4种市售黑豆及成品淡豆豉中异黄酮含量分析

目的 比较4种市售黑豆及发酵后淡豆豉中6种异黄酮的含量。方法 采用高效液相色谱法测定4种黑豆及发酵后淡豆豉中大豆苷、黄豆黄苷、染料木苷、大豆苷元、黄豆黄素、染料木素的含量。结果 4种黑豆中6种黄酮类成分的含量差异较大,黄心小黑豆中3种黄酮苷类成分含量最高,黄心大黑豆次之,绿心黑豆黄酮苷类成分含量最低;发酵后的淡豆豉中6种黄酮类成分含量也有明显差异,其中以黄心小黑豆中黄酮苷元含量最高,黄心大黑豆次之,绿心黑豆苷元含量最低。结论 黄心小黑豆为淡豆豉发酵的较优原料,该结果可为淡豆豉的生产和原料控制提供参考。     

淡豆豉不同制法其成品性状及异黄酮类化学成分含量的比较

目的:比较古法炮制与现今药典制法的淡豆豉其成品性状及异黄酮类化学成分的含量。方法:采用紫外-可见分光光度法对淡豆豉提取物中的总异黄酮进行含量测定;采用高效液相色谱法对淡豆豉提取物中异黄酮游离苷元(大豆素、染料木素)的含量进行测定。色谱柱为Prevail-C18(250 mm×10.0 mm,5μm),流动相为乙腈-水-乙酸(35∶64∶1,V/V/V),检测波长为260 nm,流速为1.0 ml/min,柱温为25℃,进样量为15μl。结果:古法炮制的淡豆豉气香,质地柔软,断面为棕褐色,表皮皱缩,总异黄酮含量为4.140 mg/g,大豆素含量为1.263 0 mg/g,染料木素含量为1.254 mg/g;现今药典制法的淡豆豉气味微弱,质地稍硬,断面为棕黄色,表皮皱缩,总异黄酮含量为3.530 mg/g,大豆素含量为0.349 mg/g,染料木素含量为0.335 mg/g。结论:古法炮制淡豆豉与现今药典制法淡豆豉相比,前者成品性状好、大豆异黄酮类含量高,古法炮制有明显优势。     

大豆异黄酮类防治骨质疏松的作用机制

从大豆异黄酮类化合物对骨形成、骨吸收以及骨髓基质细胞的成骨和成脂分化的影响3个方面,总结了近10年来用大豆异黄酮类防治骨质疏松症的作用机制,同时指出了今后可能的研究方向。     

HPLC法同时测定血脂康胶囊中3种异黄酮和洛伐他汀的含量

目的:建立血脂康胶囊中大豆苷元、黄豆黄素、染料木素和洛伐他汀的含量测定方法。方法:采用反相高效液相色谱法,YMC-C_(18)分析柱(4.6 mm×250 mm,5μm),乙腈(A)-0.1%磷酸溶液(B)梯度洗脱[0 min(25%A)→45 min(84% A)→55 min(25% A)],流速1.0 mL·min~(-1),进样量10μL,256 nm 检测。结果:大豆苷元、黄豆黄素、染料木素和洛伐他汀进样浓度分别在13.6×10~(-3)～81.6×10~(-3)μg(r=1.0000),8.0×10~(-3)～48.0×10~(-3)μg(r=1.0000),24.0×10~(-3)～144.0×10~(-3)μg(r=1.0000),4.50～8.25μg(r=0.9996)范围内与峰面积呈良好线性关系;平均加样回收率分别为99.1%,98.5%,98.2%,99.6%,RSD 分别为1.75%,1.37%,1.54%,1.49%(n=5)。结论:方法操作简便,准确,重复性好,可用于同时测定血脂康胶囊中大豆苷元、黄豆黄素、染料木素和洛伐他汀的含量。     

HPLC法同时测定葛根中5种异黄酮的含量

目的建立能同时测定葛根中5种异黄酮含量的HPLC法。方法采用Phenomenex C₁₈色谱柱(250 mm×4.6 mm,5μm);柱温:40℃;流动相A为甲醇,B为水,进行梯度洗脱;流速为1.0 mL·min^-1;检测波长为260 nm。结果葛根素、大豆苷、染料木苷、大豆苷元和染料木素的回归方程分别为y₁=35 462.65x₁+899.4(r₁=0.999 9),y₂=89 343.6x₂-720.2(r₂=0.999 9),y₃=139 651.625x₃+1 351.7(r₃=0.999 8),y₄=129 138.175x₄-1 339.5(r₄=0.999 9)和y₅=216 079.6x₅-3 679.8(r₅     

大豆异黄酮苷元滴丸的制备和含量测定

目的:研究大豆异黄酮苷元滴丸的处方、制备工艺和含量测定方法.方法:用单因素实验法确定制备工艺中的各种条件;用正交设计法优选处方组成;用HPLC法测定异黄酮苷元的含量.结果:最佳工艺条件为:冷凝剂二甲基硅油温度10℃,药液温度80℃,滴距6 cm,滴速15滴/min.染料木素在3.6～36%、6.00%和0.89%.结论:本制备工艺稳定性好、成型率高,建立的含量测定方法可控性强、专属性高.     

HPLC法同时测定中国、泰国葛根中异黄酮的含量

目的:建立HPLC法同时测定中国、泰国葛根中异黄酮大豆苷、大豆苷元和染料木素的含量,为其质量控制提供方法学参考.方法:色谱柱为AT.LICHROM ODS-C18柱(250 mm ×4.6 mm,5μm);流动相为甲醇和水,梯度洗脱;流速:0.8 ml·min-1;检测波长250 nm;柱温为25℃.结果:大豆苷、大豆苷元和染料木素分别在0.15 ～0.88 μg,0.10 ～0.63 μg,0.15 ～0.89 μg范围内呈良好线性关系,r分别为0.9997,0.9995,0.9996.其平均回收率在99.5％～100.4％范围内,RSD均小于3％.结论:建立的方法简单、准确,分离效果好,首次用于中国、泰国葛根的定量分析和比较分析,为葛根的资源研究提供参考.     

高效液相色谱法同时测定豆粕提取液中4种成分的含量

目的:高效液相色谱(HPLC)法同时测定豆粕提取液中大豆苷、染料木苷、大豆黄素、染料木素的含量.方法:高效液相色谱条件:预柱:Fusion-RP(4 mm×30 mm);分析柱:Hypersil ODS2(4.6 mm×250 mm,5 μm);流动相:甲醇(含1%醋酸的水溶液);流速:1 mL·min-1;柱温:25℃.首先用30%甲醇洗脱15 min,然后5 min内用30%甲醇梯度洗脱到48%甲醇,最后用48%甲醇洗脱维持20 min.结果:得到大豆苷、染料木苷、大豆黄素、染料木素4种成分峰面积和进样量的线性方程,测定大豆苷、染料木苷、大豆黄素、染料木素的日内精密度分别为0.52%,0.61%,0.35%,0.47%;日间精密度分别为1.52%,1.82%,1.05%,1.20%;回收率分别为98.7%,98.9%,99.1%,99.6%.结论:该方法用来测定豆粕提取液中大豆异黄酮的含量准确、简便、可靠.     

淡豆豉炮制过程中黄曲霉毒素含量的动态变化规律分析

目的: 检测淡豆豉炮制过程中不同时间点样本的黄曲霉毒素(AFTs)含量,明确AFTs的动态变化规律及其产生的关键时间点。方法: 按实验室前期已建立的规范炮制工艺制备淡豆豉,获取淡豆豉炮制过程中不同时间点的样本;采用超高效液相色谱-串联质谱法(UPLC-MS/MS)检测各样本中4种AFTs [黄曲霉毒素B₁(AFB₁),AFB₂,AFG₁,AFG₂]含量。结果: 4种黄曲霉毒素在选定的质量浓度范围内与峰面积线性关系良好(R²均大于0.99),重复性相对标准偏差(RSD)0.4%~0.9%,精密度RSD 0.7%~2.6%,稳定性RSD 0.8%~1.74%,加样回收率介于95.09%~107.20%(RSD 3.14%~12.71%);AFB₁含量在淡豆豉整个炮制过程中呈先上升后下降的趋势,在"再闷"第6天时达最高值6.95 μg·kg^-1,再闷第12天后各样本均未检测出AFTs。结论: 本研究建立了简单、快速、灵敏度高且适用于淡豆豉中4种AFTs检测的UPLC-MS/MS法;淡豆豉炮制过程中AFTs含量呈动态变化,表明淡豆豉炮制中生物拮抗作用自然存在,从安全性角度证实淡豆豉炮制中"再闷"环节的重要性和"再闷"时间的合理性。     

基于网络药理学及分子对接探讨淡豆豉抗抑郁的作用机制

目的 采用网络药理学和分子对接技术预测淡豆豉抗抑郁活性成分、作用靶点及通路,探讨其潜在作用机制。方法 利用CNKI、TCMSP、TCMIP检索并筛选淡豆豉抗抑郁潜在活性成分;运用PharmMapper服务器预测活性成分的作用靶点;利用UniProt数据库查询靶点蛋白对应的基因名;运用DrugBank、GeneCards和DisGeNET数据库检索抗抑郁作用靶基因,并将活性成分靶点相互映射获得淡豆豉活性成分抗抑郁靶点;通过String数据库下载靶蛋白相互作用数据,与获取的潜在作用靶点利用Cytoscape 3.7.2.软件构建淡豆豉–活性成分–潜在靶点相互作用网络图。运用DAVID数据库分析潜在靶点的基因本体（GO）分子功能和京都基因与基因组百科全书（KEGG）信号通路,最后利用Autodock Vina和Pymol软件对药物有效活性成分和关键靶点进行分子对接验证。结果 预测获得淡豆豉主要活性成分8个,作用靶点396个,与抑郁症交集潜在靶点334个,GO分子功能和KEGG通路涉及多种生物学过程以及脂质和动脉粥样硬化通路、磷脂酰肌醇-3激酶/蛋白激酶B（PI3K-Akt）信号通路、小分子量G蛋白（Ras）信号通路、叉头框蛋白（FoxO）信号通路等多个信号通路。将关键活性成分和靶点进行对接,其中丝氨酸-苏氨酸蛋白激酶1（AKT1）与大豆苷元结合能力较好;磷酸肌醇3-激酶调节亚基1（PIK3R1）与木犀草素、槲皮素结合能力较好;HSP90-α热休克蛋白（HSP90AA1）与黄豆黄素结合能力较好。结论 淡豆豉以多种活性成分、多个作用靶点和途径发挥其抗抑郁作用,为淡豆豉在临床上用于抑郁症的干预和治疗提供依据。     

Inhibition of the proteasome activity,a novel mechanism associated with the tumor cell apoptosis-inducing ability of genistein

Epidemiological studies have suggested that increased soy consumption is associated with reduced cancer occurrence. Genistein, a soy isoflavone, has been reported to inhibit the growth of human tumor cells although the involved molecular mechanisms are not clearly defined. Here we report that genistein inhibits the proteasomal chymotrypsin-like activity in vitro and in vivo. Computational docking studies suggest that the interaction of genistein with the proteasomal beta 5 subunit is responsible for inhibition of the chymotrypsin-like activity. Inhibition of the proteasome by genistein in prostate cancer LNCaP and breast cancer MCF-7 cells is associated with accumulation of ubiquitinated proteins and three known proteasome target proteins, the cyclin-dependent kinase inhibitor p27(Kip1), inhibitor of nuclear factor-kappa B (I kappa B-alpha), and the pro-apoptotic protein Bax. Genistein-mediated proteasome inhibition was accompanied by induction of apoptosis in these solid tumor cells. Finally, genistein induced proteasome inhibition and apoptosis selectively in simian virus 40-transformed human fibroblasts, but not in their parental normal counterpart. Our results suggest that the proteasome is a potential target of genistein in human tumor cells and that inhibition of the proteasome activity by genistein might contribute to its cancer-preventive properties.     

Identification, quantitation and biological activity of phytoestrogens in a dietary supplement for breast enhancement

A hop-based dietary supplement, marketed for natural breast enhancement, was analysed to determine the identity and biological activity of active constituents and potential biological effects in man. Extracts of the dietary supplement were analysed by LC-MS(n) and phytoestrogens identified and quantitated by reference to appropriate standards. Only hop-associated phytoestrogens were found in the dietary supplement at significant concentrations as follows (mean+/-1 S.D.); 8-prenylnaringenin 10.9+/-0.3, 6-prenylnaringenin 27.4+/-1.2, 6,8-diprenylnaringenin 0.9+/-0.1, xanthohumol 321+/-17 and isoxanthohumol 81.1+/-1.6 microg/g of dietary supplement. The oestrogenic activity of extracts in an ERalpha reporter gene assay was equivalent to 48+/-6.3 ng 17beta-oestradiol/g supplement and consistent with the 8-prenylnaringenin content. The dietary supplement extract also inhibited reductive 17beta-hydroxysteroid oxidoreductase activity, but to a greater extent than a concentration matched reference mixture of hop phytoestrogens. However, the supplement was only weakly active in mouse uterotrophic assays following administration in feed or after subcutaneous injection of extract at doses of 8-PN up to 250 times higher than that recommended for women. These preliminary findings suggest that the dietary supplement is unlikely to produce oestrogenic effects in vivo at the level of the uterus; supporting evidence is still required to demonstrate efficacy.