化学发光法测定SOD活力

超氧化物歧化酶(SOD)能清除O_2自由基。本文介绍用邻苯三酚自氧化体系的化学发光法测定SOD活力,并用该方法测定了肝炎病人血清SOD的活性及铜、锌离子取代后SOD的活性。     

化学发光法测定NK细胞杀伤活性的可行性研究

本文选择能够用化学发光法灵敏检测的金属离子 Co~(2-)为对象,研究了它与氨酸络合剂形成的络合物标记靶细胞,以化学发光法测定自然杀伤细胞(NK 细胞)活性的可行性。     

邻苯三酚法测定超氧化物歧化酶缓释片中SOD的活性

目的建立超氧化物歧化酶 (SOD)缓释片的活性测定方法。方法采用邻苯三酚自氧化法。结果在SOD浓度为 0 .0 6 0～0 . 0 0 6mg·ml-1时线性关系良好 (r=0 .9996 ) ;平均空白加样回收率为 10 1 .94 % (n=9)。结论所用方法简单、快速 ,结果准确。     

HPLC法测定清热解毒口服液中绿原酸的含量

目的:建立高效液相色谱法测定清热解毒口服液中绿原酸的含量方法。方法:色谱柱:KromasilC18(250 mm×4.6 mm,5μm),流动相:乙腈-0.4%的磷酸(20:80)检测波长:324nm,流速为1.0 ml·min-1。结果:绿原酸的回归方程为:Y=3854734.2x-73605618,г=0.9999(n=6)线性范围为:10.01μg/ml~60.07μg/ml,平均回收率为97.04%,RSD为1.01%(n=5)。结论:方法简便,结果准确,专属性强,重现性好,可将其作为清热解毒口服液的质控指标之一。     

RP-HPLC法测定芦笋口服液中芦丁的含量

目的 建立芦笋口服液中的芦丁含量测定方法.方法 用RP-HPLC法,分析色谱柱为十八烷基硅烷键合硅胶为填充剂(Shim Pack VP ODS,250 mm×4.6 mm,5 μm),柱温30℃,甲醇 2.5%冰醋酸溶液(39∶61)为流动相,检测波长为257 nm.结果 芦丁与其它各组分基线分离良好,在所试浓度3.072～76.8 mg·L-1范围内与峰面积值呈良好的线性关系,回归方程为Y=22 925X-5 176.9,r2=0.999 7(n=5);精密度试验(n=6)RSD为0.93%;重现性试验(n=6) RSD为3.81%;平均回收率(n=6)为99.21%,RSD为2.02%;三批芦笋口服液中芦丁的含量为52～76 μg/瓶.结论 该法样品处理简单,准确性高,精密度好,适合芦笋口服液中芦丁的含量测定.     

HPLC法测定额勒更口服液黄连中盐酸小檗碱含量

目的:采用HPLC法测定额勒更口服液黄连中盐酸小檗碱含量。方法:采用高效液相色谱法。结果:额利更口服液中黄连所含盐酸小檗碱线性范围0.1004μg～1.004μg,(r=1)。平均回收率为99.9%,RSD为1.96%。结论:本法操作简单,重现性好,专属性强;可作为额勒更口服液中盐酸小檗碱含量测定的一种准确、灵敏、可行的方法。     

HPLC法测定甘之肖口服液中左旋肉碱的含量

目的：建立测定甘之肖口服液中左旋肉碱含量测定的方法。方法：采用HYPERSIL ODSC18柱（250mm×4．6mm,5μm）;0．002mol·L^-1辛烷磺酸钠溶液（加入0．125％磷酸二氢钾、0．125％磷酸氢二钾,用磷酸调pH至3．0）-乙腈（92．5：7．5）为流动相;流速0．8ml·min^-1;检测波长：210nm;柱温：室温。结果：左旋肉碱的进样量在2．55～30．60g范围内呈良好的线性关系（r=0．9999）,平均加样回收率为97．1％,RSD为1．93％（n=6）。结论：本方法快速、准确、重复性好,可作为甘之肖口服液质量控制。     

流动注射化学发光法测定人尿液中尿酸的含量

目的:建立一种快速、灵敏地测定人尿液中尿酸(UA)含量的新方法。方法:利用在碱性条件下,UA可以选择性地猝灭抗坏血酸与荧光素-表面活性剂-铜离子混合物产生的化学发光,结合流动注射技术,建立了流动注射化学发光法直接测定人尿液中UA含量的方法。反应试剂的浓度分别为氢氧化钠溶液0.5mol/L、荧光素5.0×10-4mol/L、铜离子1.0×10-4mol/L、溴化十六烷基三甲铵(CTMAB)5.0×10-3mol/L、抗坏血酸25mg/L,主泵(P1)、副泵(P2)流速均为2.5ml/min,光电倍增管负高压为800V。结果:UA的检测质量浓度在0.1²0mg/L范围内线性关系良好(r=0.999 0),检出限为3.3×10-2mg/L,平均加样回收率为99.63%,精密度RSD为1.2%(n=11)。结论:本方法操作简便、灵敏度高、选择性好,为尿液中UA含量测定提供了一种新方法。     

薄层扫描法测定回生口服液中水苏碱的含量

采用醇沉及两次柱色柱的方法预处理样品，用薄层扫描法测定了样品水中苏碱的含量。λｓ＝４１５ｎｍ，λＲ＝５５０ｎｍ。测定结果表明，水苏碱含量与峰面积积分值线性范围为２。６￣８．３μｇ，ｒ＝０．９９８６，平均回收率为９７．４％，ＲＳＤ＝２．３％（ｎ＝６）。为回生口服液中水苏碱的含量测定提供了准确可靠的方法。     

HPLC法测定升白口服液中阿魏酸含量

升白口服液系在传统方“当归补血汤”的基础上增加数味药物，由党参、当归、熟地、枸杞子等十几味中药组成，利用水煮醇沉工艺提取多种水溶性成分制备而成的口服液体制剂。其中当归、黄芪、甘草等的有效成分均含有阿魏酸。本文拟订了口服液中阿魏酸含量测定的HPLC法，用于中间品容量校正及成品的质量控制，方法简便，现介绍如下。1仪器与试剂Perkin－Elmer；LC－250梯度泵；LC－290紫外-可见检测器；LCl－100数据处理器。阿魏酸对照品（773－93013），中国药品生物制品检定所；升白口服液，自制。2方法与结果2．1色谱条件ODSC18柱（1…     

连苯三酚自氧化法测定超氧化物歧化酶活性的改进

改进后的连苯三酚自氧化法测定条件是:25℃,总体积4.5ml,50mM pH8.2Tris-HCl 缓冲液,连苯三酚的自氧化速率为0.070 O.D./min。本法具有操作简便,灵敏度高,用样量少等优点。     

高纯度超氧化物歧化酶制备方法的改进

本文报道了制备超氧化物歧化酶的方法,即首先75℃下加热处理15min,然后用硫酸铵分级盐析,再用少量乙醇、氯仿处理,最后经分子筛层析,可得到高纯度的产品。从1000ml猪红细胞中,可得到46.5mg酶,比活力达每毫克蛋白14200u.1000ml码红细胞,可得到30mg酶,比活力达每毫克蛋白16600u.所得产品经disc-PAGE,SDS-PAGE,N-末端氨基酸分析等检测,均表明已达均一程度。此法可减轻环境污染,操作简便,经济实用,适用于工厂化生产。     

亚硝酸盐法测定超氧化物歧化酶活性

用亚硝酸盐、氰化钾抑制区别法测定样品中的 Cu,Zn-SOD 和 Mn-SOD,灵敏度高,重现性好,操作简便。实验表明,抑制率达50%时所需 SOD 的浓度为0.07 μg/ml,其批内和批间的变异系数分别为6.0%和5.5%,鸡肝 Cu,Zn-SOD 与 KCN 的抑制复合物的表观解离常数 K′为288.6±45.5 μmol。     

超氧化物歧化酶的分析测定

超氧化物歧化酶的测活方法很多,直接法所用仪器价格昂贵,应用受到限制,应用最广泛的是借助于氧自由基指示清除剂的间接法。本文对SOD的测活方法的原理和特点作了较全面的概述。     

PHARMACOKINETICS OF SUPEROXIDE DISMUTASE IN RATS AFTER ORAL ADMINISTRATION

The kinetic behaviour of bovine erythrocyte Cu--Zn SOD was investigated in Sprague Dawley male rats after subcutaneous and oral administrations of doses ranging from 0·5 to 20 mg kg⁻¹. Studies have been carried out with SOD and SOD encapsulated into liposomes containing or not containing ceramides. The maximum concentration (C_max) in blood cell pellets ranged from 8·65 to 11·03 U/mg haemoglobin (Hb) after subcutaneous injection, and from 4·48 to 8·23 U/mg Hb after oral administration. The maximum concentrations were reached in 5 h (t _max) for the two routes. Comparison between the areas under the curves (AUCs) obtained after subcutaneous and oral administration allowed the calculation of relative bioavailability (F ′). The maximum bioavailability after oral administration was 14% for free SOD, 22% for SOD encapsulated into liposomes, and 57% when ceramides were added to liposomes. Poor SOD bioavailability was enhanced by liposome encapsulation, and ceramide addition seemed to be beneficial for oral encapsulated SOD administration.     

EFFECTS OF ENDURANCE TRAINING ON SUPEROXIDE DISMUTASE ACTIVITY,CONTENT AND mRNA EXPRESSION IN RAT MUSCLE#

1. The purpose of the present study was to investigate the changes in superoxide dismutase (SOD) isoenzyme (Mn²⁺-SOD and Cu²⁺, Zn²⁺-SOD) activities, contents and mRNA expressions in rat skeletal muscle during endurance training and a single bout of exercise. 2. Thirty-eight male Wistar rats were divided into untrained (U) and trained (T) groups. The T group rats were treadmill-trained for 9 weeks. The activity, content and mRNA expression of Mn²⁺-SOD and Cu²⁺, Zn²⁺-SOD were determined in the soleus muscle of each rat. 3. Mn²⁺-SOD activity and content in the T group were significantly higher than in the U group, both at rest (22 and 21%, respectively) and after exercise (24 and 46%, respectively), while a single bout of exercise affected neither the activity nor content of Mn²⁺-SOD in either group. 4. The content of Cu²⁺, Zn²⁺-SOD in both groups was not different at rest and after exercise, although its activity at rest was significantly higher in the T group than in the U group (by 29%). 5. After exercise, the expression of Mn²⁺-SOD mRNA was markedly attenuated only in the U group (49%); the expression of Cu²⁺, Zn²⁺-SOD mRNA was not influenced by exercise. 6. Our results suggest that adequate endurance training increases both the activity and content of Mn²⁺-SOD and that untrained rats are rather susceptible to oxidative stress during physical exercise. It thus appears that Mn²⁺-SOD provides a reliable index of physical training. 7. The results obtained in the present study also suggest that muscle has the capacity of responding to training in such a manner as to reduce the potential harm arising from the accumulation of oxygen free radicals resulting from enhanced metabolic activity.     

评价抗癌物质活性的改良MTT方法

用于抗癌活性评价的ＭＴＴ法采用“ＳＤＳ－异丁醇－ＨＣ１”溶解液后可完全溶解甲解ｚａｎ，也可完全溶解细胞和血清蛋白等成份，得到澄明、稳定的甲ｚａｎ溶液（Ａ５７０值稳定时间不少于１２ｈ）。本法测得的抗癌活性量－效关系十分显著，与同位素核酸体参入法结果吻合。本法用于悬浮细胞的药敏测定时更为方便、可靠。     

SUPEROXIDE DISMUTASE ATTENUATED POST-ISCHAEMIC CONTRACTILE DYSFUNCTION IN A MYOCARDIAL XANTHINE OXIDASE DEFICIENT SPECIES

1. We assessed the effect of polyethylene glycol conjugated superoxide dismutase (PEG-SOD) on myocardial stunning in the rabbit heart in which xanthine oxidase level is extremely low. 2. In open-chest anaesthetized rabbits, the left marginal branch of the coronary artery was occluded for 10 min and then reperfused for 30 min. A group of rabbits (PEG-SOD group) received 1000 units/kg of PED-SOD and another group (control group) was given saline 15 min before the coronary occlusion. 3. Regional systolic thickening fraction (TF) was similarly reduced to approximately -25% of baseline value during ischaemia in both groups. However recovery of TF after reperfusion was significantly better in the PEG-SOD group (n = 9) and TF at 30 min after reperfusion was 70.1 +/- 3.9% of baseline value compared with 44.9 +/- 3.4% in the control group (n = 9; P less than 0.05). Rate-pressure products, left ventricular pressure, and LV dP/dt max were not significantly different between the PEG-SOD treated and untreated control rabbits at any time during the experiment. PEG-SOD did not modify the regional myocardial blood flow (coloured microsphere method) during ischaemia/reperfusion, which was assessed by using separate groups of rabbits. 4. These findings indicate that oxygen free radicals are important in the pathogenesis of myocardial stunning in xanthine oxidase deficient hearts.     

消化性溃疡患者血清LPO含量及SOD活性变化的初步研究

对８４例消化性溃疡患者血清ＬＰＯ含量及Ｃｕ，Ｚｎ－ＳＯＤ活性进行了测定，发现胃溃疡及复合性溃疡患者血清ＬＰＯ含明明显高于正常对照（Ｐ〈０．００１），血清Ｃｕ，Ｚｎ－ＳＯＤ活性则显低于正常对照（Ｐ〈０．０１）；十二指肠球部溃疡患者血清ＬＰＯ含量有升高趋势，而血清Ｃｕ，Ｚｎ－ＳＯＤ活性在正常范围，结果提示，胃溃疡的发生可能与氧自由基的作用有关。