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.     

HPLC法测定葛花中大豆苷和染料木苷的含量

目的建立HPLC法测定葛花中大豆苷、染料木苷的含量。方法采用C18色谱柱(250mm×4.6mm,5μm);流动相为乙腈-1.0mL·L-1磷酸溶液(15∶85);流速为1.0mL·min~(-1);检测波长为260nm;柱温为30℃。结果大豆苷回归方程为Y=166 989.8 X-1 438.6,r=0.999 9,大豆苷在0.206~1.032μg范围内线性关系良好;染料木苷回归方程为Y=142 567 X+2 543.1,r=0.999 7,染料木苷在0.204~1.018μg范围内线性关系良好。结论该方法简便、准确,重复性好,可作为葛花的质量评价依据。     

西洋参DNA导入大豆实现遗传转化的研究Ⅱ.导入后代异黄酮类成分的含？…

用高效液相色谱法测定了西洋参 DNA导入大豆后代的异黄酮类成分、大豆苷 (daidzin)、大豆黄素(daidzein)、染料木苷 (genistin)、染料木素 (genistein)的含量 ,采用 U nisil Q C1 8柱 ,4.6 mm× 2 5 0 m m,流动相乙腈 - 0 .0 0 3mol/L 磷酸氢二钠 (17∶ 83) ,流速 1.0 m L/min,检测波长 2 5 4nm,结果表明 ,4种成分分离效果好 ,成分含量发生了广泛变异 ,从中可筛选出有效成分含量较高的新材料     

酶解法转化染料木苷包合物制备染料木素的研究

 目的 采用酶解法水解染料木苷-羟丙基-β-环糊精包合物制备染料木素。方法 运用饱和水溶液法制备染料木苷-羟丙基-β-环糊精包合物,再利用蜗牛酶将其水解从而获得染料木素。以染料木素转化率为指标,通过单因素实验分别考察染料木苷包合物和染料木苷酶解过程中不同的影响因素,确定最佳的酶解条件。采用差示扫描量热法对包合物进行验证。建立高效液相色谱分析方法,比较酶解染料木苷包合物和染料木苷的转化率。采用¹H-NMR,¹³C-NMR鉴定水解产物。结果 染料木苷-羟丙基-β-环糊精包合物酶解反应最适条件为反应介质pH 5.0,温度为50 ℃,酶与底物的用量比为4∶5,反应时间为12 h,在此条件下,包合物的转化率为（90.17±2.40）%,比未经包合的染料木苷转化率高了0.95倍。结论 蜗牛酶水解染料木苷-羟丙基-β-环糊精包合物制备染料木素,方法简单,酶解时间显著缩短,适用于工业化生产。     

大豆黄卷质量控制方法研究

目的:建立大豆黄卷质量标准.方法:采用薄层色谱法,以亮氨酸、染料木苷作为对照品,对大豆黄卷进行定性鉴别;采用高效液相色谱法对大豆黄卷中大豆苷和染料木苷进行含量测定:采用C18色谱柱(4.6 mm×250 mm,5μm),以甲醇-1％乙酸溶液为流动相,进行梯度洗脱,检测波长260 nm,流速1 mL· min-1.结果:薄层色谱中斑点清晰,重复性好;大豆苷在0.042 1 ～1.011 μg呈良好的线性关系,染料木苷在0.041 3 ～0.991 μg线性关系良好.结论:所建立的方法简便快速、准确度好,可作为大豆黄卷的质量控制方法.     

西洋参DNA导入大豆实现遗传转化的研究Ⅱ.导入后代异黄酮类成分的含量测定

用高效液相色谱法测定了西洋参 DNA导入大豆后代的异黄酮类成分、大豆苷 (daidzin)、大豆黄素(daidzein)、染料木苷 (genistin)、染料木素 (genistein)的含量 ,采用 U nisil Q C1 8柱 ,4.6 mm× 2 5 0 m m,流动相乙腈 - 0 .0 0 3mol/L 磷酸氢二钠 (17∶ 83) ,流速 1.0 m L/min,检测波长 2 5 4nm,结果表明 ,4种成分分离效果好 ,成分含量发生了广泛变异 ,从中可筛选出有效成分含量较高的新材料     

基于UPLC特征图谱结合多模式识别和色度值的槐角炮制前后差异研究

目的 测定槐角炮制前后的超高效液相色谱（UPLC）特征图谱和色度值,结合不同模式识别方法比较槐角炮制前后的差异性,为槐角和蜜槐角的质量控制及评价提供参考。方法 采用UPLC法对槐角炮制前后样品进行测定,按《中药色谱指纹图谱相似度评价系统（2012版）》对色谱图进行匹配生成槐角炮制前后的UPLC特征图谱,并对炮制前后的色度值（L、a、b）进行测定,通过相似度分析、对照品比对、方差分析、聚类分析、主成分分析（principal component analysis,PCA）和正交偏最小二乘判别分析（orthogonal partial least squares discriminant analysis,OPLS-DA）对槐角炮制前后进行多模式识别研究。结果 建立了槐角炮制前后的UPLC特征图谱,槐角和蜜槐角分别确定了23、24个共有峰,指认2、15、16、20、21号峰为没食子酸、芦丁、染料木苷、槲皮苷、槐角苷,炮制后新增成分24号峰为5-羟甲基糠醛;相似度分析结果显示槐角炮制前后相似度均大于0.98,聚类分析结果显示槐角炮制前后区分不明显,PCA可以将槐角和蜜槐角大致分为2类,OPLS-DA分析可将槐角和蜜槐角明显区分为2类,且结果显示1、2、10、22、23、24号峰是引起槐角和蜜槐角间成分差异的主要标志性成分,与方差分析结果一致;色度值ΔE范围为5~13,炮制前后色差可被肉眼识别。结论 建立的多指标特征图谱和色度值鉴别方法对槐角药材及其炮制品的质量控制及整体性评价具有重要意义。     

Genistin防治增生性玻璃体视网膜病变的研究

目的：研究Genistin对外伤性增生性玻璃体视网膜病变(proliferative vitreoretinopathy, PVR)的防治作用。方法：用自体富含血小板血浆玻璃体腔内注射制备兔外伤性PVR模型,并同时将二甲基亚砜、2 μg或40 μg genistin、1mg氟尿嘧啶各0.1 mL注入兔眼玻璃体内,在不同时间点用眼底镜检查眼底情况,进行PVR分级。第28天行40 μg genistin组模型眼及正常眼ERG检查;同天将4组模型眼球取出制作病理标本,行光镜检查。结果：A组第10天出现视网膜脱离,PVR随着时间延长发展至更严重等级。40 μg genistin和氟尿嘧啶组也发生PVR,但其严重程度低于对照组(P＜0.05)。组织学检查40 μg genistin组未发现明显视网膜形态改变,视网膜电图检查也未观察到显著功能变化。结论：Genistin玻璃体腔内注射是安全的,并能有效抑制兔外伤性PVR的发生发展。     

响应面法优选葛根超微粉的制备工艺及活性成分测定研究

目的建立葛根中8种主要黄酮类成分UPLC同时测定法,研究超微粉碎技术对葛根黄酮类成分含量的影响,确定葛根超微粉的生产工艺。方法采用响应面设计方法对葛根粉碎技术中的粉碎时间、投药量和初始粒度3个因素进行考察,以粒径分布(D90、D50)和UPLC法测定的不同粒径葛根粉(10~40、40~65、300目)中8个黄酮类活性成分(3′-羟基葛根素、葛根素、3′-甲氧基葛根素、大豆苷、染料木苷、大豆苷元、染料木素和芒柄花素)的含量数据,共10个指标作为响应因子进行葛根超微粉碎技术评价研究。结果葛根中8种黄酮类化合物分别在75.8~242.7、205.6~658.0、147.3~417.3、10.2~163.3、11.3~182.0、1.2~18.8、0.25~4.00、0.35~5.37μg/mL与色谱峰峰面积呈良好的线性关系,平均回收率为98.86%、99.25%、99.90%、100.17%、100.21%、101.40%、100.73%、101.42%,RSD均小于3.0%,UPLC同时测定结果显示,相较其他粒径葛根粉,葛根超微粉(300目)黄酮类含量更高。响应面法优选结果显示超微粉的最佳工艺参数条件为在粒径为80目[(180.0±7.6)μm],通过5号筛时,投药量为247 g,粉碎时间为26 min。结论响应面法优选的葛根超微粉碎工艺简便,准确度较高,得到的超微粉黄酮含量更高。该工艺可以为葛根超微粉碎技术提供参考。     

Genistin-rich soy isoflavone extract in substrate reduction therapy for Sanfilippo syndrome: An open-label, pilot study in 10 pediatric patients

Background: Mucopolysaccharidoses (MPSs) are a group of severe metabolic disorders caused by deficiencies in enzymes involved in the degradation of glycosaminoglycans (GAGs)—long chains of sugar carbohydrates in cells that help build bone, cartilage, tendons, corneas, skin, and connective tissue. Although enzyme replacement therapy has become available for the treatment of some types of MPS, effective treatment of neurodegenerative forms of MPS has yet to be determined. Recently, genistein (4',5,7-trihydroxyisoflavone), a specific inhibitor of protein tyrosine kinase, has been found to inhibit GAG synthesis and to reduce GAG concentrations in cultures of fibroblasts of MPS patients. Therefore, a potential substrate reduction therapy has been proposed.Objective: The aim of this study was to examine urinary GAG concentration, hair morphology, and cognitive function in patients receiving genistin treatment for Sanfilippo syndrome (MPS type III).Methods:Patients aged 3 to 14 years with a biochemically confirmed diagnosis of MPS IIIA or MPS IIIB were eligible to enroll in this open-label, pilot study. Genistin-rich soy isoflavone extract 5 mg/kg/d was administered PO for 12 months. Urinary GAG concentration, hair morphology,and cognitive function (measured using a modified version of the Brief Assessment Examination [BAE] and parent observations)were measured at baseline and after 12 months of treatment.Results: Ten patients (6 girls, 4 boys; mean age, 8 years [range,3\2-14 years];mean weight, 28 kg [range, 17\2-43 kg]) were included in the study. All patients had Sanfilippo syndrome; 5 patients had MPS IIIA and 5 had MPS IIIB. After 1 year, statistically significant improvement was found in urinary GAG concentration, hair morphology, and cognitive function. Urinary GAG concentration decreased significantly in all 5 patients with MPS IIIA and in 2 patients with MPS IIIB (P = 0.028). Hair morphology improved significantly in all 5 MPS IIIA patients and in 3 MPS IIIB patients (P = 0.012). A significant increase in the BAE score (by 2-6 points) was noted in 8 patients, while the scores of 2 patients did not change after 12 months of treatment (P = 0.012). No adverse events (AEs) considered related to treatment were reported. Moreover, no AEs not related to the treatment (apart from classical symptoms of MPS III) were noted.Conclusions: This pilot study found some improvements in GAG concentration, hair morphology, and cognitive function in these pediatric patients with Sanfilippo syndrome treated with genistin-rich soy isoflavone extract for 1 year. Clinical trials are needed to evaluate the efficacy and safety of this potential treatment.