCM1g40ml130ml，加热35ml使溶解，作为供试品溶液。另取甘草对照1g1ml2mg的溶液，作为对照品溶液。照薄层色谱法（20200502）1～2μl，分别1%氢氧化钠溶液制备的硅胶G薄层板上，以乙酸乙酯-甲酸-冰醋酸-水（15∶1∶1∶2）为10%105℃（365nm）DNADNA溶液，置-20℃DNA溶液，置-20℃保存备用。反应3′2×TaqPCRMix12.5μl，通用引物（2.5μmol/l）1μl，模板DNA2μl8.5μl。将离心管置PCR仪，ITS2PCR反应参数：94℃540次（94℃30秒，56℃30秒，72℃45秒（72℃）10分钟。另取无菌超纯水，同法上述PCR反应操作，作为空0541核酸凝胶染色剂GelRedPCR5μl，DNA分子2μl（0.5μg/μl500bpDNA回收试剂盒进行纯化。以PCRDNA测序仪对目的条带进行测序。DNA条形码序列对原始测序结果应用有序列拼接功能的专业软件进行序列拼接，去除引PCR扩增正向引物方向一致。GuralensisITS2ITS2223bp10G-A变异，16位点T-CG.uralensisITS2标准序列特征如下（1，扫描二维码可读取核酸序列：T/GDATCMT/GDATCM1ITS2【检查】粒度照粒度检查法（A）10.0mm筛（GB/T6003.12012R20）2.0%5.0%。水分不得过10.0%（中国药典2020年版通则0832第二法）。总灰分不得过5.0%（20202302（20202321原子吸收分光光0.2mg/kg20mg/kg。【含量测定】照高效液相色谱法（20200512）A0.05%磷5000。70%乙醇分别制成每1ml20µg0.2mg的溶液，即得（甘草酸重量=甘草酸铵重量/1.0207供试品溶液的制备取本品粉末（过三号筛）0.2g70%70%乙醇补足减失的重量，摇匀，滤过，取续滤液，即得。10µl【性味与归经】【功能与主治】2~10g【注意】【贮藏】T/GDATCMT/GDATCM附录100g，精密称定，照粒度及粒度分布测定法（202009822.0%（12.0mm5.0%。T/GDATCMT/GDATCM附录别名美草、蜜甘、蜜草、蕗草、国老所宗【来源2020GlycyrrhizauralensisFisch.、胀果甘GlycyrrhizainflataBat.GlycyrrhizaglabraL.的干燥根和根茎。本文甘草为豆科植物甘草GlycyrrhizauralensisFisch.的干燥根和根茎。【原植物1～3cm，外皮褐色，里面淡黄色，具甜味。茎30～120cm5～20cm；托5mm2mm，两面密被白色短柔毛；叶柄密被褐色腺点和短柔毛；总状花7～14mm，密被黄色腺点及短柔毛，基部偏斜3～11，暗绿色，圆形或肾形，36～87～10月[5]。【采收加工】秋季采挖，除去芦头，茎基，枝叉须根，截成适当长短的段晒至半干，打成小捆，【制法】800ml100ml，蒸干，得干浸膏，称重，计算出膏率。T/GDATCMT/GDATCM1所示，不同规格煮散饮片的出膏率及醇溶率变化不大。0.8～10.0mm的饮片。1煮散饮片（3～10目煮散饮片（4～24目煮散饮片（3～10目煮散饮片（4～24目-【性状11【成分】根及根状茎含有甘草甜素（glycyrrhizin，为甘草的甜味成分，是一种三萜皂苷。甘草acid（glycyamarinliquiritinacid（licoricidin（glycyrolglycerolglycyrol（mannite（betulicacidT/GDATCMT/GDATCM2 910111213 151617

1J1001）J9009）J1900）4（J907）、供（J1900（J1000891199037）11107064121080451310901）YPAG00）T/GDATCMT/GDATCM3（3）DNA20209107DNA条形码分子鉴定（HiddenMarkovmodel，HMM）ITS2序列。本标准通过搜集多产地多批次甘草样本，进行多次平行试验，构建了可靠ITS2条形码标准序列，同时表明了方法适用性良好。2中国医学科学院药用植物研究所植物分类专家林余霖研究员鉴定。试验共获得甘草（甘草）ITS2序列91G.uralensisITS23种单倍型序列42（甘草）3（光果甘草）ITS242GlycyrrhizainflataITS2223bpGlycyrrhizaglabraITS2序列比对223bp43种基原的所有ITS2单倍型序列一起进行多序列比对分析（417、18位点可将甘草（GC）和胀果甘草、光果甘草（AA）区甘草T/GDATCMT/GDATCM51，ZY021-G.uralensis_Haplotype1CAGACCGTTGCCCGATGCCAATTGCCTCGCGATAGGTACTTTGGTTGTGCAGGGTGAATGTTGGCTTCCCGTGAGCATTG.uralensis_Haplotype2.........A.....................................................................G.uralensis_Haplotype3...............C............................................................... ...............CAA............................................................. ...............CAA.............................................................G.uralensis_Haplotype1CGGCCTCACGGTTGGCTCAAAACTGAGTCCATGGTAGGGTTTGGCATGATCGATGGTGGTTGAGTGACGCTCGAGACCAG.uralensis_Haplotype2...............................................................................G.uralensis_Haplotype3............................................................................... ............................................................................... ...............................................................................G.uralensis_Haplotype1TCATGTGTGACTCCACTGAGTTTGGGCTCTGTAACCAATAGGCGTCTTTGAACGCTCGTGATG.uralensis_Haplotype2..............................................................G.uralensis_Haplotype3.............................................................. .............................................................. ..............................................................43DNA条形码鉴定。ITS215Guralensis标准序列比对相一致（T/GDATCMT/GDATCMGlycyrrhizauralensisCAGACCGTTGCCCGATGCCAATTGCCTCGCGATAGGTACTTTGGTTGTGCAGGGTGAATGTTGGCTTCCCGTGAGCATT ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... ...............................................................................GlycyrrhizauralensisCGGCCTCACGGTTGGCTCAAAACTGAGTCCATGGTAGGGTTTGGCATGATCGATGGTGGTTGAGTGACGCTCGAGACCA ............................................................................... ............................................................................... ............................................................................... ............................................................................... ............................................................................... 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