HPLC-DAD法同时测定金银花中10个酚酸类成分的含量

建立HPLC-DAD法同时测定金银花中10个酚酸 (咖啡酸、绿原酸、新绿原酸、隐绿原酸、3,5-O-双咖啡酰基奎宁酸、3,4-O-双咖啡酰基奎宁酸、4,5-O-双咖啡酰基奎宁酸、3,5-O-双咖啡酰基奎宁酸甲酯、3,4-O-双咖啡酰基奎宁酸甲酯、4,5-O-双咖啡酰基奎宁酸甲酯)的含量。采用100倍量50%乙醇超声30分钟提取制备样品,选择C18色谱柱,以乙腈– 甲醇–0.1%甲酸水溶液为流动相进行梯度洗脱, 55分钟内完成10个酚酸类化合物的色谱分离 10个酚酸类成分在测试范围内线性关系良好, r2>0.999, 其平均加样回收率均在98.57%–103.22%之间, RSD均小于3%。该方法准确, 灵敏度高, 重复性好, 可用于金银花药材中10个酚酸类成分的含量测定。     

HPLC法同时测定复方双花片中10个成分的含量

目的:采用高效液相色谱(HPLC)技术,建立同时测定复方双花片中新绿原酸、绿原酸、隐绿原酸、连翘酯苷A、3,4-二-O-咖啡酰奎宁酸、3,5-二-O-咖啡酰奎宁酸、4,5-二-O-咖啡酰奎宁酸、连翘苷、穿心莲内酯和脱水穿心莲内酯10个成分含量的方法。方法:复方双花片样品用50%甲醇水超声提取,采用Agilent Zorbax SB-C₁₈(250 mm×4.6 mm, 5μm)色谱柱分析,流动相为乙腈-0.15%磷酸水溶液,梯度洗脱,流速1.0 mL·min^-1,柱温30℃,进样量10μL,检测波长327 nm(检测新绿原酸、绿原酸、隐绿原酸、连翘酯苷A、3,4-二-O-咖啡酰奎宁酸、3,5-二-O-咖啡酰奎宁酸、4,5-二-O-咖啡酰奎宁酸)和226 nm(检测连翘苷、穿心莲内酯、脱水穿心莲内酯)。结果:新绿原酸、绿原酸、隐绿原酸、连翘酯苷A、3,4-二-O-咖啡酰奎宁酸、3,5-二-O-咖啡酰奎宁酸、4,5-二-O-咖啡酰奎宁酸、连翘苷、穿心莲内酯和脱水穿心莲内酯质量浓度分别在4.071～40.71μg·mL^-1(r...     

莲菊感冒胶囊制备工艺研究

目的:优选莲菊感冒胶囊的制备工艺。方法:分别采用水煎煮、乙醇回流、水煮醇沉、水煮醇沉+大孔吸附树脂纯化、水煮醇沉羊耳菊+醇提水沉穿心莲+大孔吸附树脂纯化5种方法制备莲菊感冒胶囊样品;以3,5-O-二咖啡酰基奎宁酸和穿心莲总内酯提取率、总固体物提取率以及各样品的抗菌、解热作用为评价指标,优选制备工艺。结果:采用水煮醇沉羊耳菊+醇提水沉穿心莲+大孔吸附树脂纯化的方法制备的样品各项指标较好,3,5-O-二咖啡酰基奎宁酸和穿心莲总内酯提取率均达70%以上,总固体物提取率最低,且具有明显的抗菌和解热作用。结论:所选工艺合理、可行,可最大限度富集有效成分。     

胡蔓藤中非生物碱类成分的分离与鉴定(Ⅲ)

目的研究胡蔓藤(Gelsemium elegans Benth.)中的化学成分。方法采用硅胶柱色谱和ODS柱色谱进行分离,Sephadex LH-20及制备液相进行纯化,根据理化性质和光谱分析进行结构鉴定。结果分离鉴定了8个化合物,分别为(+)-8-hydroxypinoresinol(1)、cleomiscosin C(2)、cleomiscosinA(3)、3,4-二羟基苯甲醛(3,4-dihydroxyphenyl aldehyde,4)、咖啡酸(caffeic acid,5)、1-O-咖啡酰基奎宁酸(1-O-caffeoylquinic acid,6)、4-O-咖啡酰基奎宁酸(4-O-caffeoylquinic acid,7)、1-O-咖啡酰基奎宁酸甲酯(1-O-caffeoylquinic acid methyl ester,8)。结论化合物1-8为首次从该属植物中分离得到。     

滨蒿提取物中3种二咖啡酰奎宁酸的含量测定

目的建立滨蒿提取物中3,4-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸和4,5-二咖啡酰奎宁酸的含量测定方法。方法采用HPLC法测定,色谱条件:Shim-pack VP-ODS色谱柱(250mm×4.6mm,5μm);流动相:乙腈-0.036mol·L~(-1)磷酸二元梯度洗脱;流速:1.0mL·min~(-1);柱温:30℃,检测波长:327nm。结果 3,4-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸和4,5-二咖啡酰奎宁酸分别在0.010 3~0.309,0.009 7~0.291和0.010 5~0.315mg·mL~(-1)范围内与峰面积呈良好的线性关系(r=0.999 9);加样回收率分别为99.23%,101.30%和100.63%;RSD值分别为1.59%,0.83%和1.23%(n=6);样品溶液在32h内稳定。结论该方法操作简便,重复性好,可用于滨蒿提取物的质量控制。     

大孔吸附树脂分离精制莲菊感冒胶囊工艺

目的:探索用大孔吸附树脂分离精制莲菊感冒胶囊制备工艺。方法:以脱水穿心莲内酯、穿心莲内酯、35,-O-二咖啡酰基-奎宁酸为考察指标对6种大孔树脂进行筛选,并采用正交试验对选定的大孔树脂分离精制工艺条件进行优化,最后再对解吸附条件等进行考察。结果:选择D101大孔吸附树脂分离精制效果较好,上样吸附流速2 BV.h-1(2倍柱体积/h),依次用1BV水和3 BV的70%乙醇洗脱,洗脱流速为2 BV.h-1。结论:D101型大孔树脂适用于莲菊感冒胶囊分离精制工艺,脱水穿心莲内酯、穿心莲内酯3、5,-O-二咖啡酰基-奎宁酸的转移率均大于90%,总固体物减少60%以上,达到去除杂质、富集有效成分的目的。     

HPLC法同时测定银黄含片中6个咖啡酰奎宁酸类成分的含量

目的:建立同时测定银黄含片中6个咖啡酰奎宁酸类成分的HPLC方法。方法:采用Inertsil ODS(4.6 mm×250 mm,5μm)色谱柱,以乙腈(A)-0.4%磷酸溶液(B)为流动相,梯度洗脱(0～15 min,5%A→20%A;15～30 min,20%A→30%A;30～60min,30%A),流速1.0 mL.min-1,检测波长为327 nm。结果:3-O-咖啡酰奎宁酸、绿原酸(5-O-咖啡酰奎宁酸)、4-O-咖啡酰奎宁酸、4,5-O-二咖啡酰奎宁酸、3,5-O-二咖啡酰奎宁酸、3,4-O-二咖啡酰奎宁酸进样量分别在0.04～0.69,0.04～0.67,0.04～0.68,0.02～0.33,0.01～0.15,0.04～0.70μg范围内与峰面积呈良好的线性关系;平均回收率(n=6)分别为95.0%,99.1%,98.9%,98.9%,96.2%,103.5%。结论:本法操作简便,结果准确,灵敏度高,可用于银黄含片中多指标成分的质量控制。     

UPLC同时测定百灵安神片中七种活性成分含量

目的 建立UPLC同时测定百灵安神片中绿原酸、芒果苷、二苯乙烯苷、斯皮诺素、木犀草苷、槲皮苷、3,5-二咖啡酰奎宁酸含量的方法。方法 采用Waters Acquity UPLC BEH C18（2.1 mm×100 mm,1.7 μm）色谱柱;流动相：乙腈-0.1%磷酸水,梯度洗脱;流速：0.4 mL·min^-1;检测波长：340 nm。结果 绿原酸、芒果苷、二苯乙烯苷、斯皮诺素、木犀草苷、槲皮苷、3,5-二咖啡酰奎宁酸在检测范围内线性良好,r均〉0.999 8,方法回收率分别96.3%,98.2%,100.5%,99.3%,97.7%,97.0%,96.4%。结论 本方法准确可靠快捷,为全面控制百灵安神片质量提供了一种测定方法。     

怀菊花多成分含量测定及指纹图谱的模式识别研究

目的 建立怀菊花中多种活性成分含量的HPLC检测方法,构建怀菊花的指纹图谱识别模式。方法 采用HPLC检测10批怀菊花样品绿原酸、3,4-二咖啡酰基奎宁酸、3,5-二咖啡酰基奎宁酸、4,5-二咖啡酰基奎宁酸和木犀草素等化合物的含量,建立指纹图谱。运用主成分分析及聚类分析对指纹图谱进行模式识别研究,并采用相似度评价软件进行相似度评价。色谱柱为SWELL Chromplus C₁₈ (250 mm×4.6 mm,5 μm);流动相为乙腈-0.3%甲酸水溶液,梯度洗脱,流速为0.8?mL·min^-1;进样量：10 μL;检测波长：350 nm;柱温：25?℃。结果 绿原酸的含量为0.04%~0.27%,3,4-二咖啡酰奎尼酸的含量为0.01%~0.21%,3,5-二咖啡酰奎尼酸的含量为0.45%~1.09%,4,5-二咖啡酰奎尼酸的含量为0.32%~0.96%,木犀草素的含量为0.02%~0.18%。怀菊花的指纹图谱确定了15个共有峰,根据前3个主成分将10批样品分为5类。结论 本方法简便、准确、重现性好,为怀菊花多成分的含量测定提供参考方法,并为怀菊花的质量控制提供科学依据。     

羊耳菊药材中4, 5-O-二咖啡酰基奎宁酸的含量测定

目的：建立羊耳菊药材中4, 5-O-二咖啡酰基奎宁酸的含量测定方法,为其质量控制提供科学依据。方法：采用HPLC直接测定,色谱柱Eclipse XDB C18柱,保护柱为SecurityGuard C18柱;流动相甲醇-0.1%磷酸 (28∶72);检测波长为325 nm。结果：测定了13批不同产地市售的羊耳菊药材,4,5-O-二咖啡酰基奎宁酸的含量在0.12%～0.55%之间。结论：该法方便、快速、准确、重复性好,可以控制该药材的质量。     

A rapid method for simultaneous determination of 15 flavonoids in Epimedium using pressurized liquid extraction and ultra-performance liquid chromatography

Herba Epimedii (family Berberidaceae), Yinyanghuo in Chinese, is one of commonly used Chinese medicines. Flavonoids are considered as its active components. In this study, a rapid ultra-performance liquid chromatography (UPLC) method was developed for simultaneous determination of 15 flavonoids, including hexandraside E, kaempferol-3-O-rhamnoside, hexandraside F, epimedin A, epimedin B, epimedin C, icariin, epimedoside C, baohuoside II, caohuoside C, baohuoside VII, sagittatoside A, sagittatoside B, 2'-O-rhamnosyl icariside II and baohuoside I in different species of Epimedium. The analysis was performed on Waters Acquity UPLC system with an Acquity UPLC BEH C18 column (50 mm x 2.1mm I.D., 1.7 microm) and gradient elution of 50mM acetic acid aqueous solution and acetonitrile within 12 min. All calibration curves showed good linearity (R2>0.9997) within test ranges. The LOD and LOQ were lower than 0.13 and 0.52 ng on column, respectively. The R.S.D.s for intra- and inter-day of 15 analytes were less than 5.0% at three levels, and the recoveries were 95.0-103.7%. The validated method was successfully applied to quantitatively analyze 15 flavonoids in different species of Epimedium. The results showed there were great variations among the contents of investigated flavonoids. Hierarchical clustering analysis based on characteristics of 15 investigated compounds peaks in UPLC profiles showed that 37 samples were divided into 3 main clusters, which were in accordance with their flavonoids contents. The simulative mean chromatogram of the high content cluster was generated to compare the samples from different species and/or locations of Epimedium. Four flavonoids including epimedin A, B, C and icariin were selected as markers for quality control of the species of Epimedium used as Yinyanghuo.     

UPLC法同时测定黔产隔山消药材中3个C21甾体酯苷的含量

目的:建立同时测定民族药材隔山消中3个C21甾体酯苷(白首乌新苷F、G和H)的UPLC分析法。方法:样品经甲醇超声提取后,用UPLC-PDA进行分析检测;采用Waters Acquity UPLC系统,ACQUITY UPLC BEH C18(2.1 mm×100 mm,1.7μm)色谱柱,流动相为乙腈-水,梯度洗脱,流速为0.4 mL.min-1,检测波长220 nm,柱温30℃,进样量2μL。结果:白首乌新苷F、G和H 3个C21甾体酯苷在检测范围内均呈良好线性(r≥0.9995);平均加样回收率(n=3)在98.2%～104.1%之间,RSD均不大于1.1%。结论:UPLC分离效果及重复性好,且快速、简便,可作为隔山消药材质量控制的参考方法。     

A rapid method for chemical fingerprint analysis of Hoodia species, related genera, and dietary supplements using UPLC-UV-MS

Recently, ultra-performance liquid chromatography (UPLC) has proven to be one of the most promising developments in the area of high-speed chromatographic separations with increased sensitivity and resolution. In this work, a reverse phase chromatographic method was developed using UPLC for the chemical fingerprint analysis of 12 hoodigosides, related genera and dietary supplements. The method is also used for the quantification of P57 in Hoodia species and dietary supplements that claim to contain Hoodia. The analysis was performed on a Waters Acquity UPLC system with an Acquity UPLC BEH C18 column (100 mm x 2.1mm I.D., 1.7 microm) and a gradient elution of water and acetonitrile, both containing 0.05% formic acid with a run time of 15 min. The calibration curve of P57 showed good linearity (r(2)>0.999) within the established range (1-100 microg/mL). The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.3 and 0.9 microg/mL, respectively. The RSD for intra- and inter-day were less than 3.0%, and the recovery efficiency as 97-103%. LC-mass spectrometry coupled with electrospray ionization (ESI) interface method is described for the identification of P57. The developed method was successfully applied to the identification of 12 oxypregnane glycosides in four different species of Hoodia, 23 related genera and 35 dietary supplements that claim to contain H. gordonii. The UPLC profiles of various plant samples were compared for the presence of oxypregnane glycosides. Different sample matrices were successfully analyzed, providing the wide range of applicability of this method, including gels, capsules, tablets, sprays, tea bags, snack bars, powders and juices.     

超高效液相色谱法同时测定丹蛭降糖胶囊中丹皮酚和芍药苷含量

目的:建立一种超高效液相色谱法同时测定丹蛭降糖胶囊中丹皮酚、芍药苷的含量。方法:色谱柱为Waters Acquity UPLC BEH C18(50 mm×2.1 mm,1.7μm)柱,以乙腈-0.1%磷酸为流动相进行梯度洗脱,流速为0.25 mL.min-1,柱温为30℃,检测波长为230 nm。结果:丹皮酚和芍药苷的线性范围分别为0.653～1.959μg(r=0.999 7)和0.108～0.324μg(r=0.999 9),平均加样回收率分别为99.46%,RSD=0.46%(n=6)和99.19%,RSD=0.49%(n=6)。结论:新建方法简便、快速、准确、易行,可用于控制制剂质量。     

A rapid method for the simultaneous determination of 11 saponins in Panax notoginseng using ultra performance liquid chromatography

A rapid ultra performance liquid chromatography coupled with photo diode array detection method (UPLC-PDA) was developed for the simultaneous determination of 11 saponins, namely notoginsenoside R1, ginsenoside Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, Rb3, Rd and Rg3 in Panax notoginseng. The analysis was performed on Acquity UPLC system with Acquity UPLC BEH C(18) column and gradient elution of water and acetonitrile in 12 min. The high correlation coefficient (r(2)>0.9968) values indicated good correlations between the investigated compounds' concentrations and their peak areas within the test ranges. The LOQ and LOD were lower to 0.2-2.4 and 0.1-1.8 ng on column, respectively. The overall intra- and inter-day variations (R.S.D.) of 11 saponins were lower than 3.1%. The developed method was successfully used for the analysis of saponins in P. notoginseng with overall recovery of 93.0-101.6% for the analytes. The results show that UPLC is a powerful tool for analysis of components in Chinese medicines.     

超高速液相测定西洋参中人参皂苷Rg1、Re、Rb1的含量研究

目的:建立测定西洋参中人参皂苷Rg1、Re、Rb1含量的超高速液相色谱法.方法:采用Waters Acquity UPLC BEH C18(100 mm×2.1 mm,1.7 μm)柱分离,以甲醇-水为流动相进行梯度洗脱,流速为0.2ml·min-1,柱温为35℃,在203 nm处检测.结果:人参皂苷 Rg1、Re、Rb1在测定范围内有良好的线性关系,其平均回收率为97.36%～99.23%,RSD为1.16%～1.39%(n=6).结论:本法操作简便,准确度高,重复性好,可作为西洋参质量控制方法之一.     

连花清瘟胶囊超声逆流提取液超高效液相色谱指纹图谱研究

目的 采用超高效液相色谱（UPLC）法建立连花清瘟胶囊超声逆流提取液的指纹图谱.方法 采用Acquity UPLC@BEH C18柱（100mm×2.1 mm,1.7 μm）,流动相为乙腈-0.1％磷酸溶液梯度洗脱,流速为0.4 mL/min,柱温为40℃,检测波长为228 nm.结果 建立了连花清瘟胶囊超声逆流提取液的UPLC指纹图谱,共标定了27个共有峰,指认了其中4个共有峰,11批连花清瘟胶囊超声逆流提取液指纹图谱的相似度均在0.991 ～1.000之间.结论 该方法稳定性、重复性好,可为连花清瘟胶囊超声逆流提取液的质量评价提供依据.     

复方丹参方快速UPLC指纹图谱研究

目的:建立复方丹参方中丹参、三七指纹图谱。方法:采用Waters Acquity UPLC(色谱柱为50 mm×2.6 mm,1.7μm),流动相为0.02%磷酸(A)和0.02%磷酸的80%乙腈(B)进行梯度洗脱,柱温为40±0.15℃,流速为0.42mL·min^-1,检测波长:203nm,进样量:3μL。对10批不同丹参饮片进行分析,使用(2004A版)由国家药典委员会制定颁布的相似度软件评估其相似度。结果:建立了复方丹参方中丹参、三七提取液的指纹图谱,10批丹参饮片在此指纹图谱中的相似度均大于0.932。因人参皂苷Rg1信号强,峰形好,故以人参皂苷Rg1为参照峰,确定了15个共有峰,相对保留时间的RSD<1%,相对峰面积的RSD<3%,确定了5个特征峰,其中2个特征峰来源于组方中的丹参,3个特征峰来源于组方中的三七。结论:此色谱条件能将丹参、三七简单、方便的于同一图谱中展现出来,10批丹参饮片质量稳定。为复方丹参方的质量评价、指导临床用药、制剂设计提供科学的依据。     

超高效液相色谱法测定注射用头孢替唑钠的有关物质

目的：建立超高效液相色谱梯度洗脱法测定注射用头孢替唑钠中的有关物质。方法：采用 Waters Acquity UPLC BEH C18（100 mm ×2.1 mm,1.7μm）色谱柱,以枸橼酸溶液（取枸橼酸3 g 加水溶解并稀释至900 ml）-乙腈（90：10）为流动相 A,乙腈为流动相 B,线性梯度洗脱;流速：0.3 ml／ min;检测波长：254 nm。结果：在0.19~47.08μg／ ml浓度范围内,头孢替唑峰面积与浓度呈良好的线性关系（r ＝1.0000）。结论：本法简便快速、专属性强,可作为测定注射用头孢替唑钠有关物质的有效方法。