High-performance Liquid Chromatography System Research Articles

Development of a beetroot-based nutritional gel containing high content of bioaccessible dietary nitrate and antioxidants

Beetroot, a food rich in nitrate and antioxidants has gained attention because of its potential effect on improving cardiovascular health and exercise performance. This work had the purpose of developing a beetroot-based nutritional gel (BG) and estimating the in vitro bioaccessibility of the nitrate, total antioxidant capacity (TAC), total phenolic (TP) and potassium content, as compared to beetroot juice (BJ). Nitrate was assessed by a high-performance liquid chromatography system, TAC was assessed using the Trolox equivalent antioxidant capacity (TEAC) assay and TP was measured using the Folin–Ciocalteu method before and after an in vitro digestion. Significantly higher values of nitrate, TEAC, TP and potassium before and after digestion were observed in BG as compared to BJ. The results suggest a new nutritional strategy to give high contents of bioaccessible nutrients (nitrate, antioxidants and potassium) that are potentially relevant to improve cardiovascular health and exercise performance.

A marine algicidal Thalassospira and its active substance against the harmful algal bloom species Karenia mikimotoi.

The aim of the present study was to obtain a marine bacterium active against Karenia mikimotoi from the East China Sea and to characterize its extracellular algicidal substances. Using preparative high-performance liquid chromatography (prep-HPLC) and electrospray ionization/quadrupole-time of flight mass spectrometer coupled with a high-performance liquid chromatography (LC/MS-Q-TOF) system, we purified the alga-lysing substance produced by strain ZR-2 and determined its molecular structure. Based on morphology and l6S ribosomal DNA (rDNA) sequence analysis, the ZR-2 strain was highly homologous to Thalassospira species. Algicidal activity against K. mikimotoi was detected in the cell-free filtrate but not in bacterial cells. The alga-lysing substance produced by ZR-2 was ethanol-soluble and thermostable, with a retention time of 6.3min and a measured elemental composition of C7H5O2 ([M-H](-) ion at m/z 121.0295). The alga-lysing substance produced by ZR-2 was determined to be benzoic acid. Compared with the negative control, both purified ZR-2 bacteria-free filtrate and standard benzoic acid promoted K. mikimotoi cell disruption and induced K. mikimotoi cell content leakage. Our study is the first to report benzoic acid activity against K. mikimotoi as well as production of benzoic acid by a Thalassospira species.

Group-type characterization of crude oil and bitumen. Part II: Efficient separation and quantification of normal-paraffins iso-paraffins and naphthenes (PIN)

The SARA (saturates, aromatics, resins, and asphaltenes) separation and quantification have been widely used for characterizing the composition of crude oil and bitumen. The saturate hydrocarbons are the most valuable fraction of the oil industry and contain vital geochemical information for determining the origin and formation of petroleum fluids and for assessing the hydrocarbon potential and sedimentary environment of source rocks. It is therefore necessary to further separate the saturate fraction into normal paraffins, iso-paraffins and naphthenes (PIN fractions). Nevertheless, the convenient approach of PIN separation has never been reported on in the literature. Here, we present an efficient method that allows us to further separate the saturate fraction of crude oil or bitumen into PIN fractions through the automated multi-dimensional high performance liquid chromatography system (AMD-HPLC). In essence, the system is equipped with a “PIN1” column (Silicon: S115) packed with a 5.4Å molecular sieve and a “PIN2” column (Silicon: 50%S115+50%S130) packed with a 6.2Å molecular sieve. Each set of columns is connected in a series via multi-position valves that allow a given column to be inserted “in” or to be by-passed “out” of the flow path of the mobile phase at pre-set times in the process. When use iso-octane and n-octane or their mixture as mobile phases, the system separates the saturate hydrocarbon into PIN fractions simply through three steps under a temperature range of 25–140°C. An entire PIN analysis cycle takes nearly 60min, including system re-conditioning. The separated PIN fractions can be quantified using either gravimetric method or evaporative light scattering detector (ELSD) technique. Experimental results exhibit excellent qualitative and quantitative capability.

Comparison of a triple-quadrupole and a quadrupole time-of-flight mass analyzer to quantify 16 opioids in human plasma

The aim of this work is to study whether a quadrupole time-of-flight (QToF) mass analyzer, coupled to an ultra high performance liquid chromatography (UHPLC) system, can be a valuable alternative for a triple-quadrupole (QqQ) mass analyzer, for quantitative toxicological purposes. The case study considered was the quantification of 16 opioids (6-monoacetylmorphine, buprenorphine, codeine, dihydrocodeine, ethylmorphine, fentanyl, hydrocodone, hydromorphone, morphine, norbuprenorphine, norcodeine, norfentanyl, oxycodone, oxymorphone, pholcodine and tilidine) in human plasma. Both methods were validated in parallel in terms of selectivity, matrix effects, extraction recovery, carry-over, bias, precision and sensitivity. Accuracy-profile methodology was used to determine the optimal calibration model, and to estimate bias, repeatability, intermediate precision and total error. Selectivity was demonstrated for all opioids and deuterated analogues, except for codeine-d3 on the UHPLC-QTOF. For most compounds, extraction recoveries were in the range 60 to 80% on both systems, except for the synthetic analogues, buprenorphine, fentanyl and tilidine, where large variability is observed. Carry-over was negligible on both systems. For different opioids, the optimal calibration model was different between the systems. The accuracy profiles of the majority of the opioids indicated that, over the entire tested concentration range, for more than 5% of the future measurements, total errors are expected to exceed the a priori defined 15% acceptance limit. For some exceptions, however, the measurements even suffer from total errors above 30%, which can be attributed to the solid phase extraction procedure that was applied as sample pretreatment technique. Sensitivity was generally tenfold better on the LC-QToF system, probably due to the difference in ion choice for quantification between both systems. In conclusion, the best performing system seemed to depend on the compound, on the parameter and even on the concentration. Accuracy profiles clearly provided valuable information complementary to that obtained in classical validation tests, and therefore preferably are taken into account when deciding on a method's performance.

Short and Robust HPLC-UV Method to Determine Serum Ribavirin Concentration without Evaporation Step

Objective: Measurement of ribavirin (RBV) is important for therapeutic drug monitoring in hepatitis C patient. A simple and fast high performance liquid chromatography (HPLC) method developed and validated to measure ribavirin concentration in serum samples without an evaporation step. Design and method: About 500 μl serum sample, 50 μl internal standard and 20 mM ammonium acetate buffer (pH=8.5) were mixed for 30 seconds and centrifuged. The supernatant was transferred into preconditioned phenyl boronic acid cartridges for solid phase extraction. All cartridges were washed two times with 1 mL of 20 mM ammonium acetate buffer under vacuum not exceeding 10 psi. Ribavirin and internal standard were eluted with 300 μl of 3% formic acid. An aliquot of 100 μl was injected into HPLC system. Results: The method was linear in the range of 0.1-8.0 mg/l with 0.05 mg/l as limit of detection. The correlation coefficient of method comparison was 0.975 with p value of 0.116 showing a good reproducibility of results. The mean accuracy was checked at three different concentrations and found to be between 107-110% for all three levels. The extraction efficiency was 65.5% for ribavirin in the range of 0.1-8.0 mg/l and 71.2% for internal standard at 50 mg/l. The intra assay precisions were determined at 0.5, 2.5, and 5.0 mg/l and % CV were found to be 2.2%, 5.0%, 4.5% respectively. The injection reproducibility at three levels was 5.5%, 6.1% and 3.3%. Removal of gravity flow and evaporation step made this method faster and easy for routine analysis of ribavirin samples. Conclusion: The newly developed HPLC method was faster, accurate and sensitive. It applied for determining serum ribavirin level in hepatitis C patient in our hospital.

Emissions Reduction of Regulated and Unregulated Hydrocarbon, Gases in Gasoline Bi-mode SI/HCCI Engine by TWC Converter

A specific case of HCCI is gasoline fuelled HCCI. It is attractive due to the simplicity of implementing such a technology into existing SI engines as well as the existing fuelling infrastructure. Lean and highly diluted homogeneous charge compression ignition HCCI engines offer great potential in improving vehicle fuel economy and contribute in reducing CO2 emissions. Gasoline is a complicated mixture of many different hydrocarbons which results in rather poor auto-ignition properties. Hydrocarbons and CO emissions from HCCI engines can be higher than those from spark ignition (SI) engines, especially at low engine load when the EGR rate or the residual gas required to control NOx emission are elevated. Toxic chemicals emitted by SI engines, Carbonyl compounds and poly aromatic hydrocarbons PAH generated by V6 (SI/HCCI) gasoline engine especially in HCCI mode. A qualitative and quantitative analysis of hydrocarbon compounds, alkenes. Alkanes, aromatics and aldehydes was analysed before and after catalyst, alkanes, alkenes and aromatic were conducted using Gas Chromatography-Mass Spectrometry (GC-MS) apparatuses. Aldehydes were conducted using High Performance liquid chromatography (HPLC) on reversed phase. HPLC system, although, bi-functional after treatment, the device will be required to control the regulated and unregulated hydrocarbon, CO, and NOx emissions under lean and stoichiometric (oxygen free) engine operating conditions. This paper describes studies on the regulated and unregulated hydrocarbons, NOx, and CO emissions coming out of HCC/SI gasoline engine. Comparative study of catalyst performance will be analysed under HCCI stoichiometric and SI operation under different engine loads, analysis indicate that, the HC and CO emissions reduction over the prototype catalyst was in the range of 90-95% while the maximum NOx emissions reduction under lean engine operating conditions was in the range of 35-55%. The catalytic converter showed an excellent efficiency of eliminating unregulated hydrocarbons (alkenes, alkanes, and aromatics) and aldehydes compounds; achieved reduction efficiency was up to 92%.

Efficiency of Polyphenol Extraction from Artificial Honey Using C18 Cartridges and Amberlite® XAD-2 Resin: A Comparative Study

A comparative study of the extraction efficiency of nine known polyphenols [phenolic acids (benzoic acid, dihydroxybenzoic acid, gallic acid, trans-cinnamic acid, and vanillic acid) and flavonoids (naringenin, naringin, quercetin, and rutin)] was conducted by deliberately adding the polyphenols to an artificial honey solution and performing solid phase extraction (SPE). Two SPE methods were compared: one using Amberlite XAD-2 resin and another one using a C18 cartridge. A gradient high performance liquid chromatography system with an RP18 column and photodiode array detector was utilized to analyze the extracted polyphenols. The mean percent of recovery from the C18 cartridges was 74.2%, while that from the Amberlite XAD-2 resin was 43.7%. The recoveries of vanillic acid, naringin, and rutin were excellent (>90%); however, gallic acid was not obtained when C18 cartridges were used. Additionally, the reusability of Amberlite XAD-2 resin was investigated, revealing that the mean recovery of polyphenols decreased from 43.7% (1st extraction) to 29.3% (3rd extraction). It was concluded that although Amberlite XAD-2 resin yielded a higher number of compounds, C18 cartridges gave a better extraction recovery. The lower recovery seen for the Amberlite XAD-2 resin also cannot be compensated by repeated extractions due to the gradual decrease of extraction recovery when reused.

Ionic Liquid-Modified Copper Phosphate Electrodes for the Detection of α-Amino Acids in a Weakly Alkaline Solution

The effect of ionic liquid (IL) coating on the electrochemical properties of preoxidized Cu3(PO4)2 (ox-Cu3(PO4)2) electrodes was explored in pH-varied Na2HPO4 solutions. The IL of N-propyl-N-methylpyrrolidinium bis(trifluoromethanesulfonyl)imide (PMPy-TFSI) was found to stabilize the redox properties of ox-Cu3(PO4)2 electrodes in weakly acid and alkaline solutions. Furthermore, the amperometric response of the PMPy-TFSI/ox-Cu3(PO4)2 electrodes presented good linearity in the range of 91–500 μM for most α-amino acids (AAs). The PMPy-TFSI/ox-Cu3(PO4)2 electrodes are equipped with a high performance liquid chromatography system to perform the good selectively and specific detection of AAs. The PMPy-TFSI coating shows promise for the wider application of ox-Cu3(PO4)2 electrodes in acidic and alkaline solutions for the detection of electroactive and nonelectroacitve AAs.

Simultaneous Method for Determination of Emtricitabine, Tenofovir Disoproxil Fumarate, Elvitegravir and Cobicistat in Tablets By HPLC

The present study provides us a single analytical tool for the determination of elvitegravir, cobicistat, emtricitabine and tenofovir disoproxil fumarate in dosage forms. This method helps us to separate and detect the four active pharmaceutical ingredients in a single run. The objective is to develop and validate a simple reverse phase liquid chromatography method for the simultaneous determination of emtricitabine, tenofovir disoproxil fumarate, elvitegravir and cobicistat in dosage tablets. For this method, an Agilent 1200, high performance liquid chromatography system, Atlantis C18 column (100×4.6 mm, 5 μm) as stationary phase and a gradient mixture of 0.1% trifluoroacetic acid and acetonitrile was used as mobile phase. The gradient program was adjusted at 1.0 ml/min flow rate and 10 μl injection volumes were maintained. The eluted compounds were monitored at 240 nm. The column oven temperature was maintained at 30°. The developed chromatographic method was validated for selectivity, linearity, precision, accuracy, sensitivity, robustness, and system suitability.

Femtoliter high-performance liquid chromatography using extended-nano channels.

A high-performance liquid chromatography system with 35 fL sample volume was developed using extended-nano (10-1000 nm) fluidic channels. For many years, miniaturization and enhancement of separation performance have been important issues in separation science. Recently, we have reported an ultimate miniaturization of chromatography using extended-nano channels with extremely high separation efficiency of 7 × 106 plates per m. However, the real theoretical plate number was limited to 103 due to the short nanochannel length. In this paper, the theoretical plate number was dramatically increased by developing a new high-pressure system with a very long nanochannel. A separation experiment of two fluorescent dyes demonstrated that the theoretical plate number could be improved to 1.4 × 104, which is much higher than that with conventional HPLC. The theoretical plate number is also comparable to those of capillary monolithic columns. The extremely small sample volume of extended-nano chromatography could support innovative analytical techniques capable of analyzing a single living cell in the near future.

Extraction and Quantification of Tryptophan and Kynurenine from Cultured Cells and Media Using a High Performance Liquid Chromatography (HPLC) System Equipped with an Ultra-Sensitive Diode Array Detector.

Evidence of the involvement of tryptophan and its metabolite, kynurenine, in various biological processes including cancer is constantly expanding. Analysis of cell extracts and culture media can allow for quick snapshots of the metabolic fluctuations occurring in vitro. Here, we describe a method for metabolite extraction from mammalian cells and analysis of extracted metabolites and cell culture media by HPLC with detection using an ultra-sensitive diode array detector.

High-fat simple carbohydrate feeding impairs central and peripheral monoamine metabolic pathway triggering the onset of metabolic syndrome in C57Bl/6J mice.

Previous studies have shown disturbances in an individual monoamine pathway but have not studied metabolic pathways at the onset and progression of metabolic syndrome (MetS). Aims, Settings, and Design: The aim of this study was to investigate the effect of high-fat simple carbohydrate (HFSC) diet on central (hypothalamic) and peripheral (plasma and urine) monoamine metabolic pathways during the development of metabolic syndrome in C57BL/6J mice. Monoamines were analyzed in the 1st, 2nd, 3rd, 4th, and 5th month after feeding mice the HFSC diet or the control diet using the high performance liquid chromatography (HPLC) system (Shimadzu, Japan). Data was statistically analyzed (by Student's t-test) using Graph Pad Instat Version 3.1. Post statistical analysis, Bonferroni correction was applied to the results of 2nd, 3rd, 4th, and 5th month in order to calculate the correct error in the study. Significantly lower hypothalamic, plasma, and urine dopamine, and higher hypothalamic and plasma levels of norepinephrine and normetanephrine levels were observed in the HFSC diet fed C57BL/6J mice as compared to the control diet fed C57BL/6J mice after 5 months of feeding. No consistent changes were observed in other brain regions. The turnover ratio indicated that the lower dopamine levels in the HFSC diet fed C57BL/6J mice was due to the increased formation of norepinephrine and homovanillic acid. HFSC diet impairs the central and peripheral dopaminergic and noradrenergic pathways in mice as evidenced by the disturbances in their hypothalamic, plasma, and urine levels and this might be one of the early factors contributing towards the development of the MetS.

Development of Hydrophilic Interaction Liquid Chromatography Method for the Analysis of Moxonidine and Its Impurities.

Fast and simple hydrophilic interaction liquid chromatography (HILIC) method was developed and validated for the analysis of moxonidine and its four impurities (A, B, C, and D) in pharmaceutical dosage form. All experiments were performed on the Agilent Technologies 1200 high-performance liquid chromatography (HPLC) system using Zorbax RX-SIL, 250 mm × 4.6 mm, 5 μm column as stationary phase (T = 25°C, F = 1 mL/min, and λ = 255 nm), and mixture of acetonitrile and 40 mM ammonium formate buffer (pH 2.8) 80 : 20 (v/v) as mobile phase. Under the optimal chromatographic conditions, selected by central composite design, separation and analysis of moxonidine and its four impurities are enabled within 12 minutes. Validation of the method was conducted in accordance with ICH guidelines. Based on the obtained results selectivity, linearity (r ≥ 0.9976), accuracy (recovery: 93.66%–114.08%), precision (RSD: 0.56%–2.55%), and robustness of the method were confirmed. The obtained values of the limit of detection and quantification revealed that the method can be used for determination of impurities levels below 0.1%. Validated method was applied for determination of moxonidine and its impurities in commercially available tablet formulation. Obtained results confirmed that validated method is fast, simple, and reliable for analysis of moxonidine and its impurities in tablets.

Characterization of forced degradation products and in silico toxicity prediction of Sofosbuvir: A novel HCV NS5B polymerase inhibitor

Sofosbuvir is a direct acting antiviral medication used to treat Hepatitis C viral infection. The present study focuses on the degradation behavior of the drug under various stress conditions (hydrolysis, oxidative, thermal and photolytic) as per International Conference on Harmonization (ICH Q1A (R2)) guidelines. A high performance liquid chromatographic system (HPLC) was used to develop a selective, precise and accurate method for separating all the degradation products. The separation was achieved on a Sunfire™ C18 (150mm×4.6mm×5μm) stationary phase with a mobile phase of 10mM ammonium acetate (pH 5.0) buffer and acetonitrile in gradient elution mode. A quadrupole-time of flight mass analyzer equipped with an electrospray ionization technique was used to propose the structural information based on the MS/MS and accurate mass measurements. Seven degradation products were identified and characterised by LC-ESI-QTOF–MS/MS. In silico toxicity of the drug and its degradation products was determined using TOPKAT and DEREK toxicity prediction softwares. The proposed method was validated as per the ICH Q2 guidelines.

A new method for rapid determination of indole-3-carbinol and its condensation products in nutraceuticals using core–shell column chromatography method

Indole-3-carbinol is a natural glucosinolate known for prevention of human breast, prostate and other types of cancer and it started to be used in commercial preparations, as food supplements. However no analytical method has been proposed for quality control of nutraceuticals with this substance yet. In this paper a new high-performance liquid chromatography (HPLC) method using core–shell column for separation of indole-3-carbinol and its condensation/degradation products was developed and used for the quantitative determination of indole-3-carbinol in nutraceuticals. Separation of indole-3-carbinol, its condensation/degradation products and internal standard ethylparaben was performed on the core–shell column Kinetex 5μ XB-C18 100A (100×4.6mm), particle size 5.0μm, with mobile phase acetonitrile/water according to the gradient program at a flow rate of 1.25mLmin−1 and at temperature 50°C. The detection wavelength was set at 270nm. Under the optimal chromatographic conditions good linearity of determination was achieved. Available commercial samples of nutraceuticals were extracted with 100% methanol using ultrasound bath. A 5-μL sample volume of the supernatant was directly injected into the HPLC system. The developed method provided rapid and accurate tool for quality control of nutraceuticals based on cruciferous vegetable extracts with indole-3-carbinol content. The presented study showed that the declared content of indole-3-carbinol significantly varied in the different nutraceuticals available on the market. Two analyzed preparations showed the presence of condensation/degradation products of indole-3-carbinol which were not officially declared by the manufacturer. Moreover, further two analyzed nutraceutical preparations showed absolutely no content of declared amount of indole-3-carbinol.

고체상추출법과 HPLC/MSD/FLD를 이용한 수질중의 페놀 분석

Abstract: An analytical method for determining phenol considered priority pollutants of the US EPA andprecursor of toxic phenolic compounds by solid-phase extraction (SPE) and high performance liquidchromatographic systems (HPLC) equipped with fluorescence and mass selective detectors have been developed.The SPE process for sample preconcentration was performed on a commercially available Oasis HLB cartridgepacked with polymeric sorbents. The effect of pH, elution solvent, and elution volume on the recoveries ofthe analytes were investigated with HPLC/FLD. Average recovery of >87.0% was achieved with 60 mg sorbentsusing 5 mL of methanol as an elution solvent at pH=3.요약: Priority pollutant이며 페놀성화합물의 전구체인 페놀을 고체상추출법을 이용하여 추출 정제하여형광검출기와 질량분석기가 각각 장착된 고속액체크로마토그래프를 이용하여 분석하는 방법을 설정하여보았다. 고체상추출은 고분자성물질이 충진된 Oasis HLB cartridge를 이용하여 최적의 조건을 확립하였으며 이때의 평균회수율은 87.0% 이었다. 형광검출기를 이용한 경우가 질량분석기를 이용한 경우보다검출한계가 낮았으나 선택성의 측면에서 질량분석기가 우수하였다. Key words: phenol, hplc, solid phase extraction(spe), hplc/msd

Main Leaf Polyphenolic Components of Berry Color Variant Grapevines and Their Acclimative Responses to Sunlight Exposure

Grapevine leaf synthesizes a wide variety of bioactive secondary metabolites, including polyphenols, which are also key components in ensuring development and growth of the whole plant even under adverse environmental conditions. Our study evaluates the nonanthocyanin polyphenolic composition in grapevine leaves of three varieties of Gohér conculta (Vitis vinifera L.) native to Hungary. A high performance liquid chromatography (HPLC) system including a diode array detector (DAD) coupled to a time-of-flight mass spectrometer (q-TOFMS) was successfully applied to profile intact glycoconjugate forms in samples. In-source fragmentation was utilized in order to provide structural information on the compounds. Using this method, the presence of 16 polyphenolic metabolites were confirmed, and eight of them were subjected to further quantification in sun acclimated and half shaded leaves. Intracellular microimaging detected accumulation of flavonols in cell nuclei, cell wall and chloroplasts. Our findings demonstrated that Gohér conculta—a special grapevine taxon of our viticultural heritage with berry color variants—is a suitable model to study the interaction between genetic and environmental factors in determination of grapevine phenolic composition.

Caffeine and Creatine Content of Dietary Supplements Consumed by Brazilian Soccer Players.

Caffeine and creatine are ingredients in the most popular dietary supplements consumed by soccer players. However, some products may not contain the disclosed amounts of the ingredients listed on the label, compromising the safe usage and the effectiveness of these supplements. Therefore, the aim of this study was to evaluate the content of caffeine and creatine in dietary supplements consumed by Brazilian soccer players. The results obtained were compared with the caffeine content listed on the product label. Two batches of the supplement brands consumed by ≥ 50% of the players were considered for analysis. The quantification of caffeine and creatine in the supplements was determined by a high-performance liquid chromatography system with UV detector. Nine supplements of caffeine and 7 supplements of creatine met the inclusion criteria for analysis. Eight brands of caffeine and five brands of creatine showed significantly different values (p < .05) as compared with the values stated on the label. There were no significant differences between the two batches of supplements analyzed, except for one caffeine supplement. It can be concluded that caffeine and creatine dietary supplements consumed by Brazilian soccer players present inaccurate values listed on the label, although most presented no difference among batches. To ensure consumer safety and product efficacy, accurate information on caffeine and creatine content should be provided on all dietary supplement labels.

HPLC Method for Chemical Fingerprinting of Guggul (Commiphora wightii)--Quantification of E- and Z-Guggulsterones and Detection of Possible Adulterants.

A reversed-phase high-performance liquid chromatography method was developed for the chemical fingerprinting of Commiphora wightii gum resin (guggul). This method was also used for the quantification of E- and Z-guggulsterones in different C. wightii gum resin samples. The analysis was conducted on a high-performance liquid chromatography system with a reversed-phase column using a gradient elution comprised of water and acetonitrile with 0.1% acetic acid, and a run time of 50 min at 38 °C. The calibration curve of E- and Z-guggulsterones showed good linearity with a standard correlation coefficient >0.999, which is within the established range (0.5-250 µg/mL). Twelve marker compounds were selected and successfully analyzed by this method in 22 C. wightii gum resin samples along with 9 gum resin samples of possible adulterant species.

Beetroot juice increase nitric oxide metabolites in both men and women regardless of body mass

The nitrate (NO3−) present in beetroot juice (BJ) has been studied for its effect on the cardiovascular system by converting to nitric oxide (NO). In the present study, we evaluated the effect of BJ on the excretion of NO metabolites and its relationship with body mass in both men and women. NO metabolites – urinary NO3−, nitrite (NO2−) and NOx were analyzed by using a high-performance liquid chromatography system. There were significant increases in urinary NO3−, NO2− and NOx in BJ as compared to PLA (BJ without NO3−). No significant difference between men and women was observed in NO metabolites after BJ at any time point. There were no significant relationships between urinary NO3−, NO2− and NOx and body mass in BJ intervention for both men and women. In conclusion, urinary NO metabolites after BJ consumption increases in similar manner between sexes regardless of body mass.