AIM: To develop and validate a high performance liquid chromatography(HPLC) coupled with diode array and evaporative light scattering detectors(DAD-ELSD) method for the quantitative determination and fingerprint analy...AIM: To develop and validate a high performance liquid chromatography(HPLC) coupled with diode array and evaporative light scattering detectors(DAD-ELSD) method for the quantitative determination and fingerprint analysis of ten active constituents in three chemical classes(namely, xanthone glycosides, steroidal saponins, and alkaloids) in Zhimu-Huangbai herb pair(ZB). METHOD: Chromatographic separation was performed on a Diamonsil C18 column(4.6 mm × 250 mm, 5 μm, Dikma) by gradient elution using acetic acid in acetonitrile solution at a flow rate of 1.0 mL·min–1 at 260 nm. The drift tube temperature of ELSD was set to 60 ℃ and nebulizer gas pressure was 4.0 Bar. Method validation was performed to assure its linearity, limits of detection and quantification, precision, repeatability, stability, and accuracy. RESULTS: The HPLC-DAD-ELSD method allowed the quantification of ten compounds(phellodendrine, jatrorrhizine, palmatine, berberine, neomangiferin, mangiferin, timosaponin E-I, timosaponin B-II, timosaponin B, and timosaponin A-III), and was successfully applied to fingerprint analysis for ten batches of ZB samples. CONCLUSION: This was the first time to apply the combination of DAD and ELSD for the simultaneous determination of ten active ingredients in ZB. The results showed that the combination of quantitative analysis for marker ingredients and chemical fingerprint for the TCM herb pair provides a potentially powerful, widely introduced, and internationally accepted strategy for assessment of complex TCM formulas.展开更多
Objective： To investigate whether the dried root of Phellodendron amurense Ruprecht（Phellodendri cortex; PC） extract improves arthritic symptoms through anti-inflammatory and immune-modulatory effects in collagen-i...Objective： To investigate whether the dried root of Phellodendron amurense Ruprecht（Phellodendri cortex; PC） extract improves arthritic symptoms through anti-inflammatory and immune-modulatory effects in collagen-induced arthritis in mice. Methods： Rheumatoid arthritis（RA） was induced in male DBA/1 mice by immunization with type Ⅱ collagen（ColⅡ）. CIA mice were divided into 5 groups（n=10 per a group） with normal, CIA control, PC extract（50 mg/kg and 100 mg/kg）-treated, and meloxicam（50 mg/kg）-treated as the reference drug. The PC extract or meloxicam were administered orally in CIA mice once a day for 14 days after arthritis induction. Arthritic score, levels of anti-ColⅡ IgG2a antibody, prostaglandin E2（PGE2）, tumor necrosis factor（TNF）-α, and interleukin（IL）-17 in the sera of CIA mice were measured. Histopathological changes in the ankle joints of CIA mice were also analyzed by staining with hematoxylin and eosin（H and E）, safranin-O and immunohistochemistry using anti-TNF-α and anti-IL-17 antibodies. Results： The arthritic score was increased in CIA mice in a time-dependent manner, as were the serum levels of anti-ColⅡ IgG2a antibody, PGE2, TNF-α, and IL-17. However, the oral administration of PC extract at 50 and 100 mg/kg in CIA mice significantly decreased the arthritic scores, and the serum levels of anti-ColⅡ IgG2a, PGE2, TNF-α, and IL-17 compared with those in the CIA group（P〈0.05 or P〈0.01）. Furthermore, histopathological improvement of the joint architecture in CIA mice was observed after administration of PC extract. PC extract also significantly inhibited the expression of TNF-α and IL-17 in the joints of CIA mice by suppressing the expression of their m RNA and proteins. Conclusion： PC extract may improve the pathological progression of RA through the inhibition of joint destruction by synovial inflammation and immune-stimulation, therefore, it would be a potential anti-arthritic agent in RA.展开更多
A new sesterterpenoid phellogine was isolated from the fruits of Phellodendron chinense var. glabriusculum Schneid. Its structure was elucidated on the basis of spectral analysis.
文摘目的:研究黄柏炮制前后17种氨基酸的含量变化情况进而评价黄柏炮制前后的寒热药性的变化。方法:采用反相高效液相色潽法,以异硫氰酸苯酯(PITC)为衍生化试剂衍生后进行HPLC分析。采用C18(250mm×4.6 mm,5μm)色谱柱,柱温为43℃;流动相A为0.1 mol·L-1醋酸钠缓冲液(以醋酸调p H 6.5)-乙腈(93∶7),流动相B为乙腈-水(4∶1),梯度洗脱,流速为1.0 m L·min-1;检测波长为254 nm。结果:17种氨基酸浓度在线性范围内呈良好的线性关系(r=0.996 8~0.999 5);回收率在96.7%~104.26%之间,RSD均小于3.0%。结论:本方法灵敏、准确,具有良好的重复性和稳定性,可用于黄柏中氨基酸的检测。
基金supported by the Natural Science Foundation of Shanghai City,China(No.10411969800)the National Nature Science Foundation of China(Nos.81202866 and 81302856)
文摘AIM: To develop and validate a high performance liquid chromatography(HPLC) coupled with diode array and evaporative light scattering detectors(DAD-ELSD) method for the quantitative determination and fingerprint analysis of ten active constituents in three chemical classes(namely, xanthone glycosides, steroidal saponins, and alkaloids) in Zhimu-Huangbai herb pair(ZB). METHOD: Chromatographic separation was performed on a Diamonsil C18 column(4.6 mm × 250 mm, 5 μm, Dikma) by gradient elution using acetic acid in acetonitrile solution at a flow rate of 1.0 mL·min–1 at 260 nm. The drift tube temperature of ELSD was set to 60 ℃ and nebulizer gas pressure was 4.0 Bar. Method validation was performed to assure its linearity, limits of detection and quantification, precision, repeatability, stability, and accuracy. RESULTS: The HPLC-DAD-ELSD method allowed the quantification of ten compounds(phellodendrine, jatrorrhizine, palmatine, berberine, neomangiferin, mangiferin, timosaponin E-I, timosaponin B-II, timosaponin B, and timosaponin A-III), and was successfully applied to fingerprint analysis for ten batches of ZB samples. CONCLUSION: This was the first time to apply the combination of DAD and ELSD for the simultaneous determination of ten active ingredients in ZB. The results showed that the combination of quantitative analysis for marker ingredients and chemical fingerprint for the TCM herb pair provides a potentially powerful, widely introduced, and internationally accepted strategy for assessment of complex TCM formulas.
基金supported by the research fund from Korean Medicine R&D Center, Dongguk University, Republic of Korea
文摘Objective： To investigate whether the dried root of Phellodendron amurense Ruprecht（Phellodendri cortex; PC） extract improves arthritic symptoms through anti-inflammatory and immune-modulatory effects in collagen-induced arthritis in mice. Methods： Rheumatoid arthritis（RA） was induced in male DBA/1 mice by immunization with type Ⅱ collagen（ColⅡ）. CIA mice were divided into 5 groups（n=10 per a group） with normal, CIA control, PC extract（50 mg/kg and 100 mg/kg）-treated, and meloxicam（50 mg/kg）-treated as the reference drug. The PC extract or meloxicam were administered orally in CIA mice once a day for 14 days after arthritis induction. Arthritic score, levels of anti-ColⅡ IgG2a antibody, prostaglandin E2（PGE2）, tumor necrosis factor（TNF）-α, and interleukin（IL）-17 in the sera of CIA mice were measured. Histopathological changes in the ankle joints of CIA mice were also analyzed by staining with hematoxylin and eosin（H and E）, safranin-O and immunohistochemistry using anti-TNF-α and anti-IL-17 antibodies. Results： The arthritic score was increased in CIA mice in a time-dependent manner, as were the serum levels of anti-ColⅡ IgG2a antibody, PGE2, TNF-α, and IL-17. However, the oral administration of PC extract at 50 and 100 mg/kg in CIA mice significantly decreased the arthritic scores, and the serum levels of anti-ColⅡ IgG2a, PGE2, TNF-α, and IL-17 compared with those in the CIA group（P〈0.05 or P〈0.01）. Furthermore, histopathological improvement of the joint architecture in CIA mice was observed after administration of PC extract. PC extract also significantly inhibited the expression of TNF-α and IL-17 in the joints of CIA mice by suppressing the expression of their m RNA and proteins. Conclusion： PC extract may improve the pathological progression of RA through the inhibition of joint destruction by synovial inflammation and immune-stimulation, therefore, it would be a potential anti-arthritic agent in RA.
基金Talent Special Fund of Guizhou Province(No.200560 and No.2005247)for financial supportedprofessor Jianxin Zhang of analytical group of The Key Laboratory of Chemistry for Natural Products of Guizhou Province and Chinese Academy of Sciences for the spectral measurements.
文摘A new sesterterpenoid phellogine was isolated from the fruits of Phellodendron chinense var. glabriusculum Schneid. Its structure was elucidated on the basis of spectral analysis.
