健脾药糕的质量标准研究

目的 :完善健脾药糕的质量标准。方法 :采用薄层色谱法对健脾药糕中陈皮、山楂进行定性鉴别 ;采用高效液相色谱法对健脾药糕中橙皮苷进行含量测定。结果 :健脾药糕中陈皮、山楂的薄层色谱鉴别呈阳性 ;橙皮苷的平均含量为0 0186% ,平均加样回收率为97 75% (RSD=1 60% )。结论 :本方法准确可行 ,可用于健脾药糕的质量控制。     

二陈分散片的研制及质量控制

目的:制备二陈分散片并建立其质量控制方法。方法:采用交联羧甲基纤维素钠为崩解剂,加入聚乙烯吡咯烷酮等辅料制备分散片;采用高效液相色谱法测定制剂中橙皮苷含量,并进行成品稳定性考察。结果:橙皮苷进样量在1μg～80μg范围内线性关系良好,平均回收率为99.98%,RSD=0.98%;制剂稳定性良好。结论:该制剂处方合理,制备工艺简单,成品符合《中国药典》各项质量指标要求。     

高效液相色谱法测定小儿磨积片中橙皮苷的含量

目的:建立以高效液相色谱法测定小儿磨积片中橙皮苷含量的方法。方法:色谱柱为SpherisorbC18,流动相为甲醇-冰醋酸-水(25∶4∶71),检测波长为283nm,流速为2·0ml/min,柱温为50℃,灵敏度为0·16AUFS,进样量为20μl。结果:橙皮苷进样量在0·024μg~1·2μg范围内与峰面积积分值呈良好的线性关系(r=0·9999),平均回收率为99·1%(RSD=0·8%)。结论:本方法简便、快捷,灵敏度及准确度高,可为小儿磨积片质量控制提供依据。     

小儿止泻安颗粒质量标准研究

目的:提高小儿止泻安颗粒质量标准,更有效控制其质量.方法:采用TLC法对处方中的木香进行定性鉴别;用HPLC法测定橙皮苷的含量.结果:用TLC能检出木香;橙皮苷在0.0424～2.120μg范围内呈良好的线性关系,r=0.9999,平均回收率99.83%,RSD为0.88%.结论:方法简便,准确,重现性好,能有效控制该制剂质量.     

Effect of hesperidin and neohesperidin from bittersweet orange (Citrus aurantium var. bigaradia) peel on indomethacin-induced peptic ulcers in rats

Hesperidin and neohesperidin are the major flavanones isolated from bittersweet orange. It was recently reported that they have potent anti-inflammatory effects in various inflammatory models. In the present study, the effects of hesperidin and neohesperidin on indomethacin-induced ulcers in rats and the underlying mechanisms were investigated. Gastric ulcers were induced in rats with a single dose of indomethacin. The effects of pretreatment with hesperidin and neohesperidin were assessed in comparison with omeprazole as reference standard. Ulcer index, gene expression of gastric cyclooxygenase-2 (COX-2), tumor necrosis factor alpha (TNF-α), lipid peroxidation product, malondialdhyde (MDA), and reduced glutathione (GSH) content in stomach were measured. Furthermore, gross and histopathological examination was performed. Our results indicated that both hesperidin and neohesperidin significantly aggravated gastric damage caused by indomethacin administration as evidenced by increased ulcer index and histopathological changes of stomach.     

金果饮的质量控制研究

目的：对金果饮的质量标准进行制定。方法：对玄参、西青果、陈皮建立薄层色谱的鉴别方法;采用高效液相色谱法测定橙皮苷的含量。用Agilent SB C18柱（4.6mm×250mm,5μm）,以甲醇-6%乙酸溶液（35∶65）为流动相;检测波长为283nm,流速1mL·min-1。结果：薄层色谱鉴别中,供试品在与对照药材色谱相应的位置上,显示相同颜色的斑点,斑点清晰,分离效果好,阴性无干扰;含量测定法中,橙皮苷在36.35～726.95ng（r=0.9999,n=5）范围内呈良好的线性关系,平均加样回收率为98.58%（RSD为1.85%）。结论：建立的方法简便、准确、专属性强、重复性好,能有效用于金果饮的质量控制。     

UPLC同时测定清热止咳颗粒中7种黄酮类成分含量

目的 建立同时测定清热止咳颗粒中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷、汉黄芩苷含量的方法。方法 采用超高效液相色谱法。采用ACQUITY UPLC BEH C₁₈色谱柱（2.1 mm×100 mm,1.7 μm）,流动相乙腈-0.1%甲酸水溶液(梯度洗脱) ,流速0.3 mL·min^-1,检测波长280 nm,柱温30 ℃。结果 芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷、汉黄芩苷分别在0.324～6.480（r=0.999 9）,1.337～26.75（r=0.999 7）,0.687～13.80（r=0.999 6）,1.347～26.95（r=0.999 8）,2.770～55.40（r=0.999 2）,0.330～6.600（r=0.999 8）,0.242～4.840（r=0.999 6）μg·mL^-1范围内与峰面积呈良好的线性关系;并且芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、黄芩苷、木蝴蝶素A-7-葡萄糖醛酸苷、汉黄芩苷平均加样回收率分别为99.41%、101.80%、99.42%、98.47%、101.71%,102.27%和97.64%,RSD范围分别是2.32%、1.69%、2.38%、2.45%、1.24%、1.77%和1.91%。结论 建立的超高效液相色谱(UPLC)方法操作简便、稳定、准确且可行,可用于清热止咳颗粒的质量控制。     

多指标综合加权评分法优选消瘿散结合剂的提取工艺*

目的：优选消瘿散结合剂的提取工艺。方法：采用L9 （34）正交设计,以丹酚酸B、橙皮苷、迷迭香酸的含量及出膏率的综合评分为指标,考察料液比、提取时间、提取次数对提取效果的影响,优选消瘿散结合剂的最佳提取工艺。结果：根据综合评分的结果,优选出最佳提取工艺为加10倍水,提取2次,每次1 h。结论：该工艺准确可靠,提取率高,可行性好。     

Screening and identification of bioactive compounds from citrus against non-structural protein 3 protease of hepatitis C virus genotype 3a by fluorescence resonance energy transfer assay and mass spectrometry

BACKGROUNDHepatitis C virus genotype 3a (HCV G3a) is highly prevalent in Pakistan. Due to the elevated cost of available Food and Drug Administration-approved drugs against HCV, medicinal natural products of potent antiviral activity should be screened for the cost-effective treatment of the disease. Furthermore, from natural products, active compounds against vital HCV proteins like non-structural protein 3 (NS3) protease could be identified to prevent viral proliferation in the host.AIMTo develop cost-effective HCV genotype 3a NS3 protease inhibitors from citrus fruit extracts.METHODSFull-length NS3 without co-factor non-structural protein 4A (NS4A) and codon optimized NS3 protease in fusion with NS4A were expressed in Escherichia coli. The expressed protein was purified by metal ion affinity chromatography and gel filtration. Citrus fruit extracts were screened using fluorescence resonance energy transfer (FRET) assay against the protease and polyphenols were identified as potential inhibitors using electrospray ionization-mass spectrometry (MS)/MS technique. Among different polyphenols, highly potent compounds were screened using molecular modeling approaches and consequently the most active compound was further evaluated against HCV NS4A-NS3 protease domain using FRET assay.RESULTSNS4A fused with NS3 protease domain gene was overexpressed and the purified protein yield was high in comparison to the lower yield of the full-length NS3 protein. Furthermore, in enzyme kinetic studies, NS4A fused with NS3 protease proved to be functionally active compared to full-length NS3. So it was concluded that co-factor NS4A fusion is essential for the purification of functionally active protease. FRET assay was developed and validated by the half maximal inhibitory concentration (IC₅₀) values of commercially available inhibitors. Screening of citrus fruit extracts against the native purified fused NS4A-NS3 protease domain showed that the grapefruit mesocarp extract exhibits the highest percentage inhibition 91% of protease activity. Among the compounds identified by LCMS analysis, hesperidin showed strong binding affinity with the protease catalytic triad having S-score value of -10.98.CONCLUSIONFused NS4A-NS3 protease is functionally more active, which is effectively inhibited by hesperidin from the grapefruit mesocarp extract with an IC₅₀ value of 23.32 µmol/L.