Zinc, copper and aluminium in the corpuscular components of peripheral blood in patients with pre-terminal and terminal renal failure

Summary Trace elements play an essential role in the activation and regulation of metabolic processes. By using the flameless graphite furnace-AAS, accurate results can be obtained. In order to verify blood cells as more representative than plasma to reflect the whole body content of trace elements at uremic state, two patient groups, one with preterminal renal failure (n=40) and the second with chronic renal failure undergoing hemodialysis (n=20) were examined. The concentration levels of zinc, copper and aluminium were measured in different matrices, i.e. whole blood, plasma and corpuscular blood components. The analytical technique is eligible to isolate mono- and polymorphnuclear leucocytes for the determination of intracellular contents. In agreement with other reports, zinc and copper were decreased and aluminium was increased in the plasma of both patient groups. Different results could be found for intracellular contents with mean zinc concentration of erythrocytes, thrombocytes and leucocytes between mean and higher levels, though staying within normal values. Copper was decreased in erythrocytes and increased in thrombocytes, but not detectable in leucocytes. The results of this study point out the differences between the plasmatic and intracellular alteration of trace element contents in uremic patients. However, the concentration levels of all cellular compartments show high standard deviation, which shows the necessity of measuring additional matrices, e.g. plasma. Uremic symptoms relate to the disorders of the trace element homeostasis. Regarding the increased fragility of erythrocytes with shortened life duration, decreased copper contents could play an essential role. Increased aluminium concentration levels might also be one of the interfering factors in the uremic thrombocytopathy. Yet the measurement of the plasma content is inevitable to diagnose and reflect the whole body content of trace elements at uremic state. By keeping analysis control, important information about progression and influence of therapy can be obtained.Dedicated to Professor Dr. Wilhelm Fresenius on the occasion of his 80th birthday     

ICP-AES法测定方铅矿中多元素的方法研究

采用电感耦合等离子体原子发射光谱法(ICP-AES)进行方铅矿中多元素同时测定.通过对方铅矿样品化学处理试验建立了HCl-NH4Cl-HNO3的溶矿体系.本体系采用基体匹配、背景系数和元素干扰系数校正及元素内标法确定了最佳综合实验测试条件.本实验建立的ICP-AES法同时测定方铅矿中镉、钴、铜、铁、铟、铅、锌7种元素的方法,本方法测量相对误差RE (n=8)为1.50%～7.50%,相对标准偏差RSD (n=8)为1.7%～5.7%.经国家一级标准物质GBW 07269分析验证可以满足方铅矿单矿物样品的分析要求.     

On-line solid-phase extraction LC-MS/MS for the determination of Ac-SDKP peptide in human plasma from hemodialysis patients

We developed a high-throughput method based on on-line solid-phase extraction liquid chromatography tandem mass spectrometry (SPE-LC-MS/MS) to determine N-terminal thymosin-β fragment peptide (N-acetyl-seryl-aspartyl-lysyl-proline, Ac-SDKP) in human plasma samples. Quantification of Ac-SDKP was performed using direct injection for on-line SPE based on C(18), reversed-phase LC separation and stable isotope dilution electrospray ionization-MS/MS in multiple reaction-monitoring (MRM) mode. The Ac-SDKP-(13)C(6), (15)N(2) (m/z 496 → 137) was synthesized for the internal standard. The MRM ion for Ac-SDKP was m/z 488 → 129 (quantitative ion)/226. The limit of detection and lower limit of quantitation were 0.05 and 0.1 ng/mL in standard solution, respectively. Recovery values were 98.3-100.4% with inter-day (relative standard deviation, RSD, 0.4-14.1%) and intra-day (RSD, 0.8-19.7%) assays. This method was applied to the measurement of Ac-SDKP levels in plasma from hemodialyzed subjects. Concentrations were 0.59 ± 0.23 ng/mL (pre-hemodialyzed subjects, n = 9) and 0.44 ± 0.19 ng/mL (post-hemodialyzed subjects, n = 9). All plasma Ac-SDKP levels were decreased by dialysis. Thus, plasma Ac-SDKP was decreased through dialysis in chronic kidney disease. The findings in this study will be useful for the treatment of anemia in chronic kidney disease with dialysis.     

Determination of L-3,4-dihydroxyphenylalanine in blood by high-performance liquid chromatography after solvent extraction

The use of high-performance liquid chromatography combined with solvent extraction for sample purification is described for the determination of L-3,4-dihydroxyphenylalanine in blood plasma. It is extracted into n-hexanol via complexation of its catechol moiety with diphenyl borate and ion-pair formation of its carboxylic group with tetrapentylammonium ion in an alkaline buffer. Under optimal extraction conditions, L-3,4-dihydroxyphenylalanine and 3,4-dihydroxybenzylamine used as an internal standard are extracted from blood plasma by a simple procedure and in a short time and then separated by reversed-phase ion-pair chromatography. The analytical recovery (100.8%) and reproducibility (coefficient of variation = 2.3% for n = 6) from plasma samples are good enough for routine analysis. L-3,4-Dihydroxyphenylalanine levels in blood can be monitored by this method after oral intake of the substance.     

电感耦合等离子体原子发射光谱(ICP-AES)法测定锌锭中的微量元素

建立了电感耦合等离子体原子发射光谱(ICP-AES)法测定锌锭中的Pb、Cd、Fe、Cu、Sn 5种杂质元素的方法。样品用硝酸(1+1)溶解后,在稀硝酸介质中利用电感耦合等离子体原子发射光谱仪测定其中Pb、Cd、Fe、Cu、Sn的含量,测定的相对标准偏差(RSD,n=3)小于1.7%,对锌光谱标样BYG0505的测定结果与标准值基本一致。实现了对锌锭中多种杂质元素的简便、快速、准确的同时测定。     

Gas chromatography with nitrogen-selective detection of metoprolol from human plasma after reaction with phosgene

A method for the determination of therapeutic levels of metoprolol in human plasma is presented. Metoprolol and the internal standard are extracted from the buffered plasma sample to an organic phase containing 4 X 10(-3) M phosgene. After 10 min the organic phase is taken to dryness. The residue is dissolved in ethyl acetate and the formed oxazolidine derivatives are analyzed by gas chromatography with nitrogen-selective detection. With packed columns, rectilinear standard curves through the origin were obtained down to 80 nmoles/l of plasma. The precision of the method at 200 nmoles/l was 1.5% (n = 8). The sensitivity of the method was improved by using capillary column gas chromatography. Linear standard curves were obtained down to 10 nmoles/l of metoprolol in plasma. The precision of the method at the 50 nmoles/l level was 2.2% (n = 7). With this simple and straightforward method using extractive derivatization 30 samples can be handled in a day.     

肝素抗凝血浆与血清中微量元素测定的对比分析

目的研究肝素抗凝血浆代替血清进行微量元素测定的可行性。方法选择2014年6月在汉川市人民医院接受生化检验的272例患者,采用Beckman DXC800型全自动生化分析仪,同时检测患者的肝素抗血浆和血清中锌(Zn)、铁(Fe)、铜(Cu)3种微量元素的浓度。结果肝素抗凝血浆锌、铁与血清中的锌、铁离子浓度比较无显著差异,差异无统计学意义(P0.05);血浆中铜离子的浓度明显低于血清中铜离子的检测值,差异有统计学意义(P0.05),但通过回归方程可以校正。结论肝素抗凝检测方法能够有效避开血液的凝固过程,在短时间内分离标本并检测,从而减少放置误差。但肝素抗凝血浆铜的检验还应进行回归校正。     

亚磷酸二正丙酯的气相色谱法测定

采用气相色谱法分离和测定合成有机磷农药的中间体亚磷酸二正丙酯。试验结果表明,在 5％ OV－ 7/Chromosorb W－ AW DMCS（ 0.231～ 0.387 mm）的色谱柱上,亚磷酸二正丙酯与内标物联苯等之间具有较好的分离效果。并且,以联苯为内标物时,亚磷酸二正丙酯的质量校正因子相当稳定,fW. A=2.47± 0.09（α =0.05,n=5）。该法操作简便、快速,准确度和精密度较好,对同一试样的 5次平行独立测定的相对标准偏差（ RSD）为 1.57％;该法的标准加入回收率达 99.2％～ 101.9％。     

光疗对新生儿高胆红素血症患儿微量元素影响的临床观察

观察了４５例新生儿高胆红素血症患儿光疗（蓝光）前后血清１５种元素的变化。结果发现患儿在光疗后１２种元素含量较光疗前降低，尤其是Ｃｕ、Ｆｅ降低明显，ｔ值分别为２．０９，２．７３，Ｐ〈０．０５，Ｐ〈０．０１，有显著性差异，Ｚｎ、Ｃａ虽有所下降，但无统计学意义。推测Ｃｕ、Ｆｅ降低的原因与肠道、尿液及汗液排泄增加有关。提示对光疗后的高胆血症病人应注意及时补充Ｃｕ、Ｆｅ等微量元素，以保证小儿生长发育的需要。     

Automated analysis of oxolinic acid and flumequine in salmon whole blood and plasma using dialysis combined with trace enrichment as on-line sample preparation for high-performance liquid chromatography

The use of dialysis as sample clean-up for high-performance liquid chromatography makes fully automated determination of drugs in whole blood and plasma possible. High recoveries of the analytes oxolinic acid and flumequine and the internal standard nalidixic acid are obtained after a short time of dialysis (7.3 min). The dilute dialysates are enriched on a small column packed with polystyrene. When dialysis is discontinued, the analytes are eluted by mobile phase to the analytical column. With UV detection the limit of detection was 50 ng/ml for both oxolinic acid and flumequine. Validation showed good precision and accuracy and good correlation between determinations in plasma and whole blood.     

电感耦合等离子体发射光谱法测定稀土发火合金中铁、镁、铜和锌

采用盐酸辅以加热的方式处理稀土发火合金样品,以电感耦合等离子体原子发射光谱法测定稀土发火合金中铁、镁、铜和锌含量。采用矩阵模拟实验优化分析谱线,利用多点定标校正曲线法计算测量结果。在最佳实验条件下,稀土发火合金中的基体元素对待测元素的测定结果无明显影响,各元素在检测范围内线性关系良好,相关系数均为0.999 9,方法检出限为0.001~0.010μg/m L。测定结果的相对标准偏差为1.10%~1.75%(n=11),样品加标回收率在96.00%~104.05%之间。该方法简便、快速且具有较高的灵敏度,适用于稀土发火合金中铁、镁、铜、锌等非稀土元素的测定。     

Development of a liquid chromatography/tandem mass spectrometry assay for the quantification of Aplidin,a novel marine-derived antineoplastic agent,in human plasma

A rapid and sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) assay was developed and validated to quantify a novel marine-derived depsipeptide, Aplidin, in human plasma. The method was validated to demonstrate the specificity, recovery, limit of quantitation (LOQ), accuracy, and precision of measurements. The calibration range for Aplidin was established using Aplidin standards from 0.05-50 ng/mL in blank human plasma. The multiple reaction monitoring, based on the transition m/z 1110.7 --> 295.3, was specific for Aplidin, and that based on the transition m/z 1112.6 --> 297.3 was specific for didemnin B (the internal standard); no endogenous materials interfered with the analysis of Aplidin and didemnin B from blank human plasma. The assay was linear over the concentration range 0.05-50.0 ng/mL. The correlation coefficients for the calibration curves ranged from 0.9979 to 0.9999. The mean intra- and interday accuracies for all calibration standards (n = 12) ranged from 97 to 106% (相似文献   

微波消解-电感耦合等离子体原子发射光谱(ICP-AES)法测定含铜污泥中多种重金属

为了填补现有方法的技术空白,本方法采用微波消解和电感耦合等离子体原子发射光谱法（ ICP-AES）相结合,实现对含铜污泥中铅、锌、铬、镉、砷、镁、铝、锑量的同时测定。首先采用盐酸-硝酸-氢氟酸微波消解进行样品的前处理,消解后加入高氯酸置于电热板进行除碳并赶酸,溶样效果理想,且有效避免了高温溶样对易挥发元素砷、锑的损失,整个过程安全、高效、无损。溶样后以电感耦合等离子体发射光谱法（ ICP-AES）进行测定。对含铜污泥的分解方法进行了合理选择,并对测定时的元素分析谱线及各测定元素间干扰情况等进行了讨论。该方法的加标回收率在95.31%~107.28%%,相对标准偏差（RSD）在0.31%~2.05%之间（n=7),结果表明,该方法准确度高,操作简单快捷,可同时测定多种元素,能满足批量的测定含铜污泥中铅、锌、镍、铁、镉、铬、砷、锑含量的测定要求。     

微波消解-电感耦合等离子体质谱(ICP-MS)法测定土壤七种金属元素

实验室比对盲样测定是检验实验室能力验证、实验室资质认定、机构考核的主要手段。为研究并解决测试实验室比对土壤盲样中铍、钒、镍、铜、锌、镉、铅的含量,采用微波消解电感耦合等离子体质谱(ICP-MS)法对土壤盲样进行研究,探讨了不同消解酸体系,检出限和定量限、测试模式和干扰消除、精密度和加标回收率、质控样品进行研究。结果表明：用6 mL HNO3,2 mL HCl和1 mL HF为混合酸体系,各待测元素标准曲线相关系数大于0.9995,检出限在0.001~2.985 mg.L-1^,定量限在0.003~9.94 mg.L-1^,采用氦气碰撞模式测试钒、镍、铜、锌、镉和铅,可以有效的降低多原子离子的干扰;采用no gas模式测试铍,可以有效的提高铍的测试灵敏度。方法精密度为0.2%~6.2%(n=6),加标回收率为92.3%~110.6%,采用土壤标准样品(GSS-4)进行全过程质控研究分析,各元素结果均在标准值参考范围内。用ICP-OES法测试土壤盲样中七种待测金属元素含量与用铑为内标的ICP-MS进行比对,测量分析结果基本一致。     

Liquid chromatography with pre-column dansyl derivatisation and fluorimetric detection applied to the assay of morphine in biological samples

A simple method employing pre-column dansylation and liquid chromatography is proposed for a very sensitive and specific assay of morphine in biological samples. Nalorphine is used as an internal standard. The detection limit is 0.2 picomol of injected morphine. In the assay of human sera spiked with 150 nmol/l, the intra- and inter-assay coefficients of variation were 3.7% (n = 10) and 4.5% (n = 10), respectively. No interferences were observed from more than 70 opiate and non-opiate drugs. Urine, plasma and total blood were assayed, using different extraction methods, with negligible interference from coextractives.     

Liquid chromatography-mass spectrometry method for quantification of caspofungin in clinical plasma samples

Caspofungin [(CASPO) MK-0991] is the first broad-spectrum anti-fungal agent of the echinocandin class approved for clinical use. Measurement of CASPO levels in blood might help monitor therapy in patients who are critically ill, in particular, if high-dose regimens or combinations of CASPO with other anti-fungals are used. The objective of this study was to develop a fast method for the measurement of CASPO levels in clinical blood samples using liquid chromatography coupled to a triple-quadrupole mass spectrometer. Stock solutions were prepared in plasma to avoid CASPO adsorption to glass and plastic surfaces during processing. CASPO and the internal standard (IS) were extracted from 100 microl of plasma using acetonitrile protein precipitation. The supernatant was diluted and directly injected into an analytical column (C8; 2.1 x 30 mm). The total run time was 15 min. CASPO was ionized by electrospray in the positive mode. CASPO and IS [M + 2H]2+ parent ions (m/z 547.3 and 547.8, respectively) and specific product ions (m/z 137.1 and 62.2, respectively) were used for the ion transitions. No carry over or cross-talk was observed on the column. The mean method recovery was 90 +/- 3%. Neither blood from different individuals (n = 6) nor the presence of concomitant drugs (n = 33) in plasma samples interfered with CASPO quantification. Quantification over time of the CASPO levels in plasma and whole blood was investigated at different pre-analysis storage conditions. The calibration curve included the clinically relevant CASPO concentration range from 0.04 to 20 microg/ml. Mean intra- and inter-day accuracy was 96.1 +/- 2.2% and 102.5 +/- 2.4%, respectively. Mean intra- and inter-day precision was 7.9 +/- 3.2% and 6.3 +/- 1.8%, respectively. This simple and robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method may easily be implemented for monitoring CASPO therapy.     

Determination of mefloquine by electron-capture gas chromatography after phosgene derivatization in biological samples and in capillary blood collected on filter paper

Mefloquine is determined in 100-microliter samples of whole blood, plasma and capillary blood collected on filter paper by gas chromatography with electron-capture detection after derivatization with phosgene. Sample preparation for whole blood and plasma involves a protein precipitation step that uses a combination of zinc and acetonitrile, followed by simultaneous extraction with methylene chloride and derivatization with phosgene at pH 9.50. Filter paper spots are immersed for 12-24 h in 0.1 M hydrochloric acid, followed by simultaneous extraction with methyl tert.-butyl ether and derivatization. After evaporation of the organic phase and reconstitution with ethyl acetate, 1 microliter of the extract is injected into a megabore capillary column. Because of the high sensitivity of the method, mefloquine concentrations down to 25 nmol/l (9.5 micrograms/l) are determined in 100-microliters samples with a relative standard deviation of 12% at the 25 nmol/l level. Excellent precision was obtained over the range of concentrations tested, 0.10-3 mumol/l (45-1100 micrograms/l), in both plasma and whole blood and from filter-paper-collected capillary blood. The day-to-day relative standard deviation in plasma at the therapeutic level (1-3 mumol/l) was 4.5% (n = 8).     

原子吸收分光光度法空心阴极灯一灯多用探讨

利用原子吸收分光光度法分别用钾空心阴极灯测钠、锌空心阴极灯测铜,标准曲线线性良好,相关系数均在0.999以上。钾灯测量钠质控样结果为0.747 mg/L,在质控样标称值范围(0.712±0.049)mg/L之内。锌灯测量铜质控样结果为1.23 mg/L,在质控样标称值范围(1.19±0.05)mg/L之内。钾灯测钠的相对标准偏差为0.47%(n=6),加标回收率为99.8%。锌灯测铜的相对标准偏差为0.53%(n=6),加标回收率为103%。一灯多用在环境监测工作中是可行的。     

Determination of catecholamines in human plasma by high-performance liquid chromatography: comparison between a new method with fluorescence detection and an established method with electrochemical detection

We report a sensitive fluorimetric method, in which catecholamines are concentrated from plasma by liquid-liquid extraction and derivatized with the selective fluorescent agent 1,2-diphenylethyl-enediamine prior to chromatography. Optimal conditions for extraction, derivatization and chromatography were investigated. With alpha-methylnorepinephrine as internal standard, the chromatographic separations are complete within 6 min. Limits of detection are 0.3 pg for norepinephrine and epinephrine and 0.5 pg for dopamine. Coefficients of variation are low (3-7%). Comparison of plasma catecholamine values determined with this method and with an established method with electrochemical detection (n = 135) shows good correlation (r = 0.94-1.00), and regression lines are close to lines of identity.     

Contribution to the radionuclide X-ray fluorescence analysis of human blood and plasma

A photon-induced X-ray fluorescence method, which allows the simultaneous determination of iron, copper, zinc, bromine and rubidium in human blood and blood plasma samples is described. The method is reliable and has a good sensitivity for a wide range of elements.