Determination of didanosine in maternal plasma, amniotic fluid, fetal and placental tissues by high‐performance liquid chromatography–tandem mass spectrometry

Abstract

A rapid and efficient high-performance liquid chromatography (HPLC)-tandem mass spectrometry method for the determination of didanosine concentrations in maternal rat plasma, amniotic fluid, placental and fetal tissue samples has been developed and validated. Tissue samples were homogenized in optima water and centrifuged. The supernatant was subjected to solid-phase extraction (SPE) prior to analysis. Plasma and amniotic fluid samples were extracted without pretreatment. An Agilent 1100 Series HPLC coupled with a Micromass Quattro II triple quadrupole mass spectrometer was used for all analyses. Chromatographic resolution was achieved on a Nova-Pak phenyl analytical column (2.0 x 150 mm, 4 microm particle size) equipped with a Phenomenex Security-guard phenyl guard cartridge (2.0 x 4.0 mm) using 60% methanol in 10 mm ammonium acetate buffer mobile phase for all matrices at a flow rate of 0.15 mL/min. The method yields retention times of 2.9 min for didanosine and 3.0 min for the internal standard, stavudine. Limits of detection were 1 ng/mL for all matrices. Recoveries were 70% or greater for both compounds in the different matrices. Within- and between-run precision (%RSD) and accuracy (%error) was less than 15% for all matrices.

Nucleoside reverse transcriptase inhibitors and their phosphorylated metabolites in human immunodeficiency virus-infected human matrices

“…Clark et al utilized a C 18 SPE cartridge to extract AZT and AZDU from rat maternal plasma, amniotic fluid, placental and fetal tissue samples with recoveries ranged from 81 to 96% for AZDU and from 82 to 96% for AZT in the different matrices [25]. In another study, the same group applied a Sep-Pak-Vac C 18 SPE cartridge to extract ddI and d4T in maternal rat plasma, amniotic fluid, placental and fetal tissue [40]. The obtained recoveries were above 70% for both compounds in all four matrices.…”

“…Significant effort has been made to achieve longer chromatographic retention of the phosphorylated metabolites by using modified RP columns such as Symmetry Shield RP C 18 [48,49], Polarity dC 18 silica [84] and Microbore column [104]. Clark et al developed and validated a rapid and efficient HPLC-MS/MS method for the determination of ddI concentrations in maternal rat plasma, amniotic fluid, placental and fetal tissue samples [40]. HPLC coupled with a triple quadrupole mass spectrometer (Micromass Quattro II) was used for the sample analyses.…”

Reversed-phase liquid chromatographic method with spectrophotometric detection for the determination of antiretroviral drugs

“…In particular, MS is certainly necessary for the quantification of intracellular levels of NRTIs at concentrations in the order of magnitude of pg mL −1 . Additionally, MS detection seems to be recommendable for the determination of low concentrations of some PIs as they may present poor absorption in the UV range (e.g., lopinavir) [6,12,[20][21][22]. Apart from HPLC, capillary electrophoresis (CE) has also been used for the determination of drug mixtures in various biological matrices [23][24][25].…”

“…Solid-phase extraction (SPE) using commercial cartridges is often used for the analysis of NRTIs [6,9,10,12,13,[19][20][21], while liquid-liquid extraction has widely been applied to NNRTI and PI drugs [7,9,[14][15][16][17][18]. Alternatively, simple procedures consisting of precipitation of biomacromolecules have been also described for the analysis of PIs [22].…”

“…Most of these methods concern the analysis of plasma although other body fluids such as urine, semen and amniotic liquid have also been analyzed [6,7,20,21]. In general, high-performance liquid chromatography (HPLC) with UV detection is successful for achieving the required sensitivity and selectivity in most of the applications.…”