荧光光度法测定竹叶柴胡地上部分不同部位总黄酮含量

目的：分析竹叶柴胡不同部位总黄酮含量,研究黄酮在柴胡不同部位的分布,为中药资源合理开发利用提供参考。方法以芦丁为对照品,以5%硝酸铝与样品中黄酮成分产生络合反应,选择激发波长λex=445 nm,发射波长λem=484 nm,应用荧光光度法测样品的荧光光度值,计算总黄酮含量。结果以芦丁在0.198~3.168μg/mL范围内与荧光强度线性关系良好,y=0.9728x-0.001,r2=0.9993。竹叶柴胡花、叶、茎中总黄酮含量最分别为3.08%、1.85%、0.41%。结论竹叶柴胡花、叶中总黄酮含量较高,应在开花期采收竹叶柴胡地上部分。     

四碘荧光素钠-邻啡啰啉荧光光谱法测定水中镉

目的 建立一种荧光光谱测定水中镉的新方法. 方法 在pH =6.0的乙酸-乙酸钠（HAe-NaAc）缓冲介质中,镉（Cd2＋）与四碘荧光素钠、邻啡哆啉反应形成络合物,体系的最大激发波长为525 nm,发射波长为531 nm,在一定范围内,其荧光强度与Cd2＋的浓度成线性关系. 结果 Cd2＋浓度在3.92×10 6mol/L ～2.00 ×10-5 mol/L范围内,荧光强度变化值（△F）与镉离子浓度（cCd2＋）有良好的线性关系,线性方程为△F=58.9c（×10-5 mol/L） ＋4.8,相关系数r=0.999 2,检出限为1.91×10-6 mol/L,样品加标回收率为86.6％～101.5％.结论 本方法可直接用于水中镉的测定,选择性好,准确可靠,结果满意.     

荧光增敏法测定咽炎片中丹皮酚的含量

目的 建立测定咽炎片中丹皮酚含量的荧光分析法.方法 基于AlC13试剂对于丹皮酚具有荧光增敏作用,在pH7.0的磷酸氢二钠-磷酸二氢钠缓冲体系中,于最大发射波长λem=455 nm处（最大激发波长λex=295nm）下检测丹皮酚的含量.结果 丹皮酚的浓度在1.3 ×10-4～7.3 ×10-4μg/ml内与荧光强度增大值△F呈线性关系良好（r=0.999）,回归方程为△F=89.45p（×10-3 μg/ml）-0.2777,检出限为0.8×10-4μg/nl,回收率为95.4％～97.8％,相对标准偏差小于2.53％.结论 该方法准确可靠,可应用于咽炎片中丹皮酚含量的测定.     

岭南习用中药五指柑中木犀草素及总黄酮的含量测定

目的 建立五指柑药材中木犀草素及总黄酮的含量测定方法.方法 采用HPLC法测定木犀草素含量,色谱柱:Kromasil C18色谱柱(250 mm×4.6 mm,5μm);流动相:乙腈-0.03％(体积分数)H3 PO4(体积比25∶75);流速:1.0 mL/min;检测波长:350 nm.总黄酮测定采用Al(NO3)...     

HPLC测定三尖杉酯碱脂质体及其在大鼠组织中的含量

本文建立了用反相HPLC测定三尖杉酯碱脂质体及其大鼠组织中含量的方法。色谱柱为Shim-pack 150mm×6mm,CLC-ODS 5μm,并装有预柱20×6mm,10μm(ODS、天津);用甲醇-0.1mol/L甲酸胺缓冲液(1:1)作为流动相;在发射波长λcm325nm,激发波长λcx 290nm处,用荧光检测器测定峰高,检出量为0.78ng,标准差为.0034。     

可见及紫外分光光度法测定中药复方总黄酮的比较研究

目的：研究并比较可见分光光度法及紫外分光光度法测定中药复方总黄酮含量。方法：可见分光光度法以芦丁为参照品,显色剂为NaNO2－Al（NO3）3－NaOH 体系,在510nm 波长处测定吸光度,计算中药复方总黄酮含量;紫外分光光度法以芦丁为参照品,在359nm 波长处测定吸光度,计算中药复方总黄酮含量。结果：可见分光光度法测定的中药复方总黄酮含量比紫外分光光度法的测定值高,含量高低成正比。结论：NaNO2－Al（NO3）3－NaOH 体系可见分光光度法稳定可靠,是测定中药复方总黄酮含量的首选方法。     

同时分离富集细毡毛忍冬中总黄酮和绿原酸的工艺研究

目的:研究细毡毛忍冬总黄酮和绿原酸的聚酰胺同时富集工艺.方法:以紫外分光光度法测定其样品中总黄酮含量,HPLC测定绿原酸含量为指标,考察多个参数.结果:聚酰胺对细毡毛忍冬总黄酮和绿原酸有良好的吸附作用,其吸附洗脱工艺:上样溶液浓度0.04g生药/mL,pH 5～6(原液),以1.5 mL/min的吸附流速上样,再依次用10％、50％乙醇对吸附后的聚酰胺进行洗脱,洗脱体积200 mL,洗脱剂pH值9～ 10.结论:聚酰胺可同时用于细毡毛忍冬总黄酮和绿原酸的分离富集.     

红花及番红花色素的荧光特性分析

目的探索荧光法测定红花及番红花色素的最佳方法,方法用岛津RF-5000荧光分光光度计测定,扫描波长λex=230～440nm,λem=240～600nm。结果最佳检测波长λex340nm,λem=440nm。红花及番红花的甲醇浸泡液浓度在16.7～330μg.mL-1范围内与荧光强度呈线性关系,相关系数r(红花)=0.9997、r(番红花)=0.9996,红花色素在pH=12的碱性环境中所发出的荧光强度达到最大。结论该方法简便、快速,可直接用于红花及番红花色素的含量测定,亦可作为药材红花和番红花之间的一种鉴别手段。     

新疆两色金鸡菊中绿原酸与总黄酮含量测定

目的:建立HPLC法和UV法分别测定新疆两色金鸡菊中绿原酸和总黄酮的含量。方法:采用HPLC法测定绿原酸含量,Agilent Eclipse Plus C_(18)柱(250 mm×4.6 mm,5μm),以流动相0.1%磷酸水溶液-乙腈,梯度洗脱,流速1.0 mL/min,检测波长348 nm,柱温30℃。采用UV法测定总黄酮含量,NaNO_2-Al(NO_3)_3-NaOH方法显色,以芦丁为对照,在波长522 nm处检测。结果:方法在考察的浓度范围内线性关系良好,精密度、准确度和稳定性RSD均小于5%。不同产地新疆两色金鸡菊绿原酸含量为1.39%～5.76%,总黄酮含量为19.10%～28.40%。结论:本试验建立了含量测定的方法,简便可靠,重现性好,适用于两色金鸡菊中绿原酸和总黄酮的含量测定。     

替米沙坦片的人体药动学和生物等效性

目的:建立测定替米沙坦血浆浓度的高效液相-荧光检测法(HPLC),考察替米沙坦的药代动力学及生物等效性.方法:24例健康受试者单剂量交叉口服40 mg替米沙坦片供试制剂或参比制剂后,用HPLC测定替米沙坦血药浓度并计算药动学及相对生物利用度,进而对主要药动学参数进行统计分析.色谱柱为Diamonsil C18柱(5 μm,4.6 mm×150 mm),以乙腈-磷酸二氢钾缓冲液(61:39,磷酸调pH=3.74)为流动相,流速1.0 mL/min,坎地沙坦为内标,荧光检测波长λEx=305 nm,λEm=365 nm.结果:测定替米沙坦的线性范围为0.5～144 ng/mL,r=0.9998,绝对回收率在79.62%～85.69%(n=5),相对回收率在101.92%～108.95%(n=5),日内和日间RSD分别为4.23%～9.80%和4.03%～9.95%(n=5).供试制剂与参比制剂的AUC0～96分别为888.90±406.49和895.03±364.53.结论:两种替米沙坦制剂均具有生物等效性.     

大孔吸附树脂分离纯化元宝枫叶总多酚

目的研究大孔树脂分离纯化元宝枫叶总多酚的工艺条件。方法以元宝枫叶总多酚的静态与动态吸附量和解吸率为指标筛选大孔树脂,采用紫外分光光度法测定元宝枫叶总多酚含量,对所筛选树脂分离元宝枫叶总多酚工艺进行评价。结果H PD100型大孔树脂分离效果较好,其最佳分离条件是以浓度为5 g/L元宝枫叶提取物为供试液,上样和洗脱流速为2 mL/min,解吸液浓度为70%乙醇,解吸液体积为4BV。结论HPD100型大孔树脂在所确定的工艺条件下,树脂的吸附-解吸附性能稳定,且能较好地分离纯化元宝枫叶总多酚,多酚含量纯化前为28.55%,纯化后为39.52%。     

枫叶黄酮抑制脂多糖诱导破骨前体细胞Raw264.7细胞激活的作用

目的 探讨枫叶黄酮对脂多糖(lipopolysaccharide,LPS)诱导破骨前体细胞Raw264.7细胞炎性因子释放的抑制作用.方法 用LPS(5 mg/mL刺激Raw264.7细胞构建炎症模型,采用四甲基偶氮唑蓝比色法(MTT)检测Raw264.7细胞的活力影响;酶联免疫吸附试验(ELISA)检测一氧化氮(N0)和前列腺素E2,PGE2表达;运用免疫荧光染色方法 检测诱导型一氧化氮合酶和环氧合酶-2,COX-2、子肿瘤坏死因子-αTNF-α和白介素-1β,IL-1β、核因子-κB,NF-κB蛋白与mRNA的表达变化.结果 不同浓度的枫叶黄酮明显抑制了LPS诱导的Row246.7细胞NO、PGE2,iNOS和COX-2,TNF-α和IL-1β与NF-κB的上调,并且.NO、PGE2、iNOS、COX-2和NF-κB蛋白的表达呈剂量依赖性.结论 枫叶黄酮可抑制LPS诱导的破骨前体细胞Raw264.7细胞iNOS、COX-2和NF-κB/P65蛋白表达和炎性因子释放从而抑制破骨细胞的激活.     

RP-HPLC法对元宝枫果翅中3种黄酮苷元的研究

目的研究元宝枫果翅活性与其有效成分的相关性。方法利用反相高效液相色谱法测定不同季节元宝枫果翅中槲皮素、山柰酚和异鼠李素含量。结果不同季节样品中3种黄酮苷元的定量分析结果分别为槲皮素平均回收率99.8%,RSD=0.72%;山柰酚平均回收率为100.8%,RSD=1.09%;异鼠李素平均回收率为102.4%,RSD=1.61%。结论元宝枫果翅中含有中槲皮素、山柰酚和异鼠李素,其含量随季节变化。     

荧光光谱法研究锆离子存在下槲皮素及人血清白蛋白的相互作用

目的 采用荧光光谱法研究有无锆离子（Zr^4＋）共存时槲皮素与人血清白蛋白（HSA）的相互作用。方法 于10 mL的比色管中,依次加入0.20 mol/L Tris-HCl缓冲溶液（pH 7.40）2.50 mL,0.10 mol/L NaCl溶液1.0 mL,一定量的HSA溶液,Zr^4＋溶液,槲皮素溶液,用二次蒸馏水定容至10 mL。HSA荧光激发/发射波长为281/352 nm,激发和发射光狭缝宽度为3 nm,在波长300～500 nm内记录荧光光谱。结果 测得了有无锆离子存在时槲皮素-HSA结合反应的平衡常数和结合摩尔位点数分别为1.032×10⁶ L/mol、2.964×10⁵ L/mol,1.13、1.07;确定槲皮素-HSA的作用类型为静态猝灭过程。结论 锆离子、槲皮素对HSA荧光均有猝灭作用,而且二者共存时对HSA荧光有协同猝灭作用,其猝灭机制为槲皮素与锆离子形成配合物后再对HSA的荧光有猝灭作用,而不是槲皮素和锆离子各自猝灭的简单累加。     

元宝枫油对一次性力竭运动大鼠氧化应激和炎症反应的影响

目的 探讨元宝枫油对一次性力竭运动大鼠氧化应激和炎症反应的影响,为元宝枫油用于预防运动损伤提供科学依据.方法 30只健康SD雄性大鼠根据体质量随机分为安静对照组(A组,正常饮食+每天生理盐水灌胃),力竭组(B组,正常饮食+每天生理盐水灌胃)和元宝枫油力竭组(C组,每天元宝枫油0.21 mL/kg灌胃),6周后进行一次性力竭游泳运动,测定大鼠血清中肌酸激酶(CK)、乳酸脱氢酶(LDH)的活性及炎症因子白细胞介素(IL)-1β、IL-10和肿瘤坏死因子-α(TNF-α)的水平;测定肌肉匀浆中丙二醛(MDA)、还原性谷胱甘肽(GSH)水平及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的活性.结果 力竭运动后,B组与A组相比,血清中CK、LDH水平及IL-1β、TNF-α、IL-10水平均显著升高(P＜0.05),IL-10/TNF-α比值差异无统计学意义(P＞0.05),大鼠腓肠肌中MDA水平显著升高(P＜0.05),GSH水平与GSH-PX活性显著下降(均P＜0.05),SOD活性与CAT活性差异无统计学意义(P＞0.05);C组与B组相比,血清中CK、LDH水平及IL-1β、TNF-α显著下降(均P＜0.05),IL-10差异无统计学意义(P＞0.05),IL-10/TNF-α比值显著上升(P＜0.05),腓肠肌中MDA水平明显下降(P＜0.05),GSH水平及SOD、CAT、GSH-PX活性均显著升高(均P＜0.05).结论 元宝枫油可明显减轻力竭运动引起的氧化应激和炎症反应,对骨骼肌具有保护作用.     

Phytochemical investigation and in vitro antioxidant activity of an indigenous medicinal plant Alpinia nigra B.L. Burtt

Objective

To investigate antioxidant potential of methanol extract of Alpinia nigra leaves.

Methods

The study was done by using various in vitro methods such as 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), nitric oxide and hydrogen peroxide radical scavenging assays. Phytochemical constituents, total phenolic content and total flavonoid content of the extract at different concentrations (10-500 µg/mL) were determined.

Results

Alpinia nigra leaves showed high free radical scavenging activity as evidenced by the low IC₅₀ values in DPPH (64.51 µg/mL), in ABTS (28.32 µg/mL), in nitric oxide (80.02 µg/mL) and in H₂O₂ (77.45 µg/mL) scavenging assays. Furthermore the TPC and TFC of the extract were found to be 69.25 mg gallic acid equivalent per gram of extract and 78.84 mg quercetin equivalent per gram of extract respectively.

Conclusions

The results of present comprehensive analysis demonstrated that Alpinia nigra leaves possess high phenolic, flavonoid contents and potential antioxidant activity, and could be used as a viable source of natural antioxidants and might be exploited for functional foods and neutraceutical applications.     

Highly sensitive electrochemical determination of rutin based on the synergistic effect of 3D porous carbon and cobalt tungstate nanosheets

Rutin, a flavonoid found in fruits and vegetables, is a potential anticancer compound with strong anticancer activity. Therefore, electrochemical sensor was developed for the detection of rutin. In this study, CoWO₄ nanosheets were synthesized via a hydrothermal method, and porous carbon (PC) was prepared via high-temperature pyrolysis. Successful preparation of the materials was confirmed, and characterization was performed by transmission electron microscopy, scanning electron microscopy, and X-ray photoelectron spectroscopy. A mixture of PC and CoWO₄ nanosheets was used as an electrode modifier to fabricate the electrochemical sensor for the electrochemical determination of rutin. The 3D CoWO₄ nanosheets exhibited high electrocatalytic activity and good stability. PC has a high surface-to-volume ratio and superior conductivity. Moreover, the hydrophobicity of PC allows large amounts of rutin to be adsorbed, thereby increasing the concentration of rutin at the electrode surface. Owing to the synergistic effect of the 3D CoWO₄ nanosheets and PC, the developed electrochemical sensor was employed to quantitively determine rutin with high stability and sensitivity. The sensor showed a good linear range (5–5000 ng/mL) with a detection limit of 0.45 ng/mL. The developed sensor was successfully applied to the determination of rutin in crushed tablets and human serum samples.     

A comparative study of the antioxidant,antimicrobial, cytotoxic and thrombolytic potential of the fruits and leaves of Spondias dulcis

Objective

To investigate the antioxidant, antimicrobial, cytotoxic and thrombolytic property of the fruits and leaves of Spondias dulcis (S. dulcis).

Methods

Methanolic extracts of fruits and leaves of S. dulcis were partitioned with chloroform and dichloromethane. The antioxidant potential of the crude extract and partitioned fractions were evaluated in terms of total phenolic content, total flavonoid content, DPPH radical scavenging potential, reducing potential and total antioxidant capacity by specific standard procedures. The antimicrobial activity was evaluated using disc diffusion method. The cytotoxicity was evaluated by using brine shrimp lethality bioassay and compared with vincristine sulfate. The thrombolytic activity was compared with streptokinase.

Results

The methanolic fruit extract exhibited the highest phenolic content, flavonoid content and antioxidant capacity, among the other extracts, with the highest DPPH radical scavenging activity at a concentration of 10 µg/mL (IC₅₀: 1.91 µg/mL) and maximum reducing power at a concentration of 100 µg/mL (EC₅₀: 3.58 µg/mL). Though all extract showed moderate antimicrobial activity against the bacterial strains, weak or no activity against fungus. The range of LC₅₀ value of all extracts was 1.335-14.057 µg/mL which was far lower than the cut off index for cytotoxicity. All extracts exhibited statistically significant (P<0.001) thrombolytic activity.

Conclusions

Our study suggested that S. dulcis exhibits antimicrobial activities against a wide variety of strains while it possesses significant antioxidant, cytotoxic and thrombolytic activity.     

Evaluation of in vitro aldose reductase inhibitory potential of different fraction of Hybanthus enneaspermus Linn F. Muell

Objective

To evaluate the aldose reductase inhibitory (ARI) activity of different fractions of Hybanthus enneaspermus for potential use in diabetic cataract.

Methods

Total phenol and flavonoid content of different fractions was determined. ARI activity of different fractions in rat lens was investigated in vitro.

Results

The results showed significant level of phenolic and flavonoid content in ethyl acetate fraction [total phenol (212.15±0.79 mg/g), total flavonoid (39.11±2.27 mg/g)] and aqueous fraction [total phenol (140.62±0.57 mg/g), total flavonoid (26.07±1.49 mg/g)] as compared with the chloroform fraction [total phenol (68.56±0.51 mg/g), total flavonoid (13.41±0.82 mg/g)] and petrolium ether fraction [total phenol (36.68±0.43 mg/g), total flavonoid (11.55±1.06 mg/g)]. There was a significant difference in the ARI activity of each fraction, and it was found to be the highest in ethyl acetate fraction [IC₅₀ (49.26±1.76 µg/mL)] followed by aqueous extract [IC₅₀ (70.83±2.82 µg/mL)] and it was least in the petroleum ether fraction [IC₅₀ (118.89±0.71 µg/mL)]. Chloroform fraction showed moderate activity [IC₅₀ (98.52±1.80 µg/mL)].

Conclusions

Different fractions showed significanct amount of ARI activity, where in ethyl acetate fraction it was found to be maximum which may be due to its high phenolic and flavonoid content. The extract after further evaluation may be used in the treatment of diabetic cataract.