野生与栽培防风中升麻苷和5-O-甲基维斯阿米醇苷的含量测定及指纹图谱分析

目的:建立并比较野生和栽培品种防风中升麻苷、5-O-甲基维斯阿米醇苷HPLC测定方法和HPLC指纹图谱。方法:含量测定采用岛津C18色谱柱(4.6 mm×150 mm,5μm),流动相为甲醇-水(40∶60),检测波长254 nm,流速1.0 mL.min-1,柱温30℃。HPLC指纹图谱测定采用资生堂MGⅡC18(4.6 mm×250 mm,5μm)色谱柱,流动相为乙腈-水,梯度洗脱,检测波长254 nm,流速1.0 mL.min-1,柱温30℃。结果:升麻苷加样回收率为99.6%,RSD 0.72%(n=6);5-O-甲基维斯阿米醇苷加样回收率为102.6%,RSD 0.88%。野生与栽培品种升麻苷质量分数分别为(4.96±2.59),(3.61±1.82)mg.g-1;5-O-甲基维斯阿米醇苷质量分数分别为(3.91±2.09),(4.37±2.02)mg.g-1。野生与栽培品种升麻苷和5-O-甲基维斯阿米醇苷质量分数无显著性差异。在此条件下所建立的指纹图谱中防风各成分得到有效分离。结论:升麻苷、5-O-甲基维斯阿米醇苷HPLC测定方法简便,结果准确,HPLC指纹图谱分析可作为防风药材质量控制及分类评价的依据。     

HPLC测定防风通圣丸中2个色原酮苷类成分含量

目的:建立防风通圣丸中2个色原酮苷类成分的含量测定方法.方法:采用HPLC法,对防风通圣丸中的升麻素苷、5-O-甲基维斯阿米醇苷进行定量分析.kromasil C18柱(4.6 mm×250 mm,5μm);流动相甲醇-乙腈-0.1％磷酸(24:12:64);检测波长254 nm.结果:升麻素苷、5-O-甲基维斯阿米醇苷分别在0.0125 ～0.250,0.0108 ～0.216 μg(r ＞0.9999)线性关系良好,平均回收率＞96.75％,RSD＜ 1.65％.结论:该研究同时进行升麻素苷、5-O-甲基维斯阿米醇苷的定量分析,可作为防风通圣丸的质量控制标准.     

HPLC测定感冒清热颗粒中升麻苷和5-O-甲基维斯阿米醇苷

目的:建立反相高效液相色谱法测定感冒清热颗粒中的升麻苷及5-O-甲基维斯阿米醇苷含量;方法:采用APEXODS色谱柱,分别以醋酸钠水溶液(40 mmol·L-1,pH 6.9)-甲醇(65:35)和水-甲醇-四氢呋喃(62:38:1)为流动相,检测波长为254nm,流速为0.8mL·min-1;结果:升麻苷与5-O-甲基维斯阿米醇苷分别在0.72～6.5μg·mL-1与0.92～16.5μg·mL-1范围内峰面积与浓度线性关系良好(r=0.9999);加样回收率分别为100.3%和94.7%.测得感冒清热颗粒中含有升麻苷0.133mg·g-1和5-O-甲基维斯阿米醇苷0.167mg·g-1.结论:该法简便、快速、专属性强.     

HPLC法测定蒺藜防白缓释滴丸中升麻素苷、5-O-甲基维斯阿米醇苷和欧前胡素

目的采用高效液相梯度洗脱法测定蒺藜防白缓释滴丸(蒺藜、防风、白芷、当归等)中升麻素苷、5-O-甲基维斯阿米醇苷和欧前胡素的含有量。方法蒺藜防白缓释滴丸甲醇提取液分析在Agilent ZORBAX SB-C_(18)柱(250 mm×4.6 mm,5μm)上实施;体积流量1.0 m L/min;检测波长254 nm。升麻素苷和5-O-甲基维斯阿米醇苷的色谱流动相A为乙腈,流动相B为0.1%磷酸溶液,梯度洗脱(0~10 min,15%A;10~20 min,15%→30%A;20~25 min,30%A);柱温35℃。欧前胡素的色谱流动相为甲醇-水(68∶32,V/V);柱温30℃。结果升麻素苷和5-O-甲基维斯阿米醇苷分别在1.602 0~112.14μg/m L(r=0.999 8)、1.396~139.60μg/m L(r=0.999 9)范围内与峰面积成良好的线性关系,平均回收率分别为100.2%(RSD=0.99%)和99.9%(RSD=0.80%)。欧前胡素在0.707 2~70.72μg/m L(r=0.999 9)范围内与峰面积成良好的线性关系,平均回收率为99.4%(RSD=1.0%)。结论本法操作简便,准确可靠,可用于蒺藜防白缓释滴丸的质量评价。     

HPLC法同时测定藿香清胃胶囊中栀子苷、升麻素苷和5-O-甲基维斯阿米醇苷的含量北大核心CSCD

目的 建立HPLC波长切换法测定藿香清胃胶囊中栀子苷、升麻素苷和5-O-甲基维斯阿米醇苷含量。方法 色谱柱为Agilent HC-C18（4.6 mm×250 mm,5μm）;流动相为甲醇-水,梯度洗脱,流速为1.0m L·min-1,柱温：30℃,波长切换时间序列采样：0-12 min为238 nm;12-45 min为254 nm。结果 栀子苷进样量在0.382-3.819μg范围内线性关系良好（r=0.9999）,平均回收率为99.66%,RSD为1.5%（n=9）;升麻素苷进样量在0.011-0.108μg范围内线性关系良好（r=0.9997）,平均回收率为99.53%,RSD为1.6%（n=9）;5-O-甲基维斯阿米醇苷进样量在0.010-0.097μg范围内线性关系良好（r=0.9998）,平均回收率为100.12%,RSD为1.8%（n=9）。结论 该方法准确、快捷、可靠,为全面控制藿香清胃胶囊的质量提供参考。     

HPLC测定四季感冒片中5-O-甲基维斯阿米醇苷含量

采用高效湘相色谱法测定四季感冒片中5-O-甲基维斯阿米醇苷含量。色谱柱:DiamonsilTM(钻石)C18柱(5μm,200mm×4.6mm);流动相:乙腈-水溶液(1∶5);流速:0.65mL/min。检测波长:290nm。5-O-甲基维斯阿米醇苷对照品进样量在5.05~35.35μg/mL范围内呈现良好线性关系,r=0.9997,平均回收率为101.1%,RSD为1.224%(n=6)。本方法简便,重现性好。     

HPLC测定消痔坐浴剂中5-O-甲基维斯阿米醇苷的含量

目的:建立消痔坐浴剂剂中5-O-甲基维斯阿米醇苷的含量测定方法.方法:采用HPLC法,色谱柱:Shimadzu VP2ODS(250 mm ×4.60 mm,5μm);流动相:乙腈-0.1％磷酸水溶液梯度洗脱;流速:1.0 mL/分;检测波长:254 nm;柱温:30℃,对制剂中5-O-甲基维斯阿米醇苷的含量进行测定.结果:5-O-甲基维斯阿米醇苷的线性范围为2.56-25.62 μg/mL (r=0.9993);平均加样回收率为99.69％(RSD=1.02％).结论:本法简便准确,重现性好,可作为该产品的质量控制方法.     

HPLC法测定补虚颗粒中升麻素苷和5-O-甲基维斯阿米醇苷的含量

目的:建立补虚颗粒中升麻素苷和5-O-甲基维斯阿米醇苷含量的测定方法。方法:采用HPLC法,以Kromasil C18色谱柱,甲醇-水-三乙胺(37∶63∶0.1)为流动相,流速为1.0 mL·min-1,检测波长为254 nm,柱温为35℃。结果:升麻素苷和5-O-甲基维斯阿米醇苷的线性范围分别为2.8-224μg·mL-1(r=0.9995)和3.15-252μg·mL-1(r=0.9993);平均回收率分别为99.52%与100.6%。结论:该方法简单,结果准确、可靠,可以作为该制剂质量控制的一项重要指标。     

HPLC法测定痛泄胶囊中升麻素苷、5-O-甲基维斯阿米醇苷、芍药苷含量

目的：建立痛泄胶囊中升麻素苷、5-O-甲基维斯阿米醇苷和芍药苷含量测定的方法。方法：采用高效液相色谱法在同一色谱条件下[色谱柱：Waters Symmetry C18（250mm×4.6mm,5μm）流动相：乙腈-0.1%磷酸梯度洗脱;流速：1.0ml/min;柱温：30℃]对制剂中3种成分进行定量测定。结果：升麻素苷在47.88~718.2ng范围内线性关系良好,r=0.999 9,平均回收率为99.88%（n=9）,RSD为1.08%;5-O-甲基维斯阿米醇苷在47.52~712.8ng范围内线性关系良好,r=0.999 9,平均回收率为99.68%（n=9）,RSD为0.82%;芍药苷在51~765ng范围内线性关系良好,r=0.999 9,平均回收率为100.82%（n=9）,RSD为0.91%。结论：所建立的方法可在同一色谱条件下检测出3个成分,可用于该制剂的质量控制。     

HPLC法测定痛泄胶囊中升麻素苷、5-0-甲基维斯阿米醇苷、芍药苷含量

目的 建立痛泄胶囊中升麻素苷、5-0-甲基维斯阿米醇苷和芍药苷含量测定的方法.方法 采用高效液相色谱法在同一色谱条件下[色谱柱 Waters Symmetry C18(250mm×4.6mm,5μm)流动相 乙腈-0.1％磷酸梯度洗脱;流速 1.0mL/min;柱温 30℃]对制剂中3种成分进行定量测定.结果 升麻素苷在47.88～718.2ng范围内线性关系良好,r=0.999 9,平均回收率为99.88％ (n =9),RSD为1.08％;5-O-甲基维斯阿米醇苷在47.52～712.8ng范围内线性关系良好,r=0.999 9,平均回收率为99.68％ (n =9),RSD为0.82％;芍药苷在51～765ng范围内线性关系良好,r=0.999 9,平均回收率为100.82％ (n =9),RSD为0.91％.结论 所建立的方法可在同一色谱条件下检测出3个成分,可用于该制剂的质量控制.     

毛霉对柚皮素的硫酸酯化

利用一株毛霉属真菌Mucor sp.对柚皮素（1）进行生物转化,通过各种色谱技术得到3个O-硫酸酯化产物（2~4）,经LC-MSⁿ-IT-TOF和NMR等波谱手段鉴定出化合物2为柚皮素7-O-硫酸酯,化合物3为柚皮素4’-O-硫酸酯,化合物4为柚皮素5-O-硫酸酯。这些结果可为哺乳动物/人体对柚皮素的代谢研究提供参考。     

Analgesic and anti-inflammatory activities of 11-O-galloylbergenin

Aims of the study

Current study was designed to explore the analgesic and anti-inflammatory effects of a constituent isolated from Mallotus philippinensis, in order to validate its folk use.

Materials and methods

11-O-galloylbergenin was isolated from ethanolic extract of Mallotus philippinensis. Analgesic and anti-inflammatory activities of the test compound were assessed using formalin test and carrageenan-induced paw edema models.

Results

11-O-galloylbergenin showed significant analgesic activity at doses of 20 and 40 mg/kg against formalin test in rats. Similarly, 11-O-galloylbergenin exhibited significant anti-inflammatory activity in carrageenan-induced paw edema model at doses of 10, 20 and 30 mg/kg.

Conclusion

11-O-galloylbergenin has demonstrated its significant potential to be further investigated for its discovery as a new lead compound for management of pain and inflammation.     

HPLC测定藏药五脉绿绒蒿中甲氧基淡黄巴豆亭碱的含量

目的:建立藏药五脉绿绒蒿中甲氧基淡黄巴豆亭碱的含量测定方法.方法:Hypersil-Keystone-C18色谱柱(4.6 mm× 250 mm,5μm),柱温20℃,流动相乙腈-0.012％冰醋酸水溶液(50:50),流速1.0 mL·min-1,检测波长237 nm.结果:甲氧基淡黄巴豆亭碱在0.2～2.4 μg呈良好的线性关系(r=0.999 7);平均加样回收率为96.26％,RSD 1.2％.结论:本方法稳定易行、重复性好,可用于藏药五脉绿绒蒿药材的质量评价.     

野菊不同部位绿原酸和3,5-二咖啡酰奎尼酸的含量测定

目的:检测湖南衡阳产野菊不同部位(叶、花、花蕾、嫩茎及老茎)中绿原酸和3,5-二咖啡酰奎尼酸的含量.方法:以80％的乙醇为溶剂,用超声法提取野菊不同部位中的绿原酸和3,5-二咖啡酰奎尼酸.采用高效液相色谱法检测绿原酸和3,5-二咖啡酰奎尼酸的含量,以0.05％磷酸水溶液-乙腈为流动相,梯度洗脱,流速1.0 mL·min-1,检测波长327 nm.结果:绿原酸和3,5-二咖啡酰奎尼酸的质量浓度在10～100 mg·L-1(r =0.996,r =0.992)与蜂面积呈良好的线性关系(n=5),平均回收率(n=6)分别为98.5％,99.7％.野菊叶、花、花蕾、嫩茎及老茎中绿原酸的含量分别为1.08％,0.44％,0.39％,0.58％及0.52％;3,5-二咖啡酰奎尼酸的含量分别为1.66％,1.01％,0.82％,0.93％,0.64％,其中野菊叶中绿原酸和3,5-二咖啡酰奎尼酸的含量最高.结论:野菊不同部位均含有绿原酸和3,5-二咖啡酰奎尼酸,但它们的含量存在明显差异.     

棕榈花蕾化学成分研究

目的:研究棕榈花蕾的化学成分。方法:采用大孔吸附树脂色谱、硅胶色谱、凝胶渗透色谱等方法进行分离纯化,利用理化检验和波谱分析进行化合物结构鉴定。结果:从棕榈花蕾中分离了8个化合物,鉴定其中6个化合物的结构,分别为异香草酸(isovanillic acid,Ⅰ)、β-谷甾醇(β-sitosterol,Ⅱ)、薯蓣皂苷(dioscin,Ⅲ)、5-咖啡酰氧基莽草酸(5-O-caffeoylshikimicacid,Ⅳ)、咖啡酸(caffeic acid,Ⅴ)、芦丁(rutin,Ⅵ)。结论:化合物Ⅰ,Ⅱ,Ⅳ～Ⅵ为首次该植物中分离得到。Ⅰ,Ⅳ为首次从棕榈属植物中分离得到。     

Antinociceptive and anti-inflammatory activities of extract and two isolated flavonoids of Carthamus tinctorius L

Ethnopharmacological relevance

Safflower (Carthamus tinctorius L.) has been long used both in the traditional system and folk medicine as an analgesic anti-inflammatory agent in China. The aim of the study was to evaluate the antinociceptive and anti-inflammatory activities of hydroalcoholic extract (HE) and two isolated kaempferol glycosides of Carthamus tinctorius L. to provide experimental evidence for its traditional use.

Materials and methods

Antinociceptive effects of HE, kaempferol 3-O-rutinoside (K-3-R) and kaempferol 3-O-glucoside (K-3-G) were assessed in mice using the acetic acid-induced writhing test, formalin test and cinnamaldehyde test. The anti-inflammatory effects of HE, K-3-R and K-3-G were determined in two animal models: carrageenan-induced paw edema and xylene-induced ear edema.

Results

The HPLC analysis showed the presence of K-3-R and K-3-G in Carthamus tinctorius L. HE (500 and 1000 mg/kg) as well as K-3-R and K-3-G (150, 300 and 600 mg/kg) produced significant inhibition on nociception induced by acetic acid and formalin. Oral treatment of HE, K-3-R and K-3-G at all doses significantly reduced both the nociceptive response and cinnamaldehyde-induced paw edema, effect that was superior to aspirin. In anti-inflammatory tests, HE and K-3-G significantly inhibited the paw edema during the both phases of carrageenan-induced inflammation while K-3-G suppressed the late phase inflammation only. HE (400 and 800 mg/kg) and K-3-G (200, 400, 800 mg/kg) produced significant dose-dependent inhibition of xylene-induced ear edema development. K-3-R only suppressed ear edema formation at a high dose (800 mg/kg).

Conclusions

These results demonstrate that Carthamus tinctorius L. extract possess remarkable antinociceptive and anti-inflammatory activities which may be due to K-3-R and K-3-G at least in part, supporting the folkloric usage of the plant to treat various inflammatory and pain diseases.     

Antihepatotoxic activity of a quantified Desmodium adscendens decoction and d-pinitol against chemically-induced liver damage in rats

Ethnopharmacological relevance

The isolation of d-pinitol (or 3-O-methyl-d-chiro-inositol) from an aqueous decoction of Desmodium adscendens (Fabaceae) leaves and twigs is reported. The protective and curative effect of this decoction, in which d-pinitol was quantified, and of pure d-pinitol, against chemically-induced liver damage in rats has been evaluated.

Materials and methods

Enzyme levels of aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP), which are among the usual biomarkers for liver damage, were determined in serum samples of experimental animals. The protective effects against d-galactosamine-induced and ethanol-induced liver damage of a decoction of D. adscendens, quantified on its main constituent d-pinitol, was investigated in rats. In addition, the curative effects of pure d-pinitol and D. adscendens against chronic d-galactosamine-induced and acute acetaminophen-induced hepatotoxicity in rats was studied. Silymarin was used as positive control.

Results

In a first experiment evaluating the protective effect against acute d-galactosamine-induced liver damage in rats, a significant decrease of AST and ALT was observed for the D. adscendens decoction at a dose equivalent to 5 mg/kg/day and 20 mg/kg/day d-pinitol, as well as 20 mg/kg/day pure d-pinitol. With respect to chronic ethanol-induced liver damage in rats, the protective effects of D. adscendens at doses equivalent to 2 mg/kg/day and 10 mg/kg/day d-pinitol, were not observed for serum AST and ALT levels. However, with respect to reduced mortality of animals, statistical analysis showed a trend towards significance for the Desmodium group receiving a dose equivalent to 10 mg/kg/day d-pinitol, versus the untreated hepatotoxic animals. Additional experiments in rat models of acute acetaminophen-induced and chronic d-galactosamine-induced liver damage indicated that D. adscendens decoction and pure d-pinitol had no curative effect when given in a dose equivalent to 10 mg/kg/day d-pinitol, or up to 20 mg/kg/day as a pure compound daily, respectively.

Conclusions

The aqueous decoction of D. adscendens showed a protective effect in rats against liver damage induced by d-galactosamine and ethanol, and this effect is at least in part due to the presence of d-pinitol. However, no curative effect of D. adscendens decoction or d-pinitol on liver damage induced by the tested chemicals could be demonstrated.     

裂叶羌活中1个新的天然产物O-甲基羌活醇

目的:研究裂叶羌活Notopterygium,incisum干燥根和根茎甲醇提取物的乙酸乙酯溶性部位化学成分.方法:采用多种色谱方法对其化学成分进行分离和纯化,核磁共振波谱法和质谱法鉴定其结构.结果:从裂叶羌活甲醇提取物的乙酸乙酯溶性部位分离得到5个化合物,分别鉴定为镰叶芹二醇(1),O-甲基二氢镰叶芹酮醇(2),川白芷素(3),东莨菪素(4),O-甲基羌活醇(5).结论:化合物5为新的天然产物;化合物2～4为首次从裂叶羌活根和根茎中分离得到.     

Pharmacokinetics of 8-O-acetylharpagide and harpagide after oral administration of Ajuga decumbens Thunb extract in rats

Ethnopharmacological relevance

Ajuga decumbens Thunb is a medicinal plant native to China popularly used to treat chronic pelvic inflammation and hysteromyoma. Its main bioactive components are iridoid glycosides, such as 8-O-acetylharpagide and harpagide that had presented antibacterial, anti-inflammatory, and antiviral activities.

Aim of the study

To establish a sensitive LC–MS/MS method and compare the pharmacokinetics of 8-O-acetylharpagide and harpagide in rats after oral administration of their pure forms and from compounds obtained from Ajuga decumbens extract.

Materials and methods

Rats received orally 15 mg/kg (equivalent of 6 mg/kg 8-O-acetylharpagide and 1.5 mg/kg harpagide), 30 mg/kg and 60 mg/kg of Ajuga decumbens Thunb extract and were compared to animals that received 12 mg/kg of 8-O-acetylharpagide or 3 mg/kg of harpagide p.o. Concentrations of 8-O-acetylharpagide and harpagide in plasma were determined by LC–MS/MS method at different time points and all pharmacokinetic parameters were estimated by non-compartmental analysis.

Results

Results showed that the iridoid glycosides were quickly absorbed by oral route and showed a dose-dependence profile. Pharmacokinetic parameters of both glycosides were essentially the same except Tmax when dosed as the extract or pure forms.

Conclusion

8-O-acetylharpagide was metabolized to harpagide, which affected the pharmacokinetic profiles of harpagide when dosed as the extract. This pharmacokinetic study seems to be useful for a further clinical study of Ajuga decumbens Thunb extract.