SFC和UPLC方法用于丹参饮片中多成分研究

目的：建立SFC-PDA法同时检测丹参饮片中4个脂溶性丹参酮类成分，以及UPLC法同时测定丹参饮片中3个水溶性丹酚酸类成分，以7个成分的测定结果综合评价丹参饮片的质量。方法：采用SFC-PDA法，使用Viridis^? HSS SB C₁₈ 100A（3.0 mm×100 mm，1.8 μm）色谱柱，以二氧化碳（A）-［正庚烷-异丙醇（8：2）（B）］为流动相，梯度洗脱，流速1.5 mL·min^-1，柱温45℃，背压13.8 MPa；采用UPLC-PDA法，使用BEH Shield RP（2.1 mm×100 mm，1.7 μm）色谱柱，以乙腈（A）-0.1%磷酸溶液（B）为流动相，梯度洗脱，流速0.4 mL·min^-1，柱温30℃。结果：在建立的色谱条件下，丹参饮片中7个主要成分在各自的液相系统中分离完全，线性良好，精密度、重复性、稳定性及回收率实验结果均符合含量测定要求，同时还比较了用HPLC和SFC 2种方法测定脂溶性成分时的仪器灵敏度，结果为一个数量级。结论：SFC方法快速简便、结果准确，在节省溶剂、减少环境污染的方面具有独特优势，同时采用UPLC方法测定水溶性成分，2种方法可用于丹参饮片的质量分析，同时大大降低检测成本。

Objective: To evaluate the quality of Salvia Miltiorrhiza decoction pieces from difference sources,methods for determining 7 components were established.Among which,lipid-soluble tanshinone components were determined by(SFC-PDA),and 3 water-soluble salvianolic acid components were determined by UPLC. Methods: The SFC-PDA analysis was performed on Viridis^? HSS SB C₁₈ 100A(3.0 mm×100 mm,1.8 μm)column.The mobile phase was Carbon dioxide-n-heptane-isopropanol(8:2)with isocratic elution mode at a flow rate of 1.5 mL·min^-1.The column temperature was controlled at 45℃ and the ABPR pressure was 13.8 MPa.The UPLC-PDA analysis was performed on BEH Shield RP(2.1 mm×100 mm,1.7 μm)column,The mobile phase consisted of acetonitrile-0.1% phosphoric acid with gradient elution at a flow rate of 0.4 mL·min^-1.The column temperature was controlled at 30℃.Results: 7 main components in Salvia Miltiorrhiza decoction pieces were well separated by SFC and UPLC respectively.Results of linearity,repeatability,stability and recovery met the accept criteria of assay.The sensitivity of fat-soluble components and water-soluble components were comparable in HPLC and SFC system.Conclusion: The SFC method rapid,simple and accurate,which possesses the advantage of solvent saving and environmental friendly.At the same time,the UPLC method is used to determine the water-soluble components.The two methods can be used for analysis the quality of Salvia Miltiorrhiza decoction pieces,and reduce the detection cost remarkably.

[1] 中华人民共和国药典2015年版.一部[S].2015:76 ChP 2015.Vol Ⅰ[S].2015:76
[2] 高佳明,吕明,解微微,等.中医药心脑血管疾病同治的方剂用药规律分析[J].中国中药杂志,2019,44(1):193 GAO JM,L M,XIE WW,et al.Analysis of Chinese medicine prescriptions for common or differentially treating cardiovascular and cerebrovascular diseases[J].China J Chin Mater,2019,44(1):193
[3] 任雨贺,田静,刘淑莹,等.2015年版《中国药典》(一部)含丹参成方制剂的归类分析[J].中成药,2018,40(12):2770 REN YH,TIAN J,LIU SY,et al.Classification and analysis of formulations containing Salvia Miltiorrhiza in Chinese Pharmacopoeia(part Ⅰ)2015 edition[J].Chin Tradit Pat Med,2018,40(12):2770
[4] 李东,张纲,宗梁,等.中国药典丹参质量标准与美、欧、日药典对比分析[J].亚太传统医药,2018,14(2):3 LI D,ZhANG G,ZONG L,et al.Contrastive analysis on quality standards of Salvia Miltiorrhiza Radix Et Rhizoma in Chinese Pharmacopoeia and Pharmacopoeia of the United States,Europe,and Japan[J].Asia-Pacific Tradit Chin Med,2018,14(2):3
[5] 周军,张茉,王杰.HPLC法测定丹参胶囊中丹参素、原儿茶醛、迷迭香酸及丹酚酸B的含量[J].天津药学,2018,14(2):3 ZHOU J,ZHANG M,WANG J.Determination of the contents of tanshinol,protocatechualdehyde,rosmarinic acid and salvianolic acid B in Danshen capsules by HPLC[J].Tianjin Pharm,2018,14(2):3
[6] 张琳琳,王淳,宋志前,等.HPLC同时测定白花丹参中丹参酮ⅡA、隐丹参酮、丹参酮Ⅰ和二氢丹参酮Ⅰ的含量[J].中国实验方剂学杂志,2015,21(6):62 ZHANG LL,WANG C,SONG ZQ,et al.Simultaneous determination of tanshinone ⅡA,tanshinone Ⅰ,cryptotanshinone and dihydrotanshinoneⅠ in Salviae Albae Radix by HPLC[J].Chin J Exp Tradit Med Form,2015,21(6):62
[7] 张玉,戚莹雪,王蕾,等.山东丹参脂溶性成分的地域分布特点研究[J].中药材,2018,41(11):2271 ZHANG Y,QI XY,WANG L,et al.Geographical distribution of liposoluble constituents of Salvia Miltiorrhiza in Shandong Province[J].J Chin Med Mater,2018,41(11):2271
[8] 曾慧婷,沙秀秀,宿树兰,等.不同产地丹参茎叶UPLC指纹图谱与化学模式识别研究[J].中草药,2017,48(4):767 ZENG HT,SHA XX,SU SL,et al.UPLC fingerprint and chemical pattern recognition of Salvia Miltiorrhiza stems and leaves from different habitats[J].Chin Tradit Herb Drugs,2017,48(4):767
[9] 赵国欣,刘萌,李领川,等.超高效液相色谱-质谱联用同时测定丹参中7种成分含量[J].中国实验方剂学杂志,2017,23(1):60 ZHAO GX,LIU M,LI LC,et al.Simultaneous determination of 7 components in Salviae Miltiorrhizae Radix et Rhizoma by UPLC-MS/MS[J].Chin J Exp Tradit Med Form,2017,23(1):60
[10] 张立国,胡甜甜,张芳芳,等.丹参和紫丹参脂溶性成分的UPLC及LC-MS/MS分析[J].中国中药杂志,2019,44(6):1208 ZHANG GL,HU TT,ZHANG FF,et al.Analysis of lipophilic components of Salvia Miltiorrhiza Roots and S.yunnanensis roots by UPLC and LC-MS/MS[J].China J Chin Mater,2019,44(6):1208
[11] 翟宏宇,王海洋,单柏宇,等.丹参药材HPLC特征图谱研究及其活性成分的含量测定[J].中国卫生工程学,2018,17(2):207 ZAI HY,WANG HY,ShAN BY,et al.Studies on the specific chromatogram of Salvia Miltiorrhiza and simultaneous determination of its six active components[J].Chin J Pub Heal Eng,2018,17(2):207
[12] 刘涛,赵群,杨丹,等.一测多评法测定丹参酚酸类提取物中4个成分含量[J].成都大学学报-自然科学版,2017,36(1):7 LIU T,ZHAO Q,YANG D,et al.Determination of 4 components'content in salvianolic acid extracts by QAMS[J].J Chendu Univ(Nat Sci Ed),2017,36(1):7
[13] 翟宏宇,单柏宇,王海洋,等.一测多评法测定保心宁片中6个丹参类指标性成分的含量[J].药物分析杂志,2018,38(6):973 ZAI HY,SHAN BY,,WANG HY,et al.Simultaneous determination of six index components in Baoxinning tablets by a single marker[J].Chin J Pharm Anal,2018,38(6):973
[14] 梁从莲,张永清,李佳,等.不同株系丹参有效成分比较研究[J].山东科学,2018,31(4):31 LIANG C L,ZHANG YQ,LI J,et al.Comparative study on the active constituents of Salvia miltiorrhiza in different strains[J].Shandong Sci,2018,31(4):31
[15] 翁德会,许腊英,李佩,等.丹参饮片的外观质量与分级研究[J].亚太传统医药,2016,12(13):40 WENG DH,XU LY,LI P,et al.Exploration of the relationships of Salvia Miltiorrhiza slice in diameter,origin,color,texture between quality[J].Asia-Pacific Tradit Chin Med,2016,12(13):40
[16] 刁家葳,徐保鑫,张学兰,等.丹参药材及其饮片的规格等级与质量评价研究进展[J].山东中医杂志,2018,37(8):688 DIAO JW,XU BX,ZANG XL,et al.Research progress of specification,grade and quality evaluation of original Danshen(Salviae Miltiorrhizae)and processed Danshen(Salviae Miltiorrhizae)[J].Shandong J Tradit Chin Med,2018,37(8):688
[17] 中华人民共和国药典2015年版.四部[S].2015:64 ChP 2015.Vol Ⅳ[S].2015:64
[18] 张坤,刘家良,崔英德,等.丹参酮ⅡA的CO₂超临界流体萃取工艺研究[J].华南理工大学学报(自然科学版),1998,26(8):91 ZHANG K,LIU JL,CUI YD,et al.A study on the tanshinone ⅡA extraction with CO₂ Supercritical fluid[J].J South China Univ Technol(Nat Sci),1998,26(8):91
[19] 李霞,唐玉海,赵新锋,等.CO₂超临界流体萃取丹参中的有效成分[J].西安交通大学学报(医学版),2004,25(6):614 LI X,TANG YH,ZHAO XF,et al.An active components in the extracts of Radix Salvia Miltiorrhizae by supercritical carbon dioxide fluid[J].J Xi'an Jiaotong Univ(Med Sci),2004,25(6):614