FCLA-PPIX光触发式静态化学发光体系检测人血清白蛋白

本文对基于FCLA-PPIX触发式化学发光的人血清白蛋白检测体系进行了研究。该触发式化学发光体系利用光敏剂和活性氧探针FCLA组成了一个基本的光触发式化学发光检测体系,将待检测样品和体系充分混合后,通过脉冲光触发和延迟发光检测技术测定人血清白蛋白含量。脉冲触发式静态检测体系克服了其他化学发光体系操作复杂、试剂和样品消耗量大以及信号不稳定的缺点,灵敏度高,检测稳定。本文考察了该体系激发光强度、试剂浓度及温度等对人血清白蛋白检测结果的影响。在优化的实验条件下,测定人血清白蛋白的线性范围为2.5×10-9~6.4×10-7mol/L;方法的检出限为1.6×10-10mol/L;本法对5×10-8mol/L的标准样品平行重复测定11次相对标准偏差为4%。     

普通小球藻引发水中17β-雌二醇的光降解

研究了普通小球藻引发水中17β-雌二醇(E2)的光降解,结果表明,在250W高压汞灯(HPML,nm)的照射下,藻浓度为4．0×10^10个·L^-1时,17β-雌二醇的光降解率可达37％．藻浓度为4．2×10^10个·L^-1时,17β-雌二醇浓度在1．5×10^-5-6．0×10^-5mol·L^-1范围内,其光降解速率与初始浓度成正比,反应是假一级．另外,还研究光强、藻悬浮液浓度和17β-雌二醇初始浓度等对此反应体系的影响。     

表面活性剂增敏——牛血清白蛋白荧光猝灭法测定槐花中芦丁含量

目的：应用牛血清白蛋白荧光猝灭法建立一种测定槐花中芦丁含量的新方法。方法：牛血清白蛋白（BSA）具有很强的内源荧光性,而芦丁溶液本身不产生荧光。当芦丁与BSA结合后,会导致其荧光强度下降,表面活性剂吐温-80（T-80）对体系有促进荧光猝灭作用。BSA在λex=338nm处的荧光猝灭程度与芦丁的量在一定浓度范围内呈良好的线性关系,据此建立测定槐花中芦丁含量的新方法。结果：该结合物的最大发射波长为λmax=338nm,与芦丁摩尔浓度在6×10-7-3.0×10-5mol.L-1范围内线性关系良好。其线性回归方程为ΔF=136.36CRu（×10-5mol.L-1）-0.5454,相关系数r=0.9976,检出限为1.58×10-7mol.L-1,RSD为2.8%-4.3%,加标回收率为97.6%~101.2%。结论：本方法操作简便、快速,用于实际样本的测定,结果满意。     

表面活性剂增敏--牛血清白蛋白荧光猝灭法测定槐花中芦丁含量

目的:应用牛血清白蛋白荧光猝灭法建立一种测定槐花中芦丁含量的新方法。方法:牛血清白蛋白(BSA)具有很强的内源荧光性,而芦丁溶液本身不产生荧光。当芦丁与BSA结合后,会导致其荧光强度下降,表面活性剂吐温-80(T-80)对体系有促进荧光猝灭作用。BSA在λex=338nm处的荧光猝灭程度与芦丁的量在一定浓度范围内呈良好的线性关系,据此建立测定槐花中芦丁含量的新方法。结果:该结合物的最大发射波长为λmax=338nm,与芦丁摩尔浓度在6×10-7~3.0×10-5mol.L-1范围内线性关系良好。其线性回归方程为ΔF=136.36CRu(×10-5mol.L-1)-0.5454,相关系数r=0.9976,检出限为1.58×10-7mol.L-1,RSD为2.8%~4.3%,加标回收率为97.6%~101.2%。结论:本方法操作简便、快速,用于实际样本的测定,结果满意。     

茉莉酸甲酯对水稻幼苗生长的影响(简报)

2．5×10－7mol·L-1茉莉酸甲酯促进水稻幼苗生长,而高于2．5×10-5mol·L－1则抑制。不同水稻品种对茉莉酸甲酯的反应不同。     

花生幼苗下胚轴质膜氧化还原系统

花生幼苗下胚轴细胞膜制剂具有氧化NAD(P)H,还原Fe(CN)_6~(3-)与EDTA-Fe~(3 )的能力,当Fe(CN)_6~(3-)浓度为1mmol/L时,膜制剂氧化NADH和NAD(P)H的K_m分别为100和110μmol/L;V_(max)为1400和710nmol mg~(-1) proteinmin~(-1),最适pH为8.0。NADH 0.25 mmol/L浓度下,膜制剂还原Fe(CN)_6~(3-)的K_m为40,V_(max)为1300;还原EDTA-Fe~(3 )的K_m为125,V_(max)为180,最适pH分别为8.0与7.0。氧为该系统天然受体,鱼藤酮、抗霉素A与CN~-对其活性无影响,10μmol/L的DCCD抑制其活性约20％,10μmol/L的SHAM抑制近50％活性。     

高效液相色谱法测定大黄素在Caco-2细胞中的摄取

目的:采用反相高效液相色谱法,观察大黄素在Caco-2细胞中的摄取特点。方法:将大黄素与Caco-2细胞共同孵育,收集细胞样品,液氮反复冻融。取细胞裂解液,加入甲醇提取,提取液采用HPLC进行分析。色谱分析柱为C18柱(250mm×4．6mm,5μm,Diamonsil),流动相组成为85%乙腈及15%水(含0．1%乙酸),流速1ml·min-1,进样量20μl,柱温25℃,3D模式采集数据。结果:检测Caco-2细胞中大黄素的工作曲线的回归方程为Y=0．278x 0．148(Y=0．9996,n=5),线性范围为0．037～4．8μmol·L-1,最低检测浓度为0．018μmol·L-1。当细胞中大黄素的浓度为0．05、2和8．5μg·ml-1时,回收率分别为(101．3±7．3)%、(96．7±3．0)%和(98．7±2．1)%(n=5);相应的日内标准偏差分别为0．25%、2．9%和1．4%;相应的日间标准偏差分别为2．3%、5．6%和6．3%。大黄素在Caco-2细胞中的摄取达峰时间为10分钟,峰浓度为108．56±11．57 nmol/L·mg·protein,10分钟后Caco-2细胞中大黄素的含量迅速下降。浓度处于2-50μM之间时,Caco-2细胞对大黄素的摄取量呈线性增加,浓度达50μM后,随着剂量的增加大黄素的摄取量变化不明显。结论:大黄素可被Caco-2细胞迅速摄取,随着剂量的增加,大黄素在Caco-2细胞中的摄取存在饱和现象。     

光合细菌产辅酶Q10发酵条件的研究

利用均匀设计原理进行实验设计 ,对光合细菌R .capsulatusMT1131产辅酶Q10培养基配方及培养条件进行优化 ,结果当培养基中酵母膏质量浓度为 3 .13g·L- 1 ,硫酸铵 0 .8g·L- 1 ,Mg2 + 0 .6 4g·L- 1 ,Fe2 + 45 .2mg·L- 1 ,Mn2 + 18mg·L- 1 ,Co2 + 16mg·L- 1 ,培养基初始pH值为 7.0时 ,于 30℃ ,光照强度为 2 0 0 0Lx条件下培养 4天后 ,菌体中辅酶Q10质量浓度由 15 .2 13mg·L- 1提高至 2 0 .36 5mg·L- 1 ,产量提高约 33.87%。     

不同盐度和Fe3+浓度对小球藻生长、硝酸还原酶活性及基因表达的影响

论文以一株紫外诱变获得的蛋白核小球藻F-9-3为材料,研究了4个盐度和5个Fe3+浓度对其生长和硝酸还原酶(NR)活性及其基因表达的影响。盐度实验结果表明在0～0.30 mol L-1NaCl浓度范围内,小球藻生长较快;NR酶活性及其基因表达量都是在0.15和0.30 mol L-1NaCl较高。铁浓度实验结果表明在0.03和0.06 mmol L-1Fe3+浓度培养小球藻生长较快,NR酶活性和基因表达量较高;而高铁浓度组(0.12 mmol L-1)其生长受抑制,缺铁情况下nr基因表达量最高。因此适合该小球藻生长的最适盐度范围是0～0.30 mol L-1NaCl和0.03～0.06 mmol L-1Fe3+浓度     

Fe限制对小球藻-隆线溞食物链的影响

为了研究环境中Fe限制对小球藻-隆线溞食物链的影响,在0～10000 nmol·L-1的范围内设置5个Fe3+浓度梯度培养小球藻(Chlorella vulgaris),22 d后收集藻细胞,然后将收集到的小球藻饲喂隆线溞(Daphnia carinata)14 d,观察小球藻和隆线溞的生长状况。结果表明:Fe3+浓度为0、10、100和1000 nmol·L-1时小球藻的生物密度和叶绿素a的含量都明显低于Fe3+浓度为10000 nmol·L-1时;Fe3+浓度的下降导致小球藻对N的吸收减少,对P的吸收增加;隆线溞的体长和总产幼数随着Fe3+浓度的下降而逐渐降低,当Fe3+浓度为10000 nmol·L-1时,隆线溞最终体长和总产幼数分别为3 mm和93个,而当Fe3+浓度为0 nmol·L-1时,隆线溞最终体长和总产幼数分别为2.6 mm和60个;这说明Fe限制不仅会影响小球藻的生长和其对N、P的吸收,还会抑制以小球藻为食的隆线溞的生长和繁殖。以上结果表明,Fe限制对小球藻-隆线溞食物链的上行效应显著,这有助于进一步了解Fe对淡水浮游动植物的影响。     

Flow‐injection chemiluminescence determination of melamine in urine and plasma

A novel flow‐injection chemiluminescence method for the determination of melamine in urine and plasma was developed. It was found that melamine can remarkably enhance chemiluminescence emission from the luminol–K₃Fe(CN)₆ system in an alkaline medium. Under the optimum conditions, chemiluminescence intensity had a good linear relationship with the concentration of melamine in the range 9.0 × 10^–9–7.0 × 10^–6 g/mL, with a correlation coefficient of 0.9992. The detection limit (3σ) was 3.5 ng/mL. The method has been applied to determine the concentration of melamine in samples using liquid–liquid extraction. Average recoveries of melamine were 102.6% in urine samples and 95.1% in plasma samples. The method provided a reproducible and stable approach for the sensitive detection of melamine in urine and plasma samples. Copyright © 2011 John Wiley & Sons, Ltd.     

Determination of estradiol valerate in pharmaceutical preparations and human serum by flow injection chemiluminescence

A novel method for the detection of trace estradiol valerate (EV) in pharmaceutical preparations and human serum was developed by inhibition of luminol chemiluminescence (CL) by estradiol valerate on the zinc deuteroporphyrin (ZnDP)‐enhanced luminol‐K₃Fe(CN)₆ chemiluminescence system. Under optimized experimental conditions, CL intensity and concentration of estradiol valerate had a good linear relationship in the ranges of 8.0 × 10^‐8 to 1.0 × 10^‐5 g/mL. Detection limit (3σ) was estimated to be 3.5 × 10^‐8 g/mL. The proposed method was applied successfully for the determination of estradiol valerate in pharmaceutical preparations and human serum and recoveries were 97.0‐105.0% and 95.5‐106.0%, respectively. The possible mechanism of the CL system is discussed. Copyright © 2012 John Wiley & Sons, Ltd.     

Chemiluminescent in vitro estimation of the inhibitory constants of antioxidants ascorbic and uric acids in Fenton’s reaction in urine

The goal of this research was to measure in vitro the inhibitory constants of the antioxidants ascorbic and uric acid in urine, with lucigenin enhanced chemiluminescence (CL) in Fenton’s system. Maximum CL emission is registered in urine containing H₂O₂ (5·10⁻⁴ M), Fe²⁺ (5·10⁻⁵ M), EDTA (5·10⁻⁵ M), and chemical enhancer lucigenin (10⁻⁴ M) at pH 5.5 and 36°C. Ascorbic acid exhibits up to 4-fold stronger antioxidant effect than uric acid. The constants of antioxidant inhibition in urine were measured at concentrations 10⁻³ and 10⁻⁴ M: for ascorbic acid, 5.92 ± 0.04 and 24.05 ± 1.82 μmol·sec⁻¹; for uric acid, 1.60 ± 0.02 and 21.45 ± 0.97 μmol·sec⁻¹, respectively. Three phases of CL kinetics of urine are well observed: spontaneous CL (0–10 sec), fast flash of CL (10–50 sec), and latent period (50–300 sec). The antioxidant efficiency of ascorbic and uric acids in the final stage of catabolic processes in the body is discussed. Published in Russian in Biokhimiya, 2006, Vol. 71, No. 8, pp. 1062–1065.     

Fast repair of purine deoxynucleotide radical cations by rutin and quercetin

Repair effects of rutin and quercetin on purine deoxynucleotide radical cations were studied using pulse radiolysis technique. On electron pulse irradiation of N₂ saturated deoxynucleotide aqueous solution containing 20 mmol/L K₂S₂O₈, 200 mmol/Lt-BuOH and rutin or quercetin, the transient absorption spectra of the deoxynucleotide radical cations decayed quickly. At the same time, the spectra of flavonoid phenoxyl radicals formed within several dozen microseconds. The results indicated that deoxynucleotide radical cations can be repaired by flavonoids. The rate constants of the repair reactions were 3.8 ×10⁸-4.4 ×10⁸ mol⁻¹ · L · s⁻¹ and 1.3×10⁸-1.8×10⁸ mol⁻¹ · L · s⁻¹ for dAMP and dGMP radical cations, respectively.     

Fast repair of purine deoxynucleotide radical cations by rutin and quercetin

Repair effects of rutin and quercetin on purine deoxynucleotide radical cations were studied using pulse radiolysis technique. On electron pulse irradiation of N2 saturated deoxynucleotide aqueous solution containing 20 mmol/L K2S2O8, 200 mmol/L f-BuOH and rutin or quercetin, the transient absorption spectra of the deoxynucleotide radical cations decayed quickly. At the same time, the spectra of flavonoid phenoxyl radicals formed within several dozen microseconds. The results indicated that deoxynucleotide radical cations can be repaired by flavonoids. The rate constants of the repair reactions were 3.8 × 108-4.4×108 mol-1 · L · s-1 and 1.3×108-1.8×108 mol-1 · L · s-1 for dAMP and dGMP radical cations, respectively.     

The kinetics of oxidation of hemerythrin species—linear free energy relationships

The second-order rate constants (M^?1sec^?1, 25°C, pH 8.2, I = 0.15 M) for the oxidation to (semi-met)₀of deoxyhemerythrin from Phascolopsis gouldii (P.g.) and Themiste zostericola (T.z.) have been determined for Fe(CN)₅(4-NH₂py)^2? (3.6 × 10⁴ T.z.,2.8 × 10²P.g.),Fe(CN)₅NH₃^2?(2.4 × 10⁴ T.z.), Fe(CN)₆^3? (1.0 × 10⁵ T.z.,1.4 × 10²P.g.), Fe(CN)₅PPh₃^2? (7.3 × 10⁵T.z.), and Fe(CN)₄dipy- (~6 × 10⁶ T.z.,7.5 × 10⁴ P.g.). Corresponding rate constants for the oxidation of (semi-met)_R to met are: Fe(CN)₅(4-NH₂py)^2? (1.2 × 10³ P.g.), Fe(CN)₆^3? (3.4 × 10⁵ T.z., 4.5 × 10 Fe(CN)₅PPh₃^2? (4.4 × 10⁴P.g.), Fe(CN)₄dipy^? (1.7 × 10⁵P.g.), and Coterpy₂³⁺ (5.1 P.g.) The rates of oxidation of deoxy- and (semi-met)_R myohemetythrin by Fe(CN)₆^3? were too rapid for stopped-flow measurement. The Marcus relationship for cross-reactions was successfully applied to these data.     

A novel chemiluminescent method for determination of phloroglucinol

It was found that the inhibition and enhancement by phloroglucinol of the chemiluminescence from the luminol–K₃Fe(CN)₆ system were dependent on the pH of luminol solution and the concentration of phloroglucinol. In Na₂CO₃–NaHCO₃ buffer, phloroglucinol exhibited strong chemiluminescent enhancement at pH 9.4. On this basis, a flow injection method was developed for the determination of phloroglucinol. The method was simple, rapid, convenient and sensitive, with a detection limit of 2.0 × 10^?9 mol/L. It is effective for determining phloroglucinol in the range of 1.0 × 10^?5–5.0 × 10^?9 mol/L. The relative standard deviation is 1.3% within one day and 3.2% between days for the determination of 5.0 × 10^?7 mol/L phloroglucinol. The method has been successfully used to determine phloroglucinol in environmental water, with satisfactory results. Copyright © 2003 John Wiley & Sons, Ltd.     

A chiral,porous, organic cage‐based,enantioselective potentiometric sensor for 2‐aminobutanol

A new enantioselective potentiometric sensor containing R‐type chiral porous organic cage CC9 as the chiral selector was designed for the assay of 2‐aminobutanol. Optimized membrane electrodes displayed a linear dynamic range from 10⁻³ ~ 10⁻¹ mol·L⁻¹ with a detection limit of 2.5 × 10⁻⁴ mol·L⁻¹ and a Nernstian response of 27 ± 0.5mV·decade⁻¹ toward S‐2‐aminobutanol within the pH range 7.0–10.0. The potentiometric enantioselectivity coefficient ( ) of this sensor was −1.333, indicating that the porous organic cage‐based electrode exhibited good discrimination toward S‐2‐aminobutanol over R‐2‐aminobutanol.     

Preparation of cadmium sulfide nanoparticles and their application for improving the properties of the electrochemical sensor for the determination of enrofloxacin in real samples

An advanced electrochemical sensor for the detection of enrofloxacin (ENR) based on the use of a modified electrode containing cadmium sulfide (CdS) nanoparticles (NPs) is reported. The CdS NPs were synthesized and characterized and then coated onto the electrode to fabricate a modified electrode that exhibited a lower limit of detection of 9.5 × 10^?8 mol·L^?1. This detection limit compares with a traditional electrode that exhibited a concentration detection range of 1.0 × 10^?2 to 1.0 × 10^?7 mol·L^?1. This modified electrode demonstrated good selectivity, reproducibility, response time (<40 s), lifetime (up to 12 wk), and pH range (3.3‐7.2) for the determination of ENR in real samples (eg, pig urine).     

Effects of charged peptides on electron transfer from [Fe(CN)6]4– to cytochrome c or plastocyanin

 Interactions of charged peptides, such as aspartic acid peptides (Aspptds) and lysine peptides (Lysptds), with cytochrome c (cyt c) or plastocyanin (PC) have been studied by measuring electron transfer between [Fe(CN)₆]^4– and cyt c or PC in the presence of these peptides. Aspptds, up to penta-aspartic acid, served as competitive inhibitors of electron transfer from [Fe(CN)₆]^4– to oxidized cyt c, while Lysptds, up to penta-lysine, promoted electron transfer from [Fe(CN)₆]^4– to oxidized PC. The electron transfer inhibitory effects of Aspptds are explained as competitive inhibition due to neutralization of the positively charged amino acid residues at the surface of cyt c by electrostatic interactions, whereas the electron transfer promoting effects of Lysptds may be due to formation of PC·Lysptd or Lysptd·[Fe(CN)₆]^4– complexes subsequently forming an electron transferring complex, PC·Lysptd·[Fe(CN)₆]^4–, without repulsion of the negative charges. The inhibitory effect of Aspptds and promotional effect of Lysptds became significant as the net charge or concentration of the peptides increased. The promotional effects of Lysptds decreased as the net charge of the PC negative patch was decreased by mutagenesis. Thus, charged peptides may serve as a probe for investigation of the molecular recognition character of proteins. Received: 19 May 1998 / Accepted: 27 July 1998