A sonochemical based green synthesis method playa powerful role in nanomaterials and composite development. In this work, we developed a perovskite type of strontium titanate via sonochemical process. SrTiO3 particles were incorporated with nitrogen doped graphene oxide through simple ultrasonic irradiation method. The SrTiO3/NGO was characterized by various analytical methods. The nanocomposite of SrTiO3/NGO was modified with laser-induced graphene electrode (LIGE). The SrTiO3/NGO/LIGE was applied for electrochemical sensor towards chemotherapeutic drug detection (nilutamide). Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques have been used to examine the electrochemical performance of nilutamide (anti-cancer drug). DPV was found to be more sensitive and found to exhibit a sensitivity 8.627 µA µM-1 cm-2 for SrTiO3/NGO/LIGE with a wide linear range (0.02-892 µM) and low Limit of detection (LOD: 1.16 µM). SrTiO3/NGO/LIGE has been examined for the detection of nilutamide in blood serum and urine samples and obtained a good recovery in the range of 97.2-99.72 %. The enhanced stability and selectivity and practical application results indicates the suitability of SrTiO3/NGO/LIGE towards the detection of nilutamide drug in pharmaceutical industries.

The unabated usage of priority anthropogenic stressors is a serious concern in the global environmental context. Pharmaceutical drugs such as furazolidone (FL) and nilutamide (NL) have far-reaching repercussions due to the presence of the reactive nitroaromatic moiety. Despite the widespread awareness regarding the dangers posed by nitroaromatic drugs, the promises to alleviate the environmental consequences of drug pollution are often unmet. Accordingly, implementing practices to monitor their presence in various media is a highly desirable, but challenging undertaking. With the advent of deep eutectic solvent-assisted synthesis, it has become possible to fabricate LDH-based sensor materials with minimal energy inputs in a sustainable and scalable manner. In this work, we have framed a series of CoFe-LDH electrocatalysts utilizing deep eutectic solvent-assisted hydrothermal strategies for the simultaneous detection of FL and NL. The CoFe-LDHs intercalated with three distinct anions, namely, (i) Cl-, (ii) SO42-, and (iii) CO32- are compared so as to establish a relationship between anion intercalation and electrochemical activity. Amongst the prepared electrodes, the CF-LDH-ii/SPCE displays highly appreciable selectivity, linear response range (0.09-237.9 μM), low detetion limits (FL = 1.2 nM and NL = 3.8 nM), high sensitivity (FL = 29.71 μA μM⁻1 cm⁻2 and NL = 19.29 μA μM⁻1 cm⁻2), good reproducibility and repeatability towards FL and NL in water and urine samples. Thus, with tailored gallery anions, the proposed electrocatalyst establishes enhanced electrocatalytic performance for the real-time analysis of pharmaceutical contaminants.

The microbial model of mammalian drug metabolism, Cunninghamella elegans, has three cytochrome P450 reductase genes in its genome: g1631 (CPR_A), g4301 (CPR_B), and g7609 (CPR_C). The nitroreductase activity of the encoded enzymes was investigated via expression of the genes in the yeast Pichia pastoris X33. Whole cell assays with the recombinant yeast demonstrated that the reductases converted the anticancer drug flutamide to the nitroreduced metabolite that was also produced from the same substrate when incubated with human NADPH: cytochrome P450 reductase. The nitroreductase activity extended to other substrates such as the related drug nilutamide and the environmental contaminants 1-nitronaphthalene and 1,3-dinitronaphthalene. Comparative experiments with cell lysates of recombinant yeast were conducted under aerobic and reduced oxygen conditions and demonstrated that the reductases are oxygen sensitive.

Pharmaceuticals and personal care products, antibiotics, estrogens, and antiandrogens are found widely in aquatic environments. Monitoring studies by sampling surface water and effluents of wastewater treatment plants (WWTPs) have been conducted recently to monitor antiandrogens, which, along with estrogens, cause endocrine disruption. However, few studies have investigated antiandrogenic activity (AA) combined with a chemical analyses of emerging antiandrogens. Therefore, we analyzed the presence and persistence of 12 types of antiandrogens, atrazine, and carbamazepine using grab sampling and polar organic chemical integrative sampler (POCIS) along a river affected by WWTP discharges. Water and sediment samples were collected from the WWTP effluent (WW), as well as upstream (US) and downstream (DS) of the WWTP. We detected only tebuconazole, triclosan, propiconazole, and fluconazole during the two sampling campaigns in 2016 and 2017. Grab sampling of the site WW detected tebuconazole (7-77 ng/L), propiconazole (5-47 ng/L), and fluconazole (6-45 ng/L). However, the concentrations in the river water were below the detection limits. Nevertheless, fluconazole and triclosan were detected by POCIS in the site WW (45.7 and 26.8 ng/L, respectively) and all river samples ranges of 0.3-9.3 and 2.4-3.7, respectively. This detection was attributed to the limit of quantification of POCIS being lower than that of grab sampling. Nilutamide and triclosan were detected in the river sediment, suggesting that their concentrations in the water column were at least partly attenuated through sediment sorption. We also observed AA by analyzing POCIS extracts with the yeast androgen screen assay. The highest AA was found in the site WW and it was still observable several kilometers downstream of the point of discharge despite decreasing. Therefore, the WWTP effluent was most likely contributor to the persistent AA in the river.

Based on pioneering work by Huggins, Hodges and others, hormonal therapies have been established as an effective approach for advanced prostate cancer (PC) for the past eight decades. However, it quickly became evident that androgen deprivation therapy (ADT) via surgical or medical castration accomplishes inadequate inhibition of the androgen receptor (AR) axis, with clinical resistance inevitably emerging due to adrenal and intratumoral sources of androgens and other mechanisms. Early efforts to augment ADT by adding adrenal-targeting agents (aminoglutethimide, ketoconazole) or AR antagonists (flutamide, bicalutamide, nilutamide, cyproterone) failed to achieve overall survival (OS) benefits, although they did exhibit some evidence of limited clinical activity. More recently, four new androgen receptor signaling inhibitors (ARSIs) successfully entered clinical practice. Specifically, the CYP17 inhibitor abiraterone acetate and the second generation AR antagonists (enzalutamide, apalutamide and darolutamide) achieved OS benefits for PC patients, confirmed the importance of reactivated AR signaling in castration-resistant PC and validated important concepts that had been proposed in the field several decades ago but had remained so far unproven, including adrenal-targeted therapy and combined androgen blockade. The past decade has seen steady advances toward more comprehensive AR axis targeting. Now the question is raised whether we have accomplished the maximum AR axis inhibition possible or there is still room for improvement. This review, marking the 80-year anniversary of ADT and 10-year anniversary of successful ARSIs, examines their current clinical use and discusses future directions, in particular combination regimens, to maximize their efficacy, delay emergence of resistance and improve patient outcomes.

The therapeutic action of nitroaromatic antiandrogens nilutamide and flutamide may be complicated by their cytotoxicity, whose mechanisms are still incomprehensively understood. In particular this concerns the enzymatic redox cycling of flutamide and its metabolites, and its impact on their cytotoxicity. In this work, we examined the single-electron reduction of nilutamide, flutamide, its metabolites 2-hydroxyflutamide and 4-nitro-3-trifluorormethyl-phenylamine, and a topical antiandrogen (3-amino-2-hydroxy-2-methyl-N-(4-nitro-3-trifluoromethyl)-phenyl) propanamide by NADPH:cytochrome P-450 reductase and adrenodoxin reductase/adrenodoxin. The obtained steady-state bimolecular rate constants of oxidant reduction (kcat/Km) enabled to establish single-electron reduction midpoint potentials (E17) of compounds, -0.377 - -0.413 V, which were in line with enthalpies of formation of their free radicals, obtained by quantum mechanical calculations. Using murine hepatoma MH22a cells, the obtained cytotoxicity vs. E17 correlation based on the data of model nitroaromatic compounds shows that redox cycling and oxidative stress could be the main factor of cytotoxicity of nitroaromatic antiandrogens. Other minor cytotoxicity factors could be their redox metabolism involving NAD(P)H:quinone oxidoreductase (NQO1) and cytochromes P-450.

Attempts to obtain new cocrystals of nonsteroidal antiandrogenic drug nilutamide produced alternative polymorphic forms of the compound (Form II and Form III) and their crystal structures were elucidated by single-crystal X-ray diffraction. Apart from the cocrystallization technique, lyophilization was found to be an effective strategy for achieving polymorph control of nilutamide, which was difficult to obtain by other methods. The physicochemical properties and relative stability of the commercial Form I and newly obtained Form II were comprehensively investigated by a variety of analytical methods (thermal analysis, solution calorimetry, solubility, and sublimation), whereas for Form III, only a handful of experimental parameters were obtained due to the elusive nature of the polymorph. Form I and Form II were found to be monotropically related, with Form I being confirmed as the thermodynamically most stable solid phase. In addition, the performance of different DFT-D and semi-empirical schemes for lattice energy calculation and polymorph energy ranking was compared and analysed. Lattice energy calculations using periodic DFT at B3LYP-D3/6-31(F+)G(d,p) and PBEh-3c/def2-mSVP levels of theory were found to provide the most accurate lattice energy values for Form I against experimental data, while PIXEL and PBEh-3c/def2-mSVP were the only methods that predicted the correct order of stability of Forms I and II.

Androgen receptor (AR) is an important target for the treatment of prostate cancer, and mutations in the AR have an important impact on the resistance of existing drugs. In this work, we performed molecular dynamics simulations of the existing marketed antiandrogens flutamide, nilutamide, bicalutamide, enzalutamide, apalutamide, darolutamide, and its main metabolite ORM15341 in complex with the wild-type and F876L mutant AR. We calculated the residue-specific binding free energy contribution of the wild-type and mutant ARs with the AS-IE method and analyzed the hotspot residues and the binding free energy contributions of specific residues before and after the mutation. In addition, we analyzed the total binding obtained by adding residue binding energy contributions and compared the results with experimental values. The obtained residue-specific binding information should be very helpful in understanding the mechanism of drug resistance with respect to specific mutations and in the design of new generation drugs against possible new mutations.

Copper vanadate nanoparticles (Cu2V2O7) are synthesized by using a simple hydrothermal method and later anchored with sulfur-doped reduced graphene oxide (S-rGO) by using ultrasonication to form a hybrid nanocomposite. The synthesized composite underwent characterizations like X-ray diffraction analysis (XRD), Raman spectroscopy, Fourier transform infrared spectroscopy (FT-IR), Dynamic ray scattering-Ultra violet-visible spectroscopy (DRS-UV-visible) and X-ray photoelectron spectroscopically revealed the triclinic pattern of the P 1̅ space group of α-Cu2V2O7 and the reduced oxygen deficiency state of metal centers (Cu+ or V4+) resulting with oxides of mixed-valence oxidative states and forming of Cu-O bond. Morphological analysis was carried out by using transmission electron microscopy (TEM) and Field emission scanning electron microscopy (FE-SEM) with elemental mapping and EDX analysis. Furthermore, a novel electrochemical sensor is prepared by using the hybrid sCu2V2O7/S-rGO nanocomposite on to a disposable screen-printed carbon paste electrode (SPCE) for electrochemical sensing of antiandrogen drug nilutamide (NLT). This report reveals excellent activity in determining NLT with a low detection limit of 0.00459 nM for the linear range of 0.001-15 μM with high sensitivity of 26.2605 µA µM-1 cm-2. Further, electrode performance showed appreciable performance in real-time monitoring of biological samples like human blood serum, urine samples.

Amorphous solid dispersions (ASDs) of crystallizable drugs and polymer excipients are attractive for enhancing the solubility and bioavailability of hydrophobic drug molecules. In this study, the solution behavior of poly(N-isopropylacrylamide-co-N,N-dimethylacrylamide) (PND) and poly(vinylpyrrolidone-co-vinylacetate) (PVPVA), as polymer excipients, and nilutamide (NLT), phenytoin (PHY), and itraconazole (ITN) as model drugs, were monitored by an in vitro dissolution assay, small-angle X-ray scattering (SAXS), dynamic light scattering (DLS), cryogenic transmission electron microscopy (cryo-TEM), and polarized optical microscopy (POM). High degrees of drug supersaturation were coincident with the formation of amorphous nanoparticles in each system. The difference in particle size and kinetic stability between PND and PVPVA systems suggest a difference in how the polymers interact with the drug-rich phase. A series of scenarios are proposed based on whether the polymer interacts more strongly with the drug-rich nanoparticles or with water. Understanding the contribution of drug-rich nanoparticles to achievable supersaturation and the effect of polymer excipients on these particles will inform the design of future solid dispersion systems through a better understanding of the polymer/drug solution relationship.

Activation of epithelial-AR signaling is identified as the major cause of hyperproliferation of the cells during benign and malignant prostate conditions. However, the contribution of stromal-AR is also precarious due to its secretory actions that contribute to the progression of benign and malignant tumors. The present study was aimed to understand the influence of stromal-AR mediated actions on epithelial cells during BPH condition. The secretome (conditioned media-CM) was collected from AR agonist (testosterone-propionate-TP) and antagonist (Nilutamide-Nil) treated BPH patient-derived stromal cells and exposed to BPH epithelial cells. Epithelial cells exhibited increased cell proliferation with the treatment of CM derived from TP-treated stromal cells (TP-CM) but did not support the clonogenic growth of BPH epithelial cells. However, CM derived from Nil-treated stromal cells (Nil-CM) depicted delayed and aggressive BPH epithelial cell proliferation with increased clonogenicity of BPH epithelial cells. Further, decreased AR levels with increased cMyc transcripts and pAkt levels also validated the clonogenic transformation under the paracrine influence of inhibition of stromal-AR. Moreover, the CM of stromal-AR activation imparted positive regulation of basal/progenitor pool through LGR4, β-Catenin, and ΔNP63α expression. Hence, the present study highlighted the restricted disease progression and retains the basal/progenitor state of BPH epithelial cells through the activation of stromal-AR. On the contrary, AR-independent aggressive BPH epithelial cell growth due to paracrine action of loss stromal-AR directs us to reform AR pertaining treatment regimes for better clinical outcomes.

This study reports the sonochemical synthesis of samarium tungstate nanoparticles (SWNPs) for applications in electrochemical sensors. The synthesis process is based on a precipitation reaction, which was investigated by ultrasound and compared with the effect of stirring. A bath sonicator operated at a frequency and power of 37/100 kHz and ~60 W, respectively, was employed to prepare the material. The shock waves efficiently irradiated the reaction conditions as much as possible, resulting in the good crystallinity of the monoclinic phase of the SWNPs, which was confirmed by XRD analysis. The surface morphology and structural composition was further evaluated by HRTEM, EDS and XPS. The good crystallinity and uniform distribution of elements in the nanoparticles were confirmed. The performance of the SWNPs to electrochemically sense nilutamide (NLT) was studied, which revealed a good electrochemical signal. As a result, the SWNPs were applied to an electrode material for the detection of NLT. This study revealed the excellent activity of the SWNPs for NLT detection, resulting in a low detection limit (0.0026 µM) and good linear range (0.05-318 µM). Furthermore, the results show appreciable analytical performances, which could be applied to electrochemical anti-androgen drug nilutamide sensors.

Echinacoside (ECH) is a natural compound with an endothelium‑dependent vasodilatory effect. Nitric oxide (NO) is an important vasorelaxant released from endothelial cells. In order to examine the molecular mechanism of ECH‑induced NO production in endothelial cells, the present study investigated the involvement of androgen receptor (AR) and the phosphatidylinositol 3‑kinase (PI3K)/protein kinase B (Akt) pathway in the phosphorylation of endothelial nitric oxide synthase (eNOS) in human umbilical vein endothelial cells (HUVECs). Using the fluorescent probe DAF‑FM, the production of NO was found to be significantly increased, and eNOS was phosphorylated at Ser1177 in a concentration‑​dependent manner under 0.01‑10 µM ECH treatment in HUVECs. In addition, NO production and eNOS phosphorylation induced by ECH were diminished when pretreated with the AR antagonist nilutamide, or when transfected with AR small interfering RNAs. Furthermore, the ECH‑induced phosphorylation of the Akt at Ser473 was abrogated by 5 µM wortmannin (a PI3K inhibitor). These data indicated that ECH stimulated NO production via the AR‑dependent activation of eNOS in HUVECs, and that the PI3K/Akt pathway may be involved in eNOS phosphorylation induced by ECH.

Recent reports have concentrated on some androgens/antiandrogens and confirmed that certain chemicals have demonstrated androgenic/antiandrogenic activities in vitro. However, it is still unknown whether more chemicals in the human environment possess endocrine toxicity. 58A novel AR-mediated reporter gene assay based on LLC-MK2 cells was established by transiently co-transfecting with pARE-sv40-Luc, hAR-pcDNA3.1 and pRL-tk. pARE-sv40-Luc was constructed using a pGL3-promoter plasmid with three repeated androgen responsive elements. hAR-pcDNA3.1 was constructed using pcDNA3.1 with a hAR sequence. After transfection for 12 h, the culture medium was exposed to various concentrations of dihydrotestosterone (DHT) and other test chemicals (phthalic acid esters and dexamethasone) in order to measure the androgenic/antiandrogenic activity. The assay possessed a concentration-dependent response to DHT from 10-12 M to 10-6 M. Nilutamide concentrations of over 10-7 M completely blocked the luciferase expression induced by 10-9 M DHT. Other data showed that DBP, DEHP and MEHP possessed weak androgenic activity for certain concentration ranges, while DMP, DINP and DIBP did not show any androgenic activity. Moreover, five PAEs (DBP, DEHP, DINP, DIBP and MEHP) showed corresponding antiandrogenic activities for certain concentrations with an approximate tendency (MEHP > DBP > DEHP > DIBP > DINP). The assay is high-throughput, specific, and sensitive for the detection of androgenic/antiandrogenic chemicals. In addition, PAEs (especially transitional PAEs) exhibited corresponding androgenic/antiandrogenic activities for certain concentration ranges.

Cerium vanadate resembling the shape of a hedgehog were interconnected with carbon nanofibers to give a heterostructure (referred to as CeV/CNF) that exhibits efficient catalytic activity for the electrochemical detection of the drug nilutamide (NLT). The heterostructure material and its modification were characterized by XRD, Raman spectra, XPS, FESEM, TEM, SAED, and EDX. A glassy carbon electrode was modified with the CeV/CNF nanocomposite. Best operated at -0.52 V (vs. Ag/AgCl), it exhibits a very low detection limit (2.0 nM), wide linear range (0.01-540 μM), high sensitivity (1.36 μA μM-1 cm-2) and rapid response towards NLT. It was applied to the determination of NLT in spiked human urine. Graphical abstractSchematic presentation of cerium vanadate interconnected with carbon nanofiber heterostructure for electrochemical determination of prostate cancer drug nilutamide in biological samples.

An electrochemical sensor is described for the determination of nilutamide (NLM) in biological fluids. A flexible mat of nitrogen-doped carbon nanofibers (NCNFs) was prepared by electrospinning. This was followed by carbonization and the growth of one-dimensional vanadium tetrasulfide (VS4) nanorods in the circumference of the NCNFs by a solvothermal route. The material was used to modify a glassy carbon electrode (GCE). Cyclic voltammetry and amperometry revealed an excellent electrocatalytic activity of the VS4/NCNF mat towards NLM at a working potential of - 0.57 V (vs Ag/AgCl). The modified GCE has a wide linear range (0.001-760 μM), a low limit of detection (90 pM), good stability, and fast response. It was used to detect NLM in spiked serum and urine samples. Graphical abstract Schematic presentation of a glassy carbon electrode (GCE) composed of free-standing nitrogen-doped carbon nanofibers decorated with vanadium tetrasulfide (Patronite) nanorods (VS4/NCNF). It was fabricated by electrospinning followed by stabilization and carbonization. The GCE responds to nilutamide (NLM) with excellent selectivity and nanomolar sensitivity.

Despite the chronic effects of nandrolone decanoate (ND), the acute effects of ND on passive avoidance learning (PAL) and memory and its mechanism have not been investigated. This research examines the acute effect of ND on PAL, CA1 synaptic plasticity, testosterone and corticosterone serum levels, and the role of androgenic receptors (ARs). Adolescent male rats were treated with ND, 30 min before training and retention and after training test. AR antagonist was applied 15 min before ND. Hippocampal slices were perfused by ND. ND administration had an inverted U-shape effect on acquisition of PAL and on testosterone and corticosterone serum levels. The consolidation was only affected by high dose of ND. ND significantly decreased the retention of PAL across all doses. The magnitude of field excitatory postsynaptic potential long term potentiation was lower than that of control slices. In addition, an attenuation of field excitatory postsynaptic potential population spike coupling was also observed. Nilutamide could nullify the ND impairment effect. We concluded although a single dose of ND could affect all stages of PAL, its effects were more potent on retrieval, possibly arising from the acute effect of ND on the alterations of CA1 synaptic plasticity. In addition, ND may induce its effects directly through ARs and indirectly through plasma testosterone and corticosterone.

Human aldehyde oxidase 1 (AOX1) catalyzes the oxidation of various drugs and endogenous compounds. Recently, we found that AOX1 catalyzed the reduction of drugs such as nitrazepam and dantrolene. In this study, we aimed to clarify the substrate selectivity of human AOX1 for the reduction of nitroaromatic drugs to obtain helpful information for drug development. We investigated whether 11 nitroaromatic drugs were reduced by AOX1 using recombinant AOX1 and human liver cytosol (HLC) in the presence of N1-methylnicotinamide, an electron donor to AOX1. We found that clonazepam, flunitrazepam, flutamide, nilutamide, nimesulide, and nimetazepam were substantially reduced by recombinant AOX1 and HLC, whereas azelnidipine, nifedipine, and nimodipine were slightly reduced and metronidazole and tolcapone were not reduced. Via structural analysis, we observed that nitroaromatic drugs reduced by AOX1 possessed a relatively electron-deficient nitro group. Since the addition of NADPH to human liver microsomes (HLM) did not increase the reductase activities of the drugs that were reduced by recombinant AOX1, it was determined that NADPH-dependent enzymes in microsomes, such as cytochrome P450, were not involved in this process. Inhibition studies using known AOX1 inhibitors supported the role of AOX1 in the reduction of drugs in HLC. In conclusion, this provides new information related to the substrate selectivity of human AOX1 for the reduction of nitroaromatic drugs.

Larval zebrafish (Danio rerio) are a tool for assessing endocrine disruption during early development. Here, we investigated the extent to which a simple light/dark behavioral test at five days post fertilization could compliment current methods within the field. We exposed fertilized embryos to hormones (17β-estradiol, testosterone, dihydrotestosterone, 11-ketotestosterone, thyroxine, triiodothyronine, progesterone, and hydrocortisone) and other relevant compounds (17α ethinylestradiol, bisphenol A, bisphenol S, nonylphenol, flutamide, nilutamide, linuron, drospirenone, potassium perchlorate, mifepristone, and fadrozole) to screen for behavioral effects between 96 and 118h post fertilization (hpf). With the exception of progesterone, all the hormones tested resulted in altered behaviors. However, some inconsistencies were observed regarding the age of the larvae at testing. For example, the xenoestrogens 17α- ethinylestradiol and nonylphenol had behavioral effects at 96hpf, but not at 118hpf. Furthermore, although thyroxine exposure had pronounced effects on behavior, the thyroid disruptor potassium perchlorate did not. Finally, we were unable to demonstrate a role of nuclear receptors following testosterone and 17α- ethinylestradiol exposure, as neither the androgen receptor antagonist flutamide nor the general estrogen receptor inhibitor fulvestrant (ICI) could rescue the observed behavioral effects, respectively. Similarly, molecular markers for androgen and estrogen disruption were upregulated at concentrations below which behavioral effects were observed. These results demonstrate hormones and endocrine disruptors can alter the behavior of larval zebrafish, but the mechanistic pathways remain unclear.

Nilutamide is an oral nonsteroidal antiandrogen similar in structure to flutamide that is used widely in the therapy of prostate cancer and has been linked to rare instances of liver injury.

Prostate cancer is the most common cancer and one of the leading causes of cancer deaths in men. One of the commonly used approaches to treat metastatic prostate cancer was via first-generation nonsteroidal anti-androgens (NSAAs), namely flutamide, nilutamide, bicalutamide and topilutamide. Most prostate cancer patients who are initially responsive develop the most aggressive form of disease called castration-resistant prostate cancer. Second-generation NSAA receptor antagonists (enzalutamide, apalutamide and darolutamide) are emerging as additional new options to treat castration-resistant prostate cancer. The objective of this work was to review the literature on the bioanalytical methods for the quantification of first- and second-generation NSAA inhibitors in clinical (human plasma) and preclinical (mouse plasma, rat plasma, urine and tissue homogenates etc.) studies along with relevant case studies for some chosen drugs. Based on the review, it was concluded that the published methodologies using either HPLC or LC-MS/MS are well suited for the quantification of NSAA inhibitors in various biological fluids to delineate pharmacokinetic data.