高效液相色谱梯度洗脱法同时测定保心宁片中3种皂苷含量
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三七总皂苷中代表性皂苷含量测定方法学的建立及稳定性研究陆 燕,潘 旭(北京积水潭医院中药房,北京 100035)[摘要] 目的: 建立同时测定三七总皂苷中三七皂苷R 1、人参皂苷Rg 1和人参皂苷Rb 1三种成分的含量测定方法,并对其各种状态下的稳定性进行研究。
方法:以乙腈-水系统作为流动相,采用高效液相-梯度洗脱法,同时测定三七总皂苷中三种成分的含量,并考察方法的专属性、线性、精密度和回收率;测定四种pH 值溶液中三种皂苷成分的稳定性以及在高温、高湿和强光照射条件下,三七总皂苷粉末中三种皂苷成分的稳定性。
结果:方法学验证结果显示,在选定的梯度洗脱条件下,方法专属性良好,三种皂苷可有效分离和定量;在各自的线性范围内线性良好;精密度RSD 值均小于2%;三种皂苷的平均回收率分别为(98.98 ± 0.60)%、(98.86 ± 0.34)%和(100.19 ± 0.64)%,方法准确性良好。
稳定性研究结果显示,在溶液状态下,三种皂苷的稳定性趋势基本一致,稳定性与pH 值密切相关,在pH 1.2溶液中药物迅速降解,24 h 含量即降低了约80%;而在pH 4.5溶液、pH 6.8溶液以及水中,三种成分的稳定性均良好,24 h 内含量未发生明显变化;而在三七总皂苷粉末中,三种代表性皂苷对湿、热和光照均不敏感,10 d 内含量基本保持稳定;但是在高湿条件下,粉末有一定的吸湿性。
结论:所建立的液相方法准确、可靠,溶液状态下三七皂苷R 1、人参皂苷Rg 1和人参皂苷Rb 1的稳定性与pH 密切相关,随pH 的降低稳定性变差,而在固体状态下则稳定性良好。
[关键词] 三七皂苷R 1;人参皂苷Rg 1;人参皂苷Rb 1;稳定性;梯度洗脱[中图分类号] R917 [文献标识码] A [文章编号] 1672 – 8157(2014)02 – 0088 – 04Study on the methodology establishment of content determination and stability of representative saponins in Panax notoginseng saponinsLU Yan, PAN Xu (Dispensary of Traditional Chinese Medicine, Beijing Jishuitan Hospital, Beijing 100035, China )[ABSTRACT] Objective: The HPLC method for simultaneous determination of notoginsenoside R 1, ginsenoside Rg 1 andginsenoside Rb 1 in Panax notoginseng saponins (PNS) was established to study the stability of the saponins under different states.Methods: Acetonitrile-water system was used as the mobile phase, and the method specificity, linearity, precision, recovery werestudied. The stability of the three saponins in different pH solutions and solid state was evaluated. Results: The results of methodvalidation showed good specificity, effective separation and quantification, as well as good linearity of the three saponins in their linear range. The precision RSD values were less than 2%, average recovery rates were (98.98 ± 0.60)%, (98.86 ± 0.34)% and (100.19 ± 0.64)% for the three saponins respectively. The stability results showed that the stability of the three saponins was closely related to the pH value under the solution state, with rapid degradation about 80% in 24 h in pH 1.2 solutions and little degradation in the other test medium. In the solid powder state, the stability of saponins was good under the experimental high temperature, high humidity and highlight conditions, while a little moisture absorption was found in the high humidity conditions. Conclusion: The HPLC method was reliable. Stability of three saponins was closely related to the pH value of solution, while they were stable under solid state.[KEY WORDS] Notoginsenoside R 1; Ginsenoside Rg 1; Ginsenoside Rb 1; Stability; Gradient elution[作者简介] 陆燕,女,主管药师,主要从事临床药学工作。
反相高效液相色谱法同时测定重楼药材中4种皂苷的含量【摘要】目的成立RP HPLC法测定重楼中4种甾体皂苷的含量。
方式采纳色谱柱:Zorbax eclipse XDB C8,(150 mm× mm,5 μm);流动相:乙腈水(40∶60),流速 ml·min-1;检测波长:203 nm;柱温:40℃。
结果4种甾体皂苷范围在~μg线性良好,R= 6~ 9,回收率%~% 。
不同来源的重楼药材中4种甾体皂苷的含量不同较大。
结论该方式准确、简捷,为重楼药材的质量操纵提供了必然方式和依据。
【关键词】重楼;甾体皂苷;反相高效液相色谱法Abstract:ObjectiveA comprehensive analytical method based on high performance liquid chromatography-ultraviolet detection has been established for the simultaneous determination of four steroidal saponins in Paris column was Symmetry C8,mobile phase was acetonitrile-water (40∶60),UV detection was performed at calibration curves showed good linearity over the range of ~ g, R=~,the recoveries were %~% for the four steroidal saponins. The results showed that the contents of the four steroidal saponins in Paris polyphylla were different with different sources . ConclusionThe method issimple with good reproducibility and selectivity and high sensitivity ,which provides a method for quality control of Paris polyphylla.Key words:Paris polyphylla; Steroidal saponins; RP HPLC重楼Paris polyphylla来源于百合科(Liliaceae)重楼属植物的干燥根茎,又名蚤休、七叶一支花等。
RP-HPLC法同时测定保心宁胶囊中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的含量张婷;魏尊喜【期刊名称】《安徽医药》【年(卷),期】2010(014)005【摘要】目的采用HPLC梯度洗脱法同时测定保心宁胶囊中三七皂苷R1、人参皂苷Rg1 、人参皂苷Rb1含量.方法采用ODS-C18柱,流动相为乙腈(A)-水(B)(0~12 min,A:19%;12~60 min,19%~36%;60~70 min,36%);检测波长为203 nm;流速:1.0 ml·min-1.结果三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的线性范围分别为0.764~6.365 μg(r=0.9999)、0.633~5.275 μg(r=0.9999)、0.699~5.825 μg(r=0.9999),平均加样回收率分别为99.88%(RSD=0.55%)、99.97%(RSD=0.23%)、100.04%(RSD=0.15%).结论该方法易行、准确可靠,可用于保心宁胶囊的质量控制.【总页数】3页(P531-533)【作者】张婷;魏尊喜【作者单位】安徽省淮北市食品药品检验所,安徽,淮北,235000;安徽省淮北市食品药品检验所,安徽,淮北,235000【正文语种】中文【相关文献】1.RP-HPLC法测定血塞通胶囊中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的含量 [J], 邹丽;雷灼雨2.RP-HPLC梯度洗脱法同时测定通迪胶囊中三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1 [J], 许亚玲;廖波;周兰;罗曼3.RP-HPLC测定脑得生软胶囊中三七皂苷R1、人参皂苷Rb1及人参皂苷Rg1含量 [J], 宋恬;李桥;何再安;王光忠;刘焱文4.RP-HPLC法测定三七丹胶囊中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1的含量 [J], 辛爱玲;邢向伟5.RP-HPLC梯度洗脱法测定三七胶囊中三七皂苷R1和人参皂苷Rb1及人参皂苷Rg1的含量 [J], 陶功意;张桢因版权原因,仅展示原文概要,查看原文内容请购买。
采用HPLC法测定三七中3种皂苷的含量目的研究探讨采用高效液相色谱仪测定复三七中三种皂苷含量的临床应用价值。
方法以三七总皂苷分散片为研究对象,对其中的人参皂苷Rb1、人参皂苷Rg1、三七皂苷R1含量进行检测。
结果不同批次三七总皂苷分散片中,人参皂苷Rb1含量在17.91 mg/片~18.23 mg/片,人参皂苷Rg1含量在17.12 mg/片~17.65 mg/片,三七皂苷R1含量在4.73 mg/片~4.82 mg/片。
结论通过高效液相色谱仪(HPLC)可以同时对三七中不同皂苷的含量进行准确定量,且具有操作简单、快捷的优点,重复性好,稳定性强,可以用于三七相关药物质量控制的过程中,值得推广应用。
标签:高效液相色谱;三七;皂苷;含量三七是我国传统医学中最珍贵的药材之一,具有止血、补血、活血、抗血栓等多种功效[1]。
本文以三七总皂苷分散片为研究对象,通过高效液相色谱法对三七皂苷R1、人参皂苷Rb1、人参皂苷Rg1等三种皂苷含量进行测定,并对其具体操作方法及检测结果进行分析。
具体报告如下。
1资料与方法1.1一般资料仪器:选择日本岛津仪器生产的LC-10A高效液相色谱仪(组成部分有微体积串联双柱塞、柱温箱、紫外可见双波长检测器、超声波提取器HS 3120、中文化LC工作站)。
试剂:对照品人参皂苷Rb1(批号:110703-201420)、对照品人参皂苷Rg1 (批号:110703-201421)、对照品三七皂苷R1 (批号:110703-201423);三七总皂苷分散片(三七总皂苷含量为50 mg/片);洗脱剂乙腈(色谱纯)、甲醇(分析纯)、水(超纯水)。
其中,对照品皂苷购自中国食品药品检定研究院,三七总皂苷分散片为自制,洗脱剂购自津市科密欧化学试剂开发中心。
1.2方法分别取人参皂苷Rb1、人参皂苷Rg1、三七皂苷R1对照品进行精密称定,并使用浓度为20%的乙腈水溶液对其进行溶解,并制成1 ml的混合溶液(其中人参皂苷Rb1含量为0.5 mg、人参皂苷Rg1含量为0.4 mg、三七皂苷R1含量为0.12 m)。
紫檀素、香附烯酮和a-香附酮含量的同时测定,建立了该制剂的多指标成分质量控制模式,同时采用聚类分析、主成分分析等化学计量学手段对不同企业生产的10批复方独圣活血片质量进行了综合评价+所建立的方法操作简便、结果准确,为全面、科学地评价独圣活血片整体质量提供了参考+参考文献:国家药典委员会.中国药典!015年版:一部[S*北京:中国医药科技出版社,2015:1270-1271.徐愚聪,王曙,黄A.独圣活血片中有效成分的含量测定华西药学杂志,2005,20(4):362-363.雷波,李K.独圣活血片联合双氯芬酸钠治疗急性软组织损伤的疗效观察现代药物与临床,2018,33(11):2945-2948刘琼辉(虫圣活血片合伤科活血酊治疗软组织损伤疗效分析实用中医药杂志200723(7):415-416.陈艳辉,周昕(虫圣活血片合五子衍宗丸治疗肾虚血瘀型月经过少疗效观察实用妇科内分泌电子杂志, 20196(24):110118卢君蓉,李文兵,王世宇,等.香附醋制前后香附烯酮,圆柚酮和-香附酮的含量比较中国实验方剂学杂志, 201420(20):24-27王世宇,李文兵,卢君蓉,等.HPLC法同时测定不同产地香附药材中香附烯酮、圆柚酮和-香附酮中成药2015,37(3):588-591.陆冰,何胜利,陆飞.HPLC法测定调中四消丸中咖啡酸、芦荟大黄素、大黄酸、大黄素甲n、圆柚酮和-香附酮现代药物与临床2017,32(7):1188-1191.班小军,杨雪梅,赵翠琴•高效液相梯度洗脱法同时测定复方救必应胶囊中紫丁香昔、长梗冬青昔、圆柚酮和-香附酮含量中国中医药科技201623(4):431-433. )0*刘军民,安冉,翟明,等.鸡血藤商品药材质量评析中药新药与临床药理201223(5):573-575.)1*杨冉冉,索亚然,乔艺涵,等.HPLC同时测定鸡血藤中芒柄花素和美迪紫檀素等6个黄酮类成分)*药物分析杂志2017,37(12):21392144.)2*徐皓.HPLC法测定不同产地元胡药材中5种生物碱含量)*药物分析杂志2015,35(8):14031407.)3*陈东东,毛坤军,李祥,等.HPLC法比较延胡索炮制前后7个生物碱成分的含量)*药物分析杂志,2015,35(9):1591-1595)4*毕福钧,林彤.RP-HPLC法同时测定醋延胡索配方颗粒中7种生物碱中草药2016,47(4)=606-609.)5*李凯,原军宁,彭晶晶,等.不同规格延胡索中8种生物碱比较)*中成药2020,42(5):12441249.)6*张丹,王昌利,卜雕雕,等.高效液相色谱法同时测定延胡索中5种生物碱含量的方法学研究中南药学, 201816(12):1759-1762)7*宋洪伟,毛睿,李丽红,等.延胡索炮制前后多组分质量控制方法的研究天津中医药大学学报,2019,38(1):63-67)8*袁芮,张超,苏彤,等.香附和醋香附HPLC特征图谱及多元统计分析)*中成药2019,41(8):19911994.)9*卫昊,郭玲玲,李柳柳,等.不同海拔和光照对黄苓中7种黄酮类有效成分含量的影响)*中草药,2019,50(6):1472-1476)0*杜憬生,吴立群,马鸿雁.一测多评法测定鸡血藤中4种黄酮成分含量)*中药材2017,40(4):881883.)1*张祺嘉锂,王文静,张琳,等(向应面法优选鸡血藤总黄酮的)J*西北药学杂志201934(2):143-146(收稿日期:2020-07-22)LC-MS/MS法同时测定中药注射剂中3种成分的含量孙卫1,汪赛赛1,贾倩倩1,付佳1,李智2,韩省力1!(1.西安交通大学药学院,西安710061'.天津天士力之骄药业有限公司,天津300402)摘要:目的建立高效液相色谱-质谱联用(LC-MS/MS)法测定注射用益气复脉和生脉注射液中5-7甲基糠醛(5-HMF)、人参皂5Rd与人参皂5Rg3的含量°方法采用SHIMADZU VP-ODS色谱柱(150mmX20mm,5ym)o流动相:ImL'L1甲酸(A)乙睛(B),梯度洗脱,梯度程序:0〜6min,5%B;6〜20min,5%B〜80%B;20〜25min,80%B。
23ChinaPharmaceuticalsVol.28牞No.10牞May20牞2019第一作者:姜燕,女,大学本科,副主任中药师,主要从事药品检验工作,(电子信箱)949104094@qq.com。
·检验检测·doi:10.3969/j.issn.1006-4931.2019.10.008DrugResearch高效液相色谱法同时测定肿痛安胶囊中3种皂苷类成分含量姜燕,戴丽娜(江苏省连云港市食品药品检验检测中心,江苏连云港222006)摘要:目的建立测定肿痛安胶囊中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1含量的高效液相色谱法。
方法色谱柱为AgilentEclipseXDB-C18柱(150mm×4.6mm,5μm),流动相为乙腈-水牗梯度洗脱牘,流速为1.0mL/min,柱温为30℃,检测波长203nm,进样量为10μL。
结果三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1质量浓度线性范围分别为15.25~487.80μg/mL,11.57~370.30μg/mL,11.44~366.20μg/mL,r>0.9994牗n=5牘,精密度、重复性和稳定性试验结果的RSD均小于3%,加样回收率为99.28%~100.22%,RSD为0.98%~1.49%牗n=6牘。
结论该方法简便、快速、准确、可靠,专属性较强,可用于肿痛安胶囊的质量控制。
关键词:高效液相色谱法;肿痛安胶囊;三七皂苷R1;人参皂苷Rg1;人参皂苷Rb1;含量测定中图分类号:R932;R284.1;R286.0文献标识码:A文章编号:1006-4931牗2019牘10-0023-03SimultaneousDeterminationofThreeKindsofSaponinsinZhongtong′anCapsulesbyHPLCJIANGYan,DAILina(LianyungangFoodandDrugInspectionCenter,Lianyungang,Jiangsu,China222006)Abstract:ObjectiveToestablishanHPLCmethodforthecontentdeterminationofnotoginsenosideR1,ginsenosideRg1,ginsenosideRb1inZhongtong′anCapsules.MethodsThechromatographiccolumnwasAgilentEclipseXDB-C18column(150mm×4.6mm,5μm),themobilephasewasacetonitrile-water(gradientelution),theflowratewas1.0mL/min,thecolumntemperaturewas30℃,thedetectionwavelengthwas203nmandthesamplesizewas10μL.ResultsThenotoginsenosideR1,ginsenosideRg1,ginsenosideRb1showedgoodlinearityintherangesof15.25-487.80μg/mL,11.57-370.30μg/mL,11.44-366.20μg/mL,r>0.9994(n=5).TheRSDsoftheprecision,repeatabilityandstabilitytestswerelessthan3%,theaveragerecoveryrateswereintherangeof99.28%-100.22%,RSDswereintherangeof0.98%-1.94%(n=6).ConclusionThemethodissimple,rapid,accurateandreliable,andhasstrongspecificity.ItcanbeusedforthequalitycontrolofZhongtong′anCapsules.Keywords:HPLC;Zhongtong′anCapsules;notoginsenosideR1;ginsenosideRg1;ginsenosideRb1;contentdetermination表1流动相梯度洗脱程序(%)t牗min牘12.0040.0090.00A81.078.566.0B19.021.534.0t牗min牘90.50105.00A81.081.0B19.019.0肿痛安胶囊由三七、天麻、僵蚕、白附子(制)、防风、羌活、天南星(制)、白芷8味中药组方,有祛风化痰、行瘀散结、消肿定痛功效,临床用于治疗风痰瘀阻引起的牙痛、咽喉肿痛、口腔溃疡、痹病(症见关节肿胀疼痛、筋脉拘挛、屈伸不利),以及破伤风的辅助治疗。
高效液相色谱法测定细胞内三磷酸腺苷及其代谢物的含量2017年1月January 2017Chinese Journal of Chromatography Special issue for commemorating Professor Z O U Hanfa ( H )? A rticle Vol.35 No.154?58 D O I: 10.3724/SP.J. 1123.2016.08031Quantification of intracellular adenosine 5'-triphosphateand its metabolites by high performance liquidchromatography analysisZHU Huiyu, WU Danni, WANG Hailin**(S tate K ey Laboratory o f E n viro n m en ta l C hem istry and E cotoxicology, Research Center fo rE co-E nvironm ental S c ie n ce s, C hinese A ca d em y o f S c ie n ce s, B e ijin g, 100085, C hina)Abstract:This study was aimed to provide insight regarding the intracellular metabolites of adenosine 5'- triphosphate (A TP) and whether 2-tert-butyl- 1,4-benzoquinone (TBBQ) affects cell metabolites. A rapid high performance liquid chromatography (H PLC) protocol was developed for the separation and quantitation of A T P and its metabolites (adenosine diphosphate (A D P) and adenosine monophosphate (AMP)) in cells. Chromato-graphic separation was performed using a Shim adzu H PL C system equipped with an Agela Venusil M P C18 column; isocratic elution was adopted. The mobile phase comprised solvent A (50 mmol/L disodium hydrogen phosphate and 15 mmol/L trimethylamine (TEA); pH adjusted to 7. 88 using acetic acid (HAc)) and solvent B (methanol). The correlation coefficients of the three analytes were very high (R2^0. 999 6), and the contents of the three metabolites in the M R C-5 cells were within the linear ranges (0.1-100 pmol/L). The lim its of detection for the detected three compounds were low. Sam ples were extracted from cells (after exposure and non-exposure to quinones) using 80%(v/v) methanol aqueous solution. The method developed in this study was successfully applied to detect ATP, A D P and A M P in M R C-5 cells, and the results demonstrated that A TP, ADP, A M P levels in cells were affected by TBBQ, but the relations between the concentration of T B B Q and the level of ATP, A D P and A M P were com plex.Key words:high performance liquid chromatography (HPLC); adenosine 5’-triphosphate (ATP); quinones;metabolites; intracellularCLC number:O658 Document code:A Article IC:1000-8713(2017)01-0054-05高效液相色谱法测定细胞内三磷酸腺苷及其代谢物的含量朱会宇,吴丹妮,汪海林*(中国科学院生态环境研究中心,环境化学与生态毒理学国家重点实验室,北京100085)摘要:研究了三磷酸腺苷(ATP)及其代谢物在细胞内的含量以及2-叔丁基-1,4-苯醌(TBBQ)对ATP及其代谢产物在细胞内含量的影响。