高效液相色谱法分离与测定柴胡中的柴胡皂甙a

采用反相高效液相色谱法对北柴胡中的柴胡皂甙a进行了分离和测定。试验选出分离的最佳条件：流动相为V（乙腈）：V（水）=90：10,流速为0．8mL／min,检测波长为208nm,在0．05—1．50mg／mL范围内柴胡皂甙a的量与峰面积线性关系良好,回归方程为：Y=2258428p＋186676,相关系数r=0．9993,检出限为2．3ng／mL,回收率为98．3％-100．9％。     

三岛柴胡种子化学成分分析

应用ＧＣ／ＭＳ技术对云南省文山县引种的日本三岛柴胡种子的挥发油和脂肪酸成分进行定性定量分析,检出５３个挥发油成分和２８个脂肪酸成分,挥发油以４,４,５－三甲基－２－乙烯、２,２,４－三甲基－３－戊烯－１－醇、２,３－二甲基丁烯－３－二甲基戊烷为主;脂肪酸以十八碳烯酸,十八烷酸,壬二酸和辛二酸为主。从种子的正丁醇提取部分还分离出８个皂甙,其中３个鉴定为柴胡皂甙ｄ,ｃ和６″－Ｏ－乙酰基柴胡皂甙ｄ。     

柴胡中柴胡皂甙a的超声辅助溶剂提取

以溶剂提取法为基础,以超声波为辅助手段,对柴胡中的有效成分进行了提取.通过对提取液中柴胡皂甙a量的测定,研究了超声功率、提取温度、提取时间和样品浸渍时间等条件对提取效果的影响,优选出超声辅助溶剂提取的最佳条件为超声功率50%、提取温度50 ℃、提取时间20 min、样品浸渍时间24 h.实验结果表明,超声辅助溶剂提取比普通的溶剂提取具有更高的提取效率.     

兴安柴胡化学成分研究（Ⅱ）

从兴安柴胡根部乙醚提取部分通过层析方法分得5种化合物,经光谱方法鉴定为6,7,3′,8′-双藁本内酯(B_1),6,7,3a′,6′-双藁本内酯(B_2),顺-6,7-二羟基藁本内酯(B_3),孕甾烯醇酮(B_4),香草醛(B_5),其中B_1为新化合物。     

银洲柴胡化学成份的研究

银洲柴胡(Bupleurum ynchownse Shan et Y. Li)系伞形科柴胡属植物,在我国作为柴胡入药,但它的化学成份尚未见报道。我们对其化学成份进行了研究,分得10个已知化合物。     

不同产地柴胡中微量元素的测定分析

分别采用火焰原子吸收法测定了不同产地柴胡中微量元素钙、镁、铁、铜、锌的含量。结果表明,柴胡中富含对人体有益的钙、镁、铁、铜、锌等微量元素,且不同产地柴胡中各微量元素含量有所差异,为鉴别药材的产地特征及柴胡药效功能的研究和利用提供了依据。     

减压提取与常压提取柴胡挥发油品质比较

对比减压和常压条件下提取的柴胡挥发油在成分、含量及药效方面的差异,考察挥发油的品质,优选提取方法.采用气相色谱-质谱联用(GC-MS)法分析柴胡挥发油成分及其相对含量,再采用气相色谱法建立挥发油中己醛、庚醛、正辛醛和反式-2,4-癸二烯醛的含量检测方法,测定不同压力下提取的柴胡挥发油有效成分含量,通过背部皮下注射酵母混...     

HPLC法测定柴胡地上部分的黄酮苷元

运用HPLC法测定青海不同产地4种柴胡地上部分的黄酮, 为柴胡药材的综合有效利用提供依据. 采用C18色谱柱以V(甲醇):V(0.1% H3PO4)＝1:1为流动相, 流速1 mL/min, 检测波长360 nm. 青海产柴胡地上部分均含丰富的黄酮, 其中以秦岭柴胡黄酮质量分数最高, 槲皮素质量分数达到2.27%, 总黄酮质量分数大于6.50%. 本方法简便, 对黄酮的定量准确, 改变以往柴胡只使用地下部分而弃去地上部分的局面, 提高柴胡药材的有效利用率.     

二元环糊精体系对柴胡中柴胡皂苷a与柴胡皂苷d的毛细管电泳分离测定

以磺丁基-B-环糊精(SBE-β-CD)和β-环糊精(β-CD)组成二元手性选择体系,用毛细管电泳法对柴胡中的柴胡皂苷a及柴胡皂苷d进行分离测定.考察了缓冲液的组成和浓度、手性选择剂的组成和浓度、进样方式及样品介质等对灵敏度和分离度的影响.结果表明:采用熔融石英毛细管柱(60 cm×50 μm i.d,有效长度为53 ...     

气相色谱-串联质谱法同时测定柴胡舒肝丸10种挥发性成分

建立气相色谱-串联质谱法测定柴胡舒肝丸挥发性成分藁本内酯、α-香附酮、十六烷酸、去氢木香内酯、人参炔醇、厚朴酚、和厚朴酚、大黄酚、大黄素甲醚、β-谷甾醇含量的方法。称取5 g柴胡舒肝丸样品,以乙酸乙酯为提取溶剂,样品浸泡30 min,冰水浴超声提取10 min,以气相色谱-串联质谱法对10种挥发性成分定性,内标法定量。10种挥发性成分在对应质量浓度范围内与色谱峰面积线性关系良好,相关系数（r）均大于0.995,样品加标回收率为93.4%～102.8%,测定结果的相对标准偏差为1.2%～5.1%(n=6)。该方法重复性高、稳定性和精密度良好、专属性强、操作简便,适用于柴胡舒肝丸多组分的质量控制。     

High-performance liquid chromatographic analysis of saponin compounds in Bupleurum falcatum

A mixture of saponin compounds (saikosaponin c, a, and d) in the 70% ethanol extract of a powdered sample of Bupleuri radix are analyzed by an Inertsil ODS-3 C(18) column at a flow rate of 1.0 mL/min and detection wavelength of 203 nm. Well resolved chromatograms of saikosaponin c, a, and d are obtained with a gradient elution of acetonitrile-water from 40:60 (v/v) to 50:50 (v/v). The total time required for a single analysis is approximately 20 min. Calibration curves for saikosaponin c, a, and d are linear up to 2.5 mg/mL. The coefficient of variability values for saikosaponins in the extract are below 4%, and the recoveries for saikosaponin c, a, and d are 95.2 +/- 1.1, 96.5 +/- 0.9, and 96.2 +/- 1.0%, respectively. The changes in saikosaponin contents for a two-year growth of Bupleurum falcatum are measured by the established high-performance liquid chromatography method.     

Corticosterone secretion-inducing activity of saikosaponin metabolites formed in the alimentary tract

The corticosterone secretion-inducing activities of saikosaponin a, saikosaponin c and saikosaponin d, isolated from the root of Bupleurum falcatum L., and 27 metabolites formed in the murine alimentary tract were studied in mice. Serum corticosterone was determined by high-performance liquid chromatography (HPLC). Intraperitoneal administration of saikosaponin a and its intestinal metabolite, prosaikogenin F, showed corticosterone secretion-inducing activity at a dose of 0.1 mmol/kg, and maximally increased it at a dose of 0.4 mmol/kg. On the other hand, the genuine sapogenin, saikogenin F, was inactive. Saikosaponin b1 and saikosaponin g, gastric metabolites of saikosaponin a, and their intestinal metabolites, prosaikogenin A, prosaikogenin H, saikogenin A and saikogenin H, were also inactive. Serum corticosterone was increased by the administration of saikosaponin d and its intestinal metabolite, prosaikogenin G, at a dose of 0.04 mmol/kg, and it reached the maximal level at the dose of 0.1 mmol/kg. Saikogenin G also showed a slight activity. A gastric metabolite of saikosaponin d, saikosaponin b2, and its intestinal metabolites, prosaikogenin D and saikogenin D, were inactive. In the experiments on saikosaponin c and its metabolites, saikosaponin c was inactive but its intestinal metabolites, especially prosaikogenin E-2, showed activity almost equal to that of saikosaponin a. Saikosaponin h and saikosaponin i, gastric metabolites of saikosaponin c, were also inactive, but their prosaikogenins showed slight activities. When these compounds were orally administered, their corticosterone secretion-inducing activities were similar to those obtained in the intraperitoneal experiment. These results suggest that a proper polar balance between the sugar moiety and the aglycone is important for the corticosterone secretion-inducing activity of saikosaponins and their metabolites.     

Optimization of supercritical fluid extraction of saikosaponins from Bupleurum falcatum with orthogonal array design

Supercritical fluid extraction (SFE) was used to extract saikosaponins a, c and d from the root of Bupleurum falcatum. An orthogonal array design L₉(3)⁴ was employed as a chemometric method for the optimization of the SFE conditions. The effects of four factors including pressure (30–40 MPa), temperature (40–50°C), ethanol concentration (60–100%) and time (2.5–3.5 h) on the yields of saikosaponins were investigated by a preparative SFE system in the SFE mode. Under the optimized conditions, namely 35 MPa of pressure, 45°C of temperature, 80% of ethanol concentration and 3.0 h of time, the yields of saikosaponin c, saikosaponin a, saikosaponin d, total saikosaponins and SFE extract were 0.16, 0.12, 0.96, 1.24 and 16.48 mg/g, respectively. Determinations of the saikosaponins were performed by HPLC.     

Characterization and discrimination of raw and vinegar‐baked Bupleuri radix based on UHPLC‐Q‐TOF‐MS coupled with multivariate statistical analysis

Bupleuri Radix is a commonly used herb in clinic, and raw and vinegar‐baked Bupleuri Radix are both documented in the Pharmacopoeia of People's Republic of China. According to the theories of traditional Chinese medicine, Bupleuri Radix possesses different therapeutic effects before and after processing. However, the chemical mechanism of this processing is still unknown. In this study, ultra‐high‐performance liquid chromatography with quadruple time‐of‐flight mass spectrometry coupled with multivariate statistical analysis including principal component analysis and orthogonal partial least square‐discriminant analysis was developed to holistically compare the difference between raw and vinegar‐baked Bupleuri Radix for the first time. As a result, 50 peaks in raw and processed Bupleuri Radix were detected, respectively, and a total of 49 peak chemical compounds were identified. Saikosaponin a, saikosaponin d, saikosaponin b₃, saikosaponin e, saikosaponin c, saikosaponin b₂, saikosaponin b₁, 4′′‐O‐acetyl‐saikosaponin d, hyperoside and 3′,4′‐dimethoxy quercetin were explored as potential markers of raw and vinegar‐baked Bupleuri Radix. This study has been successfully applied for global analysis of raw and vinegar‐processed samples. Furthermore, the underlying hepatoprotective mechanism of Bupleuri Radix was predicted, which was related to the changes of chemical profiling.     

Separation of triterpenoid saponins from the root of Bupleurum falcatum by counter current chromatography: The relationship between the partition coefficients and solvent system composition

A separation method using counter current chromatography coupled with an evaporative light‐scattering detection system was developed to purify five triterpenoid saponins from the roots of Bupleurum falcatum. The methanol extract was loaded onto a Diaion® HP20 column and fractionated by a methanol and water gradient elution. The saikosaponin‐enriched fraction was obtained by elution with 100% methanol. The two‐phase solvent systems used for separation were composed of chloroform/methanol/isopropanol/water at a volume ratio of 60:60:1:60 and 6:6:1:6. The relationship between the isopropanol ratio of each phase and the partition coefficients of the target compounds was investigated by calculating partition coefficient by high‐performance liquid chromatography and measuring the accurate composition of each phase by ¹H NMR spectroscopy. Each fraction obtained was collected and dried, which yielded the following five saikosaponins from 700 mg of injected sample: saikosaponin B₁ (8.7 mg), saikosaponin A (86 mg), saikosaponin B₃ (17 mg), saikosaponin B₂ (41 mg), and saikosaponin C (33 mg). Saikosaponin A showed the most potent cytotoxicity against human cancer cells (gastric cancer, AGS cells; breast cancer, MCF‐7 cells; and hepatoma, HepG2 cells) after 24 h. The IC₅₀ values for the above three cell types were 34.6, 33.3, and 23.4 μmol/L, respectively.     

A high-performance liquid chromatographic method for saikosaponin a quantification in rat plasma

A high-performance liquid chromatographic method with UV detection has been developed for the determination of saikosaponin a in rat plasma. Saikosaponin a and internal standard jujuboside A were isolated from plasma samples by solid-phase extraction. The chromatographic separation was achieved on a reversed-phase C(18) column with the mobile phase of acetonitrile-water (35:65, v/v) at a flow rate of 1 mL/min and UV detection was set at 205 nm. The standard curve for saikosaponin a was linear over the concentration range 0.25-10 microg/mL and the limit of detection was 0.05 microg/mL. The absolute recovery was greater than 82%. The precision and accuracy ranged from 3.05 to 9.59% and 95.61 to 110.00%, respectively. The validated method was used to determine saikosaponin a in plasma samples in a pharmacokinetic study of saikosaponin a administered to Sprague-Dawley rats.     

Analysis of saikosaponins in rat plasma by anionic adducts-based liquid chromatography tandem mass spectrometry method

Saikosaponins (SSs) are a class of triterpene saponins with a wide spectrum of bioactivities. A sensitive liquid chromatography tandem mass spectrometry (LC‐MS/MS) method was developed for simultaneous determination of saikosaponin a, saikosaponin c, saikosaponin d and saikosaponin b₂ in rat plasma. Plasma samples were prepared by liquid–liquid extraction. The analytes and the internal standard (IS) digoxin were well separated on an octadecyl column using gradient elution and analyzed by monitoring the fragmentation transition pair of anionic adducts to deprotonated molecules in negative‐mode electrospray. By neutral loss of HCOOH, the transition pairs of m/z 825 → 779 for SSa, SSd, SSb₂ and the IS, and m/z 971 → 925 for SSc were sensitive for MS/MS detection with the lower limits of quantification in the range of 0.20–0.40 ng/mL. Method validation experiments were performed, including selectivity, precision, accuracy, linearity, matrix effect, recovery and stability. The validated method was further applied to determine the pharmacokinetics parameters of SSa, c and d in rats following a single oral administration of the extract of chaihu (the dried roots of Bupleurum chinense DC). Copyright © 2011 John Wiley & Sons, Ltd.     

Effects of saikosaponin metabolites on the hemolysis of red blood cells and their adsorbability on the cell membrane

The hemolytic properties and the adsorbability on red blood cells of saikosaponin a, saikosaponin d and 13 metabolites formed in the alimentary tract were investigated. Among these compounds, saikosaponin d and its intestinal product, prosaikogenin G, which possess an alpha-hydroxyl function at C16, showed the strongest hemolytic activity at the dose range of 1.0 to 5.0 micrograms/ml. Saikosaponin a and its intestinal product, prosaikogenin F, which possess a beta-hydroxyl function at C16, showed activity above 10 micrograms/ml. In this case, the monoglycoside, prosaikogenin F, showed the stronger activity than the diglycoside, saikosaponin a. Among the gastric products whose ether ring was cleaved to produce a carbinol, the monoglycosides, prosaikogenin A and prosaikogenin H, showed a slight activity above 25 micrograms/ml, and the saikogenins except saikogenin A were inactive. Saikogenin A, however, had hemolytic activity at a dose of 15 micrograms/ml. The adsorbabilities of these compounds on red blood cell membranes closely paralleled their degrees of hemolytic activity.