HPLC同时测定苦参药材中苦参碱和氧化苦参碱的含量测定方法

目的探讨高效液相色谱法在测定苦参药材中苦参碱和氧化苦参碱的含量上的具体操作方法以及可行性。方法建立高效液相色谱法对中药苦参药材中的苦参碱和氧化苦参碱的含量进行测定,并且对该实验方法的精密度,重复性,稳定性等方面进行实验考察。结果得出样品苦参中含有苦参碱平均含量为0.083%,相对标准偏差—RSD是1.79%;氧化苦参碱平均含量为2.28%,相对标准偏差—RSD是1.65%。并且使用高效液相色谱对苦参中苦参碱和氧化苦参碱进行含量测定具有良好的精密度,重复性,稳定性,以及加样回收率。结论使用高效液相色谱法对苦参药材中的苦参碱和氧化苦参碱的含量进行实验测定,实验操作简单,方便,准确度高,稳定性好,同时重复性也比较理想,在对中药苦参中苦参碱和氧化苦参碱的含量测定方面具有良好的使用价值。     

反相高效液相色谱法测定苦参药材中苦参碱的含量

目的:建立苦参药材反相高效液相色谱法(RPHPLC)含量测定方法,控制药材质量.方法:以十八烷基硅烷键合硅胶(ODS)为固定相,乙腈-水(加0.04%三乙胺,磷酸调pH7.00)(20:80)为流动相,检测波长为215 nm,进样量10μL.结果:反相高效液相色谱法(RP-HPLC)测定苦参中苦参碱含量平均加样回收率为100.1%,RSD为0.92%.结论:该方法简便、快速、准确,能有效测定苦参药材苦参碱含量.     

滴定法与高效液相色谱法测定苦参提取液中苦参总碱含量比较研究

目的筛选苦参提取工艺研究中苦参总碱含量的测定方法。方法用滴定法和高效液相色谱(HPLC)法对苦参原药材、苦参醇提取液及酸水提取液中苦参总碱的含量测定进行对比研究。结果在测定苦参原药材、苦参醇提取液及酸水提取液中苦参总碱的含量时,滳定法所得的相对标准偏差(RSD)较HPLC法低(0.92%vs.6.30%、0.72%vs.8.85%、1.53%vs.4.00%)。结论在进行含苦参原药材的提取工艺研究时,宜采用滴定法对苦参总碱进行含量测定。     

利湿合剂的定性及定量

建立了利湿合剂的药材质控方法.以高效液相色谱法测定其中的苦参碱.采用Diamonsil C18色谱柱,甲醇-水-25％氨水(52∶45∶3)为流动相,检测波长215 nm.苦参碱在0.64～3.2 μg范围内线性关系良好,回收率为95.6％,RSD为2.85％.以薄层色谱法鉴别苦参和陈皮,用高效液相色谱法鉴别金银花中的绿原酸.     

抗妇炎软胶囊质量标准研究

目的 建立抗妇炎软胶囊的质量控制标准.方法 采用薄层色谱法对抗妇炎软胶囊中连翘、黄柏、苦参进行定性鉴别,采用高效液相色谱法测定主要成分苦参碱的含量.结果 供试品溶液色谱中,在与连翘、黄柏、苦参所含有效成分对照品色谱相应位置上显相同颜色的斑点;苦参碱在0.52～2.60 μg范围内与峰面积呈良好线性关系,回归方程为A ...     

康之健洗液质量标准研究

目的 建立康之健洗液的质量标准.方法 采用薄层色谱法(TLC)对处方中苦参、黄柏进行定性鉴别;采用高效液相色谱法(HPLC)测定蛇床子素含量.结果 在TLC色谱中检出苦参、黄柏;蛇床子素进样量在0.24～1.44 μg范围内呈良好的线性关系,r=0.9990;平均回收率103.80%,RSD为0.37%.结论 所建立的...     

痢必灵片的质量标准

目的:制订痢必灵片的质量标准.方法:采用薄层色谱法对方中芍药苷进行鉴别,采用高效液相色谱法,测定方中苦参中苦参碱的含量.结果:白芍可用薄层色谱鉴别;苦参碱在0.01～0.56 mg·ml-1内线性关系良好(r=1.000 0),加样回收率为101.9%, RSD 3.0%(n=6).结论:方法简便、快速、能有效控制痢必灵片的质量.     

RP-HPLC法测定苦参中苦参碱的含量

目的建立测定苦参中苦参碱含量的RP HPLC法。方法采用反相高效液相色谱法 ,Kro masilC18柱 (2 5 0mm× 4 6mm ,5 μm) ,流动相为甲醇 水 三乙胺 (V∶V∶V =10 5 0 0∶95 0 0∶0 0 4 ) ,检测波长为 2 2 5nm ,柱温为室温。结果苦参碱质量浓度在 3 92～ 78 4mg·L-1内与峰面积呈良好线性关系 ,r =0 9998(n =6 ) ;平均回收率为 97 2 % ,RSD为 2 1% (n =9)。结论测定方法准确、可靠 ,为苦参药材质量评价提供了可靠依据     

黄柏治糜栓的制备及质量控制

目的 制备黄柏治糜栓并制订其质量控制标准。方法 以黄柏、蒲公英、苦参、血竭、紫花地丁、紫草6味中药为主药，半合成脂肪酸（38型）为栓剂基质制成栓剂，采用高效液相色谱法（HPLC法）测定苦参中氧化苦参碱的含量，同时用薄层色谱法对制剂中的黄柏、蒲公英、紫花地丁进行鉴别。结果 HPLC法的平均回收率为99．47％，RSD为1.24％。结论 该制剂制备方法简便，质量稳定，检测方法可靠。     

HPLC法测定苦参凝胶中苦参碱与氧化苦参碱的含量

目的建立苦参凝胶中苦参碱与氧化苦参碱的含量测定方法。方法采用高效液相色谱法进行测定。色谱柱为Elite hypersil氨基柱(250mm×4.6mm;5μm),乙腈-无水乙醇-3%磷酸溶液(82:10:8)为流动相,检测波长为220nm,柱温为35℃。结果苦参碱和氧化苦参碱分别在9.984～99.84μg和21.84～218.4μg范围内,进样量与峰面积呈良好的线性关系;苦参碱、氧化苦参碱的平均回收率分别为99.2%和99.8%(n=5),RSD分别为1.1%和0.8%;精密度试验RSD分别为1.1%和0.8%(n=5);重复性试验RSD分别为1.1%和1.2%(n=6)。结论本方法简便、准确、重现性好,可同时测定苦参凝胶中苦参碱与氧化苦参碱的含量。     

Characterization of alkaloids in Sophora flavescens Ait. by high-performance liquid chromatography-electrospray ionization tandem mass spectrometry

Sophora flavescens Ait., a well-known Chinese herbal medicine, is widely used in clinical practice for the treatment of viral hepatitis, cancer, gastrointestinal hemorrhage, and skin diseases. This paper is the first report on a method based on the combined use of high-performance liquid chromatography, photodiode array detection, and electrospray ionization tandem mass spectrometry for the comprehensive and systematic separation and characterization of bioactive alkaloids in Sophora flavescens Ait. A total of 22 constituents were identified on the basis of the extracted ion chromatograms for different [M+H](+) ions of the alkaloids present in S. flavescens Ait. Among these, 5 constituents were unambiguously identified by comparing the experimental data on their retention times and MS(n) spectra with those of the authentic compounds, and 17 other constituents were tentatively identified on the basis of their MS(n) fragmentation behaviors and/or molecular weight information from literatures. Furthermore, some characteristic fragmentation pathways of the alkaloids in S. flavescens Ait. were detected and examined. This information may be useful for characterizing the bioactive alkaloids present in S. flavescens Ait. and for possible applications in formulations.     

Characterization of flavonoids in the extract of Sophora flavescens Ait. by high-performance liquid chromatography coupled with diode-array detector and electrospray ionization mass spectrometry

A method coupling high-performance liquid chromatography (HPLC) with diode-array detector (DAD) and electrospray ionization mass spectrometry (ESI) was established for the separation and characterization of flavonoids in Sophora flavescens Ait. Based on the chromatographic separation of most flavonoids present in S. flavescens Ait., a total of 24 flavonoids were identified. Fourteen compounds were unambiguously identified comparing experimental data for retention time (t(R)), UV and MS spectra with those of the authentic compounds: 3',7-dihydroxy-4'-methoxy-isoflavone (13), trifolirhizin (14), kurarinol (18), formononetin (19), 7,4'-dihydroxy-5-methoxy-8-(gamma,gamma-dimethylallyl)-flavanone (22), maackiain (21), isoxanthohumol (23), kuraridine (26), kuraridinol (27), sophoraflavanone G (30), xanthohumol (31), isokurarinone (33), kurarinone (35) and kushenol D (38), and additional 10 compounds were tentatively identified as kushenol O (10), trifolirhizin-6'-malonate (15), sophoraisoflavanone A (20), norkurarinol/kosamol Q (24), kushenol I/N (25), kushenol C (28), 2'-methoxykurarinone (29), kosamol R (32), kushecarpin A (34) and kushenol A (37) by comparing experimental data for UV and MS spectra with those of literature. Furthermore, fragmentation pathways in positive ions mode of 24 flavonoid compounds of types of flavanone, flavanonol, flavonol, chalcone, isoflavone, isoflavanone and ptercocarpane were summarized. Some common features, such as CH(3)., H(2)O, CO, CO(2), C(3)O(2) and C(2)H(2)O losses, together with Retro-Diels-Alder fragmentations were observed in the prenylated flavonoids in S. flavescens Ait. The loss of the lanandulyl chain was their characteristic fragmentation, which might help deducing the structure of unknown flavonoid compounds. The present study provided an approach to rapidly characterize bioactive constituents in S. flavescens Ait.     

采用差示扫描量热技术建立苦参碱纯度测定新分析方法

目的研究采用差示扫描量热技术（DSC）,建立中药活性成分苦参碱化学样品纯度的新检测分析方法。方法针对苦参碱化学样品,考察了溶剂残留、炉体气氛、升温速率、称样量四个因素对差示扫描量热法检测结果的影响;优化并确定了最佳检测条件,测定了苦参碱化学样品的纯度值;利用高效液相色谱法（HPLC）对建立的方法及检测结果进行了验证。结果本研究建立的差示扫描量热法进行苦参碱样品纯度检测具有较高的科学性和可行性,最佳实验条件为升温速率4.0K/min,称样量为2.5mg～3.7mg,炉内气体为静态空气,经检测获得苦参碱样品的化学纯度值为99.818%;高效液相色谱法测得苦参碱样品化学纯度值为99.825%。结论利用建立的差热扫描量热分析方法可以快速、准确地测定苦参碱化学纯度,论文研究为苦参碱纯度测定提供了一种新分析技术和方法。     

Matrine-induced apoptosis in leukemia U937 cells: involvement of caspases activation and MAPK-independent pathways

It is reported that matrine, one of the major effective compounds isolated from the root of Sophora flavescens Ait., has anti-leukemia activity. In this study, we find that the treatment of leukemia U937 cells with matrine results in induction of apoptosis. Analysis of the mechanism underling this apoptotic event showed activation of caspases-9, -3, and -7, and release of cytochrome C from mitochondria to cytosol, and cleavage of poly(ADP-ribose) polymerase. Matrine did not alter the level of bcl-2 and bcl-xL as well as bax. In addition, no correlation was found between matrine administration and activation of the three major MAPK subfamilies (Erk1/2, p38, JNK/SAPK). The results indicate that matrine induces apoptosis in U937 cells via a cytochrome C-triggered caspase activation pathway.     

Anticancer effects of the Chinese medicine matrine on murine hepatocellular carcinoma cells

Matrine is a component of the traditional Chinese medical herb Sophora flavescens Ait, which is widely used to treat diseases such as viral hepatitis, cardiac arrhythmia and skin inflammations. As indicated by previous reports, the molecular mechanism of matrine's anti-cancer effect has been poorly clarified. In this study, we used both in vitro and in vivo models to investigate matrine's antitumor effect and its possible molecular mechanisms. Murine hepatocellular carcinoma H22 cells were cultured in the presence of matrine at various concentrations (0.2 - 2.0 mg/mL). A dose-dependent antiproliferation effect was observed. The 50 % inhibitory concentration (IC (50)) was 0.6 mg/mL. Antiproliferation effects of matrine were associated with an increase in cells arrested in the G (1) phase of the cell cycle. Morphological changes, flow cytometric analysis and expression of the proapoptotic protein Bax indicated that this anticancer effect was mediated via apoptosis. In vivo antitumor efficacy was evaluated following S. C. inoculation of H22 cells in BALB/c mice. Matrine administrated I. P. resulted in strong in vivo anticancer activity. Our results showed that seven doses of matrine at 50 mg/kg/dose inhibited 60.7 % of tumor growth. Transmission electron microscope (TEM) analysis and histoimmunochemical staining for Bcl-2 and Bax proteins also indicated induction of apoptosis in tumor tissues by matrine. Taken together, our results demonstrate that matrine possesses strong antitumor activities in vitro and in vivo. Inhibition of cell proliferation and induction of apoptosis are the likely mechanisms responsible for matrine's antitumor activities.     

复方苦参注射液的抗肿瘤作用研究进展

 复方苦参注射液是由苦参和白土茯苓制成的抗肿瘤中药,属非特异性细胞毒药物,具有止血、止痛、抑制肿瘤细胞扩散等功能,同时还可以改善造血功能、增加机体免疫力,且无明显毒副作用。此文就复方苦参注射液在肿瘤治疗中的临床应用研究进展作一综述。     

HPLC法测定归芪补血口服液中黄芪甲苷的含量

建立归芪补血口服液中黄芪甲苷含量测定的HPLC方法,并将结果与薄层扫描（TLCS）法进行比较。方法：色谱柱为依利特C18色谱柱（250mm×4．6mm,5μm）,以乙腈：水（31：69）为流动相,检测波长为201nm。结果：黄芪甲苷的线性范围为0．5～15．7μg（r=0．999 8,n=5）;平均加样回收率98．2％（RSD=0．73％,n=5）。HPLC法比TLCS法更简便、快速、准确。结论：本文所建立的HPLC法,准确、简便、快捷,可代替TLCS法用于制剂质量控制。     

Antiinflammatory effects of matrine in LPS-induced acute lung injury in mice

Matrine is one of the main active components of Chinese herb Sophora flavescens Ait (Kushen), which has been demonstrated to be effective in suppressing inflammation. The aim of the present study is to investigate the effect of matrine on LPS-induced lung injury. Lung injury was assessed by histological study and wet to dry weight ratios, as well as cell count and protein content in bronchoalveolar lavage fluid. We also detected MPO activity reflecting neutrophil infiltration and MDA activity examining oxidative stress in lung tissues. Cytokines and ROS production in cells were monitored by ELISA and flow cytometry, respectively. The results showed that high dose of matrine significantly reduced the mortality rate of mice with LPS administration. Treatment with matrine improved LPS-induced lung histopathologic changes, alleviated pulmonary edema and lung vascular leak, inhibited MPO and MDA activity,and reduced the production of inflammatory mediators including TNF-α, IL-6 and HMGB1. In vitro, matrine administration reduced the production of ROS and inflammatory factors, which was possibly associated with inhibition of NF-κB. In conclusion, the current study demonstrated that matrine exhibited a protective effect on LPS-induced acute lung injury by inhibiting of the inflammatory response, which may involve the suppression of ROS and tissue oxidative stress.     

苦参的化学成分

目的研究中药苦参 (SophoraflavescensAit)的化学成分。 方法采用硅胶柱色谱的手段 ,利用理化和波谱分析方法 ,对苦参中的化合物进行分离、分析、鉴定。结果从苦参根中分离得到 2个紫檀素类化合物、一个异黄酮类化合物、一个酚酸类化合物和 β 谷甾醇。确定其结构 ,分别为高丽槐素 (maackiain ,Ⅰ )、三叶豆紫檀苷 6′ 单乙酸酯 (trifolirhizin 6′ monoacetate ,Ⅱ )、芒柄花黄素 (for mononetin ,Ⅲ )、2 ,4 二羟基苯甲酸 (2 ,4 dihydroxybenzoicacid ,Ⅳ )、β 谷甾醇 (β sitosterol,Ⅴ )。 结论三叶豆紫檀苷 6′ 单乙酰酯 (trifolirhizin 6′ monoacetate)为首次从苦参中分离得到     

苦参的毛细管电泳数字化指纹图谱研究

目的 建立苦参的毛细管电泳指纹数字化图谱（CEFP），考察Jaccard相似度Sb评价苦参CEFP的可行性和特征。方法 采用胶束电动毛细管电泳法，以30mmol·L^-1硼砂＋10mmol·L^-1 SDS为背景电解质，运行电压12kV，紫外检测波长205nm，以10个不同产地药材的电泳图谱建立了苦参的对照CEFP。与双定性相似度SF和S′F、双定量相似度C％和P％相比较考察Jaccard相似度参数Sb的可行性和特征。结果 以萘普生为参照物峰，确定15个共有峰，建立了苦参CEFP，并用“中药色谱指纹图谱超信息特征数字化评价系统3．0”软件挖掘苦参CEFP的指纹特征，结合双定性双定量相似度法客观、全面地评价了不同产地苦参的质量。结论 所建立的苦参毛细管电泳数字化指纹图谱具有较好的精密度和重现性，可为苦参质量控制提供新参考，Jaccard相似度Sb参数能够作为评价中药批间相似度一个较好的指标。