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1.
The liquid Chromatographic separation of 3,3',5-triiodothyronine, 3,3',5'-triiodothyronine, and thyroxine with a nonpolar stationary phase was studied as a function of pH, temperature, organic content of the mobile phase, and ionic strength using aqueous phosphate—acetonitrile, aqueous phosphate—methanol, and aqueous phosphate— n-propanol mobile phase systems. It was demonstrated that the quality of the thyroid hormone separations, as determined by normalized peak capacity values, was unchanged with temperature, remained relatively constant with increasing ionic strength, and was affected to the greatest extent by changes in pH and organic modifier content of the mobile phase. Chromatographic behavior of the compounds studied as a function of these variables was found to be consistent with existing Chromatographic theory and/or empirical observations. Recommended conditions are aqueous phosphate—methanol mobile phase, pH 2–5 (aqueous portion), and high temperature (60–70°C).  相似文献   

2.
An analytical method for the quantification of thyroid hormones (3,5,3′,5′-tetraiodo-l-thyronine, 3,3′,5-triiodo-l-thyronine, 3,3′,5′-triiodothyronine, 3,5-diiodothyronine, 3,3′-diiodothyronine) in different water matrices has been developed. The method, consisting of solid phase extraction (SPE) and liquid chromatography–tandem mass spectrometry (LC–MS/MS), was validated for tap and surface water as well as raw and treated waste water. The limits of quantifications (LOQs) were lowest in tap water, where they ranged from 1.1 to 13.3 ng L−1, and highest in raw wastewater (10.5–84.9 ng L−1). Of the target analytes 3,5,3′,5′-tetraiodo-l-thyronine (T4) could be quantified in the influent and effluent of a waste water treatment plant (WWTP) in Finland. The study showed that despite a relatively high removal rate during treatment (66%), part of the incoming T4 will reach the aquatic environment and, due to the high endocrine activity of this compound, further studies are needed in order to assess its environmental fate and impact on natural ecosystems.  相似文献   

3.
Thyroid hormones are essential hormones for regulating growth and development in humans and wildlife. Methods to monitor precise and low levels of these hormones in serum and tissues are needed to assess overall health, whether from disease considerations or possibly from environmental contaminant exposures. Common and routine methods typically rely upon radioimmunoassays, which can be expensive, and typically only measure thyroxine and 3,3′,5-triidothyronine, which can be a limitation in fully evaluating impacts on thyroid regulation. In this study we developed a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the simultaneous analysis of five thyroid hormones—thyroxine, 3,3′,5-triidothyronine, 3,3′,5′-triiodothyronine, 3,3′-diiodothyronine, and 3,5-diiodothyronine—in serum samples. The LC-MS/MS parameters were optimized and calibrated over a wide concentration range (1.0–500 ng/mL) with on-column detection limits of 1.5–7.0 pg. With use of spiked bovine serum samples, the mean method recoveries were calculated to be 81.3–111.9% with relative standard deviations of 1.2–9.6% at spiking levels ranging from 10 to 100 ng/mL. This method was compared with measurements made by standard radioimmunoassays and with measurements made in a serum Standard Reference Material (SRM 1951b). Development of this method expands the capacity to measure thyroid hormones by including a larger suite of thyroid hormones, and has promising applications for measuring catabolism of thyroid hormones in vitro.  相似文献   

4.
Abstract

The usefulness and capabilities of gradient elution techniques with electrochemical detection are shown with a rapid separation of D,L-tyrosine (Tyr), 3-iodo-L-tyrosine (MIT), 3,5-diiodo-L-tyrosine (DIT), D,L-thyronine (T0), 3,5-diiodo-D,L-thyronine (T2), 3,3′,5-triiodo-L-thyronine (T3), and L-thyroxine (T4) by reverse phase liquid chromatography. A rapid (five minute) isocratic separation of T0, T2, T3, and T4 is also reported. Limits of detection are in the sub-nanogram range with a linear dynamic range to 500 ng for T2 and T3, and 1000 ng for T4. Analysis of levothyroxine sodium tablets and injectable intravenous samples is described.  相似文献   

5.
建立了高效液相色谱-串联质谱(HPLC-MS/MS)检测牛奶中甲状腺素3,3′,5,5′-四碘-L-甲腺原氨酸(T4), 3,3′,5-三碘-L-甲腺原氨酸(T3)和3,3′,5′-三碘-L-甲腺原氨酸(rT3)的方法。样品用乙腈提取,离心,上清液经氨水碱化和Cleanert PAX固相萃取小柱净化,在Zorbax Eclipse XDB-C18 (150 mm×2.1 mm, 3.5 μm)色谱柱上以0.1%(v/v)乙酸水溶液和甲醇为流动相等度洗脱分离,以电喷雾正离子(ESI+)模式电离,多反应监测(MRM)模式质谱检测,内标法定量。结果表明,甲状腺素的检出限(LOD)不大于0.03 ng/g,定量限(LOQ)不大于0.1 ng/g;在考察的浓度范围内线性关系良好(r2> 0.998);回收率为80.61%~101.7%,相对标准偏差(RSD)为1.48%~9.70%。室温下样品溶液中的甲状腺素保持稳定。对5个牛奶样品的测定结果显示,T3含量为0.59~1.30 ng/g, RSD为2.06%~7.70%; T4和rT3未检出。该方法具有样品处理简单,灵敏度高,重现性好,定性和定量结果可靠等特点,为牛奶中甲状腺素的测定和相关质量安全评价提供了可靠手段。  相似文献   

6.

We report here a novel approach for the extraction and analysis of thyroid hormones (TH) and their metabolites (THM) from human serum samples. Our method features a compact, 96-well micro-titre plate-based pre-analytic extraction/clean-up workflow combined with an isotope dilution LC-MS/MS-MS3 analytical method. In particular, these features make possible the detection of iodothyronines at their endogenous concentrations in serum differing by a factor of ca. 104, with potential to semi-automate the pre-analytics. The method was validated by the assessment of linearity, lower limits of quantification and detection (LLOQ and LLOD respectively), intra- and inter-day accuracy, precision, process efficiency (PE), matrix effect (ME) and relative recovery (RE). Calibration curves were linear in the concentration range in sample matrix from 0.1–250 nM for T3, rT3, T4 and 3-T1AM and from 0.005–1 nM for 3,5-T2 and 3,3′-T2. Using a 200-μL sample volume, the analyte dependant LLOQ were in the range 0.005 (3,5-T2) to 0.25 (T4) nM and LLOD were between 0.002 (3,5-T2) and 0.052 nM (T4). We applied the LC-MS/MS-MS3 method to the analysis of a cross section of patients with disorders of the thyroid hormone axis. T4, T3 and rT3 concentrations (± standard deviation) were 120 ± 18, 1.9 ± 0.4 and 0.45 ± 0.09 nM respectively. 3,3′-T2 concentrations (± standard deviation) were 0.079 ± 0.022 nM; 3,5-T2 concentrations were below the LLOQ and/or LLOD in all but a single sample (0.013 nM). This method expands the analytical spectrum to endogenous thyroid hormone metabolites such as 3,5-T2 which exert biological actions and rT3 which may act as surrogate markers for disturbed thyroid hormone metabolism.

Graphical abstract

  相似文献   

7.
The single ratio kinetic method is applied to the discrimination and quantification of the thyroid hormone isomers, 3,5,3′-triiodothyronine and 3,3′,5′-triiodothyronine, in the gas phase, based on the kinetics of the competitive unimolecular dissociations of singly charged transition-metal ion-bound trimeric complexes [MII(A)(ref*)2-H]+ (MII = divalent transition-metal ion; A=T3 or rT3; ref* = reference ligand). The trimeric complex ions are generated using electrospray ionization mass spectrometry and the ions undergo collisional activation to realize isomeric discrimination from the branching ratio of the two fragment pathways that form the dimeric complexes [MII(A)(ref*)-H]+ and [MII(ref*)2-H]+. The ratio of the individual branching ratios for the two isomers Riso is found strongly dependent on the references and the metal ions. Various sets are tried by choosing the reference from amino acids, substituted amino acids, and dipeptides in combination with the central metal ion chosen from five transition-metal ions (CoII, CuII, MnII, NiII, and ZnII) for the complexes in this experiment. The results are compared in terms of the isomeric discrimination for the T3/rT3 pair. Calibration curves are constructed by relating the ratio of the branching ratios against the isomeric composition of their mixture to allow rapid quantitative isomer analysis of the sample pair. Furthermore, the instrument-dependence of this method is investigated by comparing the two sets of results, one obtained from a quadrupole ion trap mass spectrometer and the other from a quadrupole time-of-flight mass spectrometer.  相似文献   

8.
The influence of substituents in close proximity to crown ether cavities, on the stability of complexes of the crown ethers with t-butylammonium salts, has been investigated. Crown ethers with intra-annular donor substituents (2–4) were prepared by the reaction of 2-acetylresorcinol (1) with polyethylene glycol ditosylates and subsequent modification of the acetyl group. Crown ethers with substituents above and below the plane of the crown ether 0 atoms were synthesized by the reaction of 2,2'-dihydroxy-1,1'-biphenyls with polyethylene glycol ditosylates. Chloromethylation of 5,5'-dimethyl-1,1'-biphenyl crown ethers (6) yielded 4,4'-bis(chloromethyl)-1,1'-biphenyl crown ethers (10). 3,3'-Disubstituted-1,1'-biphenyl crown ethers (13–24) were synthesized by the reaction of 3,3'-diallyl-2,2'-dihydroxy-1,1'-biphenyl (12) with polyethylene glycol ditosylates. The allyl groups of 13 were isomerized with sodium hydride to propen- 1-yl groups. Ozonolysis of 13 and 14 gave the corresponding dialdehydes (15 and 18) which were converted into other 3,3'-disubstituted biphenyl-20-crown-6 derivatives (RCH2COOMe, CH2COOH, CH2OH, CH2Cl, CH2OMe, OH and Me) by standard operations. The thermodynamic stability of the complexes of these functionalized crown ethers with t-butylammonium hexafluorophosphate has been studied in deuterochloroform in competition experiments with m-xyleno-18-crown-5 and benzo-15-crown-5 as the reference compounds. The nature of the 2-substituents in the crown ethers 2 and 3 has little effect on the stability of the complexes. The stability of the complexes of 3,3'-disubstituted biphenyl crown ethers depends of ringsize and the size and nature of the substituents. The most stable complexes are those of 24 (R = Me) and 14 (R=CH=CHMe).The Me groups in 24 represent the optimum between relief of O-O repulsion in the polyether ring and steric hindrance of complexation. The propen-1-yl substituents of 14 stabilize the complex because they provide extended π-electron donor stabilization. Substitution at the 4- and 4'-positions of the aryl groups has little effect on the stability of the complexes.  相似文献   

9.
An enzyme immunoassay based on the use of crossed-beam thermal lens detection is described. In this assay, poly-N-isopropylacrylamide, a water-soluble, thermally precipitating synthetic polymer, was used as a carrier to minimize non-specific binding. The enzyme substrate of the horseradish peroxidase that was employed was 3,3′,5,5′-tetramethylbenzidine. The color development of the enzyme–substrate reaction was stopped by SDS and Na2SO3 to achieve a stable blue solution. The background reduction and stabilization made it possible to use a crossed-beam thermal lens technique as the measurement method. This method was demonstrated to be applicable by determination of hepatitis B surface antigen in human serum. A detection limit of 0.15 ng/ml was obtained. This was more sensitive than that of the commercially available ELISA method.  相似文献   

10.
High performance liquid chromatography coupled with electrochemical detection (HPLC-EC) method has been developed for the determination of benzidine and its related congeners in wastewater at the low ng/mL level. With a µ-Bondapak C18 column, 53% acetonitrile (pH 4.7), and electrochemical detection at +1.0 V applied potential, the detection limits range from 3 ng/mL for benzidine to 10 ng/mL for 3,3′-dichlorobenzidine, assuming a 100 µL injection. The detection limits can be further lowered to the sub-ng/mL level by incorporating a short precolumn into the HPLC system for on-line sample enrichment. Recoveries are generally greater than 80% — except for benzidine and 3,3′-diaminobenzidine which give 50–70% recoveries. Industrial wastewater samples from a coke-plant and a plant producing 3,3′-dichlorobenzidine based dyes were successfully studied via this method.  相似文献   

11.
《Analytical letters》2012,45(11):1201-1215
Abstract

A simple, sensitive and specific assay method was developed for the assay of sodium liothyronine (T3Na) from tablets. Sodium liothyrorn'ne was extracted from powdered tablets with a solvent system consisting of butanol and dilute hydrochloric acid. The solvent was removed under vacuum and the residue was dissolved in ammonical methanol. An aliquot of this solution was injected on a μBondapak C18 column and the elution was carried out with a mobile phase consisting of methanol:water:phosphoric acid (55:45: 0.1). The effluent was monitored by U. V. detection at 254 nm. A linear calibration curve was obtained in the range of 10 to 250 ng on column with a precision of ±4% (C. V.). The internal standard consisted of 3, 3′, 5-triiodothyronine (T3′ or RT3). The usefulness of an alternate compound, 3, 5-diiodo-3′, 5′-dibromothyronine, which is not endogenous, was also demonstrated as an internal standard.  相似文献   

12.
An electrochemical assay used for detecting acetylcholinesterase (AChE) inhibitors has been optimised to detect pirimiphos-methyl in durum wheat. Pirimiphos-methyl is a phosphothionate insecticide and so it needs to be transformed into the corresponding oxo form to act as an effective AChE inhibitor. The inhibition assay was based on the electrochemical detection of the product of AChE, choline, via choline oxidase biosensors obtained with Prussian-Blue modified screen printed electrodes. The procedure for the oxidation of pirimiphos methyl via N-bromosuccinimide (NBS) and AChE inhibition was optimised for reagent concentrations and inhibition time in a buffer solution. A calibration of the pirimiphos-methyl (25–1,000 ng/ml) was obtained in the buffer. The intra-electrode CV ranged between 1.6 and 15.0, whereas the inter-electrode CV ranged between 4.6 and 16.0. The detection limit (LOD) was 38 ng/ml, and the I50% was 360 ng/ml. The assay conditions were then re-optimised to work with durum wheat extracts, and the calibrations were obtained under different experimental conditions, such as sample pretreatment (milled or whole grains) and extract concentration. The calibrations were slightly affected by the sample matrix, resulting in an increased LOD (65–133 ng/ml) and I50% (640–1,650 ng/ml). The LOD found for the sample, determined under optimal conditions, was 3 mg/kg. Spiked samples were prepared at the EU regulated level (5 mg/kg) and analysed with the optimised protocol, resulting in an average recovery of 70.3%.  相似文献   

13.
Zheng Y  Sun Y  Ren J 《Talanta》2006,69(1):107-112
In this paper, a new method for separation, identification and quantitation of iodotyrosines and iodothyronines [3-monoiodo-L-tyrosine (MIT), 3,5-diiodo-L-tyrosine (DIT), L-thyronine (T0), 3,5-diiodo-L-thyronine (T2), 3,5,3′-triiodo-L-thyronine (T3) and 3,3′,5,5′-tetraiodo-L-thyronine (T4)] was described by using capillary electrophoresis with photodiode-array ultraviolet-visible detection (CE-UV). The certain influence factors were systematically investigated, including the type, concentration and pH of buffer, and additive. We found that 10 mM sodium borate running buffer (pH 8.5) containing 0.10 mM β-CD as additive reagent allowed the best instrumental conditions for the optimum separation of the iodotyrosines and iodothyronines. Under optimized conditions, the analytical time was within 6 min, using an uncoated fused-silica capillary of 75 μm inner diameter with an effective length of 30 cm. The reproducibility of the migration time and peak area was less than 0.6% and 6.8%, respectively. A linear range from 10-1000 μg/mL and low limits of detection from 1.3-3.4 μg/mL were obtained at the detection wavelength of 280 nm. Our preliminary results show that the method is well suitable for determination of the hydrolysate of iodinated casein.  相似文献   

14.
Abstract

The development and application of a dual (in series) coulometric-amperometric thin-layer electrochemical detection cell system to the assay of thyroid hormones following reversed-phase chromatography is described. Using this approach the system was characterized by empirically determined electrolysis efficiencies, demonstrating adherence to the diffusion-layer approximation theory. When academic mixtures of the thyroid hormones were studied, the precision of both response and relative retention data was to within 4%. With the dual-cell approach, improvements by a factor of two in the limit of detection were noted over the same system using only amperometric detection. The electrochemical behavior of these analytes in the dual-cell system was also studied. These latter experiments demonstrated that, in principle, it is possible to assay the thyroid hormones with both cells set at a potential where mass transport control obtains, analyzing the oxidation products of the first cell at the amperometric cell.  相似文献   

15.
Summary For the title compounds 4,4-DADPM, MOCA, 3,3-DCB, 4-ADP and 4-ADPA listing on the EPA priority pollutant list, an analytical practicable, reliable, reproducible and sensitive procedure is required. Therefore a new method has been developed for the routine determination of these toxic aromatic amines in urine at the ppb level. The quantitative determination of amines is a suitable procedure of occupationally exposed persons. Urine sample preparation is done using simple liquid-liquid extraction followed by a precolumn enrichment (PRP1-material; Hamilton). Breakthrough measurements were done using an enrichment column packed with PRP1 material. The capacities of the studied amines ranged from 21.9mg/g to 96.6mg/g, while influent concentrations differed from 28.3mg/l to 332.0mg/l. The advantages of electrochemical detection regarding to selectivity and sensitivity are clearly indicated in this paper. Separation has been achieved applying reversed-phase-high-performance-liquid chromatography (LiChrosorb RP 18/5m) followed by electrochemical or UV-detection. The detection limits employing an electrochemical detector at a potential of 1 V range from 2.2ng to 12.1ng. UV detection at 254 nm and 280 nm is about 10–100 times less sensitive. Recoveries from spiked water samples at the 5ppb levels were 75% to 96% respectively. The standard deviation of the developed procedure varies from 5.3% to 14%. Day-to-day repeatability is good.  相似文献   

16.
HU  Rongzu  ZHAO  Fengqi  GAO  Hongxu  ZHANG  Jiaoqiang  ZHANG  Hai  MA  Haixia 《中国化学》2009,27(11):2145-2154
Based on reasonable hypothesis, two general expressions and their six derived formulae for estimating the critical temperature (Tb) of thermal explosion for energetic materials (EM) were derived from the Semenov's thermal explosion theory and eight non‐isothermal kinetic equations. We can easily obtain the values of the initial temperature (T0i) at which DSC curve deviates from the baseline of the non‐isothermal DSC curve of EM, the onset temperature (Tei), the exothermic decomposition reaction kinetic parameters and the values of T00 and Te0 from the equation T0i or ei=T00 or e0+a1βi+a2βi2+···+aL?2βiL?2, i=1, 2, ;···, L and then calculate the values of Tb by the six derived formulae. The Tb values for seven nitrosubstituted azetidines, 3,3‐dinitroazetidinium nitrate ( 1 ), 3,3‐dinitroazetidinium picrate ( 2 ), 3,3‐dinitroazetidinium‐3‐nitro‐1,2,4‐triazol‐5‐onate ( 3 ), 1,3‐bis(3′,3′‐dinitroazetidine group)‐2,2‐dinitropropane ( 4 ), 1‐(2′,2′,2′‐trinitroethyl)‐3,3‐dinitroazetidine ( 5 ), 3,3‐dinitroazetidinium perchlorate ( 6 ) and 1‐(3′,3′‐dinitroazetidineyl)‐2,2‐dinitropropane ( 7 ), obtained with the six derived formulae are agreeable to each other, whose differences are within 1.5%. The results indicate that the heat‐resistance stability of the seven nitrosubstituted azetidines decreases in the order 6 > 7 > 5 > 4 > 3 > 2 > 1 .  相似文献   

17.
Six isomeric tetrachlorodithizones were synthesized and their electronic and i.r. spectra were measured. Their acid dissociation constants and partition coefficients between 0.5 M NaClO4 solution and carbon tetrachloride are reported. Their extraction equilibria with Cd(II). Co(II), Cu(II), Hg(II), Ni(II), Pb(II), Tl(I), Zn(II) and Bi(III) and the spectrophotometric characteristics of the complexes formed are described. The 3,3',4,4'-tetrachloro isomer is spectrophotometrically more sensitive than dithizone for Zn, Pb, Hg(II) and Bi(III) whereas the 2,2',3,3'- and 2,2',5,5'-isomers are more sensitive for copper ions. For most of the metals tested, the tetrachlorodithizones allowed quantitative extraction at lower pH than is possible with dithizone.  相似文献   

18.
A series of novel polyimides (3ad) were prepared from 3,3′,5,5′-tetramethyl-4,4′-diaminodiphenyl-4”-tert-butyltoluene (1) with four aromatic dianhydrides via a one-step high-temperature polycondensation procedure. The obtained polyimides showed excellent solubility, with the dissolvability at a concentration of 10 wt% in most amide polar solvents and chlorinated solvents. Their films were nearly colorless and exhibited high optical transparency, with the UV cutoff wavelength in the range of 322–350 nm and the wavelength of 80% transparency in the range of 395–414 nm. They also showed low dielectric constant (2.72–2.91 at 1 MHz) and low water absorptions (0.37–0.62%). Moreover, these polyimides possessed high glass transition temperatures (Tg) (above 321 °C) and good thermal stability with 10% weight loss temperatures in the range of 526–547 °C in nitrogen atmosphere. In comparison with the analogous polyimides non-containing 3,3′,5,5′ -tetramethyl pendant groups, the resultant polyimides 3ad showed better solubility, higher optical transparency and lower dielectric constant.  相似文献   

19.
3,3′‐Dichloro‐N,N′‐biphthalimide (3,3′‐DCBPI), 3,4′‐dichloro‐N,N′‐biphthalimide (3,4′‐DCBPI), and 4,4′‐dichloro‐N,N′‐biphthalimide (4,4′‐DCBPI) were synthesized from 3‐ or 4‐chlorophthalic anhydrides and hydrazine in glacial acetic acid. The yield of 3,3′‐DCBPI (90%) was much higher than that of 4,4′‐DCBPI (33%) because of the better stability of the intermediate, 3‐chloro‐N‐aminophthalimide, and 3,3′‐DCBPI. A series of hydrazine‐based polyimides were prepared from isomeric DCBPIs and 4,4′‐thiobisbenzenethiol (TBBT) in N,N‐dimethylacetamide in the presence of tributylamine. Inherent viscosity of these polymers was in the range of 0.51–0.69 dL/g in 1‐methyl‐2‐pyrrolidinone (NMP) at 30 °C. These polyimides were soluble in 1,1,2,2‐terachloroethane, NMP, and phenols. The 5% weight‐loss temperatures (T5%s) of the polymers were near 450 °C in N2. Their glass‐transition temperatures (Tgs) determined by dynamic mechanical thermal analysis and differential scanning calorimetry increased according to the order of polyimides based on 4,4′‐DCBPI, 3,4′‐DCBPI, and 3,3′‐DCBPI. The hydrolytic stability of these polymers was measured under acid, basic, and neutral conditions and the results indicated that the order was 3,3′‐DCBPI/TBBT > 3,4′‐DCBPI/TBBT > 4,4′‐DCBPI/TBBT. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 45: 4933–4940, 2007  相似文献   

20.
An automated and sensitive sequential injection electrochemical immunoassay was developed to monitor a potential insecticide biomarker, 3,5,6-trichloro-2-pyridinol. The current method involved a sequential injection analysis (SIA) system equipped with a thin-layer electrochemical flow cell and permanent magnet, which was used to fix 3,5,6-trichloro-2-pyridinol (TCP) antibody coated magnetic beads (TCP-Ab-MBs) in the reaction zone. After competitive immunoreactions among TCP-Ab-MBs, TCP analyte, and horseradish peroxidase (HRP) labeled TCP, a 3,3′,5,5′-tetramethylbenzidine dihydrochloride and hydrogen peroxide (TMB-H2O2) substrate solution was injected to produce an electroactive enzymatic product. The activity of HRP tracers was monitored by a square wave voltammetric scanning electroactive enzymatic product in the thin-layer flow cell. The voltammetric characteristics of the substrate and the enzymatic product were investigated under batch conditions, and the parameters of the immunoassay were optimized in the SIA system. Under the optimal conditions, the system was used to measure as low as 6 ng L−1(ppt) TCP, which is around 50-fold lower than the value indicated by the manufacturer of the TCP RaPID Assay® kit (0.25 μg/L, colorimetric detection). The performance of the developed immunoassay system was successfully evaluated on tap water and river water samples spiked with TCP. This technique could be readily used for detecting other environmental contaminants by developing specific antibodies against contaminants and is expected to open new opportunities for environmental and biological monitoring.  相似文献   

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