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Bioanalytical
Services
Instrumentation and
Capabilities
Our dedicated, purpose
built bioanalytical laboratories incorporate advanced liquid
chromatography and tandem mass spectrometry instrumentation and
software. Our experienced analysts offer
expert bioanalysis capabilities across a variety of matrices,
including:
These capabilities
incorporate various sample pre-treatment processes, in 96-well
format, such as:
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Solid phase
extraction
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Protein
precipitation
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Liquid-liquid
extraction
Pharmacokinetic and
Toxicokinetic Reporting
Toxicokinetic
determinations are an essential component of pivotal toxicity
studies and we are able to offer a full services in toxicokinetic
data handling and interpretation to support these studies. In
addition, we have extensive expertise in design, conduct and data
analysis for stand-alone pharmacokinetic and bioavailability
studies. All pharmacokinetic and toxicokinetic data are analysed
using the WinNonLin® PK/PD modelling and analysis system.
Sequani’s team of
toxicokinetic/pharmacokinetic analysts, with extensive training in
Good Laboratory Practice (GLP), toxicology and compound development
fulfil these functions in every detail. Comprehensive statistical
data analysis is provided, using the fully validated networked data
handling systems. Data is interpreted and the toxicokinetic/pharmacokinetic report produced, either by itself or
as part of a toxicology report ready for regulatory submission.
In Vitro
Metabolism
In vitro metabolism
assays can provide valuable information regarding the kinetics and
metabolic fate of NCEs in development, giving an indication of
metabolic stability and alerting to the formation of major
metabolites. In vitro metabolism data provides justification for
the animal species used in pre‑clinical studies and can determine
which cytochrome P450 (CYP450) isoenzymes are involved in the
metabolism of NCEs and hence indicate the potential for drug-drug
interactions.
Cryopreserved liver
microsomes or hepatocytes from common pre-clinical species and man
are used in our studies. Microsomal preparations provide a
convenient and inexpensive source of several enzyme functions.
These are mainly associated with Phase I oxidative metabolism but
can provide some Phase II functions if appropriate cofactors are
included. Primary hepatocytes provide a more complete in vitro
system as they have intact cell membranes and physiological levels
of cofactors for complete Phase I and II metabolic processes.
In Vitro
Stability
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Determination of
enzyme kinetics (clearance and half-life, as applicable)
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Systems –
cryopreserved liver microsomes or hepatocytes Quantitative
LC-MS/MS bioanalysis
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Species available
- human, rat, dog, mouse, monkey, mini-pig
Metabolic profiling
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Major
metabolite alert
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Inter-species
comparison
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Systems -
cryopreserved liver microsomes or hepatocytes
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Qualitative HPLC-UV
analysis
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LC-MS and MS/MS
characterisation
Cytochrome P450
Inhibition Assays
Assays to detect
inhibition of the major human CYP450s are undergoing in-house
validation at Sequani so these studies can be added to our services
in the near future.
Plasma
Protein Binding
Determining the extent
to which a molecule binds to plasma (or serum) proteins is a
critical phase of NCE development since the effective dose is
related to the concentration of unbound “free” drug. The extent of
plasma-protein binding of a drug therefore influences compound
dosing, efficacy, toxicity, clearance rate and potential for
drug-drug interactions. The extent of binding to proteins can be
determined by several methods including ultra-filtration and
equilibrium dialysis.
Ultra-Filtration
Equilibrium
Dialysis
Sequani are currently performing in-house validation of assessment
of protein binding using equilibrium dialysis methodology. This may
be used in instances where the ultra-filtration method is unsuitable
due to non-specific binding.
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