Identification of potential chemical biomarkers of hexaconazole using in vitro metabolite profiling in rat and human liver microsomes and in vivo confirmation through urinary excretion study in rats.

Hexaconazole (HEX) is an azole fungicide widely used in agricultural practices across various countries and numerous studies have reported the toxic effects of HEX, such as endocrine disruption, immunotoxicity, neurotoxicity and carcinogenicity. Despite its widespread agricultural use and toxic effects, the metabolism of HEX is not completely understood, and information on urinary elimination of HEX or its metabolites is limited. Therefore, in the present study, we aimed to identify HEX metabolites in rat and human liver microsomes followed by their in vivo confirmation using a urinary excretion study in rats to identify potential candidate for exposure biomarkers for human biomonitoring studies. From the in vitro assay, a total of 12 metabolites were observed, where the single oxidation metabolites (M5 and M6) were the most abundant metabolites in both rat and human liver microsomes. The triple oxidation followed by dehydration metabolite, M8 (which could also be hexaconazole acid or hydroxy keto-hexaconazole), and the double oxidation metabolite (M9) were the major metabolites found in rat urine and were detectable in rat urine longer than the parent. These metabolites increased with decreasing concentrations of HEX in the rat urine samples. Therefore, metabolites M8, M9 and M5 could be pursued further as potential biomarkers for assessing and monitoring human exposure to HEX.

Predicting the in vivo developmental toxicity of fenarimol from in vitro toxicity data using PBTK modelling-facilitated reverse dosimetry approach.

In vitro methods are widely used in modern toxicological testing; however, the data cannot be directly employed for risk assessment. In vivo toxicity of chemicals can be predicted from in vitro data using physiologically based toxicokinetic (PBTK) modelling-facilitated reverse dosimetry (PBTK-RD). In this study, a minimal-PBTK model was constructed to predict the in-vivo kinetic profile of fenarimol (FNL) in rats and humans. The model was verified by comparing the observed and predicted pharmacokinetics of FNL for rats (calibrator) and further applied to humans. Using the PBTK-RD approach, the reported in vitro developmental toxicity data for FNL was translated to in vivo dose-response data to predict the assay equivalent oral dose in rats and humans. The predicted assay equivalent rat oral dose (36.46 mg/kg) was comparable to the literature reported in vivo BMD10 value (22.8 mg/kg). The model was also employed to derive the chemical-specific adjustment factor (CSAF) for interspecies toxicokinetics variability of FNL. Further, Monte Carlo simulations were performed to predict the population variability in the plasma concentration of FNL and to derive CSAF for intersubject human kinetic differences. The comparison of CSAF values for interspecies and intersubject toxicokinetic variability with their respective default values revealed that the applied uncertainty factors were adequately protective.

A UPLC-MS/MS method for quantification of ß-N-methylamino-L-alanine (BMAA) in Cycas sphaerica roxb. and its use in validating efficacy of a traditional BMAA removal method.

The presence of neurotoxin ß-N-Methylamino-L-alanine (BMAA) in the seeds of Cycas sphaerica is reported for first time. We developed a UPLC-MS/MS method for BMAA quantification by derivatizing with dansyl chloride. The method successfully differentiated L-BMAA from its structural isomer 2,4-diaminobutyric acid (DAB). The extracting mixture 0.1M TCA: ACN 4:1 v/v had a recovery level of >95%. The method is a high throughput sensitive chromatographic technique with 16.42 ng g-1 Limit of Quantification. BMAA was present in the endosperm of C. sphaerica, and was not detected in the leaves and pith. Washing of seeds in running cold water for 48 h reduced BMAA content by 86%. The local communities also treat the seeds under running cold water, but only for 24 h. The results of the study thus validated the traditional BMAA removal process through cold water treatment, but recommend for increase in the treatment period to 48 h or more.

Knowledge and practices of commercial banana farmers related to pesticide use in Chitwan district, Nepal; a cross-sectional study and meta-analyses.

The exposure of farmers to pesticides due to inadequate safety measures is a concern in low-income countries in Africa and Asia. However, until now, there have been limited studies on the farmers' risk due to pesticide application to fruit crops. The knowledge of farmers' exposure related to pesticide use and their safety practices was studied among 100 banana farmers in three areas (Padampur, Jagatpur, and Thimura) of Chitwan district, Nepal. More than 75% of the farmers complained about problems related to insects. Most frequently used insecticides in the area were chlorpyrifos and cypermethrin. Ten percent (10%) of the applied pesticides were highly hazardous to humans, according to the World Health Organization hazard category, with skin rash being the most common acute symptom reported by 29% of the farmers. Banned organochlorine and organophosphate insecticides, such as endosulfan and triazophos, respectively, are still being used by farmers in the aforementioned areas. Spearman's correlation analysis revealed the lack of knowledge and safety practices among farmers leading to inadequate awareness related to the negative effects of pesticide use on human health and the environment. Therefore, government extension service can play a crucial role in improving banana farmers' knowledge of the toxic effects of pesticides as well as enforcing the Nepali language in the labeling of pesticide containers and packages.

Immunopurification Reagents and Their Application in Biologics and Biomarker Quantitation Using LC-MS/MS in Drug Discovery.

The LC-MS/MS technology is one of the most utilized bio-analytical tools owing to its advantage of selectivity, sensitivity and multitasking. The advent of novel biological therapies and increasing demand for protein biomarker identification and quantitation have put the LC-MS/MS technology at the forefront. The questions which are been posed to the LC-MS/MS scientist are complex. The complexity of the question increases further with the matrices in which these questions need to be answered. To bring down the complexity of the analysis, LC-MS/MS technology is utilizing the immunopurification (IP) technique as the new sample preparation technique. The IP reagents are the most common reagents which are used to decrease the matrices' complexity and allow the LC-MS/MS system to reach greater sensitivity. The utilization of these reagents is increasing every day, but the proper utilization of these reagents is still unknown to the common analyst in drug discovery. The present review throws light on the utilization aspect of these reagents, as we have classified these reagents on basis of their utilization, which will allow the readers to gain an understanding of these reagents. This review will also talk about the merits and the demerits of each approach and the current understanding of utilizing these reagents.

Detection of bisphenols in Indian surface water, tap water, and packaged drinking water using dispersive liquid-liquid microextraction: exposure assessment for health risk.

The prevalence of bisphenols (BPs) has been well documented in the aquatic environment of many countries, but such studies from India are quite limited. The present work aimed to determine the occurrence of BPs in surface water (n = 96), tap water (n = 172), and packaged drinking water (n = 42) and estimate their exposure to humans. For this, a simple, sensitive, cost-effective, and green analytical chemistry method based on dispersive liquid-liquid microextraction (DLLME) was employed. Bisphenol A (BPA) was found as the most prevalent bisphenol (mean concentration range = 980-6470 ng/L) in all the water samples, with a % detection frequency of 17-39%. Bisphenol S (BPS) and bisphenol Z (BPZ) were also detected in all types of water samples. The mean estimated daily intake (EDI) for total BPs (tap water and packaged drinking water) was found to be 474.37 ng/kg b.w./day in adults and 665.65 ng/kg b.w./day in children, respectively. This indicated that the total exposure to all the detected BPs obtained for adults and children was lower than the temporary tolerable daily intake (t-TDI) recommended by the European Food Safety Authority (EFSA) (4 µg/kg b.w./day), thereby posing no substantial risks to humans from consuming water from the tap and/or packaged drinking water.

Co-occurrence of mycotoxins: A review on bioanalytical methods for simultaneous analysis in human biological samples, mixture toxicity and risk assessment strategies.

Mycotoxins are the toxic chemical substances that are produced by various fungal species and some of these are harmful to humans. Mycotoxins are ubiquitous in nature and humans could be exposed to multiple mycotoxins simultaneously. Unfortunately, exposure to mixed mycotoxins is not very well studied. Various studies have demonstrated the capacity of mycotoxins to show synergistic effect in the presence of other mycotoxins, thus, increasing the risk of toxicity. Hence, it is important to monitor mixed mycotoxins in human biological samples which would serve as a crucial information for risk assessment. Through this review paper, we aim to summarize the mixture toxicity of mycotoxins and the various bio-analytical techniques that are being used for the simultaneous analysis of mixed mycotoxins in human biological samples. Different sample preparation and clean-up techniques employed till date for eliminating the interferences from human biological samples without affecting the analyses of the mycotoxins are also discussed. Further, a brief introduction of risk assessment strategies that have been or could be adopted for multiple mycotoxin risk assessments is also mentioned. To the best of our knowledge, this is the first review that focuses solely on the occurrence of multiple mycotoxins in human biological samples as well as their risk assessment strategies.

Carbon nanomaterials for the detection of pesticide residues in food: A review.

In agricultural fields, pesticides are widely used, but their residual presence in the environment poses a threat to humans, animals, insects, and ecosystems. The overuse of pesticides for pest control, enhancement of crop yield, etc. leaves behind a significant residual amount in the environment. Various robust, reliable, and reusable methods using a wide class of composites have been developed for the monitoring and controlling of pesticides. Researchers have discovered that carbon nanomaterials have a wide range of characteristics such as high porosity, conductivity and easy electron transfer that can be successfully used to detect pesticide residues from food. This review emphasizes the role of carbon nanomaterials in the field of pesticide residue analysis in different food matrices. The carbon nanomaterials including carbon nanotubes, carbon dots, carbon nanofibers, graphene/graphene oxides, and activated carbon fibres are discussed in the review. In addition, the review examines future prospects in this research area to help improve detection techniques for pesticides analysis.

Cytochrome P450 isoforms contribution, plasma protein binding, toxicokinetics of enniatin A in rats and in vivo clearance prediction in humans.

Emerging mycotoxins, such as enniatin A (ENNA), are becoming a worldwide concern owing to their presence in different types of food and feed. However, comprehensive toxicokinetic data that links intake, exposure and toxicological effects of ENNA has not been elucidated yet. Therefore, the present study investigated the in vitro (rat and human) and in vivo (rat) toxicokinetic properties of ENNA. Towards this, an easily applicable and sensitive bioanalytical method was developed and validated for the estimation of ENNA in rat plasma. ENNA exhibited high plasma protein binding (99%), high hepatic clearance and mainly underwent metabolism via CYP3A4 (74%). The in-house predicted hepatic clearance (54 mL/min/kg) and observed in vivo rat clearance (55 mL/min/kg) were comparable. The predicted in vivo human hepatic clearance was 18 mL/min/kg. ENNA underwent slow absorption (Tmax = 4 h) and rapid elimination following oral administration to rats. The absolute oral bioavailability was 47%. The toxicokinetic findings for ENNA from this study will help in designing and interpreting toxicological studies in rats. Besides, these findings could be used in physiologically based toxicokinetic (PBTK) model development for exposure predictions and risk assessment for ENNA in humans.

Label-free plasma proteomics for the identification of the putative biomarkers of oral squamous cell carcinoma.

Mass spectrometry-based label-free proteomics is becoming the analytical tool of interest to identify and quantitate the biomarkers for cancer. The oral squamous cell carcinoma (OSCC) which is one of the leading cancers worldwide, lacks biomarkers for the early prognosis and diagnosis. The present study profiled plasma proteome of the Indian OSCC human patients using a label-free mass spectrometry proteomics approach. The study first time utilized the three most widely used data analysis software MaxQuant (MQ), Proteome discoverer (PD), and Trans proteomic pipeline (TPP) together for label-free quantitation of the proteins. The study identified 16 proteins which can be used as a signature protein panel for OSCC. The pathway analysis showed predominant involvement of the immune system, hemostasis as the major pathways that are indicative of the disease progression. The network analysis showed maximum interaction for the Fibronectin and C-reactive protein. The study demonstrates that plasma proteins contain signatures that can be used as putative biomarkers for OSCC. Based on the label-free quantitation and the mechanistic analysis C-reactive protein, Carbonic anhydrase-1, and Fibronectin are identified as putative biomarkers of OSCC. Once these findings are validated in the large cohorts these protein signatures can be used as a biomarker for OSCC. SIGNIFICANCE: The oral squamous cell carcinoma (OSCC) is the eighteenth most prevalent malignancy in the world and ranks second in India. There are no biomarkers that could be indicative of the diseased state. Various studies have been carried out on saliva and tumors of OSCC patients in India, but none of the studies have profiled the plasma. We utilized the label-free approach for the first time on the Indian population in generating the panel of plasma proteins which could be indicative of the diseased state. The study identified Carbonic anhydrase 1, C-reactive protein, and Fibronectin proteins as putative biomarkers for the OSCC. The study obtained the signature panel by utilizing the 3 most widely used software for the label-free quanatitation (LFQ) namely MaxQuant, Proteome Discoverer, and Trans proteomic pipeline. The utilization of 3 software for the LFQ reduced the bias and provided a comprehensive list of proteins. All the differential proteins were mechanistically studied for their biological relevance and the pathway and network analysis were carried out. The study helps us in increasing the understanding of the proteins which are involved in the progression of the diseases. Studying the panel of proteins from all biofluids along with plasma in large cohorts of the population will help in understanding the disease in greater depth and help in identifying the relevant biomarkers for the OSCC.

Iron nanoparticles decorated hierarchical carbon fiber forest for the magnetic solid-phase extraction of multi-pesticide residues from water samples.

This study describes a versatile, robust and fast sample pre-concentration novel method based on chemical vapour deposition grown iron nanoparticles dispersed hierarchical carbon fiber forest (Fe-ACF/CNF) for the determination of multi-pesticide residue in water samples. This method was developed by the implementation of Fe-ACF/CNF to magnetic solid-phase extraction method (MSPE) for the adsorption of twenty-nine pesticides of various classes using gas chromatography equipped with an electron capture detector. Fe-ACF/CNF was grown via tip growth mechanism and Fe-nanoparticles are moved to the tip of CNF. The presence of Fe-nanoparticles is responsible for the magnetic property of proposed adsorbents. The Fe-ACF/CNF is competent enough to extract twenty-nine pesticides of different physico-chemical characteristics from water samples. All the predominant parameters including the amount of sorbent desorption time, temperature, sonication effect, regeneration, and reusability of Fe-ACF/CNF were thoroughly investigated. Acceptable linearity was obtained in the range of 20-500 µg/L with a correlation coefficient value ≥ 0.990 for all pesticides. The accuracy of the developed method was evaluated and the obtained recovery of the spiked samples was within 70-120% (standard deviation ≤ 15%) and reusability up to the 4th cycle. The limit of detection and quantification values was in the range of 1.44-5.15 and 4.76-17.0 µg/L, respectively. The obtained results are also cross verified with real water samples from the Gomti river (Lucknow, India) and shown the excellent extraction efficiency of Fe-ACF/CNF.

Plausible drug interaction between cyclophosphamide and voriconazole via inhibition of CYP2B6.

The inhibitory activities of eight cytochrome P450 (CYP) isoenzymes for representative or suspected inhibitors of CYPs, including pesticides, were evaluated simultaneously using an in vitro cocktail incubation method to demonstrate the importance of systematic evaluation of CYP inhibitory risks in drug interaction (DI). Potent inhibition of CYP2B6 was noticeable for some azoles, including voriconazole. When voriconazole and cyclophosphamide were co-administered in mice, cyclophosphamide-induced alopecia and leukopenia were significantly suppressed by approximately 50% with increased blood concentrations of cyclophosphamide. The formation of an active metabolite of cyclophosphamide was suppressed effectively by voriconazole in the mouse liver microsomes. Surveys of adverse event reporting databases in Japan (JADER) and the U.S. (FAERS) showed that the proportional reporting ratios of neutropenia, hemorrhagic cystitis, and alopecia for cyclophosphamide, which is principally activated by CYP2B6 in humans, were mostly reduced, or tended to be reduced when azoles, including voriconazole, were prescribed in combination. It is highly likely that DIs between cyclophosphamide and azoles occur in the clinical setting. This study also suggests that more proper consideration of CYP2B6-mediated DIs is warranted. The combination of the in vitro cocktail method and a survey of adverse event reporting databases was a useful method to comprehensively detect pharmacokinetic DIs.

Determination of Bisphenol Analogues in Infant Formula Products from India and Evaluating the Health Risk in Infants Asssociated with Their Exposure.

Bisphenol A (BPA) is a well-recognized endocrine disruptor, and considering its adverse effects its use in infant bottles has been banned in many countries. Growing concern on the use of BPA has led to its replacement with its analogues in numerous applications. Present is the first report determining the occurrence of seven bisphenols (BPs: BPA, BPAF, BPC, BPE, BPFL, BPS, and BPZ) in Indian infant formula. A reliable and efficient UPLC-MS/MS method for their simultaneous determination was developed and validated in powdered infant formula (n = 68). The limit of quantification of the method was 0.19 ng/g for BPA, BPAF, BPE, BPS and BPZ and 0.78 ng/g for BPC and BPFL. The highest concentration was detected for BPA (mean = 5.46 ng/g) followed by BPZ and BPS. BPAF, BPFL, BPC and BPE were detected in none of the samples. The estimated daily intake (EDI) of total BPs in infants (0-12 months old infants) was determined to be 54.33-213.36 ng/kg b.w./day. BPA mainly contributed to the total intake (EDI = 92.76 ng/kg b.w./day). The dietary exposure to total BPs evaluated in the present study was approximately 1 order of magnitude lower than the reference value of BPA set by EFSA (4 µg/kg b.w./day) and, thus, may not pose considerable risks to infants.

Occurrence of Alternariol and Alternariolmonomethyl ether in edible oils: Their thermal stability and intake assessment in state of Uttar Pradesh, India.

Alternariol (AOH) and Alternariol monomethyl ether (AME) mycotoxins are found to be present naturally in various food commodities, such as barley, oats, pepper, rye, sorghum, sunflower seeds, tomatoes, and wheat. A few epidemiological studies have correlated the consumption of Alternaria-contaminated cereal grains with higher occurrence of esophageal cancer in Chinese populations. In addition, several studies have reported the toxicological properties of Alternaria mycotoxins. However, surveillance data on AOH and AME occurrence are still limited. Therefore, the goal of this study was to determine the presence of AOH and AME in various commonly consumed, edible oils using HPLC-FLD method. Thirty four percent of samples were found positive for AOH and 35% for AME. Moreover, AOH retained 80% stability, while AME retained 84% stability, after deep frying for 25 min, which is an important factor with respect to Indian cooking style. To the best of our knowledge, this is the first report on the presence of Alternaria mycotoxins in edible oils and their probable dietary intake in Indian population. This surveillance study may help in formulating guidelines for Alternaria mycotoxin levels in India, which are not yet implemented by Food Safety and Standards Authority of India. PRACTICAL APPLICATIONS: At present, no safety guidelines exist for Alternaria mycotoxins in any part of the world. This study will help the regulatory bodies to set permissible levels of Alternaria mycotoxins to safeguard the health of consumers. This study shows that Alternaria mycotoxins are heat stable even after deep frying for 25 min. The data will also help to issue guidelines against exposure of these mycotoxins, keeping in the mind the heat stability factor.

Investigating the glucuronidation and sulfation pathways contribution and disposition kinetics of Bisphenol S and its metabolites using LC-MS/MS-based nonenzymatic hydrolysis method.

Despite showing serious health consequences and widespread exposure, the toxicokinetic information required to evaluate the health risks of BPS is insufficient. Thus, we aim to describe the comprehensive toxicokinetics of BPS and its glucuronide (BPS-G) and sulfate (BPS-S) metabolites in rats. Simultaneous quantification of BPS and its metabolites (authentic standards) was accomplished using UPLC-MS/MS method. BPS displayed rapid absorption, extensive metabolism and fast elimination after oral administration. Following intravenous administration, BPS exhibited CL (8.8 L/h/kg) higher than the rat hepatic blood flow rate suggesting the likelihood of extrahepatic clearance. The CL value differed from those reported previously (sheep and piglets) and the probable reason could be attributed to dose- and/or interspecies differences. BPS was extensively metabolized and excreted primarily through urine as BPS-G (∼56%). BPS and BPS-S exhibited a high protein binding capacity in comparison to BPS-G. In in vitro metabolic stability study, BPS was predominantly metabolized through glucuronidation. The predicted in vivo hepatic clearance of BPS suggested it to be a high and intermediate clearance chemical in rats and humans, respectively. The significant interspecies difference observed in the clearance of BPS between rats and humans indicated that toxicokinetics of BPS should be considered for health risk assessment in humans.

Efficient antileishmanial activity of amphotericin B and piperine entrapped in enteric coated guar gum nanoparticles.

Amphotericin B (AmB) exhibits potential antileishmanial activity, with only a little rate of recurrence. However, low bioavailability and severe nephrotoxicity are among the major shortcomings of AmB-based therapy. Various AmB nanoformulations have been developed, which to an extent, have reduced its toxicity and increased the drug efficacy. To further reduce the nonspecific tissue distribution and the cost of the treatment, the current AmB-based formulations require additional improvements. Combination of natural bioenhancers with AmB is expected to further increase its bioavailability. Therefore, we developed a nanoformulation of AmB and piperine (Pip), a plant alkaloid, known to enhance the bioavailability of various drugs, by entrapping them in guar gum, a macrophage targeting polymer. Owing to the ease of oral delivery, these nanoparticles (NPs) were coated with eudragit to make them suitable for oral administration. The formulated eudragit-coated AmB and Pip-loaded NPs (Eu-HDGG-AmB-Pip-NPs) exhibited controlled release of the loaded therapeutic agents and protected the drug from acidic pH. These NPs exhibited effective suppression of growth of both promastigotes and amastigotes of Leishmania donovani parasite under in vitro. In vivo evaluation of these NPs for therapeutic efficacy in golden hamster-L. donovani model demonstrated enhanced drug bioavailability, non-nephrotoxic nature, and potential antileishmanial activity with up to 96% inhibition of the parasite. Graphical abstract.

Plasma protein binding, metabolism, reaction phenotyping and toxicokinetic studies of fenarimol after oral and intravenous administration in rats.

Fenarimol (FNL), an organic chlorinated fungicide, is widely used in agriculture for protection from fungal spores and fungi. Despite being an endocrine disruptor, no toxicokinetic data is reported for this fungicide. In the present work, we determined the plasma protein binding, metabolic pathways and toxicokinetics of FNL in rats. In vitro binding of FNL to rat and human plasma proteins was ∼90%, suggesting that FNL is a highly protein bound fungicide. The predicted in vivo hepatic clearance of FNL in rats and humans was estimated to be 36.71 and 14.39 mL/min/kg, respectively, indicating it to be an intermediate clearance compound. Reaction phenotyping assay showed that CYP3A4 mainly contributed to the overall metabolism of FNL. The oral toxicokinetic study of FNL in rats at no observed adverse effect level dose (1 mg/kg) showed maximum plasma concentration (C max) of 33.97 ± 4.45 ng/mL at 1 h (T max). The AUC0-∞ obtained was 180.18 ± 17.76 h*ng/mL, whereas, the t 1/2 was ∼4.74 h. Following intravenous administration, FNL displayed a clearance of 42.48 mL/min/kg which was close to the predicted in vivo hepatic clearance. The absolute oral bioavailability of FNL at 1 mg/kg dose in rats was 45.25%. FNL at 10 mg/kg oral dose exhibited non-linear toxicokinetics with greater than dose-proportional increase in the systemic exposure (AUC0-∞ 8270.53 ± 1798.59 h*ng/mL).

CVD grown carbon nanofibers: an efficient DSPE sorbent for cleanup of multi-class pesticide residue in high fat and low water commodities by QuEChERS using GC-ECD.

An inexpensive, effective, and efficient dispersive solid-phase extraction (DSPE) sorbent was developed as an alternative to traditionally used sorbents (primary secondary amine and C18) for fatty matrices using the QuEChERS method. Catalytic chemical vapor deposition grown carbon nanofibers dispersed on activated carbon fibers (Ni-ACF/CNF) having a BET specific surface area of 738 (m2/g) were for the first time evaluated as a DSPE material for sample cleanup before gas chromatographic analysis. Based on cleanup performance and recoveries, 10 mg of Ni-ACF/CNF was found optimal for the determination of twenty-seven multi-class pesticides in high fat and low water commodities/matrices (peanut, soybean, sesame, and flaxseed). The recoveries obtained for all analytes were in the range ~ 72 to ~ 117%, with relative standard deviation values ≤ 15%. The limits of detection and quantification values were 0.7-4.2 ng/g and 2.3-13.9 ng/g, respectively. The matrix match calibration curve was linear in the range 20-500 ng/g with a correlation coefficient of ≥ 0.993. The results reveal that the Ni-ACF/CNF is a competent DSPE sorbent, similar to primary secondary amines and C18 sorbent materials, for pesticide determination by QuEChERS methods in high fat and low water commodities. Graphical abstract.

Bioavailability, tissue distribution and excretion studies of a potential anti-osteoporotic agent, medicarpin, in female rats using validated LC-MS/MS method.

Medicarpin, one of the active constituents isolated from the extract of Butea monosperma, has been shown to have various pharmacological activities including potent anti-osteoporotic properties. The aim of this study was to investigate the oral pharmacokinetics, tissue distribution and excretion of medicarpin following single oral dose administration in female rats. Oral pharmacokinetics was explored at 5 and 20 mg/kg while tissue distribution, urinary and fecal excretion were studied following 20 mg/kg oral dose. Medicarpin was quantified in rat plasma, urine, feces and tissue samples using a validated LC-MS/MS method following reverse-phase HPLC separation on RP18 column (4.6 mm × 50 mm, 5.0 µm) using methanol and 10 mM ammonium acetate (pH 4.0) as mobile phase in the ratio of 80:20 (v/v) at a flow rate of 0.8 mL/min. The oral bioavailability of medicarpin was found to be low with low systemic levels. The concentration in tissues was significantly higher than plasma. Highest tissue concentrations were found in the liver followed by bone marrow. Urinary and fecal excretion of medicarpin was < 1 %. In conclusion, medicarpin was found to be highly distributed in body tissues and minimally excreted via urine or feces.