HPLC测定舒筋活络酒中升麻苷和5-O-甲基维斯阿米醇苷的含量

目的:建立舒筋活络酒中升麻苷和5-O-甲基维斯阿米醇苷的测定方法,以控制制剂的质量。方法:采用HPLC测定舒筋活络酒中升麻苷和5-O-甲基维斯阿米醇苷的含量。Kromasil C18柱(4.6 mm×250 mm,5μm);流动相乙腈-0.05%磷酸溶液为流动相梯度洗脱;检测波长254 nm;流速1.0 mL·min-1;柱温35℃,进样量为10μL。结果:升麻苷在0.030 1～1.504 5μg,5-O-甲基维斯阿米醇苷进样量在0.040 8～2.040 0μg,与升麻苷和5-O-甲基维斯阿米醇苷总量峰面积呈良好的线性关系,平均回收率为升麻苷99.46%,RSD 2.64%,5-O-甲基维斯阿米醇苷100.45%,RSD 2.54%(n=9)。结论:本方法简便、快速、准确,可作为该制剂的定量分析方法。     

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.     

肠道菌群法研究黄芪对防风中升麻素苷、5-O-甲基维斯阿米醇苷代谢的影响

目的采用肠道菌群法研究黄芪对防风中主要活性成分升麻素苷、5-O-甲基维斯阿米醇苷代谢的影响,从而探讨防风-黄芪药对的配伍机制。方法采用肠道菌群法,比较不同时间点防风组和防风-黄芪组以及防风单体组和防风单体-黄芪组中升麻素苷、5-O-甲基维斯阿米醇苷、升麻素的质量浓度。结果防风-黄芪组中升麻素苷、5-O-甲基维斯阿米醇苷的质量浓度均高于防风组,升麻素的质量浓度均低于防风组。单体-黄芪组中升麻素苷、5-O-甲基维斯阿米醇苷的质量浓度均高于单体组,升麻素的质量浓度均低于单体组。结论黄芪能抑制防风中升麻素苷和5-O-甲基维斯阿米醇苷的水解反应,减缓其代谢。     

不同生长方式防风色原酮物质含量测定及差异性分析

目的：以同时测定防风中的两种色原酮物质升麻素苷和5-O-甲基维斯阿米醇苷为目标,建立了上述两种物质的的含量测定方法。方法：色谱柱采用Agilent TC-C18（250 mm×4.6 mm,5 μm）;梯度洗脱的流动相为水、甲醇,检测波长254 nm,流速1 mL/min,柱温35 ℃,进样量10 μL。结果：不同生长方式的防风样品的5-O-甲基维斯阿米醇苷含量无统计学意义,野生和栽培防风的升麻素苷含量有差异,不同生长方式对防风样品的色原酮含量有差异。结论：野生防风质量较好,同时测定两种色原酮方法简便,结果准确,精密度、重现性、稳定性较好,可以此作为防风中升麻素苷和5-O-甲基维斯阿米醇苷的测定方法。     

玉屏风方饮片及汤剂中毛蕊异黄酮苷、升麻素苷、5-O-甲基维斯阿米醇苷的含量相关性研究

目的:建立玉屏风汤剂中毛蕊异黄酮苷、升麻素苷、5-O-甲基维斯阿米醇苷的含量测定方法,研究黄芪、防风饮片及玉屏风汤剂中此3种成分的含量相关性。方法:采用HPLC法,色谱柱为Hypersil ODS;流动相为乙腈-水(梯度洗脱);检测波长为254nm;流速为1mL·min-1;柱温为30℃。结果:黄芪中的毛蕊异黄酮苷、防风中的升麻素苷与5-O-甲基维斯阿米醇苷的提取率分别为45.58%~63.35%,45.01%~65.09%,53.62%~87.75%。结论:饮片与汤剂中的含量呈现一定相关性,但不同批次饮片的提取率差异性较大;为玉屏风汤剂物质基础的研究提供了一定的参考。     

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测定感冒清热颗粒中升麻苷和5—0—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(6535)和水-甲醇-四氢呋喃(62381)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。     

升麻素苷和5-O-甲基维斯阿米醇苷大鼠在体鼻循环吸收研究

对升麻素苷和5-O-甲基维斯阿米醇苷大鼠在体鼻黏膜吸收特性,为头痛宁鼻腔喷雾剂的给药方式提供依据。采用大鼠在体鼻腔循环灌流法,以pH为6的缓冲溶液配制的高、中、低浓度的升麻素苷、5-O-甲基维斯阿米醇苷混合溶液作为鼻腔循环液,进行大鼠在体鼻黏膜吸收试验,用HPLC测定各循环液中待测成分的浓度,并考察了不同pH条件下升麻素苷和5-O-甲基维斯阿米醇苷的吸收速率。结果显示,升麻素苷高、中、低浓度的大鼠在体吸收速率常数为(0.588±0.041)×10~(-3),(0.547±0.023)×10~(-3),(0.592±0.063)×10~(-3)min~(-1)。5-O-甲基维斯阿米醇苷高中低浓度的吸收速率常数为(0.438±0.041)×10~(-3),(0.407±0.023)×10~(-3),(0.412±0.063)×10~(-3)min~(-1)。高、中、低组间无显著性差异。两者在pH为6的情况下吸收情况较其他p H条件好。推测头痛宁鼻腔喷雾剂中的主要活性成分可经鼻黏膜吸收,制成鼻腔喷雾剂是可行的,头痛宁鼻腔喷雾剂p H控制在6是科学合理的。     

基于小粒径色谱柱的哮喘颗粒HPLC指纹图谱研究及多成分快速测定

目的采用小粒径色谱柱建立哮喘颗粒(XCG)HPLC指纹图谱,同时快速测定其中7个有效成分(莫诺苷、苦杏仁苷、马钱苷、升麻素苷、5-O-维斯阿米醇苷、木兰脂素、五味子醇甲)的方法。方法采用HPLC,色谱柱为Kromasil C18(150 mm×4.6 mm,3.5μm);流动相为甲醇-乙腈-水;体积流量1.0 m L/min;梯度洗脱;检测波长:210 nm(苦杏仁苷、木兰脂素)、240 nm(莫诺苷、马钱苷、升麻素苷、5-O-维斯阿米醇苷、五味子醇甲及指纹图谱);柱温40℃;采用UPLC-Q-TOF/MS结合对照品对共有峰进行指认。结果得到分离度和重现性良好的HPLC指纹图谱,标定了20个共有峰,10批样品相似度在0.90以上;采用UPLC-Q-TOF/MS对共有峰进行指认,11个成分经对照品比对,分别为莫诺苷、苦杏仁苷、马钱苷、升麻素苷、5-O-维斯阿米醇苷、木兰脂素、五味子醇甲、五味子醇乙、五味子酯甲、五味子甲素、五味子乙素,并对其中与功效相关的成分莫诺苷、苦杏仁苷、马钱苷、升麻素苷、5-O-甲基维斯阿米醇苷、木兰脂素、五味子醇甲进行定量分析,平均回收率在97.3%~103.8%(RSD均小于2.0%),10批次样品中7个成分量分别为莫诺苷0.51~0.69 mg/g,苦杏仁苷5.01~5.95 mg/g,马钱苷1.02~1.33 mg/g,升麻素苷0.35~0.45 mg/g,5-O-维斯阿米醇苷0.45~0.55 mg/g,木兰脂素0.38~0.48 mg/g,五味子醇甲0.89~1.08 mg/g,各成分量的RSD均小于11.0%。结论建立的指纹图谱和7个成分定量测定的方法,快速、准确、可靠,可以用于XCG的质量评价。     

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.     

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。     

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.     

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.     

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.     

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.     

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.     

筋骨草提取物有效成分在beagle犬体内药代动力学研究

乙酰哈巴苷和哈巴苷是筋骨草提取物中含量最高的2种有效成分.该文建立了LC-MS/MS法,对beagle犬口服筋骨草提取物后乙酰哈巴苷和哈巴苷的药代动力学进行研究.健康雌性Beagle犬口服不同剂量的筋骨草提取物,对不同时间点的乙酰哈巴苷和哈巴苷血药浓度进行测定,采用winnonlin 6.3软件以非房室模型进行拟合.色谱条件:安捷伦ZORBAX XDB-C₁₈柱 (2.1 mm×50 mm,3.5 μm);柱温30 ℃;流动相(A为0.1%甲酸水溶液;B为乙腈)为0~2 min 5% B;2.1~5 min 95% B;5.1~10 min 5% B.流速0.3 mL·min^-1.质谱条件:电喷雾离子源;正离子模式;选择离子分别为乙酰哈巴苷(m/z 429.2→369.2)、哈巴苷(m/z 387.2→207.2)和肉桂酸(m/z 149.2→103.1).干燥气流速10 L·min^-1;干燥气温度300 ℃;毛细管电压4 000 V.结果表明Beagle犬口服筋骨草提取物后乙酰哈巴苷和哈巴苷存在剂量依赖性,乙酰哈巴苷和哈巴苷的达峰时间分别为1.7,1.57 h左右,远大于大鼠口服筋骨草提取物后的乙酰哈巴苷和哈巴苷的达峰时间,这可能是由于种属差异造成的.