Itraconazole, Posaconazole and Voriconazole in Serum/Plasma - HPLC

Order No.: 27037, for 100 tests
Parameters:
Hydroxyitraconazole, Itraconazole, Posaconazole, Voriconazole

Itraconazole/posaconazole analysis upgraded with voriconazole
Tri-level controls available
Custom-made internal standard

Itraconazole

Hydroxyitraconazole

Posaconazole

Voriconazole

Clinical relevance

In recent years there has been an increase in invasive fungal infections due to the growing number of people with immunosuppression. Candida and Aspergillus are the most common types of infection, but previously rare fungal infections such as fusariosis or zygomycosis are also on the rise. Options for oral antimycotic therapy have improved considerably with the advent of new drugs. These include second-generation triazoles such as posaconazole and voriconazole.

Posaconazole is used primarily in case of therapy-resistant mycoses or drug intolerances. Voriconazole is also effective against some itraconazole-resistant Aspergillus species and offers fungicidal activity against emerging fungal pathogens such as Scedosporium or Fusarium. All three azoles exhibit wide inter-individual variability in bioavailability, so monitoring of drug levels is required.

 

Product advantages

  • 3PLUS1® Multilevel Calibrator Set available
  • Tri-level controls available
  • Tailor-made internal standard

This Chromsystems kit allows for reliable chromatographic determination of itraconazole, its active metabolite hydroxy-itraconazole, and posaconazole and voriconazole in only one isocratic HPLC run with fluorescence detection. Two very simple and efficient protein precipitation steps are used to separate interfering components and to obtain stable eluates. A tailor-made internal standard ensures reliable and precise quantification of the analytes.

These analytes - together with four further antimycotics - can also be determined using LC-MS/MS with the TDM Parameter Set Antimycotic Drugs from the MassTox® TDM Series A.

More Information
Number of Tests 100
Limit of quantification

Itraconazole/OH-Itraconazole: 0.03 mg/l
Posaconazole: 0.02 mg/l
Voriconazole: 0.2 mg/l

Linearity

Itraconazole/OH-Itraconazole: up to 10 mg/l
Posaconazole: up to 10 mg/l
Voriconazole: up to 20 mg/l

Recovery 101–105 %
Intraassay

CV = 1.1– 4.4 %

Interassay

CV = 2.4– 5.2 %

Analysis Time Itraconazole/OH-Itraconazole: 13 min; Posaconazole/Voriconazole: 8 min
Sample Preparation
  • Mix 100 µl serum/plasma, 25 µl Internal Standard and 25 µl Precipitation Reagent 1 in a reaction vial and vortex briefly.
  • Add 200 µl Precipitation Reagent 2 and mix 30 s (vortex).
  • Centrifuge 5 min at 15000 x g
  • Inject 20 µl of the supernatant into the HPLC system..
Sample Stability The prepared samples can be kept in sealed glass vials for 1 week at +18 °C to +30 °C and for 2 weeks at +2 °C to +8 °C. Do not store refrigerated!
Specimen Serum/Plasma
Pre-analytic Treatment Storage life is up to 2 weeks at ambient temperature, at +2 °C to +8 °C up to 4 weeks. For longer storage (max. 3 months) keep samples below -18 °C.
Injection Volume 20 µl (10 to 30 µl)
Flow rate 1.2 ml/min
Column temperature ambient (~ 25 °C)
Wavelength

EX 261 nm

EM 366 nm

Gradient isocratic
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.
Method of Analysis HPLC
Parameter Hydroxyitraconazole, Itraconazole, Posaconazole, Voriconazole
Complete Specification
Linearity:
Itraconazole/
OH-Itraconazole:
up to 10 mg/l
Posaconazole: up to 10 mg/l
Voriconazole: up to 20 mg/l
Limit of quantification:
Itraconazole/
OH-Itraconazole:
0.,03 mg/l
Posaconazole 0.02 mg/l
Voriconazole: 0.2 mg/l
Intra-assay: CV < 4,4 %
Inter-assay: CV < 5,2 %
Recovery: > 98 %
Run time:
Itraconazole/
OH-Itraconazole:
aproxx. 16 min
Posaconazole/Voriconazole: aproxx. 9 min
Stability of prepared samples: at least 7 days at ambient temperature

HPLC-parameters

For the HPLC analysis of itraconazole, posaconazole and voriconazole any isocratic HPLC system with fluorescence detector is suitable.

Injection volume: 10 to 20 µl
Flow rate: 1.2 ml/min
Detection wavelength: EX 261 nm, EM 366 nm
Column temperature: +25 °C
Autosampler rinsing solution: : 80 % water
20 % acetonitril
+0.2 % phosphoric acid (85 per cent)
Additional Info Any isocratic HPLC system with fluorescence detector is suitable.

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  3. PEEK Prefilter Housing
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  4. Precolumn Cartridge Holder 4/10
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