One-step immunoassays for free (unbound) hormones: the effect of tracer binding by serum proteins.

1986 ◽  
Vol 32 (1) ◽  
pp. 45-49 ◽  
Author(s):  
D Geiseler ◽  
P Chodha ◽  
R Ekins
Keyword(s):  
Serum Proteins ◽  
Single Step ◽  
Assay System ◽  
Measuring System ◽  
Binding Potential ◽  
Protein Distribution ◽  
Analyte Concentration ◽  
Binding Assays ◽  
One Step

Abstract In binding assays for determination of free (non-protein-bound) hormones in serum or plasma, the influence of the measuring system on the original analyte concentration in the sample must be considered. In one- or single-step free-hormone immunoassays, the labeled analyte or analog-tracer not only is bound to the antibody, it also is bound, to some extent, to serum proteins. The dependence of the assay response on two unknown variables--the concentration of free analyte and the binding potential of serum for the tracer--introduces a bias between the actual (original) and measured hormone concentrations. The significance of this protein effect is described by mathematical modeling of the analyte-protein distribution in the assay system. The theoretical consideration is validated by a clearly defined one-step assay system for measurement of free-thyroxin concentration, with a labeled thyroxin-immunoglobulin conjugate used as tracer.

Binding of labeled thyroxin analog to serum proteins evaluated after radioimmunoassay of free thyroxin.

1989 ◽  
Vol 35 (3) ◽  
pp. 475-477 ◽  
Author(s):  
G Arevalo
Keyword(s):  
Serum Proteins ◽  
Equilibrium Dialysis ◽  
Normal Serum ◽  
Thyroid Status ◽  
Nonthyroidal Illness ◽  
Serum Pool ◽  
Normal Individuals ◽  
Ambulatory Patients ◽  
One Step

Abstract In ambulatory patients, assay of free thyroxin (FT4) in serum correlates well with thyroid status and with results obtained by equilibrium dialysis. The validity of FT4 results has been questioned mainly in euthyroid patients with altered concentrations of thyroid hormone-binding proteins, as in nonthyroidal illness, hereditary analbuminemia, familial dysalbuminemic hyperthyroxinemia (FDH), and the presence of iodothyronine-binding antibodies. I present here a study of the binding of [125I]T4-derivative to serum proteins in the supernate, which is ordinarily discarded after determination of FT4 by one-step radioimmunoassay with dextran-coated charcoal used to separate the free and bound fractions. The results are expressed as a ratio, with results for a normal serum pool as reference. The average ratio was high in hyperthyroid subjects, 1.26 (SD 0.12, n = 25), and in hypoalbuminemia, 1.20 (SD 0.10, n = 15), and low in FDH, 0.62 (SD 0.11, n = 9), and hypothyroid subjects, 0.90 (SD 0.06, n = 20). In normal individuals it was 0.98 (SD 0.05, n = 30). Determination of the analog-binding rate complements the FT4 result and allows for the recognition of cases with abnormal binding by serum proteins, without recourse to other tests recommended for thyroid-function studies.

One-Step Extraction and Cleanup Procedure for Determination of p,p'-DDT, p,p'-DDD, and p,p'-DDE in Fish

1986 ◽  
Vol 69 (4) ◽  
pp. 581-586 ◽  
Author(s):  
Nazir Ahmad ◽  
Robert Slobodan Marolt
Keyword(s):  
Residue Analysis ◽  
Trisodium Citrate ◽  
Single Step ◽  
Chromatographic Column ◽  
Fish Samples ◽  
Average Coefficient ◽  
Cleanup Procedure ◽  
One Step

Abstract A simplified method that combines extraction, partitioning, and cleanup in a single step for measuring p,p'-DDT and its metabolites in fish is described. Minced fish samples are emulsified with disodium hydrogen orthophosphate and trisodium citrate, ground with sodium sulfate, and eluted from a chromatographic column prepacked with alumina and silicic acid. The fats and fatty acids are solubilized and easily extracted from the tissues and retained by the column, while p,p'-DDT and its metabolites are quantitatively eluted with 40 mL n-hexane. The eluate is directly applied to a gas chromatographic column. Average recoveries of p,p'-DDT and its metabolites added to fish in vitro are 81%. The average coefficient of variation for recoveries of p,p'-DDT and its metabolites is less than 6.5% and the detection limit is 0.001 Hg/g for p,p'-DDE, thus making this method very suitable for residue analysis.

Quantitative Determination of Plasminogen

1970 ◽  
Vol 23 (02) ◽  
pp. 191-201 ◽  
Author(s):  
H. D Bruhn ◽  
L Müller ◽  
F Duckert
Keyword(s):  
Quantitative Determination ◽  
Plasminogen Activator ◽  
Assay System ◽  
Two Stage ◽  
System Activation ◽  
Frozen Plasma

SummaryA modification of the caseinolytic assay for plasminogen is described. This assay system is characterized by the following features :1. Urokinase is used as activator achieving a complete activation of the plasminogen whereas with streptokinase caseinolytically inactive plasminogen-activator complexes are formed.2. All incubation times are reduced to the minimum which is still compatible with accuracy.3. Results are expressed in percent of a standard of ten normal plasmas.4. In this two-stage assay-system (activation of plasminogen to plasmin, digestion of casein by plasmin) both stages proceed simultaneously in the same system, thus the plasmin formed is stabilized “in statu nascendi” by the casein.5. Several conditions (stability of plasminogen in frozen plasma, use of anticoagulants, reproducibility) are defined.

Development and Validation of an LC-MS/MS Method for Simultaneous Determination of Canagliflozin and Metformin HCl in Rat Plasma and its Application

2020 ◽  
Vol 16 (6) ◽  
pp. 752-762
Author(s):  
Vivek Nalawade ◽  
Vaibhav A. Dixit ◽  
Amisha Vora ◽  
Himashu Zade
Keyword(s):  
Internal Standard ◽  
Rat Plasma ◽  
Precipitation Method ◽  
Herbal Extracts ◽  
One Step ◽  
Precision And Accuracy ◽  
Development And Validation ◽  
The Impact ◽  
Metformin Hcl

Background: Food and herbal extracts rich in Quercetin (QRT) are often self-medicated by diabetics and can potentially alter the pharmacokinetics (PK) of Metformin HCl (MET) and Canagliflozin (CNG) leading to food or herb-drug interactions and reduced therapeutic efficacy. However, the impact of these flavonoids on the pharmacokinetic behaviour of MET and CNG is mostly unknown. Methods: A simple one-step protein precipitation method was developed for the determination of MET and CNG from rat plasma. The mobile phase chosen was MeOH 65% and 35% water containing 0.1% formic acid at a flow rate of 1mL/min. Results: The retention time of MET, internal standard (Valsartan) and CNG was 1.83, 6.2 and 8.2 min, respectively. The method was found to be linear in the range of 200 - 8000 ng/mL for CNG and 100 = 4000 ng/ml for MET. Precision and accuracy of the method were below 20% at LLOQ and below 15% for LQC, MQC, and HQC. Conclusion: The method was successfully applied for the determination of PK of MET and CNG by using 100 μL of rat plasma. QRT co-administration affects the PK parameters of MET and CNG. This alteration in PK parameters might be of significant use for clinicians and patients.

Synthesis of Polycycles and Oxacycles by Tandem Metathesis of endo–norbornene Derivatives

Synthesis ◽  
2021 ◽  
Author(s):  
Sambasivarao Kotha ◽  
Sunil Pulletikurti ◽  
Ambareen Fatma ◽  
gopal dhangar ◽  
gonna somu Naidu
Keyword(s):  
Natural Products ◽  
Metal Complex ◽  
Carbonyl Group ◽  
Single Step ◽  
Time Dependent ◽  
Computational Studies ◽  
Nmr Studies ◽  
Tricyclic Compounds ◽  
One Step ◽  
Norbornene Derivatives

Here, we have demonstrated that the presence of a carbonyl group at C7 position is preventing the olefin metathesis of endo-norbornene derivatives due to the complexation of the metal alkylidene. Time-dependent NMR studies showed the presence of new proton signals in the metal alkylidene region, which indicate the formation of metal complex with the carbonyl group of the substrate. These observations were further proved by ESI-MS analysis. Whereas, computational studies provided that the catalyst was interacting with the C7 carbonyl group and aligned perpendicular to that of norbornene olefin. Later, these endo-keto norbornene derivatives were reduced to hydroxyl derivatives diastereoselectively. Ring-rearrangement metathesis (RRM) of these hydroxyl derivatives, produced the [6/5/6], and [5/6/5] carbo-tricyclic cores of the natural products in one step. Whereas the RRM of O-allyl derivatives, delivered the oxa-tricyclic compounds in a single step with excellent yields.

scholarly journals A single-step purification method for the precise determination of the antimony isotopic composition of environmental, geological and biological samples by HG-MC-ICP-MS

2021 ◽  
Author(s):  
Ferrari Colin ◽  
Resongles Eléonore ◽  
Freydier Rémi ◽  
Casiot Corinne
Keyword(s):  
Isotopic Composition ◽  
Biological Samples ◽  
Precise Determination ◽  
Single Step ◽  
Functionalized Silica ◽  
Purification Method ◽  
Silica Powder ◽  
Icp Ms ◽  
Single Step Purification

Thiol-functionalized silica powder allowed single-step purification of antimony for exploring stable Sb isotope signatures in the environment.

scholarly journals Single Step 2-Port Device De-Embedding Algorithm for Fixture-DUT-Fixture Network Assembly

Electronics ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1275
Author(s):  
Simone Scafati ◽  
Enza Pellegrino ◽  
Francesco de Paulis ◽  
Carlo Olivieri ◽  
James Drewniak ◽  
...  
Keyword(s):  
Standard Technique ◽  
Single Step ◽  
Scattering Parameters ◽  
Linear Set ◽  
S Parameter ◽  
Parameter Conversion ◽  
Before And After ◽  
One Step ◽  
Typical Measurement

The de-embedding of measurement fixtures is relevant for an accurate experimental characterization of radio frequency and digital electronic devices. The standard technique consists in removing the effects of the measurement fixtures by the calculation of the transfer scattering parameters (T-parameters) from the available measured (or simulated) global scattering parameters (S-parameters). The standard de-embedding is achieved by a multiple steps process, involving the S-to-T and subsequent T-to-S parameter conversion. In a typical measurement setup, two fixtures are usually placed before and after the device under test (DUT) allowing the connection of the device to the calibrated vector network analyzer coaxial ports. An alternative method is proposed in this paper: it is based on the newly developed multi-network cascading algorithm. The matrices involved in the fixture-DUT-fixture cascading gives rise to a non-linear set of equations that is in one step analytically solved in closed form, obtaining a unique solution. The method is shown to be effective and at least as accurate as the standard multi-step de-embedding one.

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