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The purpose of this study is to assess the aging effect of the most sensitive metabolite choline and its ratio with other metabolites N-acetyl aspartate, creatine, myoinositol and lactate in suggestive glioma brain using magnetic resonance spectroscopy. Forty-four suggestive glioma patients were scanned at 1.5 Tesla (G.E) from MRI center, Bahawalpur Victoria Hospital (BVH) Bahawalpur, Pakistan. The mean age of 44 patients was 41.40 years with S.D = 17.36 in which total 24 males (mean age = 39.1, S.D = 17.37, min = 09, max = 72) and 20 females (mean age = 44.1, S.D = 17.39, min = 15, max = 85). For chemical analysis, patients were categorized into three age groups, 16 patients in group-I (G-I) of age range 6–30, 18 patients in group-II (G-II) of age range 31–55 and 10 in group-II (G-III) of age range 56–85 in years. Single voxel spectroscopy (SVS) were employed using one STEAM technique to get 1H MRS Spectra. I.Q Lite software to demarcate the region of interest (ROI) in brain. Data analysis was performed in OrgionPro.8, China. A p-value (P ≤ 0.05) was deemed to be statistically significant in data. One-way AONVA test was used to compare the aged related variation of five metabolites in three age groups G-I, G-II and G-III. High peak of choline with lactate peak verified the malignancy in the suggestive glioma cases. The p-value (p > 0.5) of Ch/Cr was observed in G-I compare to the p-value (p > 0.05) of Ch/Cr for G-II and G-III during study. Statistically, results demonstrated no significant difference in three proposed age groups in the region of suggestive glioma brain. Statistically, it was concluded that age‐related metabolic changes in three proposed groups remained insignificant (p > 0.05) in region of glioma brain. However, higher choline ratio and reduced ratio of N-acetylaspartate revealed the higher progression of malignancy in G-II that might be monitoring marker of aging effect of metabolites.

Background: Ilexsaponin A1, one of the most representative triterpene saponin components in the roots of I. pubescens, showed its effects in anticoagulation and antithrombosis, attenuating ischemia-reperfusion-induced myocardial, angiogenesis and inhibiting phosphodiesterase. Objective: Reveal the key intestinal bacterial strains responsible for ilexsaponin A1 metabolism, and clarify their metabolic behavior. Methods: An accurate and sensitive LC-MS/MS method for the determination of “ilexsaponin A1 in General Anaerobic Medium (GAM) broth” was established and systematically validated. Then it was applied to screen and study the metabolic potential of the intestinal bacterial strains in an anaerobic incubation system. Results: Quantitation of ilexsaponin A1 could be performed within an analytical run time of 14.5 min, in the linear range of 2 - 2000 ng/ml. Enterobacter sakazakii, Bifidobacterium breve, Bifidobacterium adolescentis, Bifidobacterium catenulatum, and Bifidobacterium angulatum were identified to have a potential effect to metabolize ilexsaponin A1 to different extents; and further bacterial metabolic studies were performed to clarify their metabolic capacity and behavior. Conclusion: This paper contributes to a better understanding of the intestinal bacterial metabolism of ilexsaponin A1 and provides scientific evidence for its clinical application. Additionally, the importance of intestinal bacterial strains in the disposition of natural products was also highlighted.

A 78-year-old woman presented with an atypical femoral fracture of her right femur associated with a left thigh stress fracture following 9 years of bisphosphonate therapy. The fracture was managed with femoral nailing. However, she represented 3 years later with peri-prosthetic infection, worsening of the left-side stress fracture, and low bone turnover in a bone metabolic study, consistent with an ongoing bisphosphonate effect. This case highlights the increased rate of postoperative complications in atypical femoral fractures even years after surgery as well as the challenging management issues for contralateral fracture and the risk of missing bilateral fractures.

Dingkun Dan (DKD), a reputable traditional Chinese medicine formula, has been used to treat gynecological diseases and showed significant clinical effects since ancient times. However, the application and development of DKD are seriously hampered by the unclear active substances. Structural characterization of compounds absorbed in vivo and their corresponding metabolites is significant for clarifying the pharmacodynamic material basis. In this study, an integrated strategy using ultra-performance liquid chromatography, coupled with quadrupole time-of-flight mass spectrometry and UNIFI™ software, was used to identify prototypes and metabolites after oral administration of DKD in rats. As a result, a total of 261 compounds, including 140 prototypes and 121 metabolites, were tentatively characterized in rat plasma, urine, and feces. The metabolic pathways of prototypes have been studied to clarify their possible transformation process in vivo. Moreover, an in vitro metabolism study was applied for verifying the metabolites under simulating the metabolic environment in vivo. This first systematic metabolic study of DKD is important for elucidating the metabolites and metabolic pathways and could provide a scientific basis for explaining the integrative mechanism in further pharmacology study.

The main objective of this study was to perform quantitative genetic analyses of tibia and foot ash traits, which might serve as proxy traits to improve phosphorus utilization (PU) in a breeding program. Additionally, data for ash concentration in tibia and foot were compared with data for total amount of ash. Heritabilities for bone ash traits and genetic and phenotypic correlations between bone ash traits and PU were estimated. A total of 887 F2 birds, established from 2 Japanese quail lines divergently selected on social reinstatement behavior, were provided a P deficient diet. In a metabolic study, feed consumption was measured and total excreta collected for each bird separately. Afterwards, birds were euthanized, the bones obtained and incinerated. Bone ash data showed a heritability of 0.230 (amount of tibia ash) to 0.342 (amount of foot ash), which was higher than estimated for PU, P retention, calcium utilization (0.120–0.174), and performance traits (0.088–0.114). The strongest genetic and phenotypic correlations between PU and bone ash traits were detected for the amount of foot ash with 0.549 and 0.527, respectively. Genetic and phenotypic correlations were stronger between PU and ash amount than between PU and ash percentage, irrespective of bone. Therefore, ash amount was considered a better trait than ash percentage to reflect PU. Strong genetic and phenotypic correlations were detected between the amount of foot and tibia ash (0.887 and 0.901, respectively). Phenotypic and genetic correlations between ash amount and PU or calcium utilization were almost identical, irrespective of bone. Foot ash is as suitable as tibia ash, but easier to determine. Bone ash data, especially the amount of foot ash, seem to be suitable indirect selection criteria for P efficiency breeding.

Neurosyphilis is a chronic central nervous system infectious disease caused by Treponema pallidum. Our aim was to study the metabolic profiling in the cerebrospinal fluid of neurosyphilis patients and identify specific potential biomarkers. Fifteen cerebrospinal fluid samples from neurosyphilis patients and 14 non-neurosyphilis samples were analyzed by liquid chromatography-mass spectrometer (LC-MS). The LC-MS data were preprocessed by supervised pattern recognition to obtain diagnostic models. Both orthogonal projections to a latent structures discriminant analysis (OPLS-DA) and a t test were used to obtain specific metabolites for neurosyphilis. LC-MS data showed that the metabolites in cerebrospinal fluid (CSF) from neurosyphilis are different from the non-neurosyphilis group. The OPLS-DA model parameters R2Y and Q2Y are both more than 0.7 and indicated a satisfactory diagnostic performance. Bilirubin, L-histidine, prostaglandin E2, alpha-kamlolenic acid, and butyryl-L-carnitine and palmitoyl-L-carnitine were identified as novel potential biomarkers for neurosyphilis. The metabolic study of CSF may provide a new way to explore the pathogenesis of neurosyphilis.

Nephrolithiasis: how the laboratory of clinical pathology can play a pivotal role in the prevention of lithiasic eventsWe here describe our 15-year experience and report both on methodological problems (proposing a new datasheet and biochemical techniques) and on organizational ones (we discuss our patient approach and useful reporting for the physician).

The use of the ketogenic diet (KD) as an adjuvant therapy in high-grade gliomas (HGG) is supported by preclinical studies, but clinical data on its effects on metabolism are currently lacking. In this study, we describe the effects of a KD on glucose profile, ketonemia, energy metabolism, and nutritional status, in adults affected by HGG. This was a single-arm prospective study. An isocaloric 3:1 KD was administered for 1 mo. Glucose profile was assessed by using fasting glycemia, insulin, and glycated hemoglobin. To evaluate ketonemia changes, a hand-held ketone meter was used from home. Energy metabolism was assessed by indirect calorimetry. Nutritional status was evaluated through changes in body composition and in lipid and hepatic profile. No changes in fasting glycemia were observed; however, insulinemia dropped to half of baseline levels. The KD shifted the metabolism, rising ketonemia and decreasing glucose oxidation rate to a quarter of the initial values. Moreover, the KD was generally safe. One-month intervention with the KD was able to act upon key metabolic substrates potentially involved in HGG metabolism. The lack of a significant reduction in fasting glycemia should be investigated in future studies.

1. Introduction The Department of Ergometry, Exercise, Nuclear Cardiology and Cardiovascular Rehabilitation (DERC), through its commission on Health and Diagnosis of Cardiovascular Diseases in Women and in association with the Sports Cardiology Study Group (GECESP) and the Cardiopulmonary Rehabilitation and Metabolic Study Group (GERCPM), has prepared this position paper in accordance with Brazilian Society of Cardiology norms to guide health professionals regarding the prescription of physical exercise during pregnancy and the postpartum period, as well as to strengthen the relationship [...]

A metabolic study comparing people who lost weight through dieting or through bariatric surgery shows that the effects on diabetes are due to weight loss.

The aim of this study was to report normal measurements of green-emitting fluorophores in the macula of healthy young individuals and to assess the repeatability of these quantitative metrics. To do so, healthy young volunteers were imaged twice (7 &plusmn, 3 days apart) using a confocal blue-light fundus autofluorescence (FAF) device with a shorter excitation wavelength (peak at 450 nm) and the capability for separately detecting the red and green components of the emission spectrum. The main outcome measure was the percentage of area occupied by green-emitting fluorophores in the macula. In addition, this measure was performed in separate regions providing a topographical assessment in the foveal, parafoveal and perifoveal regions. Furthermore, the level of agreement between repeated measurements was evaluated. Thirty eyes from 30 healthy volunteers were included in this analysis. Mean age was 26.2 &plusmn, 2.8 years (median: 25.0 years, range: 23.0&ndash, 32.0 years). Median (interquartile range&mdash, IQR) area occupied by green-emitting fluorophores was 3.6% (1.9&ndash, 4.7%) in the macular region. In the topographical analysis, this percentage was higher in the foveal area (median = 33.3%, IQR = 21.9&ndash, 41.2%), as compared with both the parafoveal (median = 5.3%, IQR = 2.4&ndash, 8.1%, p &lt, 0.0001) and perifoveal (median = 0.5%, IQR = 0.2&ndash, 0.8%, 0.0001) regions. The coefficient of variation (CV, ranging from 1.1% to 1.7% in the analyzed regions) and the intraclass correlation coefficient (ICC, ranging from 0.93 to 0.97) indicated high levels of repeatability. In conclusion, the assessment of green-emitting fluorophores is repeatable. The distribution of these fluorophores is highest in the foveal region. Assuming that flavin adenine dinucleotide (FAD) emits in the green autofluorescence spectrum, this variability could be secondary to an increased quantity of mitochondria in the foveal region.

Six Leuconostoc mesenteroides strains isolated from camel (3) and sheep (1) milk, silage (1) and honey (1) have been investigated. All showed probiotic properties due to their resistance to gastrointestinal tract stresses, antibiogram profiles, hydrophobicity levels and antimicrobial activities against Staphylococcus aureus and Escherichia coli. Six had proteolytic, lipolytic, acidifying, and coagulation activities and five produced dextran. Metabolic flux analysis during growth in a sucrose-containing medium of two representative dextran-producing strains, and a non-producing strain revealed differences in the levels of the sucrose metabolites: fructose, glucose, lactic acid and mannitol, and a correlation between sucrose consumption and dextran synthesis for the producing strains. CM9 from camel milk showed high dextran production and the best pattern of intestinal tract colonization of gnotobiotic zebrafish embryos. These facts together with CM9’s technological and probiotic properties indicate that this strain may be useful for the production of functional dairy food.

Introduction Mauritia flexuosa is a dioecious species native to the Amazon, and the gender identification can currently be determined through the differentiation between female and male flowers in the first flowering. Objectives To identify variations in the metabolome profiles of pooled and individual samples of leaves from female and male plants of M. flexuosa in reproductive phase, and the plant in its vegetative phase, using two extraction systems. Material and methods Extractions of leaves were separately conducted using deuterated water and methanol. The extracts were evaluated by NMR spectroscopy and chemometric methods. Results Different NMR spectroscopic profiles were observed for females, males, and plants in the vegetative phase. Significant variations were found in the carbohydrate and fatty acid contents for the aqueous and methanolic extracts, respectively. Although principal component analysis (PCA) has not been efficient to distinguish the genders, orthogonal signal correction/partial least squares-discriminant analysis (OPLS-DA) was able to successfully differentiate male and female plants in the reproductive phase, independently on the sampling approach. For plants in the vegetative phase, OPLS-DA method from aqueous extracts of M. flexuosa leaves also distinguished female samples from the others and showed a small overlap between male and young plants, while the classification model was not able to be used for prediction. Conclusions NMR spectroscopy and chemometric-based approach was demonstrated to be useful in the metabolic study of dioecy in M. flexuosa. The extracts of the leaves allowed for differentiation between male and female plants; however, for plants in the vegetative phase, the identification of the gender was not effective. Copyright © 2018 John Wiley & Sons, Ltd.

Troxipide is widely used to treat gastric ulcer (GU) in the clinic. However, a lack of systematic metabolic, pharmacokinetic and pharmacological studies limits its clinical use. This study aimed to firstly explore the metabolic, pharmacokinetic and pharmacological mechanisms of troxipide in rats with GU compared to normal control (NC) rats. First, metabolic study was perormed by a highly selective, high-resolution mass spectrometry method. A total of 45 metabolites, including 9 phase I metabolites and 36 phase II metabolites, were identified based on MS/MS spectra. Subsequently, the pharmacokinetics results suggested that the Cmax, Ka, t1/2, AUC(0−t) and AUC(0−∞) of troxipide were significantly increased in rats with GU compared with NC rats. The Vz, K10 and absolute bioavailability of troxipide were obviously decreased in rats with GU compared with NC rats, and its tissue distribution (in the liver, lung and kidney) was significantly different between the two groups of rats. Additionally, the pharmacodynamic results suggested that the levels of biochemical factors (IL-17, IL-6, TNF-α, IFN-γ, AP-1, MTL, GAS, and PG-II) were significantly increased, the PG-Ӏ level was obviously decreased, and the protein expression levels of HSP-90, C-Cas-3 and C-PARP-1 were markedly increased in rats with GU compared with NC rats. The above results suggested that the therapeutic mechanisms underlying the metabolic, pharmacokinetic and pharmacological properties of troxipide in vivo in rats deserve further attention based on the importance of troxipide in the treatment of GU in this study, and these mechanisms could be targets for future studies.

Ketones provide an alternative brain fuel and may be neuroprotective in older people. Little is known of how to optimize the ketogenic effect of C8:0–C10:0 medium chain triglyceride supplement (kMCT). Metabolic switching (MS) from glucose to ketones as a fuel may have metabolic benefits but has not been extensively studied in humans. The objective of the present study was to use an 8 h metabolic study day protocol to assess the influence of typical components of MS, including a kMCT supplement, low-carbohydrate meal and meal timing, on blood ketones, glucose, insulin and free fatty acids (FFA). In one test, the effect of age was also investigated. Over the 8 h metabolic study day, two 10 g doses of the kMCT increased the plasma ketone response by 19% while reducing overall glycemia by 12% without altering insulin or FFA levels. Moreover, a single early meal (breakfast but no lunch) potentiated the ketogenic effect of MS over 8 h, compared to a single delayed meal (lunch but no breakfast). Age and the low carbohydrate meal did not affect the ketones response. We conclude that an 8-h test period can be used to assess metabolic changes during short-term MS. kMCT provide a robust short-term increase in ketones and might enhance the metabolic effectiveness of short-term or intermittent fasting as a component of MS.

Synthetic cathinones are new psychoactive substances that represent a health risk worldwide. For most of the 130 reported compounds, information about toxicology and/or metabolism is not available, which hampers their detection (and subsequent medical treatment) in intoxication cases. The principles of forensic analytical chemistry and the use of powerful analytical techniques are indispensable for stablishing the most appropriate biomarkers for these substances. Human metabolic fate of synthetic cathinones can be assessed by the analysis of urine and blood obtained from authentic consumers; however, this type of samples is limited and difficult to access. In this work, the metabolic behaviour of three synthetic cathinones (4-CEC, 4-CPrC and 5-PPDi) and one amphetamine (3-FEA) has been evaluated by incubation with pooled human hepatocytes and metabolite identification has been performed by high-resolution mass spectrometry. This in vitro approach has previously shown its feasibility for obtaining excretory human metabolites. 4-CEC and 3-FEA were not metabolised, and for 4-CPrC only two minor metabolites were obtained. On the contrary, for the recently reported 5-PPDi, twelve phase I metabolites were elucidated. Up to our knowledge, this is the first metabolic study of an indanyl-cathinone. Data reported in this paper will allow the detection of these synthetic stimulants in intoxication cases, and will facilitate future research on the metabolic behaviour of other indanyl-based cathinones., Graphical abstract Image 1, Highlights • In vitro metabolic behaviour of 4-CEC, 4-CPrC, 5-PPDi and 3-FEA has been evaluated. • Metabolite elucidation has been performed by high-resolution mass spectrometry. • 4-CEC and 3-FEA were not metabolised. Two minor metabolites were obtained for 4-CPrC. • Twelve phase I metabolites were elucidated for the indanyl-cathinone 5-PPDi. • Up to our knowledge, this is the first metabolic study of an indanyl-cathinone.

The present study aimed to assess the effect of Zn supplementation on trace element levels in the liver, serum, and hair of rats with dietary-induced non-alcoholic fatty liver disease (NAFLD). A total of 26 3-month-old female Wistar rats were divided into four groups: control, NAFLD, Zn-supplemented (227 mg/L zinc as Zn sulfate Zn(SO)4 dissolved in a drinking water), and NAFLD-Zn-supplemented. NAFLD was verified by histological assessment of liver samples. The serum was examined for routine biochemical parameters. Trace elements content was assessed using inductively coupled plasma mass spectrometry (ICP-MS). Zn treatment resulted in an improvement in liver weight and morphology. Dietary supplementation with Zn prevented NAFLD-induced decrease liver Co. The tendency to increase liver Fe in the Zn-treated group was observed. Zn treatment decreased hepatic Al and serum V levels. However, Zn administration did not affect NAFLD-induced I, Mn, and Se depletion in the liver. Hair Zn levels raised in Zn-supplemented groups. Conclusively, the results of the study indicate that Zn supplementation could have a beneficial effect in modulation of the altered trace element status and liver morphology. •Zn treatment improved liver weight and morphology in rats with NAFLD. •Zn supplementation decreased liver Al in NAFLD. •Treatment by Zn prevented depletion of liver Co. •Zn decreased serum V and increased hair Zn levels. •No effect of Zn on NAFLD-induced hepatic I, Mn and Se depletion was observed.

Background: Medium chain triglycerides (MCT) are ketogenic but the relationship between the change in plasma ketones and the change plasma medium chain fatty acids (MCFA)—octanoate, decanoate, or dodecanoate—after an oral dose of MCT is not well-known. An 8 h metabolic study day is a suitable model to assess the acute effects on plasma ketones and MCFA after a dose of tricaprylin (C8), tricaprin (C10), trilaurin (C12) or mixed MCT (C8C10).Objective: To assess in healthy humans the relationship between the change in plasma ketones, and octanoate, decanoate and dodecanoate in plasma total lipids during an 8 h metabolic study day in which a first 20 ml dose of the homogenized test oil is taken with breakfast and a second 20 ml dose is taken 4 h later without an accompanying meal.Results: The change in plasma acetoacetate, β-hydroxybutyrate and total ketones was highest after C8 (0.5 to 3 h post-dose) and was lower during tests in which octanoate was absent or was diluted by C10 in the test oil. The plasma ketone response was also about 2 fold higher without an accompanying meal (P = 0.012). However, except during the pure C10 test, the response of octanoate, decanoate or dodecanoate in plasma total lipids to the test oils was not affected by consuming an accompanying meal. Except with C12, the 4 h area-under-the-curve of plasma β-hydroxybutyrate/acetoacetate was 2–3 fold higher when no meal was consumed (P < 0.04).Conclusion: C8 was about three times more ketogenic than C10 and about six times more ketogenic than C12 under these acute metabolic test conditions, an effect related to the post-dose increase in octanoate in plasma total lipids.

Introducción:la urolitiasis es una enfermedad frecuente de la cual en Colombia se han publicado estudios previos; sin embargo, estos no comparan las características sociodemográficas y clínicas de los pacientes con las comorbilidades y los factores predisponentes de litiasis como hiperuricemia, hipertensión arterial (HTA), obesidad y enfermedad renal crónica (ERC). Objetivo: caracterizar clínica y metabólicamente los pacientes con diagnóstico de urolitiasis atendidos en una clínica de cuarto nivel de Barranquilla, Colombia, en el año 2019. Materiales y métodos: se realizó un estudio observacional, descriptivo y transversal en 49 pacientes con base en el estudio de fichas clínicas. Resultados:el 53,1 % de los participantes eran hombres y las medianas de edad y de índice de masa corporal (IMC) fueron 58 años y 26,4 kg/m2, respectivamente. Algunas de las comorbilidades identificadas fueron, en orden de frecuencia, HTA (69,4 %), ERC (36,7 %), infección de vías urinarias recurrente (24,5 %), hiperuricemia (44,9 %), hipercalcemia (16,3 %) e hiperfosfatemia (12,2 %). Los tipos de cristal encontrados fueron oxalato (20,4 %), urato (12,2 %), mezcla de oxalato y urato (4,1 %), fosfato (4,1 %), hipercalciuria e hiperoxaluria (38,8 %), hiperuricosuria e hipocalciuria (18,4 %) y hipofosfaturia o hipofosfaturia (4,1 %). Asimismo, la hiperuricemia se asoció a edad (p=0,028), ERC (p=0,026), medicamentos antihipertensivos (p=0,022), posición del cálculo en cáliz renal (p=0,012), hiperparatiroidismo (p=0,007), depuración de creatinina (p=0,046) e hipercalciuria (p=0,049). El IMC ? 30 se asoció con ERC estadio 5 (p=0,025), diálisis (p=0,025) e hiperoxaluria (p=0,021). Conclusión:en la población analizada se evidenció una frecuencia significativa de ERC, hiperuricemia, obesidad e HTA.

Anyone who works on plants should be dazzled by the complexity and versatility of plant metabolism. In fact, why restrict this to plant biologists? We all love plant metabolites, whether we’re enjoying the caffeine in our morning cup of tea or the capsaicin heat in the pepper flakes on our

RESUMO O conhecimento sobre modelos animais para estudo metabólico representa a base da pesquisa nessa área. Este trabalho tem por objetivo revisar os principais modelos animais a serem utilizados no estudo da obesidade e da síndrome metabólica. Para isso, pesquisa no banco de dados Pubmed foi realizada usando as palavras-chave “animal models”, “obesity”, "metabolic syndrome”, e “bariatric surgery”. Várias espécies de animais podem ser usadas para o estudo de distúrbios metabólicos, no entanto, os roedores, tanto modelos monogênicos quanto modelos de obesidade induzida por dieta (DIO), são os animais mais utilizados nessa área. Animais monogênicos são a melhor escolha se apenas um aspecto estiver sendo avaliado. Animais DIO tendem a demonstrar melhor a interação entre doença, ambiente e gene. No entanto, eles ainda não são totalmente eficazes para a compreensão de todos os mecanismos dessa doença.

Poria cocos are extensively used as nutritious food, dietary supplements, and oriental medicine in Asia. However, the effect substances are still not very clear. In this study, a targeted screening approach was developed to systematically identify absorbed constituents of Poria cocos in vivo using ultrahigh performance liquid chromatography tandem mass spectrometry combined with UNIFI software. First, incubation reactions in vitro with rat intestinal microflora and rat liver microsomes were conducted to sum up metabolic rules of main constituents. Second, the absorbed constituents in vivo were picked out and identified based on the results of metabolic study in vitro. Finally, the absorbed active constituents in the treatment of Alzheimer's disease were screened by targeted network pharmacology analysis. A total of 62 absorbed prototypes and 59 metabolites were identified and characterized in dosed plasma. Thirty potential active constituents were screened, and 86 drug-targets shared by absorbed constituents and Alzheimer's disease were discovered by targeted network pharmacology analysis. In general, this proposed targeted strategy comprehensively provides new insight for active ingredients of Poria cocos.

Vardenafil, a remedy for erectile dysfunction, is easily modified, facilitating the creation of analogues that have been illegally added to functional foods and counterfeit medications. However, the medical profile of these analogues, including their safety, efficacy, safe drug combinations, metabolism and excretion, has not been completely evaluated, which could cause serious health problems. In this study, two representative vardenafil analogues, pseudovardenafil and hydroxyvardenafil, were metabolized with in-vitro model (human liver microsome) and in-vivo model (rats). The metabolized samples were extracted and characterized, using liquid chromatography quadrupole-time of flight mass spectrometry (LC-Q-TOF-MS). Some imprecise interpretations were evaluated with tandem mass spectrometry (LC-Q-TOF-MS/MS) for mass fragmentation analysis. A total of 11 metabolites of pseudovardenafil and 13 metabolites of hydroxyvardenafil that were identified have never been reported. These new metabolites could be usefully applied to forensic science and other metabolic fields. Furthermore, they could serve as principal references for the toxicity, danger, and side effects of unlawful vardenafil counterfeits.