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Sanidad Militar

Print version ISSN 1887-8571

Abstract

AGUILAR-ROMERO, A et al. Validation of a method to determine the serum concentration of voriconazole by HPLC/UV-Vis. Sanid. Mil. [online]. 2022, vol.78, n.3, pp.146-150.  Epub Jan 30, 2023. ISSN 1887-8571.  https://dx.doi.org/0.4321/s1887-85712022000300004.

Background:

The high increase of invasive fungal infections has increased the use of voriconazole as prophylaxis and treatment, being necessary to monitor its serum concentrations.

Objective:

To standardize and validate a simple method with high efficacy and specificity for the determination of voriconazole.

Method:

For the quantification of voriconazole, a Shimadzu high performance liquid chromatography equipment was used, coupled to an ultraviolet-visible diode array detector, performing the chromatographic separation with a Brisa LC2 C18 column. The chromatographic conditions defined were: column temperature, 35ºC; wavelength, 256 nm; injection volume, 20 µl; flow rate, 1.5 ml/min; analysis time, 9 min, mobile phase water with formic acid 0.5 % / acetonitrile 65/35. Prior to chromatographic injection, the samples underwent a treatment consisting of protein precipitation with acetonitrile and subsequent centrifugation, and the supernatant was injected The SPSS v. 25 statistical program was used, considering a p<0.05 as statistically significant.

Results:

The method developed is selective and linear (r2 =1), with a coefficient of variation ≤ 5%. In terms of accuracy and precision, the coefficients of variation were ≤ 5 %, thus complying with the requirements established for the concentration range 0.1 µg/ml-10 µg/ml.

Conclusion:

The selectivity and the simplicity of the sample treatment make it an effective, fast and simple method for the determination of voriconazole in serum and with a higher sensitivity than the immunoassays used.

Keywords : Voriconazole; fungical infections; HPLC; validation.

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