阔叶十大功劳叶与花总黄酮提取及含量测定

目的:研究阔叶十大功劳叶与花总黄酮的提取方法和含量测定。方法:比较以索氏提取法、超声波提取法和微波辅助提取法3种方法提取阔叶十大功劳叶与花中总黄酮,用亚硝酸钠-硝酸铝-氢氧化钠比色法测定总黄酮含量。结果:3种提取方法中,微波法提取效率最高,结果重现性好,并测得叶与花中总黄酮含量分别为2.182%和8.155%。结论:阔叶十大功劳总黄酮含量较高,总黄酮在花中的含量明显高于叶。     

高通量检测大样本桑黄发酵液总黄酮含量方法的建立

建立了大样本的桑黄发酵液样品中总黄酮的高通量测定方法。通过L9(34)正交实验设计优化,确定了基于亚硝酸钠-硝酸铝比色法的微孔板检测方法,优化后总黄酮含量测定的最佳反应条件:5%亚硝酸钠30μL,10%硝酸铝30μL,10%氢氧化钠50μL,室温反应10 min,检测波长为510 nm。检测结果表明,针对大样本检测,该法稳定性高,平均回收率为100.7%,相对标准偏差为0.607%,线性范围为0~50μg/m L,线性相关系数r2=0.9994。比较分析结果显示,和多种传统总黄酮测定方法相比,实验所建立的高通量测定方法检测大样本快速、准确、用药量小,且稳定性高。     

高通量检测大样本桑黄发酵液总黄酮含量方法的建立北大核心CSCD

建立了大样本的桑黄发酵液样品中总黄酮的高通量测定方法。通过L9（3^4）正交实验设计优化,确定了基于亚硝酸钠．硝酸铝比色法的微孔板检测方法,优化后总黄酮含量测定的最佳反应条件：5％亚硝酸钠30μL,10％硝酸铝30μL,10％氢氧化钠50μL,室温反应10min,检测波长为510nm。检测结果表明,针对大样本检测,该法稳定性高,平均回收率为100．7％,相对标准偏差为0．607％,线性范围为0—50μg／mL,线性相关系数r^2=0．9994。比较分析结果显示,和多种传统总黄酮测定方法相比,实验所建立的高通量测定方法检测大样本快速、准确、用药量小,且稳定性高。     

迎春花植物茎叶中黄酮类物质初步研究

采用硝酸铝——亚硝酸钠比色法对迎春花茎叶内的黄酮类物质含量进行了初步测定,确定了总黄酮含量提取的适宜溶剂浓度和提取的适宜时间。结果表明,用60％甲醇溶剂提取5h总黄酮含量最高,茎杆中的含量为10．42mg/g DW,叶片中的含量为18．31mg／g DW。此外,还对迎春花叶提取液进行了各种黄酮定性颜色反应,初步确定其中的黄酮类物质主要为双氢黄酮类及查尔酮类。     

四妙勇安汤改良后治疗心脑血管有效部位中总黄酮的含量测定

采用紫外可见分光光度法,以芦丁为参照品,以硝酸铝为显色剂,在510 nm波长处测定总黄酮的含量。结果显示四妙勇安汤加石斛、牛膝治疗心脑血管有效部位中的总黄酮平均含量为37.76%。研究结果表明该方法操作简单,重现性好,可用于的质量控制,适合用于四妙勇安汤加石斛、牛膝治疗心脑血管有效部位中总黄酮含量的测定。     

大孔树脂纯化卫矛总黄酮的工艺研究

筛选纯化卫矛总黄酮的最佳大孔树脂及其最佳工艺条件,采用正交设计法考察树脂种类、洗脱液浓度、pH值等因素对纯化的影响。用紫外分光光度法测定总黄酮的含量,计算吸附量、解吸率和浸膏总黄酮含量,最后确定最佳工艺条件。AB-8树脂最适于纯化卫矛总黄酮;最佳吸附量和解吸率分别是7.32 mg·g^-1和90.98%;浸膏的总黄酮含量从7.64%提高到了52.12%。该树脂可以用于精制卫矛总黄酮,提高提取物中总黄酮的含量。     

尖叶胡枝子总黄酮含量测定方法(比色法)的优化

采用NaOH-Al(NO₃)₃-NaNO₂比色法对尖叶胡枝子的总黄酮含量进行测定。以芦丁为对照品,检测波长为510 nm。结果表明,在０～1 g·L^-1范围内,芦丁吸光度与其质量浓度呈良好的线性关系（r＝0.999 8）,对测定方法的稳定性、精密度、重现性及回收率进行试验,相对标准偏差（RSD）为0.199%～1.023%。根据拟合的线性回归方程对尖叶胡枝子不同提取液中总黄酮进行定量,所得测定数据分别为0.029 0、0.024 0、0.315 0、0.379 0和0.012 4 ｇ/Ｌ。该方法简便、快速、准确,可作为尖叶胡枝子中总黄酮含量的测定方法。     

沙棘不同部位总黄酮含量的测定及比较

考察了引种于俄罗斯的大果沙棘不同部位总黄酮含量的差异。用超声提取作为沙棘总黄酮的制备方法;以芦丁为对照品,采用分光光度法作为沙棘总黄酮的测定方法。实验结果表明,沙棘叶、果肉和果皮中总黄酮含量分别为1.71%、0.76%和0.55%,叶中总黄酮含量分别是果肉和果皮的2.3倍和3.1倍,因此引种于俄罗斯的大果沙棘叶具有较大的开发价值和应用前景。     

不同季节新疆圆柏总黄酮含量的变化

目的:确定乌鲁木齐南山地区新疆圆柏总黄酮含量最高的时期及其季节变化。方法:采用溶剂法萃取不同季节新疆圆柏叶、果实与枝干的总黄酮,以芦丁作为标准品,用比色法进行含量测定。结果:新疆圆柏的果实中黄酮含量最高,为0.62%～1.13%,以10月份的含量最高;叶中黄酮含量变化为0.28%～0.83%,4月份最高;枝干中的黄酮含量最少,变化不明显。结论:新疆圆柏黄酮含量的季节变化与其生长习性及乌鲁木齐南山气候有关。     

广西不同产地番石榴叶总黄酮含量测定

目的对广西不同产地番石榴叶中总黄酮含量进行测定。方法应用紫外分光光度法,以芦丁为对照品,于357nm下测定吸光度,计算番石榴叶中总黄酮的含量。结果在1.7~14.3范围内总黄酮含量与吸光度A值线性关系良好,平均回收率=99.28%,RSD=3.69%,R2=0.9993。结论该方法简便可靠、快速、重现性好,适用于番石榴叶总黄酮含量测定。采收地区不同,番石榴叶中总黄酮的含量也不同。     

不同炮制方法对玉竹提取物得率及体外抗氧化作用的影响

以多糖、总黄酮、醇溶物和水溶物的得率及体外抗氧化活性为考察指标,研究了酒蒸和蜜蒸两种炮制方法对玉竹品质的影响。结果表明：酒蒸炮制玉竹的多糖、醇溶物得率最高,蜜蒸炮制玉竹总黄酮、水溶物的得率最高,比未炮制的玉竹（生品玉竹）中相应成分的得率分别提高了43.86%、29.53%、49.46%和34.66%。将多糖、水溶性浸出物、醇溶性浸出物及总黄酮四者得率相加的和进行比较,蜜蒸最好,蜜蒸为111.069%,酒蒸为107.309%,生品玉竹为80.926%。酒蒸炮制玉竹的多糖、水溶物对DPPH自由基的清除率均高于蜜蒸玉竹和生品玉竹,其DPPH_IC50分别为0.345±0.019和0.441±0.022 mg·mL^-1;蜜蒸炮制玉竹的总黄酮、醇溶物对DPPH自由基的清除率均高于酒蒸玉竹和生品玉竹,其DPPH_IC50分别为0.047±0.011和0.199±0.036 mg·mL^-1;在浓度为1 mg·mL^-1时,蜜蒸玉竹总黄酮对DPPH自由基的清除率最大,为90.29%,超过了浓度为0.05 mg·mL^-1的芦丁和槲皮素标品对DPPH自由基的清除率。两种炮制方法均提高了多糖、水溶物、总黄酮3种提取物的还原能力,但是降低了醇溶物的还原能力。     

HPLC法测定甘薯叶片中的叶黄素

建立甘薯叶片中有效成分叶黄素含量的高效液相色谱测定方法,以寻求含量高的甘薯品种。采用Waters SunFire^TM C₁₈(150 mm×4.6 mm,5 μm )色谱柱;以甲醇—水为95:5(V/V)为流动相,流速为0.8 mL·min^-1;检测波长为445 nm;外标法定量。此色谱条件下,叶黄素含量在5~100 μg·mL^-1范围内时,与峰面积呈线性关系;样品平均回收率为99.5%;相对标准偏差 (RSD)为1.9%(n=5)。该方法灵敏、准确、专属性强,适用于甘薯叶片中叶黄素的测定;大部分供试品种间存在显著差异,其中以苏薯8号的含量最高。     

Nitric oxide induces [Ca2+]i oscillations in pituitary GH3 cells: involvement of IDR and ERG K+ currents

The role of nitric oxide (NO) in the occurrence of intracellular Ca²⁺ concentration ([Ca²⁺]_i) oscillations in pituitary GH₃ cells was evaluated by studying the effect of increasing or decreasing endogenous NO synthesis with L-arginine and nitro-L-arginine methyl ester (L-NAME), respectively. When NO synthesis was blocked with L-NAME (1 mM) [Ca²⁺]_i, oscillations disappeared in 68% of spontaneously active cells, whereas 41% of the quiescent cells showed [Ca²⁺]_i oscillations in response to the NO synthase (NOS) substrate L-arginine (10 mM). This effect was reproduced by the NO donors NOC-18 and S-nitroso-N-acetylpenicillamine (SNAP). NOC-18 was ineffective in the presence of the L-type voltage-dependent Ca²⁺ channels (VDCC) blocker nimodipine (1 µM) or in Ca²⁺-free medium. Conversely, its effect was preserved when Ca²⁺ release from intracellular Ca²⁺ stores was inhibited either with the ryanodine-receptor blocker ryanodine (500 µM) or with the inositol 1,4,5-trisphosphate receptor blocker xestospongin C (3 µM). These results suggest that NO induces the appearance of [Ca²⁺]_i oscillations by determining Ca²⁺ influx. Patch-clamp experiments excluded that NO acted directly on VDCC but suggested that NO determined membrane depolarization because of the inhibition of voltage-gated K⁺ channels. NOC-18 and SNAP caused a decrease in the amplitude of slow-inactivating (I_DR) and ether-à-go-go-related gene (ERG) hyperpolarization-evoked, deactivating K⁺ currents. Similar results were obtained when GH₃ cells were treated with L-arginine. The present study suggests that in GH₃ cells, endogenous NO plays a permissive role for the occurrence of spontaneous [Ca²⁺]_i oscillations through an inhibitory effect on I_DR and on I_ERG. voltage-gated potassium channels; ether-à-go-go-related gene potassium channels; slow-inactivating outward currents; fast-inactivating outward currents     

Ammonia Content in Mitochondria Isolated from Detached Pea Shoots. Effect of Light and Inhibitors of Photorespiratory Nitrogen Cycle

Mitochondria isolated from detached pea shoots which had beengrown in water for 1.5 h in the dark contained about 1.64 nmolNH₄^$ per mg of protein. Light treatment caused approximately50% increase in the mitochondrial NH₄^$. Methionine sulfoximineslightly reduced the mitochondrial NH₄^$, although total NH₄^$in the shoot extracts was greatly increased by the inhibitor. (Received June 17, 1985; Accepted September 24, 1985)     

野生和栽培马蹄金抗旱性比较及其抗旱机制初探

 为了鉴别野生和栽培马蹄金（Dichondra repens）的抗旱性,探讨其抗旱适应性的生理机制,对野生和栽培马蹄金进行了土壤水分胁迫处理,系统测定了野生和栽培马蹄金叶片内超氧化物歧化酶（SOD）、过氧化物酶（POD）、硝酸还原酶的活性和游离脯氨酸、可溶性糖、可溶性蛋白、NO₂^- /NO₃^-含量以及总DNA片断化程度。结果表明野生和栽培马蹄金在抗旱适应性上存在显著差异。随水分胁迫强度的增加,野生马蹄金叶片内抗氧化酶活性及其升高幅度,渗透调节物质的积累量及积累速度均高于栽培马蹄金,而叶片含水量下降的程度、DNA片断化程度低于栽培马蹄金,其中野生和栽培马蹄金叶片内NO₂^- /NO₃^-含量,硝酸还原酶的活性变化差异尤其显著,野生马蹄金叶片内的NO₂^- /NO₃^-含量、硝酸还原酶的活性明显高于栽培马蹄金,二者最大含量分别相差10和2.2倍。研究结果说明了野生马蹄金对干旱环境的适应性强于栽培品种;在干旱逆境下马蹄金叶片内的NO₂^－ /NO₃^－含量的变化在一定程度上可能反映了内源NO的变化,内源NO浓度的高低可能是野生和栽培马蹄金不同抗性的真正原因。     

The Uptake of Gaseous Ammonia by the Leaves of Italian Ryegrass

Lockyer, D. R. and Whitehead, D. C. 1986. The uptake of gaseousammonia by the leaves of Italian ryegrass.—J. exp. Bot.37: 919–927. Plants of Italian ryegrass (Lolium multiflorum Lam.) grown insoil with two rates of added ¹⁵N-labelled nitrate were exposed,in chambers, for 40 d to NH₃ in the air at concentrations of16, 118 and 520 µg m^–3. At the highest concentrationof NH₃, this source provided 47?3% of the total nitrogen inplants grown with the lower rate of nitrate addition (100mgN kg^–1 dry soil) and 35?2% with the higher rate (200mgN kg^–1 dry soil) At the intermediate concentration ofNH₃, the contributions to total plant N were 19?6% and 10?8%,respectively, at low and high nitrate while, at the lowest concentrationof NH₃, they were 5?1% and 32%. Most of the N derived from theNH₃ remained in the leaves, but some was transported to theroots. The amount of N derived from the NH³ that was presentin the leaves was not reduced by washing the leaves in waterat pH 5?0 before harvesting, indicating that the N was assimilatedby the plant and not adsorbed superficially. Rates of uptakeof NH₃ per unit leaf area ranged from 1?7 µg dm^–2h^–1 at a concentration of 16 µg m^–3 to 29?0µg dm^–2 h^–1 at a concentration of 520 µgm^–3 and with the lower rate of nitrate addition. Increasingthe supply of nitrate to the roots slightly reduced the rateof uptake of NH₃ per unit leaf area. Uptake of N from the higherrate of nitrate was reduced at the highest concentration ofNH₃ in the air. Key words: Ammonia, nitrogen, leaf sorption, Lolium multiflorum     

Pulmonary diffusing capacities for oxygen-labeled CO2 and nitric oxide in rabbits

Heller, Hartmut, Gabi Fuchs, and Klaus-DieterSchuster. Pulmonary diffusing capacities foroxygen-labeled CO₂ and nitric oxide in rabbits.J. Appl. Physiol. 84(2): 606-611, 1998.We determined the pulmonary diffusing capacity(DL) for¹⁸O-labeledCO₂(C¹⁸O₂)and nitric oxide (NO) to estimate the membrane component of therespective gas conductances. Six anesthetized paralyzed rabbits wereventilated by a computerized ventilatory servo system. Single-breath maneuvers were automatically performed by inflating the lungs with gasmixtures containing 0.9%C¹⁸O₂or 0.05% NO in nitrogen, with breath-holding periods ranging from 0 to1 s forC¹⁸O₂and from 2 to 8 s for NO. The alveolar partial pressures of C¹⁸O₂and NO were determined by using respiratory mass spectrometry. DL was calculated from gasexchange during inflation, breath hold, and deflation. We obtainedvalues of 14.0 ± 1.1 and 2.2 ± 0.1 (mean value ± SD)ml · mmHg¹ · min¹forDL_C¹⁸O2and DL_NO,respectively. The measured DL_C¹⁸O2/DL_NOratio was one-half that of the theoretically predicted value accordingto Graham's law (6.3 ± 0.5 vs. 12, respectively).Analyses of the several mechanisms influencing the determination ofDL_C¹⁸O2and DL_NOand their ratio are discussed. An underestimation of the membranediffusing component for CO₂ isconsidered the likely reason for the lowDL_C¹⁸O2/DL_NOratio obtained.

     

Light-dependent potassium uptake by Pisum sativum leaf fragments

A net K⁺ influx into chopped pea leaves bathed in 5 mM KCl,0.26 M sucrose and illuminated with 4000 lux amounted to about7.5 µmoles/g fresh weight-hr, while essentially no netflux occurred in the dark. This light-dependent K⁺ uptake waslinear with time for nearly 2 hr and continuously increasedas the light intensity was raised to 9000 lux. Over half ofthe K⁺ uptake was balanced by H⁺ release into the bathing solution,possibly by a mechanism in which bicarbonate was the anion accompanyingK⁺. The replacement of Cl^– by HCO₃^– increased thelight-dependent K⁺ uptake to 56 µmoles/g fresh weight-hr.About 23% of the light-dependent K⁺ uptake in 5 mM KCl was accompaniedby a Cl^– uptake. This net Cl^– influx was less sensitiveto the uncoupler tri-Fl-CCP and more sensitive to DCMU in thebathing solution than was the K⁺ uptake. The remaining net K⁺influx into pea leaf fragments was balanced by effluxes of sodium(accounting for 5%), magnesium (8%) and calcium (1%). (Received March 31, 1969; )     

Regulation of intracellular Ca2+ release in corpus cavernosum smooth muscle: synergism between nitric oxide and cGMP

Tonic contraction of corpus cavernosum smooth muscle cells (SMCs) maintains the flaccid state of the penis, and relaxation is initiated by nitric oxide (NO), leading to erection. Our aim was to investigate the effect of NO on the smooth muscle cellular response to adrenergic stimulation in corpus cavernosum. Fura-2 fluorescence was used to record intracellular Ca²⁺ concentration ([Ca²⁺]_i) from freshly isolated SMCs from rat and human. Phenylephrine (PE) transiently elevated [Ca²⁺]_i in the presence and absence of extracellular Ca²⁺, indicating release from intracellular stores. Whereas the NO donor S-nitroso-N-acetylpenicillamine (SNAP) with sildenafil citrate (SIL) caused no change in basal [Ca²⁺]_i, the PE-induced rise of [Ca²⁺]_i was reversibly inhibited by 27 ± 7% (n = 21, P < 0.005) in rat and by 55 ± 15% (n = 9, P < 0.01) in human SMCs. SNAP and SIL also reduced the contractile response to PE. To investigate the mechanism, we applied mediators alone or in combination. The soluble guanylyl cyclase inhibitor ODQ reduced the effect of SNAP and SIL. SIL, cGMP analogs, and NO donors without SIL did not reduce the PE-induced rise of [Ca²⁺]_i. However, the combination of 8-bromo-cGMP with SNAP reduced the Ca²⁺ peak by 42 ± 9% (n = 22, P < 0.01). Our results demonstrate that NO and cGMP act synergistically to reduce Ca²⁺ release from intracellular stores. Reduction of intracellular Ca²⁺ release may contribute to relaxation of the corpus cavernosum, leading to erection. calcium stores; nitric oxide; sildenafil citrate; inositol 1,4,5-trisphosphate receptor     

Effects of NO donors and synthase agonists on endothelial cell uptake of L-Arg and superoxide production

It is commonly believed thatthe activity of NO synthase (NOS) solely controls NO production fromits substrates, L-Arg and O₂. The Michaelis-Menten constant(K_m) of NOS forL-Arg is in the micromolarrange; cellular levels of L-Argare much higher. However, evidence strongly suggests that cellularsupply of L-Arg may becomelimiting and lead to reduced NO and increased superoxide anion(O₂·) formation, promotingcardiovascular dysfunction. Uptake ofL-Arg into cells occursprimarily (~85%) through the actions of aNa⁺-independent, carrier-mediatedtransporter (system y⁺). We haveexamined the effects of NOS agonists (substance P, bradykinin, and ACh)and NO donors(S-nitroso-N-acetyl-penicillamine and dipropylenetriamine NONOate) on transport ofL-Arg into bovine aorticendothelial cells (BAEC). Our results demonstrate that NOS agonistsincrease y⁺ transporter activity.A rapidly acting NO donor initially increases L-Arg uptake; however, afterlonger exposure, L-Arg uptake is suppressed. Exposure of BAEC withoutL-Arg to substance P and aCa²⁺ ionophore (A-23187) increasedO₂· formation, which was blockedwith concurrent presence ofL-Arg or the NOS antagonistN-nitro-L-arginine methyl ester.We conclude that factors including NO itself controly⁺ transport function and theproduction of NO and O₂·.