ABSTRACT
A simple and fast liquid chromatography-tandem mass spectrometry method was established and validated for the simultaneous determination of tenofovir alafenamide (TAF) and tenofovir (TNF) in human plasma. A simple protein precipitation procedure was employed to extract analytes from plasma. Chromatographic separation was performed on an Eclipse Plus C18 column utilizing a fast gradient elution starting with 2% of 2 mM ammonium acetate-formic acid (100/0.1, v/v) followed by increasing the percentage of acetonitrile. Detection was performed on a tandem mass spectrometer equipped with an electrospray ionization source operated in the positive ionization mode, using the transitions m/z 477.2 â m/z 346.1 for TAF and m/z 288.1 â m/z 176.1 for TNF. TAF-d5 and TNF-d7 were used as the internal standard of TAF and TNF, respectively. The method was validated in the concentration ranges 1.25-500 ng/mlfor TAF and 0.300-15.0 ng/ml for TNF with acceptable accuracy and precision.
Subject(s)
Adenine , Tandem Mass Spectrometry , Alanine , Chromatography, High Pressure Liquid , Chromatography, Liquid/methods , Humans , Reproducibility of Results , Spectrometry, Mass, Electrospray Ionization , Tandem Mass Spectrometry/methods , Tenofovir/analogs & derivatives , Tenofovir/analysisABSTRACT
A novel method was developed for the simultaneous estimation of the doravirine, lamivudine, and tenofovir disoproxil fumarate in the pharmaceutical dosage form. The chromatogram was run through an Ascentis C18 column (150 × 4.6 mm, 2.7 µm), with the mobile phase consisting of a phosphate buffer and acetonitrile in the ratio of 50:50 (v/v). The mobile phase was pumped through the column at a flow rate of 1 mL/min. The column temperature was maintained at 30°C. The optimized wavelength for doravirine, lamivudine, and tenofovir disoproxil fumarate was 230.0 nm. The retention times for doravirine, lamivudine, and tenofovir disoproxil were 2.222, 2.764, and 3.403 min, respectively; the relative standard deviation (%) values of method precision for doravirine, lamivudine, and tenofovir disoproxil were 0.6, 0.6, and 0.1, respectively. The % recovery was 100.20%, 100.15%, and 100.36% for doravirine, lamivudine, and tenofovir disoproxil fumarate, respectively. The limit of detection and limit of quantification values were obtained from regression equations of doravirine, lamivudine, and tenofovir disoproxil fumarate, and were 0.24 and 0.73 ppm, 0.53 and 1.60 ppm, and 0.47 and 1.43 ppm, respectively. The regression equations of doravirine, lamivudine, and tenofovir disoproxil fumarate were y = 17,541x + 117,303, y = 15,555x + 10,791, and y = 15,250x + 31,663, respectively. The method developed was accurate, simple, precise, sensitive, and economical. Hence, it could be adopted for regular quality control for estimation of doravirine, lamivudine, and tenofovir disoproxil fumarate in pharmaceutical industries.
Subject(s)
Chromatography, High Pressure Liquid/methods , Lamivudine/analysis , Pyridones/analysis , Tenofovir/analysis , Triazoles/analysis , Drug Stability , Lamivudine/chemistry , Limit of Detection , Linear Models , Pyridones/chemistry , Reproducibility of Results , Tablets , Tenofovir/chemistry , Triazoles/chemistryABSTRACT
Tenofovir disoproxil fumarate (TDF) in combination with emtricitabine (FTC) is the backbone for both human immunodeficiency virus (HIV) treatment and pre-exposure prophylaxis (PrEP) worldwide. Tenofovir alafenamide (TAF) with FTC is increasingly used in HIV treatment and was recently approved for PrEP among men-who-have-sex-with-men. TDF and TAF are both metabolized into tenofovir (TFV). Antiretrovirals in plasma are taken up into hair over time, with hair levels providing a long-term measure of adherence. Here, we report a simple, robust, highly sensitive, and validated high-performance liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS)-based analytical method for analyzing TFV and FTC from individuals on either TDF/FTC or TAF/FTC in small hair samples. TFV/FTC are extracted from ~5 mg hair and separated on a column using a gradient elution. The lower quantification limits are 0.00200 (TFV) and 0.0200 (FTC) ng/mg hair; the assay is linear up to 0.400 (TFV) and 4.00 (FTC) ng/mg hair. The intra-day and inter-day coefficients of variance (CVs) are 5.39-12.6% and 6.40-13.5% for TFV and 0.571-2.45% and 2.45-5.16% for FTC. TFV concentrations from participants on TDF/FTC-based regimens with undetectable plasma HIV RNA were 0.0525 ± 0.0295 ng/mg, whereas those from individuals on TAF/FTC-based regimens were 0.0426 ± 0.0246 ng/mg. Despite the dose of TFV in TDF being 10 times that of TAF, hair concentrations of TFV were not significantly different for those on TDF versus TAF regimens. Pharmacological enhancers (ritonavir and cobicistat) did not boost TFV concentrations in hair. In summary, we developed and validated a sensitive analytical method to analyze TFV and FTC in hair and found that hair concentrations of TFV were essentially equivalent among those on TDF and TAF.
Subject(s)
Adenine/analogs & derivatives , Anti-HIV Agents/analysis , Emtricitabine, Tenofovir Disoproxil Fumarate Drug Combination/analysis , Emtricitabine/analysis , Hair/chemistry , Tenofovir/analysis , Adenine/analysis , Adenine/pharmacokinetics , Anti-HIV Agents/administration & dosage , Anti-HIV Agents/pharmacokinetics , Chromatography, High Pressure Liquid/methods , Cobicistat/administration & dosage , Dose-Response Relationship, Drug , Emtricitabine/pharmacokinetics , Emtricitabine, Tenofovir Disoproxil Fumarate Drug Combination/administration & dosage , Emtricitabine, Tenofovir Disoproxil Fumarate Drug Combination/pharmacokinetics , HIV Infections/drug therapy , Hair Analysis , Humans , Ritonavir/administration & dosage , Tandem Mass Spectrometry/methods , Tenofovir/pharmacokinetics , Tissue DistributionABSTRACT
Objective methods of measuring antiretroviral adherence are limited. We assessed the relationship between tenofovir disoproxil fumarate (TDF) hair concentrations, self-reported adherence, and virological outcomes in HIV-infected adolescents in Harare, Zimbabwe. HIV-infected adolescents on atazanavir/ritonavir-based second-line treatment for >6 months with viral load (VL) ≥1,000 copies/mL were randomized to either modified directly administered antiretroviral therapy (mDAART) or standard of care. Hair and VL samples were collected at baseline and after 90 days. Treatment outcome was defined as TDF concentrations in hair. Virological suppression was defined as VL <1,000 copies/mL. Thirty-four adolescents had TDF concentrations measured at baseline and follow-up. Mean (median); range age was 16 (16); 13-18 years and 53% were females. Nineteen (56%) were randomized to mDAART. Mean (SD); range TDF concentrations were 0.03 (0.04); 0-0.17 ng/mg hair and 0.06 (0.06); 0-0.3 ng/mg hair at baseline and follow-up, respectively. Higher TDF concentrations were associated with decreased VL [regression coefficient (RC) 0.8; 95% confidence interval (CI) 0.7-1.0; p = .008] and mDAART (RC 0.5; 95% CI 0.3-1.0; p = .04), but were not associated with self-reported adherence and virological suppression (VL <1,000 copies/mL). Higher TDF hair concentrations were observed with virological decrease and an adherence intervention. Hair antiretroviral concentrations could be useful in triggering adherence interventions among adolescents with second-line virological failure.
Subject(s)
Anti-HIV Agents , HIV Infections , Hair/chemistry , Tenofovir , Viral Load , Adolescent , Anti-HIV Agents/analysis , Anti-HIV Agents/therapeutic use , Female , HIV Infections/drug therapy , Humans , Male , Medication Adherence , Self Report , Tenofovir/analysis , Tenofovir/therapeutic use , Treatment Failure , ZimbabweABSTRACT
Prevention of HIV infection and unintended pregnancies are public health priorities. In sub-Saharan Africa, where HIV prevalence is highest, depot medroxyprogesterone acetate (DMPA) is widely used as contraception. Therefore, understanding potential interactions between DMPA and antiretrovirals is critical. Here, we use a macaque model to investigate the effect of DMPA on the pharmacology of the antiretroviral tenofovir alafenamide (TAF). Female rhesus macaques received 30 mg of DMPA (n = 9) or were untreated (n = 9). Macaques received a human equivalent dose of TAF (1.5 mg/kg) orally by gavage. Tenofovir (TFV) and TFV-diphosphate (TFV-DP) were measured in blood, secretions, and tissues over 72 h. The median area under the curve (AUC0-72h) values for TFV-DP in peripheral blood mononuclear cells were similar in DMPA-treated (6991 fmol*h/106 cells) and untreated controls (5256 fmol*h/106 cells) (P = 0.174). Rectal tissue TFV-DP concentrations from DMPA+ animals [median: 20.23 fmol/mg of tissue (range: 4.94-107.95)] were higher than the DMPA- group [median: below the limit of quantification (BLOQ-11.92)], (P = 0.019). TFV-DP was not detectable in vaginal tissue from either group. A high-dose DMPA treatment in macaques was associated with increased rectal TFV-DP levels, indicating a potential tissue-specific drug-drug interaction. The lack of detectable TFV-DP in the vaginal tissue warrants further investigation of PrEP efficacy with single-agent TAF products. DMPA did not affect systemic TAF metabolism, with similar PBMC TFV-DP in both groups, suggesting that DMPA use should not alter the antiviral activity of TAF.
Subject(s)
Anti-HIV Agents/administration & dosage , Contraceptive Agents, Female/administration & dosage , Medroxyprogesterone Acetate/administration & dosage , Tenofovir/administration & dosage , Animals , Anti-HIV Agents/analysis , Anti-HIV Agents/blood , Anti-HIV Agents/pharmacology , Drug Interactions , Female , HIV Infections/drug therapy , Leukocytes, Mononuclear/drug effects , Macaca mulatta , Medroxyprogesterone Acetate/analysis , Medroxyprogesterone Acetate/blood , Medroxyprogesterone Acetate/pharmacology , Models, Animal , Tenofovir/analysis , Tenofovir/blood , Tenofovir/pharmacologyABSTRACT
BACKGROUND: Transgender women (TGW) and men who have sex with men (MSM) in sub-Saharan Africa have high HIV acquisition risks and can benefit from daily pre-exposure prophylaxis (PrEP). We assessed PrEP adherence by measuring tenofovir-diphosphate (TFV-DP) levels and explore motives for PrEP persistence in TGW and MSM. METHODS: Participants were enrolled in a one-year PrEP programme and made quarterly visits irrespective of whether they were still using PrEP. At their month 6 visit, participants provided a dried blood spot to test for TFV-DP levels; protective levels were defined as those compatible with ≥4 pills per week (700-1249 fmol/punch). Before TFV-DP levels were available, a sub-set of these participants were invited for an in-depth interview (IDI). Semi-structured IDI topic guides were used to explore motives to uptake, adhere to, and discontinue PrEP. IDI data were analyzed thematically. RESULTS: Fifty-three participants (42 MSM and 11 TGW) were enrolled. At month 6, 11 (20.7%) participants (8 MSM and 3 TGW) were lost to follow up or stopped taking PrEP. Any TFV-DP was detected in 62.5% (5/8) of TGW vs. 14.7% of MSM (5/34, p = 0.01). Protective levels were detected in 37.5% of TGW (3/8), but not in any MSM. Nineteen IDI were conducted with 7 TGW and 9 MSM on PrEP, and 1 TGW and 2 MSM off PrEP. Unplanned or frequent risky sexual risk behaviour were the main motives for PrEP uptake. Among participants on PrEP, TGW had a more complete understanding of the benefits of PrEP. Inconsistent PrEP use was attributed to situational factors. Motives to discontinue PrEP included negative reactions from partners and stigmatizing healthcare services. CONCLUSION: While MSM evinced greater adherence challenges in this PrEP programme, almost 40% of TGW were protected by PrEP. Given high HIV incidences in TGW these findings hold promise for TGW PrEP programming in the region.
Subject(s)
Anti-HIV Agents/therapeutic use , HIV Infections/prevention & control , Homosexuality, Male/psychology , Pre-Exposure Prophylaxis , Transgender Persons/psychology , Adolescent , Adult , Anti-HIV Agents/analysis , Dried Blood Spot Testing , Health Risk Behaviors , Humans , Interviews as Topic , Kenya , Male , Medication Adherence , Tenofovir/analysis , Tenofovir/therapeutic use , Young AdultABSTRACT
Understanding in utero transfer of antiretrovirals is critical for interpreting safety. Hair levels measure cumulative exposure. We measured tenofovir (TFV) concentrations in hair at delivery among women living with human immunodeficiency virus receiving TFV disoproxil fumarate-based treatment and their infants, using liquid chromatography-tandem mass spectrometry. Among 103 mother-infant pairs, the mean log10 ratio of infant-to-maternal TFV levels was 1.08 (95% confidence interval, .97-1.20). TFV transfer was 60% lower from mothers who had preterm compared with term deliveries and 42% lower from mothers who had cesarean compared with vaginal deliveries. Like prior studies assessing transfer via short-term measures (plasma, cord blood, amniotic fluid), we found high cumulative transfer using hair.
Subject(s)
Anti-HIV Agents/analysis , Anti-HIV Agents/pharmacokinetics , Fetus/metabolism , Hair/chemistry , Tenofovir/analysis , Tenofovir/pharmacokinetics , Adult , Delivery, Obstetric , Female , HIV Infections/drug therapy , Humans , Infant, Newborn , Mothers , Pregnancy , Pregnancy Complications, Infectious/drug therapy , Pregnancy Trimesters , Premature Birth , Prospective Studies , United States , Young AdultABSTRACT
OBJECTIVE: We evaluated peripartum tenofovir (TFV) exposure via hair measures among women living with HIV in the United States. DESIGN: Observational cohort study. METHODS: Hair samples were collected at or shortly after childbirth among mothers enrolled in the Surveillance Monitoring for Antiretroviral Therapy Toxicities Study of the Pediatric HIV/AIDS Cohort Study between 6/2014 and 7/2016. Among mothers receiving TFV disoproxil fumarate (TDF)-based regimens during pregnancy, TFV hair concentrations were analyzed using liquid chromatography/tandem mass spectrometry. Weight-normalized TFV concentrations were log10 transformed. Multivariable linear regression assessed correlates of TFV concentrations. RESULTS: Overall, 121 mothers on TDF-based antiretroviral therapy during pregnancy had hair specimens tested for TFV concentrations and were included in the analysis. Median age at delivery was 31 years [interquartile range (IQR) 26-36]; 71% self-identified as non-Hispanic black, and 10% had unsuppressed viral loads in late pregnancy (HIV RNAâ≥â400 copies/ml). Median time from birth to hair collection was 3 days (IQR 1-14) and median TFV hair concentration was 0.02âng/mg (IQR 0.01-0.04). In multivariable models, an unsuppressed viral load in late pregnancy was associated with 80% lower adjusted mean peripartum TFV concentrations than pregnancies with viral suppression (95% confidence interval: -90% to -59%, Pâ<â0.001). Use of TDF only in the first trimester and attaining high school graduation were also associated with lower TFV hair concentrations. CONCLUSION: Unsuppressed viral load during late pregnancy was strongly associated with lower maternal TFV hair concentrations at birth, though viremia was rare. Efforts to improve maternal virological outcomes and eliminate vertical HIV transmission could incorporate drug exposure monitoring using hair or other metrics.
Subject(s)
Anti-HIV Agents , HIV Infections , Tenofovir/analysis , Adult , Anti-HIV Agents/therapeutic use , Child , Cohort Studies , Female , HIV Infections/drug therapy , Hair/chemistry , Humans , Infectious Disease Transmission, Vertical , Pregnancy , RNA/therapeutic use , Tenofovir/therapeutic use , United StatesABSTRACT
Fixed dose combination (FDC) of tenofovir disoproxil fumarate (TDF) and lamivudine (3TC) is one of the most preferred FDC for the treatment of acquired immunodeficiency syndrome (AIDS)/human immunodeficiency virus (HIV) infection. To the best of authors' knowledge there are no reported methods for chiral purity estimation of both drugs simultaneously from a FDC. The current study was focused on the development of a single chiral method uisng supercritical fluid chromatography (SFC) for separation of stereoisomers of TDF and 3TC combination employing design of experiment (DoE) approach. Method development was planned in three steps by using different experimental designs for each step. I-optimal, Taguchi orthogonal array and face-centred central composite designs (CCD) were employed for primary parameter selection, secondary parameter screening and final method optimization, respectively. All six stereoisomers were separated in a 10 minute run on Chiralpak IA column with carbon di-oxide /methanol (containing 0.5 % v/v n-butylamine) as mobile phase at 1.5 mL/min in gradient mode. The optimized method was verified for performance through establishing specificity, precision, linearity, accuracy, limit of quantification, and solution stability. Resolution between each isomeric pair was more than 1.5. The method was found to be linear from 1.5 µg/mL to 7.5 µg/mL for 3TC and 7.5 µg/mL to 37.5 µg/mL for TDF stereoisomers. The R2 values for all the linearity curves for undesired isomers were greater than 0.995. The method proved to be rapid, reproducible and efficient to quantify stereoisomers of both drugs in a single run.
Subject(s)
Chromatography, Supercritical Fluid/methods , Lamivudine/analysis , Tenofovir/analysis , Lamivudine/chemistry , Reference Standards , Reproducibility of Results , Stereoisomerism , Tenofovir/chemistryABSTRACT
Antiretroviral fixed-dose-combination drugs are best assayed with high-performance liquid chromatography, or liquid chromatography-tandem mass spectrometry. However, most scientists in developing nations have no access to these expensive instruments. A more affordable quantitative technique is the use of ultraviolet-visible spectroscopy-where often the absorption spectra of these antiretrovirals are overlapping; thus complex derivative methodologies are required for quantification. A simple, rapid, and accurate thin layer chromatography-ultraviolet spectrophotometric method for the quantification of binary mixtures of lamivudine, zidovudine, and tenofovir-disoproxil-fumarate in tablet formulations was developed. Lamivudine/tenofovir-disoproxil-fumarate and lamivudine/zidovudine were extracted and separated on glass thin-layer chromatography plates. Drugs were identified in ultraviolet light at 254 nm and quantified in acidic medium using ultraviolet spectrophotometry. The retardation factors were 0.43, 0.79, and 0.81 for lamivudine, tenofovir-disoproxil-fumarate, and zidovudine, respectively, with corresponding absorption maxima at 270, 260, and 265 nm. Linearity ranged from 1 to 40 µg/mL for all drugs (R = 0.9998-0.9999), while recovery studies were 95.10-102.11% and amount in formulations ranged from 97.99 ± 0.63 to 101.47 ± 2.39%. The paired t-test (n = 5) indicated no significant difference between the proposed and high-performance liquid chromatography methods, hence comparable and can be used as an alternative method in routine quality determination of antiretroviral medicines.
Subject(s)
Anti-HIV Agents/analysis , Lamivudine/analysis , Tenofovir/analysis , Zidovudine/analysis , Chromatography, Thin Layer , Drug Combinations , Drug Compounding , Molecular Structure , Spectrophotometry, Ultraviolet , Tablets/analysisABSTRACT
BACKGROUND: Antiretroviral adherence is essential to HIV treatment efficacy. Various self-reported measures are commonly used for assessing antiretroviral adherence. Limited data are available regarding the validity of those self-reported measures in comparison with long-term objective biomarkers of adherence measures such as hair measures. METHODS: Self-reported adherence (frequency, percentage, and visual analog scale [VAS]) and hair tenofovir concentration were evaluated at a single time point from 268 people living with HIV in China. The responses to each of three self-reported measures were converted into percentage and then dichotomized as "optimal" (100%) vs. "suboptimal" (less than 100%) adherence. Two composite adherence scores (CAS) were created from the three self-reported measures: (1) an overall adherence was the average percentage of the three self-reported measures; (2) responses were termed optimal adherence if participants reporting optimal adherence in all three self-reported measures, while were termed suboptimal adherence. Hair tenofovir concentration was also dichotomized as "optimal" (above the limit of quantitation, 36 pg/mg) vs. "suboptimal" adherence (blow 36 pg/mg). Spearman correlation, kappa statistics, and logistic regression analysis were used to calculate the correlations, agreements, and predictions of self-reported measures with hair measure, respectively. RESULTS: Overall adherence, but any of the three self-reported adherence, was correlated with hair tenofovir concentration (r = 0.13, p < 0.05). Self-reported optimal adherence in VAS and CAS measures were agreed with and predicted optimal adherence assessed by hair measure (Kappa = 0.107, adjusted OR = 1.88, 95% CI 1.03-3.45; Kappa = 0.109, adjusted OR = 1.80, 95% CI 1.02-3.18; all p < 0.05, respectively). CONCLUSION: VAS may be a good individual self-reported measure for antiretroviral adherence, and CAS may be a good composite self-reported measure for antiretroviral adherence.
Subject(s)
Anti-HIV Agents/analysis , Anti-HIV Agents/therapeutic use , HIV Infections/drug therapy , Hair/chemistry , Medication Adherence/statistics & numerical data , Adult , China/epidemiology , Female , HIV Infections/epidemiology , Humans , Male , Middle Aged , Self Report , Surveys and Questionnaires , Tenofovir/analysis , Tenofovir/therapeutic use , Treatment OutcomeABSTRACT
RATIONALE: Tenofovir (TFV) is a first-line antiviral agent against hepatitis B virus (HBV) and is recommended for the prevention of mother-to-infant transmission of HBV. To study the distribution of TFV in umbilical cord plasma and amniotic fluid of HBV-infected pregnant women, a rapid and sensitive method for TFV determination was developed and validated. METHODS: The quantification method was developed using liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS). The analytes were separated on an Acquity UPLC HSS T3 column under gradient elution with methanol and 0.01% ammonia solution in 10 mM ammonium acetate/water. This is the first reported method for the determination of TFV using alkaline rather than acidic mobile phases. Linearity, accuracy, precision, limit of quantification, specificity and stability were assessed. RESULTS: Detection of TFV was achieved within 4 min. The calibration curves for TFV quantification showed excellent linearity in the range of 1-500 ng/mL. The intra- and interbatch precision and accuracy ranged from -4.35% to 6.92%. This method was successfully applied to determination of samples from 50 HBV mono-infected women undergoing tenofovir disoproxil fumarate therapy. The mean concentrations of TFV in the umbilical cord and amniotic fluid samples were 29.2 (4.6-86) and 470.9 (156-902) ng/mL, respectively, which showed a moderate positive correlation (r = 0.5299, P<0.001). CONCLUSIONS: A simple, rapid but sensitive bioanalytical method to determine TFV concentration in both umbilical cord plasma and amniotic fluid using LC/MS/MS was developed and applied to HBV-infected women during labor who were undergoing TDF therapy, which will help us understand the efficacy and safety of tenofovir during pregnancy.
Subject(s)
Amniotic Fluid/chemistry , Antiviral Agents/analysis , Fetal Blood/chemistry , Tandem Mass Spectrometry/methods , Tenofovir/analysis , Animals , Antiviral Agents/blood , Chromatography, High Pressure Liquid/economics , Chromatography, High Pressure Liquid/methods , Drug Monitoring/economics , Drug Monitoring/methods , Female , Hepatitis B/blood , Hepatitis B/drug therapy , Humans , Limit of Detection , Pregnancy , Pregnancy Complications, Infectious/blood , Pregnancy Complications, Infectious/drug therapy , Tandem Mass Spectrometry/economics , Tenofovir/blood , Umbilical Cord/blood supplyABSTRACT
This work describes a simple and sensitive method for the simultaneous isolation, enrichment, identification and quantitation of selected antiretroviral drugs; emtricitabine, tenofovir disoproxil and efavirenz in aqueous samples and plants. The analytical method was based on microwave extraction and hollow fibre liquid phase microextraction technique coupled with ultra-high pressure liquid chromatography-high resolution mass spectrometry. A multivariate approach via a half-fractional factorial design was used focusing on six factors; donor phase pH, acceptor phase HCl concentration, extraction time, stirring rate, supported liquid membrane carrier composition and salt content. The optimal enrichment factors for emtricitabine, tenofovir disoproxil and efavirenz from aqueous phase were 78, 111 and 24, respectively. The analytical method yielded recoveries in the range of 86 to 111%, and quantitation limits for emtricitabine, tenofovir disoproxil and efavirenz in wastewater were 0.033, 0.10 and 0.53⯵gâ¯L-1, respectively. The drugs were detected in most samples with concentrations up to 37.6⯵gâ¯L-1 recorded for efavirenz in wastewater effluent. Roots of the water hyacinth plant had higher concentrations of the investigated drugs ranging from 7.4 to 29.6⯵gâ¯kg-1. Overall, hollow fibre liquid phase microextraction proved to be an ideal tool for isolating and pre-concentrating the selected antiretroviral drugs from environmental samples.
Subject(s)
Anti-Retroviral Agents/analysis , Benzoxazines/analysis , Emtricitabine/analysis , Tenofovir/analysis , Wastewater/analysis , Water Pollutants, Chemical/analysis , Alkynes , Chromatography, High Pressure Liquid , Cyclopropanes , Eichhornia/chemistry , Environmental Monitoring , Liquid Phase Microextraction , Plant Roots/chemistry , Tandem Mass SpectrometryABSTRACT
Tenofovir disoproxil fumarate (TDF) is a prodrug of tenofovir, and after being administered orally, it converts to tenofovir in the blood. With the increasing use of TDF in women for treatment and prevention of mother-to-child transmission (MTCT) of both human immunodeficiency virus (HIV) and hepatitis B virus (HBV), or the pre-exposure prophylaxis (PrEP) for HIV, many nursing mothers have to understand the risk of exposure to tenofovir via breastmilk and make the decision about breastfeeding while on TDF treatment. Despite the safety record of TDF in pregnancy, some guidelines recommend against its use during breastfeeding. In this paper, we compared the dosage levels of tenofovir exposure in fetuses, breastfed infants, and children receiving tenofovir treatment. We found that breastfed infants were exposed to only 0.5%-16% of the tenofovir dosage that fetuses experienced via placental transfer, and 0.01-0.04% of the recommended weight-adjusted therapeutic dose. The assessment of toxicity risk from the dose perspective is an important and natural way of addressing safety concerns about exposure to tenofovir via breastfeeding. Based on the safety data from fetuses and children with tenofovir exposure, and the comparatively negligible exposure dosage from breastfeeding, our study supports mothers on TDF treatment should be encouraged to breastfeed.
Subject(s)
Anti-HIV Agents/therapeutic use , HIV Infections/prevention & control , Infectious Disease Transmission, Vertical/prevention & control , Pregnancy Complications, Infectious/prevention & control , Tenofovir/therapeutic use , Animals , Anti-HIV Agents/analysis , Antiviral Agents/analysis , Antiviral Agents/therapeutic use , Female , HIV Infections/drug therapy , Hepatitis B/prevention & control , Hepatitis B/virology , Humans , Male , Milk, Human/chemistry , Pregnancy , Pregnancy Complications, Infectious/virology , Tenofovir/analysisABSTRACT
BACKGROUND: Antiretroviral treatment (ART) adherence is often suboptimal in the perinatal period. We measured hair tenofovir (TFV) concentrations as a metric of adherence in postpartum women to understand patterns and predictors of adherence throughout this critical period. In addition, we examined the association between hair TFV concentrations and virologic outcomes. METHODS: Between 12/2012 and 09/2016, hair samples were collected longitudinally from delivery through breastfeeding from women on ART in the Promoting Maternal and Infant Survival Everywhere study (NCT01061151) in sub-Saharan Africa. Hair TFV levels were measured using validated methods. Using generalized estimating equations, we estimated the association between hair TFV levels and virologic suppression (<400 copies/ml) over time and assessed predictors of hair TFV levels. RESULTS: Hair TFV levels were measured at 370 visits in 71 women from delivery through a median of 14 months (interquartile range 12-15) of breastfeeding. Levels ranged from below detection (0.002) to 1.067âng/mg (geometric mean: 0.047). After at least 90 days on ART, 69 women had at least one viral load measured (median 5 measures, range 1-9); 18 (26%) experienced viremia at least once. Each doubling of TFV level more than doubled odds of concurrent virologic suppression [odds ratio 2.35, 95% confidence interval (CI): 1.44-3.84, Pâ=â0.0006] and was associated with 1.43 times the odds of future suppression (95% CI: 0.75-2.73, Pâ=â0.28). Relative to the first 3 months after delivery, hair levels were highest in months 6-12 (1.42-fold higher, 95% CI: 1.09-1.85, Pâ=â0.01). CONCLUSION: Hair TFV levels strongly predicted concurrent virologic suppression among breastfeeding women. Objective adherence metrics can supplement virologic monitoring to optimize treatment outcomes in this important transition period.
Subject(s)
Anti-HIV Agents/administration & dosage , Anti-HIV Agents/analysis , HIV Infections/drug therapy , Hair/chemistry , Sustained Virologic Response , Tenofovir/administration & dosage , Tenofovir/analysis , Adult , Africa South of the Sahara , Breast Feeding , Female , Humans , Longitudinal Studies , Medication Adherence/statistics & numerical data , Postpartum Period , Pregnancy , Treatment Outcome , Viral Load , Young AdultABSTRACT
Combination antiretroviral therapy (cART) regimens are recommended for HIV patients to better achieve and maintain plasma viral suppression. Despite adequate plasma viral suppression, HIV persists inside the brain, which is, in part thought to result from poor brain penetration of antiretroviral drugs. In this study, a simple and ultra-sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of tenofovir, emtricitabine, and dolutegravir in cell lysates of an immortalized human brain microvascular endothelial cell line (hCMEC/D3) was developed and validated. Analytes were separated on a reverse phase C18 column using water and 0.1% formic acid in acetonitrile as mobile phases. The analytes were detected using positive electrospray ionization mode with multiple reaction monitoring (MRM). The assay was linear in the concentration range of 0.1-100â¯ngâ¯mL-1 for all analytes. Intra- and inter-assay precision and accuracy were within ±13.33% and ±10.53%, respectively. This approach described herein was used to determine the intracellular accumulation of tenofovir, emtricitabine, dolutegravir simultaneously in hCMEC/D3 cells samples.
Subject(s)
Anti-HIV Agents/analysis , Brain/cytology , Chromatography, Liquid/methods , Endothelial Cells/cytology , Intracellular Space/chemistry , Tandem Mass Spectrometry/methods , Analytic Sample Preparation Methods , Anti-HIV Agents/isolation & purification , Cell Line , Emtricitabine/analysis , Emtricitabine/isolation & purification , Heterocyclic Compounds, 3-Ring/analysis , Heterocyclic Compounds, 3-Ring/isolation & purification , Humans , Linear Models , Oxazines , Piperazines , Pyridones , Reproducibility of Results , Tenofovir/analysis , Tenofovir/isolation & purificationABSTRACT
OBJECTIVES: Women likely require higher adherence than men to preexposure prophylaxis (PrEP) with tenofovir disoproxil fumarate (TDF)/emtricitabine (FTC) for similar efficacy. Pharmacologic metrics of adherence predict efficacy better than self-report, but expected drug levels (adherence benchmarks) must be established using directly observed therapy. We sought to evaluate whether tenofovir hair concentrations differ between women and men receiving directly observed TDF/FTC. METHODS: We assessed tenofovir hair concentrations in HIV-uninfected volunteers randomized to receive 100%, 67%, or 33% of daily dosing of TDF/FTC for 12 weeks (DOT-DBS, NCT02022657). Hair samples were collected at dosing weeks 4, 8, and 12 and every 3 weeks during a 12-week washout. Tenofovir concentrations in the proximal 1.5 cm of hair (representing â¼6 weeks of exposure) were analyzed using liquid chromatography/tandem mass spectrometry. Linear regression was used to model tenofovir hair concentrations in terms of sex, doses over the prior 6 weeks, and number of days since last dose. RESULTS: A total of 264 hair samples were analyzed from 23 female and 24 male participants. Female participants had similar tenofovir hair concentrations to men (estimated fold-difference 0.92, 95% CI 0.75-1.13, Pâ=â0.43). The estimated fold-difference in tenofovir levels for female versus male participants did not appreciably change when age (0.93, 95% CI 0.76-1.15), weight (0.89, 95% CI 0.71-1.11), or race/ethnicity (0.95, 95% CI 0.77-1.17) were added to the model. CONCLUSION: Women and men have similar adherence benchmarks for tenofovir in hair samples. As pharmacokinetic metrics are increasingly used for PrEP monitoring, these findings provide guidance for assessing adherence via hair concentrations.
Subject(s)
Anti-HIV Agents/administration & dosage , Anti-HIV Agents/pharmacokinetics , Emtricitabine/administration & dosage , Hair/chemistry , Tenofovir/administration & dosage , Tenofovir/pharmacokinetics , Adult , Anti-HIV Agents/analysis , Chemoprevention/methods , Chromatography, Liquid , Disease Transmission, Infectious/prevention & control , Female , HIV Infections/prevention & control , Healthy Volunteers , Humans , Male , Middle Aged , Pre-Exposure Prophylaxis/methods , Sex Factors , Tandem Mass Spectrometry , Tenofovir/analysis , Young AdultABSTRACT
In the present study, the electrochemical behavior of the antiviral drug tenofovir (tnf) on a boron-doped diamond electrode (BDDE) has been studied using square-wave voltammetry (SWV). The experimental conditions such as the supporting electrolyte composition and the SWV parameters were optimized. Under the optimized conditions, a simple and sensitive SWV procedure for tnf determination was developed in the concentration range of 5.0â¯×â¯10-6 to 1.0â¯×â¯10-4â¯molâ¯L-1. The calculated limit of detection and limit of quantification were equal to 5.6â¯×â¯10-7 and 1.9â¯×â¯10-6â¯molâ¯L-1, respectively. The biological significance of the developed method was demonstrated by a quantitative analysis of the pharmaceutical formulations Viread and Tenofovir disoproxil Teva. Moreover, both voltammetric and spectroscopic techniques were used to study the interaction between tnf and DNA. Changes in the electrochemical and spectroscopic behavior of tnf in the presence of DNA were used to calculate the binding constants of the formed complexes. The estimated values of Gibbs free energy revealed that the formation of the drug-DNA complexes was a spontaneous and favorable process. Moreover, for free and bound tenofovir, kinetic parameters such as heterogeneous rate constant, electron transfer coefficient, and diffusion coefficient were determined.
Subject(s)
Antiviral Agents/metabolism , DNA/metabolism , Tenofovir/metabolism , Animals , Antiviral Agents/analysis , Boron/chemistry , Diamond/chemistry , Electrochemical Techniques/instrumentation , Electrodes , Limit of Detection , Salmon , Spectrum Analysis , Tenofovir/analysis , ThermodynamicsABSTRACT
A liquid chromatography with triple quadrupole mass spectrometry method was developed and validated for the determination of tenofovir and tenofovir alafenamide concentrations in human plasma and cerebrospinal fluid. Tenofovir and tenofovir alafenamide were extracted from matrix by solid phase extraction. The dried extraction eluents were dissolved in water for LC-MS/MS analysis. Separation was achieved with a Phenomenex Synergi 4⯵m Polar-RP 80A column (50â¯×â¯2â¯mm) with a gradient elution of 0.1% formic acid in water and acetonitrile. The total run time was 5â¯min. Detection of analytes was achieved using electrospray ionization (positive mode) and triple quadrupole selected reaction monitoring. Standard curve concentrations ranged from 0.5 to 500â¯ng/mL for the plasma assay and 0.1-50â¯ng/mL for the cerebrospinal fluid assay. The intra- and inter-day accuracy and precision were less than 12% in low, medium, and high quality control samples for both matrices. The validated methods were applied to the analysis of plasma and cerebrospinal fluid samples of a patient undergoing tenofovir therapy which involved the switch from Stribild® (elvitegravir 150â¯mg/cobicistat 150â¯mg/emtricitabine 200â¯mg/tenofovir disoproxil fumarate 300â¯mg) to Genvoya® (elvitegravir 150â¯mg/cobicistat 150â¯mg/emtricitabine 200â¯mg/tenofovir alafenamide 10â¯mg).
Subject(s)
Adenine/analogs & derivatives , Anti-HIV Agents/analysis , HIV Infections/drug therapy , Tenofovir/analysis , Adenine/analysis , Adenine/therapeutic use , Alanine , Chromatography, High Pressure Liquid , Cobicistat/analysis , Cobicistat/therapeutic use , Drug Combinations , Elvitegravir, Cobicistat, Emtricitabine, Tenofovir Disoproxil Fumarate Drug Combination/analysis , Elvitegravir, Cobicistat, Emtricitabine, Tenofovir Disoproxil Fumarate Drug Combination/therapeutic use , Emtricitabine/analysis , Emtricitabine/therapeutic use , HIV Infections/blood , HIV Infections/cerebrospinal fluid , Humans , Quinolones/analysis , Quinolones/therapeutic use , Reproducibility of Results , Solid Phase Extraction , Tandem Mass Spectrometry , Tenofovir/analogs & derivatives , Tenofovir/therapeutic useABSTRACT
OBJECTIVE: We describe the third case report of seroconversion with multidrug resistant (MDR)-HIV despite pre-exposure prophylaxis (PrEP) with emtricitabine (FTC) and tenofovir (TFV) disoproxil (TDF). DESIGN: Case report. METHODS: PrEP adherence was assessed via self-report, pharmacy records, and measuring TFV/FTC levels with liquid-chromatography/tandem-mass-spectrometry in plasma and hair. Segmental hair analysis was performed to assess PrEP adherence over prior months. Genotypic resistance was assessed. RESULTS: A 34 year-old white MSM started daily FTC/TDF in February 2016 after being provided 11 refills. In March 2017, he developed fevers, chills, myalgias and was assessed, but no HIV test was sent. In April 2017, an antigen/antibody HIV test was reactive (day 0). On day 2, HIV-1 RNA was 27â316 copies/ml, genotyping revealed M184V, K70T, K65R, and K103N mutations, plasma TFV and FTC concentrations were consistent with recent dosing. To evaluate adherence over preceding months, a hair sample was collected at day 27 and segmental analysis of TFV/FTC levels performed in one-centimeter segments from the scalp. Hair drug levels (0.0434-0.0520 ng TFV/mg hair) were commensurate with consistently high PrEP adherence over the prior 3 months. CONCLUSIONS: This study employs segmental analysis of PrEP drug levels in hair for the first time to assess adherence over preceding months in the setting of an HIV seroconversion on PrEP. High adherence over the period of likely acquisition makes MDR-HIV infection the most likely scenario in this case. Adequate adherence assessment when examining PrEP failures and ensuring best practices in PrEP prescribing and follow-up are important.