ADME/TOX

Service ADME/TOX is a part of the research-service group of biochemical pharmacology. The service carries out in-vitro testing of potential medicines which should minimalize the sizeable testing on mice and other organisms. The ADME services we offer are listed below (they are free of charge):

• Kinetic solubility

• Plasmatic protein binding

• Plasmatic stability

• Microsomal stability

• PAMPA

• CACO-2 assay


All compounds are measured in “MRM mode” (multiple ion monitoring) on ECHO-MS. The ECHO-MS consists of TRIPLE QUAD 6500+ mass spectrometer (SCIEX) and ECHO module (analyte sampler based on acoustic wave system) and is able to analyze samples with very high speed (around 1 sample per second). The measurements can be performed on 384 and 1536 well plates (to date, 384 well plates are routinely used for guaranteed data). 

Details

We offer an investigation of kinetic solubility in PBS buffer (pH 7.2) and in solutions simulating gastric acid (pH 1.7) and intestinal environment (pH 6.5). Good solubility of compound in gut is crucial for its absorption to the organism.

Plasmatic protein binding is another important parameter determining how fast will the compound be eliminated from the body (by renal excretion). High binding to the proteins means slower elimination. In order to obtain relevant data, the compound must be stable in plasma.
Plasmatic stability expresses how fast is a compound metabolized in plasma and is defined by a parameter “half-life”. Compounds unstable in plasma are usually not suitable as medicines. We offer the plasma stability investigation in human, mouse and rat plasma.
Microsomal stability determines how fast is a compound metabolized in liver and is also defined by “half-life”. Many detoxification reactions occur in liver, which metabolize the potential drug before it reaches the desired destination cells. We offer the stability investigation in human, mouse and rat microsomes.
PAMPA (Parallel artificial membrane permeability assay) is an assay that determines the permeation of compounds from donor to acceptor wells. In this case, passive diffusion through the synthetic lipid membrane is investigated. The permeation declares how the compound is penetrating the lipid membrane without the active transport and can also reveal the sticking of compounds on the plastic well walls. This can be particularly useful in evaluating CACO-2 results.
CACO-2 assay is simulating the absorption of compounds through small intestinal cells. CACO-2 cell line is used, which are immortalized human adenocarcinoma cells. This assay determines the permeation through cell from apical (donor) to basolateral (acceptor) well and both passive and active cell transport are taking part in it. Important parameter is also “recovery” defining the amount of compound that got metabolized or stuck within the cells.
For sample registration for ADME, please use the form available on this website.

Contacts

Helena Mertlíková Kaiserová (sample submission, consultation)

Karel Chalupský (sample submission, consultation)

Timotej Strmeň (technical issues, MS analysis)