HPLC-DAD法测定血满草中熊果酸和齐墩果酸的含量(英文)

建立了HPLC-DAD法测定血满草中熊果酸和齐墩果酸含量,并进行方法学考察。采用HPLC-DAD进行分析,fusion-RP C18柱(4.6 mm×250 mm,4μm),甲醇-0.2%磷酸水溶液(90∶10)为流动相,检测波长210 nm,体积流量1.0 mL/min。同时采用微波辅助提取、回流提取、索氏提取、冷浸提取、超声提取五种方法对血满草中熊果酸和齐墩果酸含量进行测定并比较不同方法所得结果的差异,还比较了血满草不同部位中熊果酸和齐墩果酸的含量差异。测定结果表明熊果酸进样量在3.6～8.4μg范围内,齐墩果酸进样量在3.2～16μg范围内,呈良好线性关系。血满草中熊果酸和齐墩果酸平均回收率分别为98.3%和101.4%(n=5),相对标准偏差分别为1.13%和0.72%(n=5)。五种方法比较得出索氏提取得熊果酸和齐墩果酸含量最高;血满草花中熊果酸和齐墩果酸含量最高,而根中含量最低。该方法使血满草中熊果酸和齐墩果酸达到基线分离,操作简便,结果稳定可靠。

HPLC法测定枇杷止咳胶囊中齐墩果酸与熊果酸的含量

目的:建立枇杷止咳胶囊中齐墩果酸与熊果酸的HPLC方法。方法:采用十八烷基硅烷键合硅胶色谱柱(5μm,4.6mm×250mm),流动相为乙腈:甲醇:0.5%醋酸铵溶液(671221),检测波长210nm,流速0.8mL/min,柱温40℃,进样量10μL。结果:齐墩果酸与熊果酸分别在进样量范围为(0.026～0.130)μg和(0.102～0.510)μg时,线性关系良好,在选定的条件下,齐墩果酸与熊果酸获得较好分离。结论:其方法简便、灵敏,可准确测定枇杷止咳胶囊中齐墩果酸和熊果酸的测定。

夏枯草中熊果酸与齐墩果酸提取工艺研究

主要研究了响应面优化超声辅助提取夏枯草中熊果酸与齐墩果酸工艺。以熊果酸与齐墩果酸含量为指标,通过响应面分析法确定响应面优化超声辅助提取夏枯草中熊果酸与齐墩果酸工艺的最佳工艺条件,即液料比值为19.8(mL/g),超声波时间为29 min,超声波功率为260 W,乙醇体积分数为78%。在此条件下熊果酸与齐墩果酸含量为2.74 mg/g。

Box-Behnken-RSM法优化宣木瓜最佳产地加工工艺的研究

目的 Box-Behnken-RSM法优选宣木瓜最佳产地加工工艺。方法以齐墩果酸和熊果酸的总量为指标,采用Box-Behnken法实验设计考察煮制时间、干燥温度和切片厚度对宣木瓜饮片的影响,优选最佳加工工艺参数。结果宣木瓜最佳工艺为煮制4min,干燥80°C,切片厚度为2.5mm。结论优选的宣木瓜加工工艺合理可行。

败酱片质量标准研究

目的：对败酱片（黄花败酱）建立质量标准。方法：以齐墩果酸、熊果酸为对照品,对方中败酱建立TLC鉴别;用高效液相色谱法对方中齐墩果酸、熊果酸进行测定。色谱条件：迪马Kr C18;流动相：乙腈-甲醇-水-乙酸铵（70∶10∶20∶0.5）;流速：1.0 mL/min;柱温：30℃;检测波长：215 nm。供试品溶液的制备：取样品约4 g,加入乙醇30 mL,置85℃水浴中加热回流提取,分取乙醇提取液15 mL,蒸干,残渣加盐酸溶液（2→10）10 mL水解1 h,用三氯甲烷液萃取,将萃取液蒸干,残渣加甲醇使溶解并定容,即得。结果：齐墩果酸在0.476 8～9.536 0μg范围内呈线性关系,平均加样回收率为99.5%;熊果酸在0.651 0～13.020 0μg范围内呈线性关系,平均加样回收率为103.3%。结论：本法操作简便,结果准确,重现性好,可以控制败酱片的质量。

Effects of ursolic acid and oleanolic acid on human colon carcinoma cell line HCT15

AIM: Ursolic acid (UA) and oleanolic acid (OA) are triperpene acids having a similar chemical structure and are distributed wildly in plants all over the world. In recent years, it was found that they had marked anti-tumor effects. There is little literature currently available regarding their effects on colon carcinoma cells. The present study was designed to investigate their inhibitory effects on human colon carcinoma cell line HCT15. METHODS: HCT15 cells were cultured with different drugs. The treated cells were stained with hematoxylin-eosin and their morphologic changes observed under a light microscope. The cytotoxicity of these drugs was evaluated by tetrazolium dye assay. Cell cycle analysis was performed by flow cytometry (FCM). Data were expressed as means +/-SEM and Analysis of variance and Student' t-test for individual comparisons. RESULTS: Twenty-four to 72 h after UA or OA 60 micromol/L treatment, the numbers of dead cells and cell fragments were increased and most cells were dead at the 72nd hour. The cytotoxicity of UA was stronger than that of OA. Seventy-eight hours after 30 micromol/L of UA or OA treatment, a number of cells were degenerated, but cell fragments were rarely seen. The IC(50) values for UA and OA were 30 and 60 micromol/L, respectively. Proliferation assay showed that proliferation of UA and OA-treated cells was slightly increased at 24h and significantly decreased at 48 h and 60 h, whereas untreated control cells maintained an exponential growth curve. Cell cycle analysis by FCM showed HCT15 cells treated with UA 30 and OA 60 for 36 h and 72 h gradually accumulated in G(0)/G(1) phase (both drugs P【0.05 for 72 h), with a concomitant decrease of cell populations in S phase (both drugs P【0.01 for 72 h) and no detectable apoptotic fraction. CONCLUSION: UA and OA have significant anti-tumor activity. The effect of UA is stronger than that of OA. The possible mechanism of action is that both drugs have an inhibitory effect on tumor cell proliferation through cell-cycle arrest.

Analysis of Oleanolic Acid and Ursolic Acid, Potential Antifertility Agents in Moringa （Moringa oleifera） Seed

Oleanolic acid （OA） and ursolic acid （UA） are isomeric tritcrpenes that are difficult to separate chromatographically. Studies have indicated that both OA and UA have antifertility properties in several livestock species, suppressing attainment of puberty, gonad function and gamete production. Being able to determine the presence and quantity of the OA and UA in moringa and pawpaw seeds will allow for calculation of optimal inclusion levels of these acids in animal diets for antifertility-activity. The purpose of the study was thus to quantify the levels of OA and UA in Moringa and pawpaw seed powder by means of ultra-high performance liquid chromatography with electrospray ionization tandem mass spectrometry （UHPLC-ESI-MS/MS）. Only OA was detected in moringa seeds at a concentration level of 0.508 p.g/g ＋ 0.032 pg/g dry weight. UA and OA could not be detected in pawpaw seeds used in this study. The presence of OA in moringa seeds suggest that the seeds can be included in tilapia diets as a potential antifertility treatment to control precocious breeding in tilapia culture in small scale farming systems in Sub-Saharan Africa. Further studies are warranted to determine the levels of OA and UA composition in moringa seeds from other parts of the Sub-Saharan Africa （other than Zambia）.

木瓜片质量控制

为了提高木瓜片的质量标准,建立木瓜片质量控制的定性与定量方法。方法:采用薄层色谱法对处方木瓜中含有的齐墩果酸和熊果酸进行色谱鉴别;采用醇溶性浸出物测定法中的热浸法测定;采用高效液相色谱法对木瓜中齐墩果酸和熊果酸进行定量分析。结果:薄层色谱图中能清晰检出齐墩果酸和熊果酸;齐墩果酸和熊果酸进样量分别在0.476~1.902 (g(R=0.9991)和0.520~2.079 (g (R=0.9999)线性关系良好;平均回收率分别为98.62%和99.68%,RSD分别为2.18% (n = 9)和2.24% (n = 9)。并初步确定每片含齐墩果酸(C30H48O3)和熊果酸(C30H48O3)的总量不得少于1.25mg。结论:本方法专属性强,操作简便、结果准确可靠,可作为木瓜片的木瓜质量控制标准。