夏枯草高效液相色谱指纹图谱的研究

目的:建立夏枯草的指纹图谱分析方法,并对不同产地的夏枯草进行指纹图谱的比较研究。方法:以齐墩果酸为参照,采用高效液相色谱法,色谱柱为HypersilC18,流动相为甲醇:水:醋酸(86:14:0.1),检测波长为208nm,流速为1ml/min。结果:标出10个主要共有峰,方法学考察表明,本研究建立的分析方法有较好的重现性,比较了不同产地夏枯草药材与标准药材指纹图谱的相似性。结论:方法稳定、可靠、简便,为提高夏枯草药材质量控制提供依据。     

夏枯草的HPLC指纹图谱鉴别研究

目的 建立夏枯草HPLC指纹图谱分析方法.方法 采用高效液相色谱法,BIOSIL U546250-1型色谱柱(4.6 mm×250 mm,5 μm),流动相:甲醇-水-醋酸-三乙胺(84:16:0.05:0.03),流速0.8 ml·min-1,柱温30℃;检测波长215 nm.结果 建立了夏枯草的指纹图谱,确定了12...     

夏枯草HPLC融合指纹图谱研究

目的:建立夏枯草的融合指纹图谱,实现同分异构体熊果酸和齐墩果酸完全色谱分离和同步含量测定,对夏枯草质量进行评价。方法:采用高效液相色谱法,建立夏枯草的指纹图谱,并为互为同分异构体的熊果酸和齐墩果酸单独建立分析方法,使其达到完全色谱分离,使用"MATLAB R2012a版"将两个色谱图融合部分进行基线融合计算,使用"中药色谱指纹图谱相似度评价系统2004A版"进行图谱融合。结果:建立了可单独、分段操作的"模块式"融合指纹图谱,共标定了18个共有色谱峰,结合质谱和对照品对部分共有峰进行了推断,其中1,3~8,17,18号峰分别推测为丹参素、咖啡酸、芦丁、金丝桃苷、异槲皮苷、异迷迭香酸苷、迷迭香酸、齐墩果酸和熊果酸。同时对15批夏枯草样品中熊果酸和齐墩果酸的含量进行了测定,熊果酸含量0.19%~0.36%,齐墩果酸含量0.055%~0.11%。结论:建立的熊果酸和齐墩果酸的分析方法分离度高、分析速度快、结果准确可靠;建立的"模块式"融合指纹图谱可以更准确全面反映夏枯草中的化学成分,可为夏枯草质量控制研究提供参考。     

不同采收期苍耳草的指纹图谱研究

目的:研究不同采收期苍耳草的指纹图谱,建立稳定、可重复的苍耳草质量控制手段。方法:运用HPLC色谱技术建立不同采收期不同产地苍耳草的指纹图谱。结果:通过比较14批苍耳草和2批苍耳子药材样品HPLC图谱,建立其对照指纹图谱,并以共有模式为参照,通过《中药指纹图谱相似度计算软件》计算,得到色谱指纹图谱模拟图谱及相似度计算结果。除1号、15号和16号与其他批次样品相似度不高外,其他批次相似度较大,但是其主要成分在含量方面存在一定的差异,与药材采自不同采收期有关。结论:本文建立了苍耳草的HPLC指纹图谱,为苍耳草质量标准的建立提供了试验依据。     

不同产地桑白皮HPLC的指纹图谱的研究

目的 建立简单可行的桑白皮药材指纹图谱方法.方法 采用高效液相色谱法,Agilent Extend-C18(4.6 mm×250 mm,5μm)色谱柱,流动相:以乙腈-0.2%甲酸进行梯度洗脱,流速:1 mL·min-1,柱温:30℃,检测波长:280 nm,进样量为10μL.结果 用梯度洗脱得到的色谱图中各色谱峰分离较好,达到指纹图谱分析的效果.结论 该方法重复性好、检测方法简便,可为桑白皮指纹图谱的质量控制提供依据.     

不同产地黄芪总黄酮HPLC指纹图谱研究

目的:建立黄芪总黄酮的HPLC指纹图谱分析方法,评价不同来源药材的质量.方法:采用二极管阵列检测器,色谱柱为AglientExtend C18(250mm×4.6mm,5μm),流动相为乙腈(A)-0.2％甲酸水溶液(B),梯度洗脱,流速为1.0mL/min,检测波长为280nm.结果:建立14个共有特征峰的HPLC指纹图谱,方法学考察结果符合指纹图谱技术要求.结论:该方法稳定可靠、重复性好,可结合含量测定用于全面控制黄芪的质量.     

指纹图谱法鉴别红参产地

目的建立控制红参药材产地和质量的指纹图谱。方法采用十八烷基硅烷键合硅胶为填充剂,乙腈 0.05%磷酸（99：400）为流动相;检测波长为203 nm;柱温为25 ℃;流速1.0 mL·min 1,进样10 μL。运用指纹图谱相似度计算软件计算相似度。结果以人参皂苷Rg1为参照物峰,确定了其中的13个峰为特征峰,经过计算第10产地的红参相似度最好。结论采用高效液相色谱法建立红参的特征指纹图谱,能够更好的保证成品的质量,控制药材的来源。     

不同产地山合欢皮HPLC指纹图谱的研究

建立山合欢皮正丁醇部位的指纹图谱。方法：采用HPLC法,色谱柱：Kromasil C18（250mm×4．6mm,5μm）;流动相：甲醇-乙腈-0．4％磷酸溶液,梯度洗脱;流速：1．0ml·min^-1;柱温：23℃;检测波长：210nm;参照物：icarside E5。结果：确定了25个共有峰,得到了以淫羊藿次苷E5为参照物的保留时间和相对峰面积,并利用《中药色谱指纹图谱相似度评价系统》计算相似度。结论：采用HPLC指纹图谱技术,可作为山合欢皮药材的质量控制。     

咳喘颗粒HPLC指纹图谱研究

目的 建立咳喘颗粒的HPLC指纹图谱方法,为科学地评价咳喘颗粒的质量提供依据.方法 Agilent ZOR-BAX Eclipse XDB-C18色谱柱(250 mm×4.6 mm,5μm)以乙腈-0.1％磷酸水为流动相梯度洗脱,检测波长237 nm,流速1.0 ml/min,柱温:30℃.结果 确定了16个共有峰,10批咳喘颗粒指纹图谱的相似度均＞0.96.结论 建立了分离效果好、简单、灵敏、准确的方法,可用于咳喘颗粒质量评价的方法.     

夏枯草中苯丙素和三萜的分离和鉴定

从夏枯草中用色谱法分离得到1个新的苯丙素类化合物1和5个已知的三萜类化合物2－6,经波谱法和化学法鉴定为3,4,α-三羟基苯丙素丁酯（1）,2α,3α,24－三羟基乌苏－12,20（30）－二烯－28－酸（2）、2α,3α,24－三羟基齐墩果－12－烯－28－酸（3）、2α,3α,24－羟基乌苏－12－烯－28－酸（4）、2α,3β－二羟基齐墩果－12－烯－28－酸（5）和2α,3β－二羟基乌苏－12－烯－28－酸（6）,5个三萜类化合物均为首次从夏枯草中分离得到。     

Isolation and characterization of an anti-HSV polysaccharide from Prunella vulgaris

A water soluble substance was isolated from a Chinese herb, Prunella vulgaris, by hot water extraction, ethanol precipitation and gel permeation column chromatography. Chemical tests showed that the substance was an anionic polysaccharide. Using a plaque reduction assay, the polysaccharide at 100 microg/ml was active against the herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), but was inactive against cytomegalovirus, the human influenza virus types A and B, the poliovirus type 1 or the vesicular stomatitis virus. The 50% plaque reduction dose of the polysaccharide for HSV-1 and HSV-2 was 10 microg/ml. Clinical isolates and known acyclovir-resistant (TK-deficient or polymerase-defective) strains of HSV-1 and HSV-2 were similarly inhibited by the polysaccharide. Pre-incubation of HSV-1 with the polysaccharide at 4, 25 or 37 degrees C completely abrogated the infectivity of HSV-1, but pre-treatment of Vero cells with the polysaccharide did not protect cells from infection by the virus. The addition of the polysaccharide at 0, 2, 5.5 and 8 h post-infection of Vero cells with HSV-1 at a multiplicity of infection (MOI) of five reduced the 20 h-yield of intracellular infectious virus by 100, 99, 99 and 94%, respectively. In contrast, a similar addition of heparin showed 85, 63, 53 and 3% reduction of intracellular virus yield, respectively. These results suggest that the polysaccharide may inhibit HSV by competing for cell receptors as well as by some unknown mechanisms after the virus has penetrated the cells. The Prunella polysaccharide was not cytotoxic to mammalian cells up to the highest concentration tested, 0.5 mg/ml and did not show any anti-coagulant activity. In conclusion, the polysaccharide isolated from P. vulgaris has specific activity against HSV and its mode of action appears to be different from other anionic carbohydrates, such as heparin.     

Anti-allergic and anti-inflammatory triterpenes from the herb of Prunella vulgaris

The activity-guided fractionation of the extract of the herb of Prunella vulgaris (Labiatae) led to the isolation of four triterpenes, i.e., betulinic acid, ursolic acid, 2 alpha,3 alpha-dihydroxyurs-12-en-28-oic acid, and 2 alpha-hydroxyursolic acid. One of these compounds, 2 alpha,3 alpha-dihydroxyursolic acid, demonstrated significant inhibition on the release of beta-hexosaminidase from the cultured RBL-2H3 cells in a dose-dependent manner; the IC50 value was calculated to be 57 microM. When the isolated compounds were tested for their effects on the production of nitric oxide from cultured murine macrophages, RAW 264.7 cells, ursolic acid and 2 alpha-hydroxyursolic acid exhibited strong inhibitory activities (IC50 values, 17 and 27 microM, respectively).     

夏枯草中水溶性多糖的分离纯化及化学研究

目的研究夏枯草中水溶性多糖组分.方法采用DEAD琼脂糖凝胶柱色谱进行纯化,化学和光谱方法进行结构特征分析.结果从夏枯草沸水提取物中分离得到5个多糖部位,其中XKC00、XKC02-A、XKC02-B均为均一分子量多糖,分子量分别为2.6x 10(4),1.36x 10(5),5 600 Da.结论XKC00为首次从夏枯草中分离得到的一个中性杂多糖.     

夏枯草化学成分研究

采用柱色谱分离技术,从夏枯草全草70%乙醇浸膏的乙酸乙酯和正丁醇部位中分离得到14个化合物,经MS和NMR分析鉴定为:白桦脂酸(1)、乌苏酸(2)、2α,3α-二羟基-12-烯-28-乌苏酸(3)、2α,3β-二羟基-12-烯-28-乌苏酸(4)、2α,3β,24-三羟基-12-烯-28-乌苏酸(5)、2α,3α,19-三羟基-12-烯-28-乌苏酸(6)、齐墩果酸(7)、槲皮素-3-O-β-D-葡萄糖苷(8)、槲皮素-3-O-β-D-半乳糖苷(9)、山奈酚-3-O-β-D-葡萄糖苷(10)、芦丁(11)、丹参素甲酯(12)、丹参素乙酯(13)、迷迭香酸乙酯(14).其中化合物5、6、10、13为首次从该植物中分离得到,化合物5、13为首次从该属植物中分离得到.     

夏枯草中齐墩果酸和熊果酸的含量测定方法研究

目的通过采用高效液相色谱法,测定不同产地夏枯草中齐墩果酸和熊果酸的含量,比较夏枯草药材质量的优劣。方法色谱柱:Kromasil-C_(18)(250 mm×4.6 mm,5μm);流动相:乙腈-甲醇-0.2%磷酸水(5∶82∶13)等度洗脱;流速:1.0 mL·min~(-1);检测波长:210 nm;柱温:25℃。结果齐墩果酸在0.041~0.410 mg·mL~(-1)与峰面积呈现良好的线性关系(r=0.9999),平均回收率为99.1%,RSD为1.8%;熊果酸在0.054~0.540 mg·mL~(-1)与峰面积呈现良好的线性关系(r=0.9998),平均回收率为98.1%,RSD为2.4%。结论该方法简单、准确、可靠,为夏枯草的质量控制与评价提供了有效的手段。     

HPLC测定夏枯草中的芦丁和槲皮素

目的 采用HPLC法测定夏枯草中芦丁和槲皮素的含量.方法 色谱柱为Alltima C_(18)(250MM×4.6 mm,5 μm),流动相为甲醇-0.1%冰醋酸,检测波长350 nm,流速1.0 mL·min~(-1),柱温30℃.结果 芦丁、槲皮素的线性范围分别为0.0149～0.2380 mg·mL~(-1)(r=0.9999)、0.0019～0.0306 mg·mL~(-1)(r=0.9998),平均回收率分别为97.78%(RSD=0.83%)、101.18%(RSD=0.83%).安徽、湖北、贵州、江西、广西五产地中芦丁含量分别为0.3527、0、0.2337、2.1164、0.9578 mg·g~(-1)生药,槲皮素含量分别为0.0872、0.0473、0.0721、0.5219、0.1755 mg·g~(-1)生药.结论 所建方法操作简便、准确,可作为夏枯草中芦丁和槲皮素的含量测定方法.     

Changes in bioactive components related to the harvest time from the spicas of Prunella vulgaris

Context: Prunella vulgaris L. (Labiatae) is a perennial plant common in China and Europe and is rich in rosmarinic acid (RA), ursolic acid (UA), and oleanolic acid (OA). The dried spica of P. vulgaris has been used as traditional medicine in China for over a hundred years. To our best knowledge, no study has been conducted to determine the influence of harvesting time on concentrations of bioactive compounds of P. vulgaris. Objective: In the current study, changes in the bioactive compounds present in spicas were investigated at five harvest times over 2 months. Materials and methods: Plant material were collected at five fixed dates: 5th May, 20th May, 7th June, 15th June, and 25th June and assayed for chemical contents by high-performance liquid chromatography (HPLC). Results: Among the different harvest times, the highest levels of RA (56.81 mg·g(-1)), UA (2.77 mg·g(-1)), and OA (0.91 mg·g(-1)) were found on 5th May, whereas the lowest levels of RA (1.66 mg·g(-1)), UA (2.27 mg·g(-1)), and OA (0.43 mg·g(-1)) were observed on 25th June. Discussion and conclusion: As each medicinal product has its own content requirement for different bioactive components, the optimum harvest time might be determined according to the accumulation dynamics of target compound in dried spicas of P. vulgaris. These results may be useful for determining the optimal harvest time when bioactive components are at the maximum level, which is in early May.     

Changes in bioactive components related to the harvest time from the spicas of Prunella vulgaris

Context: Prunella vulgaris L. (Labiatae) is a perennial plant common in China and Europe and is rich in rosmarinic acid (RA), ursolic acid (UA), and oleanolic acid (OA). The dried spica of P. vulgaris has been used as traditional medicine in China for over a hundred years. To our best knowledge, no study has been conducted to determine the influence of harvesting time on concentrations of bioactive compounds of P. vulgaris.

Objective: In the current study, changes in the bioactive compounds present in spicas were investigated at five harvest times over 2 months.

Materials and methods: Plant material were collected at five fixed dates: 5th May, 20th May, 7th June, 15th June, and 25th June and assayed for chemical contents by high-performance liquid chromatography (HPLC).

Results: Among the different harvest times, the highest levels of RA (56.81 mg·g^?1), UA (2.77 mg·g^?1), and OA (0.91 mg·g^?1) were found on 5th May, whereas the lowest levels of RA (1.66 mg·g^?1), UA (2.27 mg·g^?1), and OA (0.43 mg·g^?1) were observed on 25th June.

Discussion and conclusion: As each medicinal product has its own content requirement for different bioactive components, the optimum harvest time might be determined according to the accumulation dynamics of target compound in dried spicas of P. vulgaris. These results may be useful for determining the optimal harvest time when bioactive components are at the maximum level, which is in early May.     

夏枯草中熊果酸含量的UPLC测定

目的：建立夏枯草药材中熊果酸含量的超高效液相色谱法（UPLC）测定方法。方法采用UPLC进行测定,色谱柱为ACQUITY UPLC BEH C18（2.1 mm×100 mm,1.7μm）,以甲醇-0.5%冰乙酸水为流动相进行等度洗脱,流速为0.3 ml/min,柱温30℃,检测波长210 nm。结果熊果酸在0.2461～0.4922 mg范围内呈良好的线性关系（r^2=0.9999）,平均回收率为99.29%,R SD=1.74%,熊果酸在夏枯草中的含量为0.28%～0.53%。结论该方法快速、高效、重现性好,可以为夏枯草药材的质量控制提供更有效的手段。