HPLC法测定蜂胶中高良姜素的含量

目的:建立蜂胶中高良姜素的含量测定方法。方法:采用HPLC法,Diamonsil(钻石)C18(46mm×250mm,5μm)色谱柱,甲醇-0.4%的磷酸水(65∶35)为流动相,流速为1.0ml/min,检测波长为256nm。结果:高良姜素在0.00412～0.02472mg/ml范围内呈良好的线性关系(R2=0.9998)。结论:本方法简便、可靠,能用于蜂胶中高良姜素的含量测定。     

HPLC法测定高良姜中高良姜素的含量

目的以高良姜中的专属性成分高良姜素为标准品,用HPLC法测定高良姜素的含量,制定高良姜的质量标准.方法用高效液相法测定了10个不同地区的高良姜样品中高良姜素的含量,色谱柱:Hypersil ODS;检测波长:267 nm;柱温:40℃;流动相:甲醇-0.5%磷酸(58:42);流速:1 mL/min.结果与结论本方法简便、可靠、重现性好,可作为高良姜质量控制的方法.据高良姜素含量的离散程度,提出了高良姜素的限量标准为不少于6.0mg/g,提高了高良姜的质量控制标准.     

高良姜饮片HPLC指纹图谱及高良姜素含量测定研究

目的:采用高效液相色谱法建立高良姜饮片的HPLC指纹图谱,同时测定样品中高良姜素的含量,为其质量控制提供新方法。方法:以Phenom enex C18柱(4.6 mm×250 mm,5μm)为色谱柱,流动相为乙腈-0.2%磷酸溶剂系统,线性梯度洗脱,检测波长为280 nm(高良姜素含量测定为235 nm);采用指纹图谱相似度软件进行数据分析。结果:确定了高良姜饮片9个共有峰,建立了14批药材的共有图谱,并比较了样品中高良姜素的含量。结论:指纹图谱信息结合有效成分含量测定,可较为全面评价高良姜饮片的质量。     

HPCL法测定高良姜中高良姜素的含量

目的：以高良姜中的专属性成分高良姜素为标准品 ，用HPLC法测定高良姜素的含量，制定高良姜的质量标准。方法：用高效液相法测定了10个不同 地区的高良姜样品中高良姜素的含量，色谱柱：HypersilOSD；检测波长：267nm；柱温：40℃；流动相：甲醇－0．5％磷酸（58:42)；流速：1mL/min。结果与结论：本方法简便、可靠、重现性好，可作为高良姜质量控制的方法。据高良姜素含量的离散程度，提出了高良姜素的限量标准为不少于6．0mg/g，提高了高良姜的质量控制标准。     

疏肝和胃颗粒质量标准研究

目的:建立疏肝和胃颗粒的质量标准。方法:采用薄层色谱对元胡、大黄、香附进行定性鉴别;采用高效液相色谱测定制剂中高良姜素的含量,色谱柱为inertsil C18(150mm×4.6mm,5μm);流动相为甲醇-0.2%磷酸溶液(60∶40);检测波长为266nm;柱温31℃,理论塔板数按高良姜素计算应不低于6000。结果:薄层色谱斑点清晰,分离度良好,阴性无干扰,专属性强,重复性良好;高良姜素在0.081~1.62μg/ml(r=0.9999)区间内线性关系良好,平均回收率为97.93%,RSD为1.18%。结论:本方法方便、可靠、准确,可用于疏肝和胃颗粒的质量控制。     

HPLC法测定维药罗补甫克比日丸中高良姜素的含量

目的:高效液相色谱法测定罗补甫克比日丸中高良姜素的含量。方法:甲醇-0.2%磷酸溶液(65∶35);流速lml/min;检测波长266nm;柱温:25℃;进样量:l0μl;分析方法:外标法。结果:高良姜素在0.19~9.76μg范围内线性良好(r=1.0),方法精密度RSD为0.7%(n=5),平均回收率为98.65%,RSD为1.6%(n=6)。结论:该方法简便、准确,测得结果稳定性、重现性好,可作为该制剂中高良姜素的含量测定方法。     

基于UPLC-Triple TOF MS/MS技术比较高良姜根中髓及皮层间的差异

目的:建立高良姜主要成分的UPLC-Triple TOF MS/MS定性与定量检测方法。方法:利用UPLC-Triple TOF MS/MS技术对8批高良姜根中的髓与皮层进行测定分析。在电喷雾负离子及TOF MS触发TOF MS/MS的模式下检测,以一级离子质量准确度、一级离子同位素分布、二级碎片与文献匹配等手段确保准确定性分析,以TOF MS提取离子的峰面积定量分析。结果:高良姜根中髓及皮层间均含黄酮类与二苯基庚烷类成分,但皮层含二苯基庚烷类成分种类较少。髓中高良姜素含量(18.78 mg/g)约为皮层含量(9.01 mg/g)的2倍;二苯基庚烷A含量(3.56 mg/g)约为皮层含量(2.25 mg/g)的1.5倍。结论:该检测方法具有准确、高效的特点;同时可为高良姜药材商品规格等级划分及标准制订提供理论依据。     

高效液相色谱法测定良附微乳口服液中高良姜素的含量

目的:建立高效液相色谱法(HPLC)测定良附微乳口服液中高良姜素含量的方法。方法:采用高效液相色谱法,色谱柱为Diamonsil C18(5μm,150 mm×4.6 mm),以甲醇-0.5%H3PO4(58∶42)为流动相,流速为1.0 mL.min-1,检测波长为360 nm,柱温40℃。结果:高良姜素在(0.005~0.04)mg.mL-1范围内呈良好的线性关系,r=0.999 4,平均回收率为99.0%,RSD为2.0%(n=5)。结论:该方法简便、准确、可靠,可用于良附微乳口服液的质量控制。     

不同地区蜂胶原料药的质量评价

目的:建立不同产地蜂胶的质量评价方法。方法:采用薄层色谱法,以白杨素和高良姜素为对照品,甲苯-乙酸乙酯-甲酸(10∶4∶3)为展开剂,进行定性鉴别研究;采用高效液相色谱法,色谱柱Dikma Technologies(250 mm×4.6mm,5μm);流动相甲醇-0.15%磷酸水溶液(62∶38);流速1.0 mL·min-1;柱温25℃;检测波长268 nm。结果:28个产地蜂胶原料药中白杨素和高良姜素的含量均符合《中国药典》(2010年版)的规定的仅有6个地区,大部分地区蜂胶原料药中高良姜素含量符合标准,而白杨素含量不达标。结论:不同产地蜂胶原料药中白杨素和高良姜素的含量不同,建立的定性、定量方法准确可靠,专属性强,适合不同产地蜂胶原料药的质量控制。     

生长年限及环境因素对高良姜中高良姜素动态积累的影响

目的： 对采自广东、海南等地区的53批次代表性样品中高良姜素的含量进行测定,探讨生长年限及环境因素对高良姜中高良姜素动态积累的影响。 方法： 采用高效液相色谱法测定,ODS-C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相甲醇-0.2%磷酸（55：45）,流速1 mL·min^-1,柱温30℃,检测波长266 nm。 结果： 海南产样品中含量明显高于广东产样品;4年生药材中高良姜素的含量明显高于2,3,5年生样品;在4年生药材中夏末秋初采收样品中高良姜素的含量明显高于初春采收的样品。 结论： 海南栽培高良姜优于广东栽培品,4年生长期为高良姜最佳栽培周期,采收时间应在夏末秋初。     

Chemical composition of the ethanolic propolis extracts and its effect on HeLa cells

Ethnopharmacological relevance

Propolis is a resinous hive product collected by honeybees from various plant sources. It is widely used in traditional medicine and is reported to have a broad spectrum of pharmacological effects (antibacterial, antihepatoxic, antioxidative, anti-inflammatory, etc.). Thus the aim of this study was to assess cytotoxic effect of various ethanol propolis extractions on the cervical tumor cell line (HeLa) and compare it with their phenolic acids and flavonoids composition.

Materials and methods

Twenty samples of raw propolis were collected from 17 localities of Croatia (I-XVII), 2 of Bosnia and Hercegovina (XVIII, XIX) and 1 of Macedonia (XX). Reverse phase HPLC was used to determine the chemical composition of polyphenols. Biological experiments were carried out in vitro on cervix adenocarcinoma cell line (HeLa).

Results

Phenolic acids (ferulic acid, p-coumaric acid, caffeic acid) and flavonoids (tectochrysin, galangin, pinocembrin, pinocembrin-7-methylether, chrysin, apigenin, kaempferol, quercetin) have been determined using HPLC analysis in 20 ethanolic propolis extracts. All samples contain tectochrysin in ranges of 0.1988 mg/g (XVIII) to 1.2004 mg/g (III), while caffeic acid and quercetin have not been found. Flavonoid that is most abundant is galangin in ranges from 0.3706 mg/g (XVII) to 47.4879 mg/g (IX). The samples of propolis numbers I, VI and X applied in the investigated concentration range manifested significant reduction of cell growth. GI₅₀ value as indicator of cytotoxicity among propolis samples showed that propolis number VII is the most effective (GI₅₀ = 76 μg/ml) followed by propolis nos. XV, XVIII and I.

Conclusion

Antiproliferative and cytotoxic effect of propolis on the HeLa cells is not correlating with the concentration of particular components but on establishing the possible synergistic antiproliferative activity of individual phenolic acid and flavonoids. Differences in the chemical composition lead to diversity in biological activity of propolis samples.     

基于UHPLC-MS-MS大通量分析的蜂胶影响PXR/CYP3A4调控血脂质量标志物筛选和评价

目的采用精准、可量化的方法筛选蜂胶影响PXR/CYP3A4通路调控血脂的质量标志物。方法采用人结肠癌LS174T细胞给予一定浓度的咪达唑仑注射液,同时给予不同浓度的蜂胶已知成分对照品溶液,孵育后提取样品,采用UHPLC-MS法测定生成的1′-羟基咪达唑仑含量,根据测定结果筛选出对PXR/CYP3A4通路有显著调控作用的成分,以这些成分为指标,建立同时定量方法,根据结果初步确定蜂胶中影响PXR/CYP3A4通路调控血脂代谢的质量标志物。结果所评价的蜂胶成分中,白杨素、高良姜素、异绿原酸A、槲皮素、咖啡酸苯乙酯均能够显著提高或降低1′-羟基咪达唑仑的量(与对照组和阳性对照组比较),提示上述成分能够显著影响PXR/CYP3A4表达;UHPLC-MS-MS含量测定结果显示,蜂胶中上述成分除异绿原酸A和乔松素外,均具有适宜的含量以达到显效。结论白杨素、高良姜素、咖啡酸苯乙酯和槲皮素可能是蜂胶影响PXR/CYP3A4通路调控血脂的质量标志物,且均具有抑制相关靶标表达,进而调节血脂水平的活性;同时,本研究所用质量标志物筛选、评价方法快速、高效、可量化,能够适应基于PXR/CYP3A4的大通量活性成分筛选研究。     

Comparison of the free radical-scavenging activity of propolis from several regions of Argentina

Propolis is extensively used in Argentine folk medicine. Alcoholic extracts of propolis from different regions of Argentina were prepared. The extracts were analysed for the determination of total flavonoid content (from 13.3 to 42.6 mg/g of propolis) by using the aluminum nitrate method, UV spectrophotometry and thin layer chromatography. All of them contained high total flavonoid content. It was also observed that all samples of ethanolic extracts of propolis showed free radical-scavenging activity in terms of scavenging of the radical DPPH but the highest activities were found for samples from Tucumán and Santiago del Estero. In all cases with 20 microg/ml of soluble principles, the percentage of DPPH degradation was different (Banda Oeste: 67.5%; Verónica: 45%; Forres: 35%; Saenz Pe?a: 20% and Juan José Castelli: 55%). These results may justify their use as a source of natural antioxidants.     

蜂胶毛胶中总黄酮、高良姜素和白杨素含量分析

目的：建立蜂胶毛胶中总黄酮以及高良姜素、白杨素含量测定方法,测定不同基原毛胶中上述各指标的含量。方法：以芦丁为对照品,采用uV法测定总黄酮含量,检测波长为415nm;用HPLC同时测定高良姜素、白杨素含量,采用Welchrom C18柱,以甲醇-0．15％磷酸溶液梯度洗脱,检测波长为268nm。结果：总黄酮在0～1．248mg线性关系良好,r=0．9999（n=6）,平均回收率为98．82％,RSD=1．45％。高良姜素、白杨素分别在0．089～0．4450,0．115～0．580μg线性关系良好,r均达到1．000,平均回收率分别为99．03％、99．32％,RSD分别为1．10％、1．72％。结论：该方法准确、可靠、简便易行,可用于蜂胶毛胶的定量分析,为蜂胶毛胶、其提取物以及蜂胶制品的质量评价和控制提供了依据。     

HPLC法测定不同产地蜂胶中白杨素的含量

目的：测定国内不同产地蜂胶中白杨素的含量。方法：采用HPLC法,Diamomil（钻石）C18（46mm×250mm,5μm）色谱柱,甲醇-0．4％的磷酸水（65：35）为流动相,流速为1．0mL／min,检测波长为2．56nm。结果：白杨素在0．00592—0．03552mg／mL范围内呈良好的线性关系（R^2=0．9999）;白杨素的含量,山东最高,为23．054mg·g^-1,其次是内蒙和河南,分别为14．170mg·g^-1,13．656mg·g^-1,甘肃最低为6．697mg·g^-1。结论：不同产地蜂胶中白杨素的含量有一定差异。     

反相高效液相色谱法测定寻骨风中马兜铃酸A的含量

目的:应用高效液相色谱法测定寻骨风中马兜铃酸A的含量。方法:以Diamonsil C18为色谱柱(250 mm×4.6 mm,5μm)。流动相:甲醇∶水(4∶1);流速:1.0 ml/min;柱温:35℃;检测波长:315 nm。结果:该方法的线性范围为0.005~0.1 mg,进样量与其峰面积之间呈良好的线性关系,r=0.9998,RSD为0.94%(n=4),表明进样精密度良好。结论:本方法操作简便,结果准确,专属性强,适用于寻骨风中马兜铃酸A的含量测定和质量控制。     

HPLC测定尼美舒利含量

 目的：用高效液相色谱法测定尼美舒利的含量。方法：以ODS-C₁₈反相柱、水（1g/LNa₂HP0₄溶液)-甲醉(30：70)为流动相，以二苯甲酮作内标，检测波长254 nm。结果：尼美舒利在10?60/xg/ml的浓度范围内，线性关系良好。回归方程:Y=0.0094+0.0203X,r=0.9998。结论：方法精密，专一，灵敏，且在此条件下，可将尼美舒利与其 含有的杂质较好地分离。     

Analysis of propolis from Baccharis dracunculifolia DC. (Compositae) and its effects on mouse fibroblasts

This paper confirms Baccharis dracunculifolia DC. (Compositae) as the main botanical source of the propolis from southeastern Brazil (state of São Paulo) investigated to ascertain specific biological activity in relation to mouse NIH-3T3 fibroblasts, skin cells directly involved in the cicatrization processes. Flavonoid and total phenolic compounds were determined by spectrophotometry, and chemical composition by HPLC; the chromatographic profile, characterized largely by flavonoids and aromatic acids, was found to be qualitatively similar to that of Baccharis dracunculifolia DC. The adsorption of phenolic compounds in the propolis to skin powder was also investigated, and 68% of these compounds adsorbed to the skin powder. At concentrations from 0.12 to 7.81 μg/ml, the propolis revealed no statistical significant differences from its control solutions; however, at concentrations of 31.25 μg/ml or more, the propolis was toxic to NIH-3T3 cells. Thus, the propolis from Baccharis dracunculifolia DC. (Compositae) presents an in vitro concentration-dependent toxicity on mouse NIH-3T3 fibroblasts.     

菱角皮提取物中鞣酸类化合物的含量测定及其指纹图谱研究

目的 建立菱角皮醋酸乙酯提取物中鞣质类化合物的HPLC含量测定方法,并对提取物HPLC指纹图谱条件进行研究分析.方法 采用反相C18柱(4.6 mm×200 mm,5 μm),甲醇-0.5%乙酸水溶液为流动相,梯度洗脱,柱温为25℃,流速为0.8 ml/min,在276 nm下进行检测.结果 菱角皮醋酸乙酯提取物中鞣质类化合物1,2,6-三没食子酰-β-D-葡萄糖的含量为85.7 mg/g,加样回收率为92.49%;1,2,4,6-四没食子酰-β-D-葡萄糖的含量为244 mg/g,加样回收率为96.41%;初步探讨了菱角皮提取物的指纹图谱条件.结论 采用高效液相法测定菱角皮醋酸乙酯提取物中鞣质类化合物的含量,简便、快速、准确;初步得到分离菱角皮提取物各特征峰的色谱条件,为以后建立指纹图谱和制定菱角皮药材及其活性提取物的质量标准提供了依据.