Benzodiazepines and Tricyclic Antidepressants in Serum/Plasma - HPLC

Order No.: 49000, for 100 tests
Parameters:
Alprazolam, Amitriptyline, Bromazepam, Chlordiazepoxide, Clobazam, Clomipramine, Clonazepam, Clozapine, Desipramine, Diazepam, Doxepin, Flunitrazepam, Imipramine, Maprotiline, Nitrazepam, Norclobazam, Norclomipramine, Norclozapine, Nordiazepam, Nordoxepin, Nortriptyline, Oxazepam, Trimipramine

All important benzodiazepines, TCA & metabolites
One sample preparation for both substance groups
Identical HPLC conditions, one HPLC column and mobile phases

Alprazolam

Amitriptyline

Bromazepam

Chlordiazepoxide

Clobazam

Norclobazam

Clomipramine

Norclomipramine

Clonazepam

Clozapine

Norclozapine

Desipramine

Diazepam

Nordiazepam

Doxepin

Nordoxepin

Flunitrazepam

Imipramine

Maprotiline

Nitrazepam

Nortriptyline

Oxazepam

Trimipramine

Clinical relevance

Benzodiazepines are a large group of psychotropic drugs with a similar chemical structure. They are the most important representatives of sedatives and sleeping pills. Some are also used as antiepileptic drugs, muscle relaxants, and as analgesics in administering general anaesthesia. In the case of alcohol withdrawal treatments they facilitate weaning, although the danger of an alternative dependence exists. The various benzodiazepines are metabolised at different rates, and in some cases it is not until the metabolites form (e.g. desalkylflurazepam) that they become therapeutically effective.

Tricyclic antidepressants (TCAs) are used to treat psychological conditions such as anxiety, pain syndromes, and phobias, and produce a positive mood enhancing effect. Their pharmacological effect is based on increasing the concentration of dopamine, noradrenalin and serotonin in the synaptic cleft by inhibiting the degradation or reuptake of these neurotransmitters.

By regularly determining the plasma levels side effects can be minimised and overdoses can be avoided. Also, patient compliance can be checked to help ensure optimal therapy outcomes.

 

Product advantages

  • Covers all important benzodiazepines, TCA & metabolites
  • Simultaneous sample preparation for both substance groups
  • Identical HPLC column and mobile phase for all parameters

This kit is designed for the determination of the main therapeutic benzodiazepines and tricyclic antidepressants and their relevant metabolites with a single sample preparation. The two substance groups are assayed in two chromatographic runs on an isocratic HPLC system with UV detector. The analytes are quantified by the inclusion of two specifically synthesized internal standards which themselves are not used as medication and therefore do not lead to interferences. By means of selective solid phase extraction, the sample preparation eliminates interfering co-eluants and ensures reproducible and reliable results.

More Information
Method of Analysis HPLC
Number of Tests 100
Limit of quantification

1.5–27 ng/ml; The limit of quantification depends on the detector employed.

Linearity

Entire therapeutic range

Recovery 86–122 %
Intraassay CV = 0.7–4.5 %
Interassay CV = 1.9–6 %
Analysis Time 28 min (BZD), 25 min (TCA)
Specimen Serum/Plasma
Pre-analytic Treatment The patient samples are stable for at least 24 hours if stored in a dark and cool (+4 °C) place. For longer storage keep samples below -20 °C. Use polypropylene vials for transport.
Sample Preparation

Extraction

  • Precondition the Sample Clean Up Columns with 2 x 1 ml of each Equilibration Buffers 1 and 2.
  • Add 1 ml serum/plasma (calibrator/control) and 100 µl Internal Standard to the Sample Clean Up Column, mix briefly.
  • Draw complete sample through Sample Clean Up Column by centrifugation or suction, discard effluent.

Washing

  • Draw 2 x 1 ml Wash Buffer through Sample Clean Up Column by centrifugation or suction, discard effluents.

Elution

  • Elute first the benzodiazepines and then the tricyclic antidepressants with 400 µl of appropriate Elution Buffer.
  • Add 200 µl ultrapure water (HPLC grade) to each eluate.
  • Inject 50 µl of the diluted eluate into the HPLC system.
Sample Stability Prepared samples can be kept at room temperature for 3 days or at +2 to +8 °C for 5 days, light protected in glass vials. For longer storage keep samples below -18 °C (up to 2 weeks).
Injection Volume 50 µl
Flow rate 0.6 ml/min
Column temperature ambient (~ 25 °C)
Wavelength 210 nm
Gradient isocratic
Additional Info

Any isocratic HPLC system with UV detector is suitable.

Please note The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.
Parameter Alprazolam, Amitriptyline, Bromazepam, Chlordiazepoxide, Clobazam, Clomipramine, Clonazepam, Clozapine, Desipramine, Diazepam, Doxepin, Flunitrazepam, Imipramine, Maprotiline, Nitrazepam, Norclobazam, Norclomipramine, Norclozapine, Nordiazepam, Nordoxepin, Nortriptyline, Oxazepam, Trimipramine
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