不同产地黄芩样品中主要有效成分含量的对比研究

目的：对不同产地黄芩样品中的主要有效成分进行对比研究。方法分别选取河北、内蒙古、甘肃三产地的黄芩样品进行分析,选用HPLC法对三组样品进行重复性、稳定性、回收率、线性关系考察及样品含量测定。结果三个产地的黄芩样品都具有很好的重复性、稳定性、回收率,黄芩样品中黄芩苷、汉黄芩苷的含量河北＞内蒙古＞甘肃,而黄芩素、汉黄芩素的含量内蒙古＞河北＞甘肃。结论不同产地的黄芩样品中的主要成分不同,在制剂中应根据不同作用选择合适产地的黄芩入药。     

基于黄酮类成分含量构成特征和体外抗病毒活性的不同产地黄芩的质-效评价研究

目的:评价黄芩道地药材与非道地药材中主要黄酮类成分含量构成特征及其抗病毒活性,以探讨黄芩药材的质-效关系及药效物质基础。方法:制备8批不同产地黄芩(S1~S8)的水提物冻干粉,采用超高效液相色谱法测定水提物中黄芩苷、汉黄芩苷、黄芩素、汉黄芩素等4种黄酮类成分的含量并计算其含量构成比。按不同产地黄芩的黄酮类成分含量构成比,制备上述4种成分混合物作为对应的黄酮类成分模拟样品(E1~E8)。以利巴韦林为阳性对照,采用MTT法和细胞病变程度法,考察8批黄芩水提物样品及其对应的模拟样品对人喉癌细胞Hep-2的半数有毒浓度(TD50)和对呼吸道合胞病毒(RSV)的半数抑制浓度(IC50),并计算治疗指数(TI)。采用SPSS 17.0软件对黄芩苷、汉黄芩苷、黄芩素、汉黄芩素含量与其抗RSV活性(IC50值)进行Pearson相关性分析。结果:样品S4(产地为河北承德)的黄芩苷、汉黄芩苷含量最高,样品S6(产地为内蒙古-2)的黄芩素、汉黄芩素含量最高;上述4种成分含量最低的分别为样品S6、S6、S7(产地为北京)、S4。样品S4中黄酮苷类成分含量较高,相应的苷元类成分含量较低,其黄芩苷、汉黄芩苷、黄芩素、汉黄芩素的含量构成比为1∶0.224∶0.111∶0.013;样品S6中黄酮苷类成分含量较低,相应的苷元类成分含量较高,上述4种成分的含量构成比为1∶0.241∶0.713∶0.106。不同产地黄芩水提物对Hep-2细胞的TC50均高于50μg/mL,对RSV的IC50为11.11~51.74μg/m L,TI为1.86~5.20;其对应的黄酮类成分模拟样品的TC50为23.11~52.23μg/mL,对RSV的IC50为4.87~14.61μg/mL,TI为1.85~4.75。道地产区(河北承德)黄芩(S4)的水提物及其对应模拟样品(E4)的抗RSV作用最强。相关性分析显示,4种黄酮类成分的含量与其抗病毒活性的相关系数均小于0.5,两者无显著相关性。结论:当黄芩中4种主要黄酮类成分的含量构成比为1∶0.224∶0.111∶0.013时,其对RSV的抑制作用较强。道地产区黄芩样品具有较优的黄酮类成分构成特征,该特征可能是道地药材发挥最佳抗病毒疗效的重要物质基础。     

自身酶解法测定黄芩药材中黄芩素和汉黄芩素的含量

目的 探讨一种黄芩药材中黄酮苷元类成分黄芩素和汉黄芩素定量测定方法.方法 通过黄芩自身所含的酶在适宜温度条件下将黄芩药材中的黄芩苷、汉黄芩苷等黄酮苷类成分酶解成黄芩素和汉黄芩素等黄酮苷元,再利用高效液相色谱法进行含量测定.结果 黄芩素酶解完全,色谱峰分离良好,且同一流动相下可同时测定黄芩素和汉黄芩素的含量.黄芩素的平均回收率为96.45%,RSD为1.34%(n=9);汉黄芩素的平均回收率为97.86%,RSD为1.89%(n=9).结论 所用测定方法简便、准确,为黄芩素和汉黄芩素的充分提取、分离提供了真实的基础含量数据.     

自身酶解水提取黄芩中黄芩素及汉黄芩素的方法

目的建立黄芩中黄芩素及汉黄芩素的绿色环保提取方法。方法在适宜温度下,采用自身酶解法,以水为提取溶剂,借助于苷元类、黄芩药材碎块和黄芩苷的粒度和溶解度之差,分离得到黄芩苷元总提物。结果总提物中黄芩总苷元含量在70％以上,其中黄芩素含量为63％以上,汉黄芩素含量为7％以上。结论所用方法简便、快捷、成本低,是一条绿色的无污染环保提取工艺。     

骨疏康胶囊联合利塞膦酸钠治疗绝经后骨质疏松症的临床研究

目的 比较黄芩酒炙前后化学成分变化,为建立全面的黄芩饮片质量评价方法提供参考。方法 HPLC法建立黄芩片、酒黄芩的特征图谱,并生成对照特征图谱,标定共有峰并进行相似度评价。采用UPLC-Q-TOF/MS定性分析黄芩酒炙前后化学成分,利用UPLC-TQMS对2种饮片中11种黄酮类成分（黄芩苷、黄芩素、汉黄芩苷、汉黄芩素、千层纸素A、千层纸苷、野黄芩苷、芹菜素、高车前素、木犀草苷、白杨素）进行定量分析,并以含量为变量进行多元统计分析。结果 建立了黄芩片、酒黄芩的特征图谱,标定9个共有峰,2种饮片相似度均在0.947以上。在黄芩饮片中,共发现50种成分,通过多级质谱数据分析,保留时间匹配,并结合对照品及数据库检索,鉴定了其中44种成分。UPLC-TQMS定量结果显示酒炙后黄芩中黄芩苷、汉黄芩苷等黄酮苷类成分含量略有下降,黄芩素、汉黄芩素等黄酮苷元类成分含量稍有增加。经多元统计学分析,2种饮片有明显的分离趋势,载荷图结果表明黄芩苷、汉黄芩苷、千层纸苷的含量差异可能是引起黄芩酒炙前后化学成分变化的主要因素。结论 黄芩酒炙后没有新增或消失成分,但成分含量有所变化。所建立黄芩饮片的定性、定量方法重现性好,分析快速、准确,可用于黄芩饮片酒炙前后的质量评价。     

不同产地黄芩中6种化学成分含量测定

目的：采用HPLC色谱法同时测定不同产地黄芩中黄芩苷、黄芩素、汉黄芩苷、汉黄芩素、去甲汉黄芩苷、千层纸素A苷的含量。方法：采用Agilent Zorbax SB-C₁₈色谱柱（250 mm×4.6 mm,5 μm）,以乙腈-0.1%磷酸水溶液梯度为流动相,流速1.0 mL·min^-1,检测波长280 nm,柱温30℃。结果：黄芩苷、黄芩素、汉黄芩苷、汉黄芩素、去甲汉黄芩苷、千层纸素A苷的线性关系良好,平均回收率在98.70%~99.17%之间。结论：3批样品测定结果表明,采用该方法可以更简便、更准确定量黄芩总黄酮部位中6种主要化学成分。     

基于一测多评法定量的四季三黄片质量评价研究

目的 建立同时测定四季三黄片中黄芩苷、去甲汉黄芩素苷、千层纸素A-7-O-β-D葡萄糖醛酸苷、汉黄芩苷、黄芩素、汉黄芩素、千层纸素A等7种成分的一测多评法。方法 采用HPLC,岛津VP-ODS色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.1%三氟乙酸溶液为流动相,梯度洗脱;流速1.0 mL·min^-1;柱温30℃;检测波长276 nm。以黄芩苷为内参物,建立去甲汉黄芩素苷、千层纸素A-7-O-β-D葡萄糖醛酸苷、汉黄芩苷、黄芩素、汉黄芩素和千层纸素A的相对校正因子,同时采用一测多评法和外标法测定四季三黄片中7个成分的含量,并将2种方法的测定结果进行比较。结果 7种成分在各自范围内线性关系良好(r>0.999 8),平均加样回收率97.0%～102.1%,RSD为1.2%～2.5%。共测定了4个企业29批四季三黄片中7种成分的含量,结果一测多评法与外标法测定结果无明显差异。不同企业样品间各成分含量存在明显差异。结论 建立的含量方法能够反映出投料用黄芩饮片的质量状况,进而全面有效地评价四季三黄片的质量,为其质量标准的提升奠定基础。     

高效液相色谱法定量分析黄芩苷粗品中4种黄酮类成分

目的：建立黄芩苷粗品中黄芩苷、黄芩素、汉黄芩苷和汉黄芩素含量的高效液相色谱（HPLC）测定方法，以便控制其质量。方法：固定相用Agilent ZORBAX SB-C18柱（5μm，4．6mm×250mm），苷流动相为甲醇-水-冰醋酸（49：51：0．2），苷元流动相为甲醇-水-冰醋酸（61：39：0．2），检测波长为275nm。结果：黄芩苷、黄芩素、汉黄芩苷和汉黄芩素线性范围分别是0．102～1．020μg（r=0．9997），0．051～0．510μg（r=0．9994），0．01～0．51μg（r=0．9995），0．02～0．20μg（r=0．9999）；平均回收率（n=5）分别为99．12％（RSD为1．55％），101．60％（RSD为2．67％），102．27％（RSD为1．92％／40）和98．51％（RSD为3．13％）。结论：本方法简便、准确，可用于黄芩苷粗品的质量控制。     

黄芩及其有效成分的药理学研究进展

综述了黄芩及其有效成分黄芩苷、黄芩苷元、汉黄芩素、汉黄芩苷、黄芩新素Ⅰ和黄芩新素Ⅱ等黄酮类化合物的抗氧化、抗炎、抗变态、抗菌、抗病毒以及抗肿瘤等药理作用。     

黄芩药材的高效液相色谱指纹图谱及黄芩苷、黄芩素、汉黄芩素含量的同时测定

目的: 建立黄芩药材的HPLC指纹图谱,并同时测定不同产地商品中黄芩苷、黄芩素和汉黄芩素的含量,全面评价黄芩药材质量。方法: 采用Aglient TC-C₁₈(4.6 mm×250 mm,5 μm)色谱柱。采用梯度洗脱,流速为1.0 ml·min^-1;检测波长280 nm;柱温30℃。指纹图谱共有模式的建立采用国家药典委员会"中药色谱指纹图谱相似度评价系统2004A版"进行处理分析。结果: 在指纹图谱研究中,标定了9个共有峰,建立了对照指纹图谱,10批药材与共有模式之间相似性良好,相似度均在0.9以上;在所建立的分析条件下,黄芩苷、黄芩素、汉黄芩素均达到了基线分离,且在线性范围内线性关系良好,3种成分的加样回收率分别为99.44%,99.71%和99.83%。结论: 本法灵敏、准确、简便,可用于黄芩药材的综合质量评价。     

Identification and quantification of baicalein, wogonin, oroxylin A and their major glucuronide conjugated metabolites in rat plasma after oral administration of Radix scutellariae product

The current study aims to identify and quantify three flavones (baicalein, wogonin and oroxylin A) and their major metabolites (baicalin, wogonoside and oroxylin A-7-O-glucuronide) in rat plasma after oral administration of Radix scutellariae product. A simple HPLC/UV method has been developed to simultaneously determine the three flavones and their major metabolites in rat plasma. The chromatographic separation of the six analytes was achieved by a Thermo C(18) column with linear gradient elution of a mobile phase containing acetonitrile and 20mM sodium dihydrogen phosphate buffer (pH 4.6). All the tested analytes were detected by PDA detector at a wavelength of 320nm. The intra-day and inter-day precision for the current assay of the six analytes was within the range of -2.23% to 15.13% and -10.83% to 6.42%, respectively. All the studied analytes could be efficiently extracted from the rat plasma using HLB cartridge with extraction recoveries above 70% and were stable under different storage conditions. The developed assay method was successfully applied to the pharmacokinetic study of baicalin, wogonoside after oral administration of a commercially available Radix scutellariae containing capsule at a dose of 3.2g/kg to Sprague-Dawley rats. In addition to wogonoside, a new metabolite of wogonin has been identified using LC/MS/MS for the first time.     

一测多评法测定复方黄白胶囊中四种黄酮成分的含量

目的建立一测多评测定复方黄白胶囊中4种黄酮成分（黄芩苷、汉黄芩苷、黄芩素和汉黄芩素）的方法。方法采用斜率校正法,以黄芩苷作为内参物,计算汉黄芩苷、黄芩素和汉黄芩素与黄芩苷的相对校正因子;分别用一测多评法和外标法测定复方黄白胶囊中4种黄酮成分的含量。结果黄芩苷、汉黄芩苷、黄芩素及汉黄芩素的色谱保留时间分别为14．80、28．58、37．76及47．52。阴性对照在相应位置处未见色谱峰,样品色谱图中黄芩苷、汉黄芩苷、黄芩素及汉黄芩索色谱峰的纯度因子分别为998．8、995．2、989．6及988．5。汉黄芩苷、黄芩素及汉黄芩素与黄芩苷间的相对校正因子分别为1．396、1．808和2．010。复方黄白胶囊中4种黄酮成分采用校正因子计算的含量值与外标法实测值之间无明显差异。黄芩苷、汉黄芩苷、黄芩素及汉黄芩素的加样回收率平均值分别为99．5％、99．7％、100．5％、99．4％,相对标准偏差分别为1．3％、0．9％、1．6％及1．1％（n=6）。本研究表明一测多评法测定黄芩苷、汉黄芩苷、黄芩素及汉黄芩素方法专属性、精密度、稳定性、重复性和回收率均良好。结论一测多评法可用于复方黄白胶囊中4种黄酮成分的定量分析。     

Urinary pharmacokinetics of baicalein,wogonin and their glycosides after oral administration of Scutellariae Radix in humans

Baicalin, baicalein, wogonoside and wogonin are flavone constituents of Scutellariae Radix with various beneficial biological activities. The purpose of this study was to investigate the urinary pharmacokinetics of these flavones after oral administration of Scutellariae Radix commercial powder. Ten healthy male volunteers received a dose of 5.2 g commercial powder (comparable to 9 g crude drug), respectively. The concentrations of baicalin, baicalein and wogonin in the commercial powder as well as their metabolites in urine were assayed by HPLC method. The glucuronides and sulfates of baicalein and wogonin in urine were hydrolyzed with beta-glucuronidase and sulfatase, respectively. Our results showed that the mean cumulated renal excretion of baicalein glucuronides and sulfates were 43.1+/-4.5 micromol (2.9% of dose) and 64.8+/-6.3 micromol (4.3% of dose), respectively, whereas wogonin glucuronides and sulfates were 21.6+/-2.0 micromol (5.9% of dose) and 20.7+/-1.7 micromol (5.7% of dose), respectively. The result indicated that the renal excretion of conjugated metabolites of wogonin (11.6% of dose) were higher than that of baicalein (7.2% of dose). The baicalein sulfates was predominant than the corresponding glucuronides, whereas wogonin sulfates was comparable to the corresponding glucuronides.     

利胆消炎颗粒的质量标准

目的：建立利胆消炎颗粒质量标准。方法：采用薄层色谱法对制剂中黄芩、大黄、枳实、延胡索进行鉴别;采用高效液相色谱法对制剂中黄芩苷进行含量测定。结果：薄层图谱斑点清晰、分离度好、专属性强、阴性无干扰。黄芩苷进样量在30．20～181．20μg·ml^-1范围内与峰面积积分值呈良好线性关系（r=0．9994）;平均加样回收率为98．80％,RSD：1．62％（n=9）。结论：所建标准可用于利胆消炎颗粒的质量控制。     

Different effects of baicalein, baicalin and wogonin on mitochondrial function, glutathione content and cell cycle progression in human hepatoma cell lines

The effects of the flavonoids from Scutellaria baicalensis Georgi (baicalein, baicalin and wogonin) in cultured human hepatoma cells (Hep G2, Hep 3B and SK-Hep1) were compared by MTT assay and flow cytometry. All three flavonoids dose-dependently decreased the cell viabilities accompanying the collapse of mitochondrial membrane potential and the depletion of glutathione content. However, the influence of baicalein, baicalin or wogonin on cell cycle progression was different. All three flavonoids resulted in prominent increase of G2/M population in Hep G2 cells, whereas an accumulation of sub G1 (hypoploid) peak in Hep 3B cells was observed. In SK-Hep1 cells, baicalein and baicalin resulted in dramatic boost in hypoploid peak, but wogonin made mainly in G1 phase accumulation. These data, together with the previous findings in other hepatoma cell lines, suggest that baicalein, baicalin and wogonin might be effective candidates for inducing apoptosis or inhibiting proliferation in various human hepatoma cell lines.     

Flavonoid pharmacokinetics and tissue distribution after repeated dosing of the roots of Scutellaria baicalensis in rats

Scutellariae Radix (root of Scutellaria baicalensis, SR) contains numerous flavonoids such as baicalin, baicalein, and wogonin. This study investigated the pharmacokinetics and tissue distribution of flavonoids and their metabolites in rats after repeated dosing of a SR decoction. Sprague-Dawley rats were orally administered SR at 2 g/kg for seven doses. After the 7th dose, blood samples were withdrawn at specific times and organs, including the liver, kidney, lung, and brain, and collected. The concentrations of baicalein and wogonin in the serum and various tissues were assayed by HPLC before and after hydrolysis with glucuronidase and sulfatase. Baicalein and wogonin were not detected in the serum, and the molecules found were their glucuronides/sulfates. In tissues, the free forms of baicalein and wogonin appeared in the liver, kidney, and lung in addition to their glucuronides/sulfates. Baicalein was the major form in the lung, whereas baicalein glucuronides/sulfates were the major forms in the liver and kidney. Wogonin was the major form in the liver, kidney, lung, and traces of wogonin glucuronides/sulfates were detected in the kidney and liver. Neither baicalein and wogonin nor their glucuronides/sulfates were detected in the brain. In conclusion, the glucuronides/sulfates of baicalein and wogonin were exclusively present in the circulation, whereas their free forms appeared in the lung, liver, and kidney.     

Pharmacokinetics of baicalein,baicalin and wogonin after oral administration of a standardized extract of<Emphasis Type="Italic">Scutellaria baicalensis</Emphasis>, PF-2405 in rats

The pharmacokinetics of active components such as baicalein, wogonin and oroxylin A were evaluated after oral administration of a purified extract of Scutellaria baicalensis GEORGI (PF-2405) containing the high contents of baicalein, wogonin and oroxylin A to rats. Following oral administration of PF-2405 at 10, 20 and 40 mg/kg dose (equivalent to 4.5, 9.0 and 18 mg/kg baicalein), a major constituent baicalein and its active metabolite baicalin showed dose-linear pharmacokinetics as evidenced by unaltered dose-normalized AUC, dose-normalized Cmax, Ae(0-30h) and GI(30h) values. Following oral administration of PF-2405 at three doses (equivalent to 0.4, 0.8 and 1.6 mg/kg wogonin), dose-normalized Cmax and dose-normalized AUC were comparable between the 20 and 40 mg/kg PF2405 doses, but plasma concentrations of wogonin at 10 mg/kg of PF-2405 were not measurable as they were below limit of quantitation (LOQ; 18 pmol/mL). Following oral administration of PF-2405 at the three doses (equivalent to 1.5, 3.0 and 6.0 mg/kg oroxylin A), the concentrations of oroxylin A in plasma, urine and gastrointestine samples were below the assay LOQ (18 pmol/mL). Significant differences in AUCs, Ae(0-30h) and GI(30h) values for baicalein and baicalin were observed after oral administration of pure baicalein (18 mg/kg) and PF-2405 (40 mg/kg). The increases in AUCs of baicalein and baicalin after oral administration of PF-2405 may have been due to the significant decrease in GO(30h) values for baicalein.     

HPLC法同时测定黄芩中黄芩苷、黄芩素、汉黄芩素的含量

目的:建立反相高效液相色谱法同时测定黄芩中黄芩苷、黄芩素、汉黄芩素的含量.方法:采用Agilent Eclipse XDB-C18(250 mm×4.6 mm,5μm)色谱柱,以甲醇-乙腈-0.1％磷酸溶液为流动相,梯度洗脱;检测波长为277 nm;流速1.0 ml·min -1;进样量10 μl;柱温35℃.结果:黄芩苷、黄芩素、汉黄芩素的线性范围分别为0.059 ～ 1.175 μg,r=1.000 0(n =7)、0.010～0.204μg,r=1.000 0(n =7)和8.210×10-3～1.642×10-1 μg,r =1.000 0(n =7);提取回收率分别为99.08％,RSD=0.5％(n=6)、98.66％,RSD =0.5％ (n =6)和98.31％,RSD =0.7％ (n =6).结论:本法操作简便、快速、结果准确,可用于黄芩的质量控制.