Abstract
Therapeutic drug monitoring (TDM) is pivotal to improve the management of HIV infection. Here, a HPLC-UV method has been developed to quantify simultaneously seven HIV protease inhibitors (amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir, and saquinavir; PIs), seven nucleoside reverse transcriptase inhibitors (abacavir, didanosine, emtricitabine, lamivudine, stavudine, zalcitabine, and zidovudine; NRTIs), and two non-nucleoside reverse transcriptase inhibitors (efavirenz and nevirapine; NNRTIs) in human plasma. The volume of the plasma sample was 600 μL. This method involved automated solid-phase extraction with Oasis HLB Cartridge 1 cc (divinylbenzene and N-vinylpyrrolidone) and evaporation in a water bath under nitrogen stream. The extracted samples were reconstituted with 100 μL methanol. Twenty microliters of these samples were injected into a HPLC-UV system, the analytes were eluted on an analytical C18 Symmetry™ column (250 mm × 4.6 mm I.D.) with a particle size of 5 μm. The mobile phase (0.01 M KH2PO4 and acetonitrile) was delivered at 1.0 mL/min with linear gradient elution. The total run time for a single analysis was 35 min, the anti-HIV drugs were detected by UV at 240 and 260 nm. The calibration curves were linear up to 10 μg/mL. The absolute recovery ranged between 88 and 120%. The in vitro stability of anti-HIV drugs (0.005-10 μg/mL) in plasma has been studied at 24.0°C. On these bases, a two to four analyte method has been tailored to the individual needs of the HIV-infected patient. The HPLC-UV method here reported has been validated and is currently applied to monitor PIs, NRTIs, and NNRTIs in plasma of HIV-infected patients. It allows to monitor the largest number of anti-HIV drugs simultaneously, appearing useful in a routine laboratory, and represents an essential step to elucidate the utility of a formal therapeutic drug monitoring for the optimal follow-up of HIV-infected patients.
| Original language | English |
|---|---|
| Pages (from-to) | 258-266 |
| Number of pages | 9 |
| Journal | Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences |
| Volume | 831 |
| Issue number | 1-2 |
| DOIs | |
| Publication status | Published - Feb 2 2006 |
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Keywords
- HIV non-nucleoside reverse transcriptase inhibitors
- HIV nucleoside reverse transcriptase inhibitors
- HIV protease inhibitors
- HPLC-UV
- Therapeutic drug monitoring
ASJC Scopus subject areas
- Biochemistry
Cite this
Simultaneous determination of 16 anti-HIV drugs in human plasma by high-performance liquid chromatography. / Notari, Stefania; Bocedi, Alessio; Ippolito, Giuseppe; Narciso, Pasquale; Pucillo, Leopoldo Paolo; Tossini, Gianna; Donnorso, Raffaele Perrone; Gasparrini, Francesco; Ascenzi, Paolo.
In: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol. 831, No. 1-2, 02.02.2006, p. 258-266.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Simultaneous determination of 16 anti-HIV drugs in human plasma by high-performance liquid chromatography
AU - Notari, Stefania
AU - Bocedi, Alessio
AU - Ippolito, Giuseppe
AU - Narciso, Pasquale
AU - Pucillo, Leopoldo Paolo
AU - Tossini, Gianna
AU - Donnorso, Raffaele Perrone
AU - Gasparrini, Francesco
AU - Ascenzi, Paolo
PY - 2006/2/2
Y1 - 2006/2/2
N2 - Therapeutic drug monitoring (TDM) is pivotal to improve the management of HIV infection. Here, a HPLC-UV method has been developed to quantify simultaneously seven HIV protease inhibitors (amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir, and saquinavir; PIs), seven nucleoside reverse transcriptase inhibitors (abacavir, didanosine, emtricitabine, lamivudine, stavudine, zalcitabine, and zidovudine; NRTIs), and two non-nucleoside reverse transcriptase inhibitors (efavirenz and nevirapine; NNRTIs) in human plasma. The volume of the plasma sample was 600 μL. This method involved automated solid-phase extraction with Oasis HLB Cartridge 1 cc (divinylbenzene and N-vinylpyrrolidone) and evaporation in a water bath under nitrogen stream. The extracted samples were reconstituted with 100 μL methanol. Twenty microliters of these samples were injected into a HPLC-UV system, the analytes were eluted on an analytical C18 Symmetry™ column (250 mm × 4.6 mm I.D.) with a particle size of 5 μm. The mobile phase (0.01 M KH2PO4 and acetonitrile) was delivered at 1.0 mL/min with linear gradient elution. The total run time for a single analysis was 35 min, the anti-HIV drugs were detected by UV at 240 and 260 nm. The calibration curves were linear up to 10 μg/mL. The absolute recovery ranged between 88 and 120%. The in vitro stability of anti-HIV drugs (0.005-10 μg/mL) in plasma has been studied at 24.0°C. On these bases, a two to four analyte method has been tailored to the individual needs of the HIV-infected patient. The HPLC-UV method here reported has been validated and is currently applied to monitor PIs, NRTIs, and NNRTIs in plasma of HIV-infected patients. It allows to monitor the largest number of anti-HIV drugs simultaneously, appearing useful in a routine laboratory, and represents an essential step to elucidate the utility of a formal therapeutic drug monitoring for the optimal follow-up of HIV-infected patients.
AB - Therapeutic drug monitoring (TDM) is pivotal to improve the management of HIV infection. Here, a HPLC-UV method has been developed to quantify simultaneously seven HIV protease inhibitors (amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir, and saquinavir; PIs), seven nucleoside reverse transcriptase inhibitors (abacavir, didanosine, emtricitabine, lamivudine, stavudine, zalcitabine, and zidovudine; NRTIs), and two non-nucleoside reverse transcriptase inhibitors (efavirenz and nevirapine; NNRTIs) in human plasma. The volume of the plasma sample was 600 μL. This method involved automated solid-phase extraction with Oasis HLB Cartridge 1 cc (divinylbenzene and N-vinylpyrrolidone) and evaporation in a water bath under nitrogen stream. The extracted samples were reconstituted with 100 μL methanol. Twenty microliters of these samples were injected into a HPLC-UV system, the analytes were eluted on an analytical C18 Symmetry™ column (250 mm × 4.6 mm I.D.) with a particle size of 5 μm. The mobile phase (0.01 M KH2PO4 and acetonitrile) was delivered at 1.0 mL/min with linear gradient elution. The total run time for a single analysis was 35 min, the anti-HIV drugs were detected by UV at 240 and 260 nm. The calibration curves were linear up to 10 μg/mL. The absolute recovery ranged between 88 and 120%. The in vitro stability of anti-HIV drugs (0.005-10 μg/mL) in plasma has been studied at 24.0°C. On these bases, a two to four analyte method has been tailored to the individual needs of the HIV-infected patient. The HPLC-UV method here reported has been validated and is currently applied to monitor PIs, NRTIs, and NNRTIs in plasma of HIV-infected patients. It allows to monitor the largest number of anti-HIV drugs simultaneously, appearing useful in a routine laboratory, and represents an essential step to elucidate the utility of a formal therapeutic drug monitoring for the optimal follow-up of HIV-infected patients.
KW - HIV non-nucleoside reverse transcriptase inhibitors
KW - HIV nucleoside reverse transcriptase inhibitors
KW - HIV protease inhibitors
KW - HPLC-UV
KW - Therapeutic drug monitoring
UR - http://www.scopus.com/inward/record.url?scp=31544476582&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=31544476582&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2005.12.016
DO - 10.1016/j.jchromb.2005.12.016
M3 - Article
C2 - 16406832
AN - SCOPUS:31544476582
VL - 831
SP - 258
EP - 266
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
SN - 1570-0232
IS - 1-2
ER -