A Novel Approach to Integrate Human Biomonitoring Data with Model Predicted Dietary Exposures: A Crop Protection Chemical Case Study Using Lambda-Cyhalothrin.

The appropriate use of human biomonitoring data to model population chemical exposures is challenging, especially for rapidly metabolized chemicals, such as agricultural chemicals. The objective of this study is to demonstrate a novel approach integrating model predicted dietary exposures and biomonitoring data to potentially inform regulatory risk assessments. We use lambda-cyhalothrin as a case study, and for the same representative U.S. population in the National Health and Nutrition Examination Survey (NHANES), an integrated exposure and pharmacokinetic model predicted exposures are calibrated to measurements of the urinary metabolite 3-phenoxybenzoic acid (3PBA), using an approximate Bayesian computing (ABC) methodology. We demonstrate that the correlation between modeled urinary 3PBA and the NHANES 3PBA measurements more than doubled as ABC thresholding narrowed the acceptable tolerance range for predicted versus observed urinary measurements. The median predicted urinary concentrations were closer to the median measured value using ABC than using current regulatory Monte Carlo methods.

Voltammetric measurement of entacapone in the presence of other medicines against Parkinson's disease by a screen-printed electrode modified with sulfur-tin oxide nanoparticles.

A screen-printed electrode (SPE) is described modified with sulfur-tin oxide nanoparticles (S@SnO2NP) for the determination of entacapone (ENT) in the presence of other medicines against Parkinson's disease (PD). The S@SnO2NP was synthesized through the hydrothermal method and used in the modification of the SPE. The smart utilization of the S@SnO2NP and the SPE provided excellent properties such as high surface area and current density amplification by embedding an efficient sensing interface for highly selective electrochemical measurement. Under optimized experimental conditions, the anodic peak current related to the ENT oxidation onto the sensor surface at 0.46 V presented a linear response towards different ENT concentration sin the range 100 nM to 75 µM. The limit of detection (LOD) and electrochemical sensitivity were estimated to be 0.010 µM and 2.27 µA·µM-1·cm-2, respectively. The applicability of the sensor was evaluated during ENT determination in the presence of other conventional medicines againts, including levodopa (LD), carbidopa (CD), and pramipexole (PPX). The results of the analysis of human urine and pharmaceutical formulation as real samples using the developed sensor were in good agreement withre sults of high-performance liquid chromatography (HPLC) as a standard method. These findings demonstrated that the strategy based on the SPE is a cost-effective platform creating a promising candidate for practical determination of ENT in routine clinical testing.Graphical abstract.

Metabolic studies of selective androgen receptor modulators RAD140 and S-23 in horses.

This paper describes the studies of the in vitro biotransformation of two selective androgen receptor modulators (SARMs), namely, RAD140 and S-23, and the in vivo metabolism of RAD140 in horses using ultra-high performance liquid chromatography-high resolution mass spectrometry. in vitro metabolic studies of RAD140 and S-23 were performed using homogenised horse liver. The more prominent in vitro biotransformation pathways for RAD140 included hydrolysis, hydroxylation, glucuronidation and sulfation. Metabolic pathways for S-23 were similar to those for other arylpropionamide-based SARMs. The administration study of RAD140 was carried out using three retired thoroughbred geldings. RAD140 and the majority of the identified in vitro metabolites were detected in post-administration urine samples. For controlling the misuse of RAD140 in horses, RAD140 and its metabolite in sulfate form gave the longest detection time in hydrolysed urine and could be detected for up to 6 days post-administration. In plasma, RAD140 itself gave the longest detection time of up to 13 days. Apart from RAD140 glucuronide, the metabolites of RAD140 described herein have never been reported before.

Investigation of the metabolism of the selective androgen receptor modulator LGD-4033 in equine urine, plasma and hair following oral administration.

LGD-4033 is one of a number of selective androgen receptor modulators (SARMs) that are being developed by the pharmaceutical industry to provide the therapeutic benefits of anabolic androgenic steroids, without the less desirable side effects. Though not available therapeutically, SARMs are available for purchase online as supplement products. The potential for performance enhancing effects associated with these products makes them a significant concern with regards to doping control in sports. The purpose of this study was to investigate the metabolism of LGD-4033 in the horse following oral administration, in order to identify the most appropriate analytical targets for doping control laboratories. LGD-4033 was orally administered to two Thoroughbred horses and urine, plasma and hair samples were collected and analysed for parent drug and metabolites. LC-HRMS was used for metabolite identification in urine and plasma. Eight metabolites were detected in urine, five of which were excreted only as phase II conjugates, with the longest detection time being observed for di- and tri-hydroxylated metabolites. The parent compound could only be detected in urine in the conjugated fraction. Seven metabolites were detected in plasma along with the parent compound where mono-hydroxylated metabolites provided the longest duration of detection. Preliminary investigations with hair samples using LC-MS/MS analysis indicated the presence of trace amounts of the parent compound and one of the mono-hydroxylated metabolites. In vitro incubation of LGD-4033 with equine liver microsomes was also performed for comparison, yielding 11 phase I metabolites. All of the metabolites observed in vivo were also observed in vitro.

Identification of Patients with Preeclampsia by Measuring Fluorescence of an Amyloid-Binding Aryl Cyano Amide in Human Urine Samples.

Preeclampsia (PE) is a hypertensive disorder of pregnancy and one of the leading contributors to both maternal and perinatal morbidity and mortality. Reliable diagnostic parameters unique to the disorder that accurately define and diagnose PE are currently unavailable. Recent studies have revealed that PE is accompanied by the accumulation of amyloidogenic deposits in the placenta and the presence of congophilic amyloid-like protein aggregates in the urine. Here, we evaluate the capability of an amyloid-targeting aryl cyano amide (ARCAM-1) fluorophore to identify PE patients from analysis of urine samples. Our results reveal that this probe can distinguish patients with PE from gestationally healthy patients and patients suffering from non-PE hypertension, highlighting the potential for amyloid-targeting fluorophores to help identify PE patients during pregnancy.

Human in vivo metabolism study of LGD-4033.

Selective androgen receptor modulators (SARMs) are an emerging class of therapeutics targeted to cachexia, sarcopenia, and hypogonadism treatment. LGD-4033 is a SARM which has been included on the Prohibited List annually released by the World Anti-Doping Agency (WADA). The aim of the present work was the investigation of the metabolism of LGD-4033 in a human excretion study after administration of an LGD-4033 supplement, the determination of the metabolites' excretion profiles with special interest in the determination of its long-term metabolites, and the comparison of the excretion time of the phase I and phase II metabolites. The results were also compared to those derived from previous LGD-4033 studies concerning both in vitro and in vivo experiments. Supplement containing LGD-4033 was administered to one human male volunteer and urine samples were collected up to almost 21 days. Analysis of the hydrolyzed (with ß-glucuronidase) as well as of the non-hydrolyzed samples was performed using liquid chromatography-high resolution mass spectrometry (LC-HRMS) in negative ionization mode and revealed that, in both cases, the two isomers of the dihydroxylated metabolite (M5) were preferred target metabolites. The gluco-conjugated parent LGD-4033 and its gluco-conjugated metabolites M1 and M2 can be also considered as useful target analytes in non-hydrolyzed samples. The study also presents two trihydroxylated metabolites (M6) identified for the first time in human urine; one of them was recently reported in an LGD-4033 metabolism study in horse urine and plasma.

[Determination of deltamethrin and its toxicity biomarkers in rabbit urine by high performance liquid chromatography-tandem mass spectrometry].

A method was developed for the determination of biomarkers related to toxicity of deltamethrin in rabbit urine by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The target analytes in this method are as follows:deltamethrin and its two metabolites (1R-cis)-3-(2,2-dibromoethenyl)-2,2-dimethylcyclopropane carboxylic acid (dibromochrysanthemic acid) and 3-phenoxybenzoic acid (3-PBA), and five toxic biomarkers, viz. serotonin hydrochloride (5-HT), 5-hydroxyindole-3-acetic acid (5-HIAA), 3-nitropropionic acid (3-NPA), 8-hydroxy-2'-deoxyguanosine (8-OHdG), and 6-methoxyguanine. Urine samples were cleaned by matrix solid-phase dispersion extraction (MSPD) with diatomite; and protein was precipitated with trichloroacetic acid; and then the sample solutions were purified with hydrophilic-lipophilic balance (HLB) solid-phase extraction cartridges. The biomarkers were analyzed with electrospray ionization (ESI) in a positive and negative switching scan mode, in which the positive scan mode was used for deltamethrin, 5-HT, 5-HIAA, 8-OHdG, and 6-methoxyguanine, and the negative scan mode was used for (1R-cis)-3-(2,2-dibromoethenyl)-2,2-dimethylcyclopropane, 3-PBA, and 3-NPA. The target compounds were quantified with the external standard using matrix calibration curves. The linear regression curves of the eight target compounds were linear with correlation coefficients no less than 0.9914. The LOD and LOQ of 5-HIAA were 20 µg/L and 50 µg/L, respectively, and the LODs and LOQs of the other analytes were 0.2-5.0 µg/L and 0.5-10 µg/L, respectively. The average recoveries of the analytes spiked in rabbit urine ranged from 74.2% to 98.7% at three levels, with relative standard deviations (RSDs) no more than 12%. The method was simple, fast, accurate, sensitive, and suitable for the detection for the exposure evaluation of deltamethrin.

Determination of Urinary Metabolites of the Emerging UV Filter Octocrylene by Online-SPE-LC-MS/MS.

Octocrylene (OC) is an emerging UV filter, which is used in the majority of sunscreens as well as other personal care products (PCP) and consumer products. Its presence in various environmental matrices has been reported. However, information on the internal OC exposure in humans is not available, due to the lack of appropriate biomarkers of exposure and analytical methods. Here, we describe a rugged, precise, and accurate analytical method for the determination of three OC metabolites (ester hydrolysis and alkyl chain oxidation products) in human urine by stable isotope dilution analysis. Urine samples are incubated with ß-glucuronidase (E. coli K12) and then analyzed by liquid chromatography-electrospray ionization-triple quadrupole-tandem mass spectrometry with online turbulent flow chromatography for sample cleanup and analyte enrichment (online-SPE-LC-MS/MS). Syntheses of analytical standards, including deuterium-labeled internal standards, are also described. In a pilot study, we investigated the applicability of the metabolites as biomarkers of exposure in urine samples from the general population (n = 35). OC metabolites were detected in 91% of the samples, with the highest concentrations for three individuals having used sunscreen within 5 days prior to sample collection. We will apply the method in future human biomonitoring studies for OC exposure and risk assessment.

Screening-level Biomonitoring Equivalents for tiered interpretation of urinary 3-phenoxybenzoic acid (3-PBA) in a risk assessment context.

3-Phenoxybenzoic acid (3-PBA) is a common metabolite of several pyrethroid pesticides of differing potency and also occurs as a residue in foods resulting from environmental degradation of parent pyrethroid compounds. Thus, 3-PBA in urine is not a specific biomarker of exposure to a particular pyrethroid. However, an approach derived from the use of Biomonitoring Equivalents (BEs) can be used to estimate a conservative initial screening value for a tiered assessment of population data on 3-PBA in urine. A conservative generic urinary excretion fraction for 3-PBA was estimated from data for five pyrethroid compounds with human data. Estimated steady-state urinary 3-PBA concentrations associated with reference doses and acceptable daily intakes for each of the nine compounds ranged from 1.7 µg/L for cyhalothrin and deltamethrin to 520 µg/L for permethrin. The lower value can be used as a highly conservative Tier 1 screening value for assessment of population urinary 3-PBA data. A second tier screening value of 87 µg/L was derived based on weighting by relative exposure estimates for the different pyrethroid compounds, to be applied as part of the data evaluation process if biomonitoring data exceed the Tier 1 value. These BE values are most appropriately used to evaluate the central tendency of population biomarker concentration data in a risk assessment context. The provisional BEs were compared to available national biomonitoring data from the US and Canada.

Documenting the kinetic time course of lambda-cyhalothrin metabolites in orally exposed volunteers for the interpretation of biomonitoring data.

Lambda-cyhalothrin is a pyrethroid pesticide largely used in agriculture. Exposure assessment can be performed by measuring key urinary metabolites. For a proper use of biomonitoring data, it is however important to gain information on the toxicokinetics of these key biomarkers of exposure. A human volunteer study was performed to document the plasma and urinary time courses of major lambda-cyhalothrin metabolites. Seven volunteers ingested 0.025mgkg-1 body weight of lambda-cyhalothrin. Blood samples were withdrawn prior to dosing and at fixed time periods over the 72 h-period following ingestion and complete urine voids were collected pre-exposure and at pre-established intervals over 84h post-dosing. The cis-3-(2-chloro-3,3,3-trifluoroprop-1-en-1-yl)-2,2-dimethylcyclopropanecarboxylic acid (CFMP) and 3-phenoxybenzoic acid (3-PBA) metabolites were quantified in these samples. Plasma concentrations of CFMP and 3-PBA increased rapidly after ingestion, with average peak values at 3.1 and 4.0h post-dosing, respectively; subsequent elimination phase showed a rapid decay with a mean half-life (t½) of ≈5.3 and 6.4h for CFMP and 3-PBA, respectively. Urinary rate time courses displayed a profile similar to the plasma concentration-time curves with corresponding mean t½ of ≈4.2 and 5.9h. In the 84-h period post-treatment, on average 21% of lambda-cyhalothrin dose were excreted in urine as CFMP as compared to 30% as 3-PBA. Overall, CFMP and 3-PBA metabolites were confirmed to be major metabolites of lambda-cyhalothrin and exhibited similar kinetics with short half-lives; they thus both appear as useful biomarkers of exposure to lambda-cyhalothrin in humans.

Magnetic iron oxide nanoparticles grafted N-isopropylacrylamide/chitosan copolymer for the extraction and determination of letrozole in human biological samples.

Magnetic iron oxide nanoparticles are used for the extraction of a drug from an aqueous solution. In the current study, the magnetic iron oxide nanoparticles were synthesized via a facile coprecipitation approach, and then modified by (3-mercaptopropyl)trimethoxysilane followed by grafting thermosensitive polymer N-isopropylacrylamide and biopolymer chitosan. Structure, morphology, size, thermal resistance, specific surface area, and magnetic properties of the grafted nanosorbent were characterized by using Fourier transform infrared spectroscopy, field emission scanning electron microscopy, transmission electron microscopy, elemental analysis, thermogravimetric analysis, specific surface area analysis and vibrating sample magnetometry. The effective parameters on sorption/desorption of letrozole on grafted magnetic nanosorbent were evaluated. The best sorption of letrozole via the grafted nanosorbent occurred at 20°C at an optimum pH of 7. The extraction of trace letrozole in human biological fluids is investigated and revealed 89.1 and 97.8% recovery in plasma and urine, respectively.

Selective androgen receptor modulators: comparative excretion study of bicalutamide in bovine urine and faeces.

Besides their development for therapeutic purposes, non-steroidal selective androgen receptor modulators (non-steroidal SARMs) are also known to impact growth-associated pathways as ligands of androgenic receptors (AR). They present a potential for abuse in sports and food-producing animals as an interesting alternative to anabolic androgenic steroids (AAS). These compounds are easily available and could therefore be (mis)used in livestock production as growth promoters. To prevent such practices, dedicated analytical strategies should be developed for specific and sensitive detection of these compounds in biological matrices. The present study focused on Bicalutamide, a non-steroidal SARM used in human treatment of non-metastatic prostate cancer because of its anti-androgenic activity exhibiting no anti-anabolic effects. To select the most appropriate matrix to be used for control purposes, different animal matrices (urine and faeces) have been investigated and SARM metabolism studied to highlight relevant metabolites of such treatments and establish associated detection time windows. The aim of this work was thus to compare the urinary and faecal eliminations of bicalutamide in a calf, and investigate phase I and II metabolites. The results in both matrices showed that bicalutamide was very rapidly and mainly excreted under its free form. The concentration levels were observed as higher in faeces (ppm) than urine (ppb); although both matrices were assessed as suitable for residue control. The metabolites found were consistent with hydroxylation (phase I reaction) combined or not with glucuronidation and sulfation (phase II reactions). Copyright © 2016 John Wiley & Sons, Ltd.

A novel way for detection of antiparkinsonism drug entacapone via electrodeposition of silver nanoparticles/functionalized multi-walled carbon nanotubes as an amperometric sensor.

Silver (Ag) nanoparticles were electrochemically deposited on the film of a metformin functionalized multi-walled carbon nanotube modified glassy carbon electrode (Met-MWCNT/GCE), which fabricated an Ag@Met-MWCNT nanocomposite sensor (Ag@Met-MWCNT/GCE) to detect entacapone (ENT). The Ag@Met-MWCNT nanocomposite was characterized by field emission scanning electrochemical microscopy (FESEM), X-ray diffraction (XRD) analysis, FT-IR and electrochemical tests. The modified electrode showed a large electrocatalytic activity for reduction of ENT. This improved activity indicates that Met@MWCNT plays a crucial role in the dispersion and stabilization of Ag nanoparticles on GCE. Under the optimized conditions the linear range for the detection of the ENT was obtained to be 0.05 to 70.0µM with a low detection limit of 15.3nM. The proposed sensor can effectively analyse ENT concentration in pharmaceutical formulations and human urine samples, avoiding interference, and is a promising ENT sensor due to good sensitivity, stability and low cost.

In vivo dermal absorption of pyrethroid pesticides in the rat.

Exposure to pyrethroid pesticides is a potential cause for concern. The objective of this study was to examine the in vivo dermal absorption of bifenthrin, deltamethrin, and permethrin in the rat. Dorsal hair on adult male Long-Evans rats was removed. The next day, the skin was dosed with 1750 nmol (312.5 nmol/cm(2)) of radiolabeled (5 µCi) bifenthrin, deltamethrin, or permethrin in acetone. A nonoccluding plastic cover was glued over the dosing site. The animals were placed in metabolism cages to collect excreta. At 24 h postdosing, the skin was washed with soap and water, and rats in one group were euthanized and their tissues were collected. The skin was removed and tape stripped. The remaining animals were returned to the metabolism cages after the wash for 4 d. These rats were then euthanized and handled as already described. Excreta, wash, tape strips, tissues, and carcass were analyzed for pyrethroid-derived radioactivity. The wash and tape strips removed >50% of the dose and skin retained 9-24%. Cumulative radioactivity in excreta was 0.5-7% at 24 h and 3-26% at 120 h. Radioactivity in tissues was <0.3% of the dose, while carcass retained 2 to 5%. Assuming absorption equals cumulative recovery in skin (washed and tape stripped), excreta, tissues, and carcass, absorption was permethrin ~ bifenthrin > deltamethrin at 24 h and permethrin > deltamethrin > bifenthrin at 120 h. Using the parallelogram approach with published in vitro data, human dermal absorption of these pyrethroids was estimated to be <10% of the dose.

Pesticide exposures to migrant farmworkers in Eastern NC: detection of metabolites in farmworker urine associated with housing violations and camp characteristics.

BACKGROUND: The purpose of this paper is to present and evaluate descriptively bivariate associations between urinary metabolites of pesticides and herbicides and migrant camp conditions, violations, and personal worker behaviors at home for farmworkers who do not apply pesticides. METHODS: We studied 183 migrant farmworker camps in eastern North Carolina in 2010. Data and urine samples were collected from 371 men. Predictor measures included violations in six domains of housing regulations and nonviolation characteristics and personal behaviors that might impact urinary metabolites. RESULTS: Cockroaches and bathroom violations were predictive of increased exposure to pyrethroids and cyfluthrin/chlorpyrifos, respectively. Changing and storing clothing and shoes in sleeping rooms increased the number of detects for the diazinon metabolite. CONCLUSIONS: Farmworkers had exposures to multiple chemicals. No single housing domain was identified as critical to mitigating housing-related exposure; specific attention should be paid to changing and storing soiled clothing in sleeping rooms, and insect infestations.

Synthesis and characterization of poly[1-(N,N-bis-carboxymethyl)amino-3-allylglycerol-co-dimethylacrylamide] grafted to magnetic nano-particles for extraction and determination of letrozole in biological and pharmaceutical samples.

In this paper, a new method is reported for the surface grafting of poly[1-(N,N-bis-carboxymethyl)amino-3-allylglycerol-co-dimethylacrylamide] onto magnetic nano-particles modified by 3-mercaptopropyltrimethoxysilane. The grafted nano-sorbent was characterized by Fourier transform infrared spectroscopy, elemental analysis, thermogravimetric analysis, and scanning electron microscopy. Agglomerated nano-particles with multi-pores were used for extraction and determination of trace letrozole in human biological fluids and pharmaceutical samples. The profile of the letrozole uptake by the magnetic nano-sorbent reflected good accessibility of the active sites in the grafted polymer. Scatchard analysis revealed that the sorption capacity of the functionalized nano-sorbent was 6.27 µmol g(-1) at an optimum pH of 4. The equilibrium adsorption data of letrozole by grafted magnetic nano-sorbent were analyzed by Langmuir, Freundlich, Temkin and Redlich-Peterson models. Conformation of the experimental data in the Langmuir isotherm model indicated the homogeneous binding site of functional polymer-grafted magnetic nano-sorbent surface. Nearly 89% of letrozole was released in simulated gastric fluid, pH 1.2, in 2h and 79% in simulated intestinal fluid, pH 7.4, in 30 h. These results show the utility of the letrozole loaded- polymer grafted magnetite nano-particles for enteric drug delivery.

Control of konzo by detoxification of cassava flour in three villages in the Democratic Republic of Congo.

Three villages in Boko Health Zone, Bandundu Province, Democratic Republic of Congo (DRC), had 61 konzo cases and konzo prevalences of 2.5%, 4.1% and 7.5% respectively. Konzo cases occurred every year for 10 years and every month, peaking in July. The high mean cyanide content of cassava flour of 50 ppm was due to short soaking of cassava roots for 1-2 days instead of 3-4 days. Konzo cases were examined and village women taught the wetting method that removes cyanogens from flour. The villages were visited every month for 1 year following previous methodology. No new konzo cases occurred during the intervention, mean flour cyanide levels reduced from 50 to 14 ppm and mean urinary thiocyanate levels of school children reduced from 930 to 150 µmole/L. The percentage of children with urinary thiocyanate levels of >350 µmole/L was reduced from a maximum of 80 in Ikialala before the intervention to 0 in Ikusama, Ikialala and 3 in Imboso Mwanga 1 year later. This is the second time that konzo has been controlled and success depends on regular use of the wetting method by village women. The methodology is now being used in other villages in DRC with financial support of AusAID.

Monitoring phosphodiesterase-4 inhibitors using liquid chromatography/(tandem) mass spectrometry in sports drug testing.

RATIONALE: The recent discovery of resveratrol's capability to inhibit cAMP-specific phosphodiesterases (PDEs) and, as a consequence, to enhance particularly the activity of Sirt1 in animal models has reinforced the interest of preventive doping research organizations, especially in PDE4 inhibitors. Among these, the archetypical PDE4-inhibitor rolipram significantly increased the number of mitochondria in laboratory rodents, which further demonstrated a performance increase in a treadmill-test (time-to-exhaustion) of approximately 40%. Besides rolipram, a variety of new PDE4-inhibiting substances including cilomilast, roflumilast, and numerous additional new drug entities were described, with roflumilast being the first-in-class having received clinical approval for the treatment of chronic obstructive pulmonary disease (COPD). Due to the availability of these substances, and the fact that a misuse of such compounds in sport cannot be excluded, it deems relevant to probe for the prevalence of these compounds in sports drug testing programs. METHODS: Known urinary phase-I metabolites of rolipram, roflumilast, and cilomilast were generated by in vitro incubations employing human liver microsomal preparations. The metabolites obtained were studied by liquid chromatography with high-resolution/high-accuracy tandem mass spectrometry (LC/MS/MS) and the reference product ion mass spectra of established and most relevant metabolites were utilized to provide the information necessary for comprehensive doping controls. The analytical procedure was based on conventional routine doping control assays employing enzymatic hydrolysis followed by liquid-liquid extraction and subsequent LC/MS/MS measurement. RESULTS: Structures of diagnostic product ions and dissociation pathways of target analytes were elucidated, providing the information required for implementation into an existing test method for routine sports drug testing. The established method allowed for detection limits for the intact drugs of 1-5 ng/mL, and further assay characteristics (intraday precision 1.5-13.7%, interday precision 7.3-18.6%, recovery 20-100%, ion suppression/enhancement, and specificity) were determined. In addition, proof-of-concept analyses concerning roflumilast were conducted with a urine sample obtained from a COPD patient under roflumilast treatment.

Rapid determination of letrozole, citalopram and their metabolites by high performance liquid chromatography-fluorescence detection in urine: Method validation and application to real samples.

This work reports the validation of a high precision and accuracy method for the simultaneous determination of letrozole, citalopram and their metabolites in urine by high performance liquid chromatography with fluorescence detection. Dilution (urine:mobile phase, 1:2, v/v) was the only sample preparation step. The separation was carried out in a Kromasil C(18) (150mm×4.6mm) column, and the mobile phase was phosphate buffer 80mM (pH 3.0) and acetonitrile (65:35, v/v) at a flow rate of 1.0mL/min. The analytes were detected at 295nm after excitation at 230nm. Linearity was observed in the range of 1.0-1000ng/mL for letrozole and its metabolite and 2.5-1000ng/mL for citalopram and their metabolites, with limits of detection and quantification between 0.09-1.0 and 0.27-1.65ng/mL, respectively. The precisions were satisfactory with RSDs between 0.17 and 5.71%. The accuracy was studied by spiking three urines from healthy female volunteers, and the recoveries were from 85 to 103%. The method was applied to urine samples from women under treatment for breast cancer and depression diseases.