Simultaneous quantification of four antiretroviral drugs in breast milk samples from HIV-positive women by an ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method

Abstract

The primary strategy to avoid mother-to-child transmission of human immunodeficiency virus (HIV) through breastfeeding is administration of highly active antiretroviral therapy (HAART) to HIV-positive pregnant women. Because significant changes in the pharmacokinetics of antiretroviral (ARV) drugs occur during pregnancy, quantifying HAART and the viral load in breast milk in this population is essential. Here, we developed an analytical assay for the simultaneous quantification of four ARV drugs in breast milk using ultra-performance liquid chromatography coupled to tandem mass spectrometry. We validated this method following Mexican and international guidelines. ARV drugs. We extracted the ARV drugs from 200 μL samples of breast milk and detected these drugs in a triple quadrupole mass spectrometer with positive electrospray ionization. The validated concentration ranges (ng/mL) for zidovudine, lamivudine, lopinavir, and ritonavir were 12.5–750, 50–2500, 100–5000 and 5 to 250, respectively. Additionally, the absolute recovery percentages (and matrix effects) were 91.4 (8.39), 88.78 (28.75), 91.38 (11.77) and 89.78 (12.37), respectively. We determined that ARV drugs are stable for 24 h at 8°C and 24°C for 15 days at –80°C. This methodology had the capacity for simultaneous detection; separation; and accurate, precise quantification of ARV drugs in human breast milk samples according to Mexican standard laws and United States Food and Drug Administration guidelines.

Detection and quantification of Covid-19 antiviral drugs in biological fluids and tissues

“…Moreover, in some cases, acetonitrile [ 45 , 63 , 64 ] acidified with 0.01% HCl [ 65 ] or water adjusted with 8% (v/v) trichloroacetic acid (TCA) [ 66 ] and methanol alone [ 67 ] have been used too. In addition to plasma samples, PP has been applied to other matrices, such as seminal plasma [ 42 ], cell lysates [ 68 ], cerebrospinal fluid [ 67 ], mice vaginal lavage [ 69 ], saliva [ 64 ], urine [ 46 ] and breast milk [ 70 ]. Yamada et al [ 64 ] used protein precipitation for extracting darunavir, tenofovir and other antiviral drugs from saliva samples, through a three-step protocol, including the addition of ACN, drying of the supernatant and its regeneration with the mobile phase.…”

“…Moreover, ACN was used also for the extraction from urine samples assisted by sonication in an ultrasonic bath, in a method developed by Gumustas et al [ 46 ] for the quantification of emtricitabine, rilpivirine and tenofovir. For mice vaginal lavage, the addition of ACN is used to precipitate mucins and dilute salts [ 69 ], while in the case of breast milk, PP is conducted after lipid removal through hexane treatment [ 70 ]. When applied to samples different from plasma, protein precipitation gave satisfactory results; it should be noted however that, in some cases, to prevent carryover effects during liquid chromatography, but keeping the same LLOQ as in plasma, an extra dilution of the residue obtained after evaporation was needed [ 42 ].…”

Mitochondrial DNA Parameters in Blood of Infants Receiving Lopinavir/Ritonavir or Lamivudine Prophylaxis to Prevent Breastfeeding Transmission of HIV-1

“…In addition to in utero exposure to both HIV and the antiretroviral drugs (ARVs), children who are HIV-exposed but uninfected (CHEU) continue to be exposed to HIV and the ARVs during breastfeeding as well as for 6–12 weeks of recommended pre-exposure prophylaxis (PrEP) with either nevirapine (NVP) alone or in combination with zidovudine (ZDV) [ 1 ]. Furthermore, maternal ART results in significant ARV drug concentrations in breast milk to which the baby is also exposed for several months [ 2 , 3 , 4 , 5 ]. In 2019, 15.2 million children were CHEU worldwide, thus exposed to HIV and ARV [ 6 ].…”

On-Site Therapeutic Drug Monitoring

“…Pioneering multiplex approaches have been reported for antiretroviral agents (ARVs) [11], antifungals [12], antineoplastics [13], antibiotics [6,7,14], antidepressants [15], and immunosuppressive drugs [16] in the past decade. More recently, ultra-performance liquid chromatography (UPLC)-MS/MS has been used for simultaneous quantification of antibiotics [17] and ARVs from plasma [11] and breast milk samples [18]. Another focus is the extension of TDM studies toward unconventional samples and sampling.…”

“…Another focus is the extension of TDM studies toward unconventional samples and sampling. Several LC-MS/MS methods have been developed and applied for hair [11,18], dried blood spots [19], urine [20], sweat [21], saliva [22,23], and tissue biopsies [24].…”