2024

Jump to: 2023, 2022, 2021, 2020, 2019, 2018

15 pages, 2918 KiB

Open AccessArticle

HPLC and LC–MS/MS-Based Quantitative Characterization of Related Substances Associated with Sotalol Hydrochloride

by Pengyan Zhu, Xiaojing Shen, Xinting Wang, Xinlan Liu, Yingshuang Zhang, Ke Wang, Wenfen Gao, Xuanjun Wang and Wenjuan Yuan

Molecules 2024, 29(3), 588; https://doi.org/10.3390/molecules29030588 - 25 Jan 2024

Viewed by 705

Abstract

In total, three related substances (RS) associated with sotalol hydrochloride (STHCl) were herein identified with a novel gradient high-performance liquid chromatography (HPLC) protocol. Further characterization of these substances was then performed via liquid chromatography–mass spectroscopy (LC–MS/MS) and nuclear magnetic resonance (NMR) approaches. For [...] Read more.

In total, three related substances (RS) associated with sotalol hydrochloride (STHCl) were herein identified with a novel gradient high-performance liquid chromatography (HPLC) protocol. Further characterization of these substances was then performed via liquid chromatography–mass spectroscopy (LC–MS/MS) and nuclear magnetic resonance (NMR) approaches. For these analyses, commercial STHCl samples were used for quantitative HPLC studies and the degradation of STHCl under acidic (1M HCl), alkaline (1M NaOH), oxidative (30% H₂O₂), photolytic (4500 Lx), and thermal stress conditions (100 °C) was assessed. This approach revealed this drug to be resistant to acidic, alkaline, and high-temperature conditions, whereas it was susceptible to light and oxidation as confirmed through long-term experiments. The putative mechanisms governing RS formation were also explored, revealing that RS3 was derived from the manufacturing process, whereas RS2 was generated via oxidation and RS1 was generated in response to light exposure. The cytotoxicity of these RS compounds was then assessed using MTT assays and acute toxicity test. Overall, this study provides details regarding the characterization, isolation, quantification, and toxicological evaluation of STHCl and associated RS compounds together with details regarding the precise, specific, and reliable novel HPLC technique, thus providing the requisite information necessary to ensure STHCl purity and safety. Full article

► Show Figures

Figure 1

2023

Jump to: 2024, 2022, 2021, 2020, 2019, 2018

10 pages, 1956 KiB

Open AccessArticle

A Novel Chromatographic Method to Assess the Binding Ability towards Dicarbonyls

by Angelica Artasensi, Emanuele Salina, Laura Fumagalli and Luca Regazzoni

Molecules 2023, 28(14), 5341; https://doi.org/10.3390/molecules28145341 - 11 Jul 2023

Viewed by 831

Abstract

Human exposure to dicarbonyls occurs via ingestion (e.g., food), inhalation (e.g., electronic cigarettes) and dysregulation of endogenous metabolic pathways (e.g., glycolysis). Dicarbonyls are electrophiles able to induce carbonylation of endogenous substrate. They have been associated with the onset and progression of several human [...] Read more.

Human exposure to dicarbonyls occurs via ingestion (e.g., food), inhalation (e.g., electronic cigarettes) and dysregulation of endogenous metabolic pathways (e.g., glycolysis). Dicarbonyls are electrophiles able to induce carbonylation of endogenous substrate. They have been associated with the onset and progression of several human diseases. Several studies have advocated the use of dicarbonyl binders as food preservatives or as drugs aimed at mitigating carbonylation. This study presents the setup of an easy and cheap assay for the screening of selective and potent dicarbonyl binders. The method is based on the incubation of the candidate molecules with a molecular probe. The activity is then determined by measuring the residual concentration of the molecular probe over time by liquid chromatography (LC). However, the naturally occurring dicarbonyls (e.g., glyoxal, methylglyoxal) are not appealing as probes since they are hard to separate and detect using the most popular LC variants. Benzylglyoxal (BGO) was therefore synthesized and tested, proving to be a convenient probe that allows a direct quantification of residual dicarbonyls by reversed phase LC without derivatization. The method was qualified by assessing the binding ability of some molecules known as binders of natural occurring dicarbonyls, obtaining results consistent with literature. Full article

► Show Figures

Figure 1

2022

Jump to: 2024, 2023, 2021, 2020, 2019, 2018

13 pages, 1309 KiB

Open AccessArticle

A Rapid and Sensitive Liquid Chromatography-Tandem Mass Spectrometry Bioanalytical Method for the Quantification of Encorafenib and Binimetinib as a First-Line Treatment for Advanced (Unresectable or Metastatic) Melanoma—Application to a Pharmacokinetic Study

by Mohamed M. Hefnawy, Mohammed M. Alanazi, Abdullah M. Al-Hossaini, Abdulaziz I. Alnasser, Adel S. El-Azab, Yousef A. Bin Jardan, Mohamed W. Attwa and Manal A. El-Gendy

Molecules 2023, 28(1), 79; https://doi.org/10.3390/molecules28010079 - 22 Dec 2022

Cited by 4 | Viewed by 2121

Abstract

The combination regimen targeting BRAF and MEK inhibition, for instance, encorafenib (Braftovi^™, ENF) plus binimetinib (Mektovi^®, BNB), are now recommended as first-line treatment in patients with unresectable or metastatic melanoma with a BRAF V600-activating mutation. Patients treated with combination [...] Read more.

The combination regimen targeting BRAF and MEK inhibition, for instance, encorafenib (Braftovi^™, ENF) plus binimetinib (Mektovi^®, BNB), are now recommended as first-line treatment in patients with unresectable or metastatic melanoma with a BRAF V600-activating mutation. Patients treated with combination therapy of ENF and BNB demonstrated a delay in resistance development, increases in antitumor activity, and attenuation of toxicities compared with the activity of either agent alone. However, the pharmacokinetic profile of the FDA-approved ENF and BNB is still unclear. In this study, a rapid and sensitive LC-MS/MS bioanalytical method for simultaneous quantification of ENF and BNB in rat plasma was developed and validated. Chromatography was performed on an Agilent Eclipse plus C18 column (50 mm × 2.1 mm, 1.8 µm), with an isocratic mobile phase composed of 0.1% formic acid in water/acetonitrile (67:33, v/v, pH 3.2) at a flow rate of 0.35 mL/min. A positive multiple reaction monitoring (MRM) mode was chosen for detection and the process of analysis was run for 2 min. Plasma samples were pre-treated using protein precipitation with acetonitrile containing spebrutinib as the internal standard (IS). Method validation was assessed as per the FDA guidelines for the determination of ENF and BNB over concentration ranges of 0.5–3000 ng/mL (r² ≥ 0.997) for each drug (plasma). The lower limits of detection (LLOD) for both drugs were 0.2 ng/mL. The mean relative standard deviation (RSD) of the results for accuracy and precision was ≤ 7.52%, and the overall recoveries of ENF and BNB from rat plasma were in the range of 92.88–102.28%. The newly developed approach is the first LC–MS/MS bioanalytical method that can perform simultaneous quantification of ENF and BNB in rat plasma and its application to a pharmacokinetic study. The mean result for C_max for BNB and ENF was found to be 3.43 ± 0.46 and 16.42 ± 1.47 µg/mL achieved at 1.0 h for both drugs, respectively. The AUC_0-∞ for BNB and ENF was found to be 18.16 ± 1.31 and 36.52 ± 3.92 µg/mL.h, respectively. On the other hand, the elimination half-life (t_1/2kel) parameters for BNB and ENF in the rat plasma were found to be 3.39 ± 0.43 h and 2.48 ± 0.24 h, and these results are consistent with previously reported values. Full article

► Show Figures

Figure 1

15 pages, 2830 KiB

Open AccessArticle

Structural Elucidation of Alkali Degradation Impurities of Favipiravir from the Oral Suspension: UPLC-TQ-ESI-MS/MS and NMR

by Ravi Patel, Abhishek Dube, Ravisinh Solanki, Dignesh Khunt, Shalin Parikh, Vijayabhaskarreddy Junnuthula and Sathish Dyawanapelly

Molecules 2022, 27(17), 5606; https://doi.org/10.3390/molecules27175606 - 31 Aug 2022

Cited by 6 | Viewed by 2029

Abstract

A novel stability-indicating, reversed-phase, high-performance liquid chromatography (RP-HPLC) method was developed and validated for the determination of favipiravir in an oral suspension. The effective separation of favipiravir and its degradation products was achieved on a Zorbax Eclipse Plus C18 column (5 μm particle [...] Read more.

A novel stability-indicating, reversed-phase, high-performance liquid chromatography (RP-HPLC) method was developed and validated for the determination of favipiravir in an oral suspension. The effective separation of favipiravir and its degradation products was achieved on a Zorbax Eclipse Plus C18 column (5 μm particle size, 150 mm length × 4.6 mm diameter). The mobile phase was prepared by mixing 5 mM of phosphate buffer (pH 3.5) and methanol in a 75:25 v/v ratio delivered at a 1.0 mL/min flow rate. The eluents were monitored using a photodiode array detector at a wavelength of 322 nm. The stability-indicating nature of this method was evaluated by performing force degradation studies under various stress conditions, such as acidic, alkali, oxidative, thermal, and photolytic degradation. Significant degradation was observed during the alkali stress degradation condition. The degradation products generated during various stress conditions were well separated from the favipiravir peak. In addition, the major degradation product formed under alkali stress conditions was identified using UPLC-ESI-TQ-MS/MS and NMR. Method validation was performed according to the ICH Q2 (R1) guideline requirements. The developed method is simple, accurate, robust, and reliable for routine quality control analysis of favipiravir oral suspensions. Full article

► Show Figures

Graphical abstract

21 pages, 2737 KiB

Open AccessArticle

Development of the Validated Stability-Indicating Method for the Determination of Vortioxetine in Bulk and Pharmaceutical Formulation by HPLC-DAD, Stress Degradation Kinetics Studies and Detection of Degradation Products by LC-ESI-QTOF-MS

by Karol Wróblewski, Małgorzata Szultka-Młyńska, Daria Janiszewska, Anna Petruczynik and Bogusław Buszewski

Molecules 2022, 27(6), 1883; https://doi.org/10.3390/molecules27061883 - 14 Mar 2022

Cited by 2 | Viewed by 2975

Abstract

Vortioxetine (VOR) is a new antidepressant drug used to treat major depressive disorder. In this work, a novel, simple, rapid, accurate, precise, selective, stability-indicating, and fully validated high-performance liquid chromatography method with diode array detection (HPLC-DAD) was developed to determine VOR in bulk [...] Read more.

Vortioxetine (VOR) is a new antidepressant drug used to treat major depressive disorder. In this work, a novel, simple, rapid, accurate, precise, selective, stability-indicating, and fully validated high-performance liquid chromatography method with diode array detection (HPLC-DAD) was developed to determine VOR in bulk and pharmaceutical formulations. A Polar-RP column was used, with a mobile phase consisting of acetonitrile (ACN), methanol (MeOH), acetate buffer pH 3.5, and addition of diethylamine (DEA) in the isocratic elution mode. Assessing the stability of the VOR is fundamental to guarantee the efficacy, safety, and quality of drug products. In this study, the VOR active pharmaceutical ingredient (API) and tablets were subjected to a detailed study of forced degradation, using several degrading agents (acid, alkaline, water, heat, light, and oxidation agents). The developed HPLC-DAD method allows the collection of all the essential data to determine degradation kinetics. It was found that the decomposition of vortioxetine is fragile towards oxidative conditions and photolysis, yielding the first-order and second-order kinetic reaction in the above stress conditions, respectively. The degradation products (DPs) were identified by the high-resolution liquid chromatography coupled with electrospray ionization-quadrupole-time of flight-mass spectrometry (LC-ESI-QTOF-MS) method. The HPLC-DAD method was successfully applied for the quantification of VOR in tablets. Additionally, in silico toxicity prediction of the DPs was performed. Full article

► Show Figures

Graphical abstract

14 pages, 1298 KiB

Open AccessArticle

Comparison Studies on Several Ligands Used in Determination of Cd(II) in Rice by Flame Atomic Absorption Spectrometry after Ultrasound-Assisted Dispersive Liquid–Liquid Microextraction

by Qian Sun, Xinyu Cui, Yanfeng Wang, Pingping Zhang and Wenjuan Lu

Molecules 2022, 27(3), 590; https://doi.org/10.3390/molecules27030590 - 18 Jan 2022

Cited by 5 | Viewed by 1550

Abstract

Ligands plays an important role in the extraction procedures for the determination of cadmium in rice samples by using flame atomic absorption spectrometry (FAAS). In the present study, comparative evaluation of 10 commercially available ligands for formation of Cd(II)-ligand complex and determination of [...] Read more.

Ligands plays an important role in the extraction procedures for the determination of cadmium in rice samples by using flame atomic absorption spectrometry (FAAS). In the present study, comparative evaluation of 10 commercially available ligands for formation of Cd(II)-ligand complex and determination of cadmium in rice samples by ultrasound-assisted dispersive liquid–liquid microextraction (UADLLME) combined with FAAS was developed. Sodium diethyldithiocarbamate (DDTC) provided a high distribution coefficient as well as a good absorbance signal, therefore DDTC was used as a ligand in UADLLME. A low density and less toxic solvent, 1-heptanol, was used as the extraction solvent and ethanol was used as the disperser solvent. In addition, the experimental conditions of UADLLME were optimized in standard solution first and then applied in rice, such as the type and volume of extractant and dispersant, pH, extraction time, and temperature. Under the optimal experimental conditions, the detection limit (3σ) was 0.69 μg/L for Cd(II). The proposed method was applied for the determination of Cd(II) in three different rice samples (polished rice, brown rice, and glutinous rice), the recovery test was carried out, and the results ranged between 96.7 to 113.6%. The proposed method has the advantages of simplicity, low cost, and accurate and was successfully applied to analyze Cd(II) in rice. Full article

► Show Figures

Figure 1

2021

Jump to: 2024, 2023, 2022, 2020, 2019, 2018

10 pages, 726 KiB

Open AccessArticle

A Rapid and Simple UHPLC-MS/MS Method for Quantification of Plasma Globotriaosylsphingosine (lyso-Gb3)

by Alessandro Perrone, Susan Mohamed, Vincenzo Donadio, Rocco Liguori and Manuela Contin

Molecules 2021, 26(23), 7358; https://doi.org/10.3390/molecules26237358 - 03 Dec 2021

Cited by 3 | Viewed by 2201

Abstract

Fabry disease (FD) is a rare X-linked lysosomal storage disorder caused by α-galactosidase A gene (GLA) mutations, resulting in loss of activity of the lysosomal hydrolase, α-galactosidase A (α-Gal A). As a result, the main glycosphingolipid substrates, globotriaosylceramide (Gb3) and globotriaosylsphingosine (lyso-Gb3), accumulate [...] Read more.

Fabry disease (FD) is a rare X-linked lysosomal storage disorder caused by α-galactosidase A gene (GLA) mutations, resulting in loss of activity of the lysosomal hydrolase, α-galactosidase A (α-Gal A). As a result, the main glycosphingolipid substrates, globotriaosylceramide (Gb3) and globotriaosylsphingosine (lyso-Gb3), accumulate in plasma, urine, and tissues. Here, we propose a simple, fast, and sensitive method for plasma quantification of lyso-Gb3, the most promising secondary screening target for FD. Assisted protein precipitation with methanol using Phree cartridges was performed as sample pre-treatment and plasma concentrations were measured using UHPLC-MS/MS operating in MRM positive electrospray ionization. Method validation provided excellent results for the whole calibration range (0.25–100 ng/mL). Intra-assay and inter-assay accuracy and precision (CV%) were calculated as <10%. The method was successfully applied to 55 plasma samples obtained from 34 patients with FD, 5 individuals carrying non-relevant polymorphisms of the GLA gene, and 16 healthy controls. Plasma lyso-Gb3 concentrations were larger in both male and female FD groups compared to healthy subjects (p < 0.001). Normal levels of plasma lyso-Gb3 were observed for patients carrying non-relevant mutations of the GLA gene compared to the control group (p = 0.141). Dropping the lower limit of quantification (LLOQ) to 0.25 ng/mL allowed us to set the optimal plasma lyso-Gb3 cut-off value between FD patients and healthy controls at 0.6 ng/mL, with a sensitivity of 97.1%, specificity of 100%, and accuracy of 0.998 expressed by the area under the ROC curve (C.I. 0.992 to 1.000, p-value < 0.001). Based on the results obtained, this method can be a reliable tool for early phenotypic assignment, assessing diagnoses in patients with borderline GalA activity, and confirming non-relevant mutations of the GLA gene. Full article

► Show Figures

Figure 1

11 pages, 1007 KiB

Open AccessArticle

Characterization of the Trans-Epithelial Transport of Green Tea (C. sinensis) Catechin Extracts with In Vitro Inhibitory Effect against the SARS-CoV-2 Papain-like Protease Activity

by Carmela Maria Montone, Sara Elsa Aita, Anna Arnoldi, Anna Laura Capriotti, Chiara Cavaliere, Andrea Cerrato, Carmen Lammi, Susy Piovesana, Giulia Ranaldi and Aldo Laganà

Molecules 2021, 26(21), 6744; https://doi.org/10.3390/molecules26216744 - 08 Nov 2021

Cited by 9 | Viewed by 1969

Abstract

This work describes an untargeted analytical approach for the screening, identification, and characterization of the trans-epithelial transport of green tea (Camellia sinensis) catechin extracts with in vitro inhibitory effect against the SARS-CoV-2 papain-like protease (PLpro) activity. After specific catechin extraction, a [...] Read more.

This work describes an untargeted analytical approach for the screening, identification, and characterization of the trans-epithelial transport of green tea (Camellia sinensis) catechin extracts with in vitro inhibitory effect against the SARS-CoV-2 papain-like protease (PLpro) activity. After specific catechin extraction, a chromatographic separation obtained six fractions were carried out. The fractions were assessed in vitro against the PLpro target. Fraction 5 showed the highest inhibitory activity against the SARS-CoV-2 PLpro (IC₅₀ of 0.125 μg mL⁻¹). The untargeted characterization revealed that (−)-epicatechin-3-gallate (ECG) was the most abundant compound in the fraction and the primary molecule absorbed by differentiated Caco-2 cells. Results indicated that fraction 5 was approximately 10 times more active than ECG (IC₅₀ value equal to 11.62 ± 0.47 μg mL⁻¹) to inhibit the PLpro target. Overall, our findings highlight the synergistic effects of the various components of the crude extract compared to isolated ECG. Full article

► Show Figures

Graphical abstract

19 pages, 4054 KiB

Open AccessArticle

High-Throughput Identification of Organic Compounds from Polygoni Multiflori Radix Praeparata (Zhiheshouwu) by UHPLC-Q-Exactive Orbitrap-MS

by Shaoyun Wang, Xiaozhu Sun, Shuo An, Fang Sang, Yunli Zhao and Zhiguo Yu

Molecules 2021, 26(13), 3977; https://doi.org/10.3390/molecules26133977 - 29 Jun 2021

Cited by 14 | Viewed by 2349

Abstract

Polygoni Multiflori Radix Praeparata (PMRP), as the processed product of tuberous roots of Polygonum multiflorum Thunb., is one of the most famous traditional Chinese medicines, with a long history. However, in recent years, liver adverse reactions linked to PMRP have been frequently reported. [...] Read more.

Polygoni Multiflori Radix Praeparata (PMRP), as the processed product of tuberous roots of Polygonum multiflorum Thunb., is one of the most famous traditional Chinese medicines, with a long history. However, in recent years, liver adverse reactions linked to PMRP have been frequently reported. Our work attempted to investigate the chemical constituents of PMRP for clinical research and safe medication. In this study, an effective and rapid method was established to separate and characterize the constituents in PMRP by combining ultra-high performance liquid chromatography with hybrid quadrupole-orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap-MS). Based on the accurate mass measurements for molecular and characteristic fragment ions, a total of 103 compounds, including 24 anthraquinones, 21 stilbenes, 15 phenolic acids, 14 flavones, and 29 other compounds were identified or tentatively characterized. Forty-eight compounds were tentatively characterized from PMRP for the first time, and their fragmentation behaviors were summarized. There were 101 components in PMRP ethanol extract (PMRPE) and 91 components in PMRP water extract (PMRPW). Simultaneously, the peak areas of several potential xenobiotic components were compared in the detection, which showed that PMRPE has a higher content of anthraquinones and stilbenes. The obtained results can be used in pharmacological and toxicological research and provided useful information for further in vitro and in vivo studies. Full article

► Show Figures

Graphical abstract

14 pages, 7386 KiB

Open AccessArticle

Development and Validation of a Chiral Liquid Chromatographic Assay for Enantiomeric Separation and Quantification of Verapamil in Rat Plasma: Stereoselective Pharmacokinetic Application

by Mostafa S. Mohammed, Mohamed M. Hefnawy, Abdulrhman A. Al-Majed, Haitham K. Alrabiah, Nasser A. Algrain, Ahmad J. Obaidullah, Abdulmalik S. Altamimi, Yousef A. Bin Jardan and Abdullah M. Al-Hossaini

Molecules 2021, 26(7), 2091; https://doi.org/10.3390/molecules26072091 - 06 Apr 2021

Cited by 5 | Viewed by 2241

Abstract

A novel, fast and sensitive enantioselective HPLC assay with a new core–shell isopropyl carbamate cyclofructan 6 (superficially porous particle, SPP) chiral column (LarihcShell-P, LSP) was developed and validated for the enantiomeric separation and quantification of verapamil (VER) in rat plasma. The polar organic [...] Read more.

A novel, fast and sensitive enantioselective HPLC assay with a new core–shell isopropyl carbamate cyclofructan 6 (superficially porous particle, SPP) chiral column (LarihcShell-P, LSP) was developed and validated for the enantiomeric separation and quantification of verapamil (VER) in rat plasma. The polar organic mobile phase composed of acetonitrile/methanol/trifluoroacetic acid/triethylamine (98:2:0.05: 0.025, v/v/v/v) and a flow rate of 0.5 mL/min was applied. Fluorescence detection set at excitation/emission wavelengths 280/313 nm was used and the whole analysis process was within 3.5 min, which is 10-fold lower than the previous reported HPLC methods in the literature. Propranolol was selected as the internal standard. The S-(−)- and R-(+)-VER enantiomers with the IS were extracted from rat plasma by utilizing Waters Oasis HLB C18 solid phase extraction cartridges without interference from endogenous compounds. The developed assay was validated following the US-FDA guidelines over the concentration range of 1–450 ng/mL (r² ≥ 0.997) for each enantiomer (plasma) and the lower limit of quantification was 1 ng/mL for both isomers. The intra- and inter-day precisions were not more than 11.6% and the recoveries of S-(−)- and R-(+)-VER at all quality control levels ranged from 92.3% to 98.2%. The developed approach was successfully applied to the stereoselective pharmacokinetic study of VER enantiomers after oral administration of 10 mg/kg racemic VER to Wistar rats. It was found that S-(−)-VER established higher C_max and area under the concentration-time curve (AUC) values than the R-(+)-enantiomer. The newly developed approach is the first chiral HPLC for the enantiomeric separation and quantification of verapamil utilizing a core–shell isopropyl carbamate cyclofructan 6 chiral column in rat plasma within 3.5 min after solid phase extraction (SPE). Full article

► Show Figures

Figure 1

2020

Jump to: 2024, 2023, 2022, 2021, 2019, 2018

12 pages, 994 KiB

Open AccessCommunication

Establishment of a UPLC-PDA/ESI-Q-TOF/MS-Based Approach for the Simultaneous Analysis of Multiple Phenolic Compounds in Amaranth (A. cruentus and A. tricolor)

by Won Tea Jeong, Jun-Hyoung Bang, Seahee Han, Tae Kyung Hyun, Hyunwoo Cho, Heung Bin Lim and Jong-Wook Chung

Molecules 2020, 25(23), 5674; https://doi.org/10.3390/molecules25235674 - 01 Dec 2020

Cited by 6 | Viewed by 3351

Abstract

We used ultraperformance liquid chromatography coupled with a photodiode-array detector and electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-PDA/ESI-Q-TOF/MS) to rapidly and accurately quantify 17 phenolic compounds. Then, we applied this method to the seed and leaf extracts of two Amaranthus species to identify [...] Read more.

We used ultraperformance liquid chromatography coupled with a photodiode-array detector and electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-PDA/ESI-Q-TOF/MS) to rapidly and accurately quantify 17 phenolic compounds. Then, we applied this method to the seed and leaf extracts of two Amaranthus species to identify and quantify phenolic compounds other than the 17 compounds mentioned above. Compounds were eluted within 30 min on a C18 column using a mobile phase (water and acetonitrile) containing 0.1% formic acid, and the specific wavelength and ion information of the compounds obtained by PDA and ESI-Q-TOF/MS were confirmed. The proposed method showed good linearity (r² > 0.990). Limits of detection and quantification were less than 0.1 and 0.1 μg/mL, respectively. Intra- and interday precision were less than 2.4% and 1.8%, respectively. Analysis of amaranth seed and leaf extracts using the established method showed that the seeds contained high amounts of 2,4-dihydroxybenzoic acid and kaempferol, and leaves contained diverse phenolic compounds. In addition, six tentatively new phenolic compounds were identified. Moreover, seeds potentially contained 2,3-dihydroxybenzaldehyde, a beneficial bioactive compound. Thus, our method was an efficient approach for the qualitative and quantitative analysis of phenolic compounds, and could be used to investigate phenolic compounds in plants. Full article

► Show Figures

Figure 1

15 pages, 2836 KiB

Open AccessArticle

Untargeted Characterization of Chestnut (Castanea sativa Mill.) Shell Polyphenol Extract: A Valued Bioresource for Prostate Cancer Cell Growth Inhibition

by Nunzio Antonio Cacciola, Andrea Cerrato, Anna Laura Capriotti, Chiara Cavaliere, Maria D’Apolito, Carmela Maria Montone, Susy Piovesana, Giuseppe Squillaci, Gianfranco Peluso and Aldo Laganà

Molecules 2020, 25(12), 2730; https://doi.org/10.3390/molecules25122730 - 12 Jun 2020

Cited by 18 | Viewed by 3084

Abstract

Chestnut seeds are used for fresh consumption and for the industrial preparation of derivatives, such as chestnut flour. During industrial processing, large amounts of by-products are generally produced, such as leaves, flowers, shells and burs. In the present study, chestnut shells were extracted [...] Read more.

Chestnut seeds are used for fresh consumption and for the industrial preparation of derivatives, such as chestnut flour. During industrial processing, large amounts of by-products are generally produced, such as leaves, flowers, shells and burs. In the present study, chestnut shells were extracted by boiling water in order to obtain polyphenol-rich extracts. Moreover, for the removal or non-phenolic compounds, a separation by preparative reverse phase chromatography in ten fractions was carried out. The richest fractions in terms of phenolic content were characterized by means of untargeted high-resolution mass spectrometric analysis together with a dedicated and customized data processing workflow. A total of 243 flavonoids, phenolic acids, proanthocyanidins and ellagitannins were tentatively identified in the five richest fractions. Due its high phenolic content (450.03 µg GAE per mg of fraction), one tumor cell line (DU 145) and one normal prostate epithelial cell line (PNT2) were exposed to increasing concentration of fraction 3 dry extract for 24, 48 and 72 h. Moreover, for DU 145 cell lines, increase of apoptotic cells and perturbation of cell cycle was demonstrated for the same extract. Those outcomes suggest that chestnut industrial by-products could be potentially employed as a source of bioresources. Full article

► Show Figures

Graphical abstract

2019

Jump to: 2024, 2023, 2022, 2021, 2020, 2018

13 pages, 1049 KiB

Open AccessArticle

Peptidomic Approach for the Identification of Peptides with Potential Antioxidant and Anti-Hyperthensive Effects Derived From Asparagus By-Products

by Carmela Maria Montone, Riccardo Zenezini Chiozzi, Nicola Marchetti, Andrea Cerrato, Michela Antonelli, Anna Laura Capriotti, Chiara Cavaliere, Susy Piovesana and Aldo Laganà

Molecules 2019, 24(19), 3627; https://doi.org/10.3390/molecules24193627 - 08 Oct 2019

Cited by 25 | Viewed by 3882

Abstract

Asparagus waste represents products of great interest since many compounds with high biological value are located in the lower portion of the spears. The extraction of bioactive compounds from asparagus by-products is therefore crucial for the purpose of adding value to these by-products. [...] Read more.

Asparagus waste represents products of great interest since many compounds with high biological value are located in the lower portion of the spears. The extraction of bioactive compounds from asparagus by-products is therefore crucial for the purpose of adding value to these by-products. In this paper, bioactive peptides from asparagus waste were extracted, digested, purified and identified. In particular, Alcalase^® was chosen as the enzyme to use to obtain protein hydrolysate due to its low cost and, consequently, the possibility of implementing the method on a large scale. In order to simplify the peptide extract to reach better identification, the hydrolysate was fractionated by reversed-phase chromatography in 10 fractions. Two tests were carried out for antioxidant activity (ABTS-DPPH) and one for antihypertensive activity (ACE). Fractions with a higher bioactivity score were identified by peptidomics technologies and screened for bioactivity with the use of bioinformatics. For ACE-inhibitor activity, two peptides were synthetized, PDWFLLL and ASQSIWLPGWL, which provided an EC₅₀ value of 1.76 µmol L⁻¹ and 4.02 µmol L⁻¹, respectively. For the antioxidant activity, by DPPH assay, MLLFPM exhibited the lowest EC50 value at 4.14 µmol L⁻¹, followed by FIARNFLLGW and FAPVPFDF with EC50 values of 6.76 µmol L⁻¹ and 10.01 µmol L⁻¹, respectively. A validation of the five identified peptides was also carried out. The obtained results showed that peptides obtained from asparagus by-products are of interest for their biological activity and are suitable for being used as functional ingredients. Full article

► Show Figures

Graphical abstract

12 pages, 1855 KiB

Open AccessArticle

Identification of Potential Auxin-Responsive Small Signaling Peptides through a Peptidomics Approach in Arabidopsis thaliana

by Weigui Luo, Yuan Xiao, Qiwen Liang, Yi Su and Langtao Xiao

Molecules 2019, 24(17), 3146; https://doi.org/10.3390/molecules24173146 - 29 Aug 2019

Cited by 9 | Viewed by 3719

Abstract

Small signaling peptides (SSPs) are a class of short peptides playing critical roles in plant growth and development. SSPs are also involved in the phytohormone signaling pathway. However, identification of mature SSPs is still a technical challenge because of their extremely low concentrations [...] Read more.

Small signaling peptides (SSPs) are a class of short peptides playing critical roles in plant growth and development. SSPs are also involved in the phytohormone signaling pathway. However, identification of mature SSPs is still a technical challenge because of their extremely low concentrations in plant tissue and complicated interference by many other metabolites. Here, we report an optimized protocol to extract SSPs based on protoplast extraction and to analyze SSPs based on tandem mass spectrometry peptidomics. Using plant protoplasts as the material, soluble peptides were directly extracted into phosphate buffer. The interference of non-signaling peptides was significantly decreased. Moreover, we applied the protocol to identify potential SSPs in auxin treated wild type and auxin biosynthesis defective mutant yuc2yuc6. Over 100 potential SSPs showed a response to auxin in Arabidopsis thaliana. Full article

► Show Figures

Figure 1

19 pages, 2078 KiB

Open AccessArticle

An Improved HILIC HPLC-MS/MS Method for the Determination of β-ODAP and Its α Isomer in Lathyrus sativus

by Andreia Bento-Silva, Letice Gonçalves, Elsa Mecha, Filipe Pereira, Maria Carlota Vaz Patto and Maria do Rosário Bronze

Molecules 2019, 24(17), 3043; https://doi.org/10.3390/molecules24173043 - 22 Aug 2019

Cited by 7 | Viewed by 3856

Abstract

β-N-Oxalyl-l-α,β-diaminopropionic acid (β-ODAP) is a non-protein amino acid present in Lathyrus sativus (grass pea) and other Lathyrus species, in parallel with its nontoxic isomer, α-ODAP. When consuming grass pea for several months as staple food, β-ODAP may cause neurolathyrism, [...] Read more.

β-N-Oxalyl-l-α,β-diaminopropionic acid (β-ODAP) is a non-protein amino acid present in Lathyrus sativus (grass pea) and other Lathyrus species, in parallel with its nontoxic isomer, α-ODAP. When consuming grass pea for several months as staple food, β-ODAP may cause neurolathyrism, a motor neuron degeneration syndrome. Therefore, the independent quantification of both ODAP isomers instead of only the total amount in grass pea allows the identification of less toxic varieties and the development of tools to support breeding for improving grass pea quality. In this work, a simple and fast HPLC-MS/MS method was developed without sample derivatization, using a hydrophilic interaction chromatography (HILIC) column and an isocratic gradient of eluents for 18 min, which allowed the determination of both α- and β-ODAP. The proposed method was fully validated and applied to the determination of α- and β-ODAP contents in a diverse collection of 107 grass pea accessions representative of the main grass pea-growing geographical regions in the world, with the prompt identification of contrasting accessions. β-ODAP content in the analyzed grass pea samples ranged from 0.45 ± 0.02 to 6.04 ± 0.45 mg g⁻¹. The moderate correlation found between α- and β-ODAP contents (0.65) in this collection reinforces the importance of the independent quantification of both ODAP isomers. Full article

► Show Figures

Graphical abstract

13 pages, 1600 KiB

Open AccessArticle

Xylopia aethiopica Seeds from Two Countries in West Africa Exhibit Differences in Their Proteomes, Mineral Content and Bioactive Phytochemical Composition

by Xiaojian Yin, María A.S.C. Chávez León, Richard Osae, Loveth O. Linus, Lian-Wen Qi and Raphael N. Alolga

Molecules 2019, 24(10), 1979; https://doi.org/10.3390/molecules24101979 - 23 May 2019

Cited by 17 | Viewed by 4033

Abstract

Aside from its multiple medicinal uses, the fruit of Xylopia aethiopica is widely used in Africa as food. Herein, we characterize the protein profiles, mineral content and bioactive phytochemical composition of the seeds of this plant sourced in Ghana and Nigeria. Using label-free [...] Read more.

Aside from its multiple medicinal uses, the fruit of Xylopia aethiopica is widely used in Africa as food. Herein, we characterize the protein profiles, mineral content and bioactive phytochemical composition of the seeds of this plant sourced in Ghana and Nigeria. Using label-free proteomics, a total of 677 proteins were identified, with 260 found in the Ghana-sourced samples while 608 proteins were detected in the samples from Nigeria. However, 114 proteins were common between the samples from the two countries, among which 48 were significantly changed. Bioinformatics and functional analyses revealed that the differential levels of the proteins were mainly linked to pathways involved amino acids metabolism and biosynthesis. The significantly changed proteins related mainly to catalytic activity and carbon metabolism. The samples from Nigeria also exhibited superior qualities in terms of their antioxidant effects, and total phenolic and flavonoid content. Finally, only the content of Na varied to a statistically significant level. This study lends support to its culinary use and hints towards the impact of location of cultivation on the quality of the seeds. There is however need for further mechanistic investigations to unravel the underlying reasons for the observed differences. Full article

► Show Figures

Figure 1

16 pages, 1461 KiB

Open AccessArticle

Comparative Analysis of Carbohydrates, Nucleosides and Amino Acids in Different Parts of Trichosanthes kirilowii Maxim. by (Ultra) High-Performance Liquid Chromatography Coupled with Tandem Mass Spectrometry and Evaporative Light Scattering Detector Methods

by Huang-qin Zhang, Pei Liu, Jin-ao Duan, Ling Dong, Er-xin Shang, Da-wei Qian, Zhen-hua Zhu, Hui-wei Li and Wei-wen Li

Molecules 2019, 24(7), 1440; https://doi.org/10.3390/molecules24071440 - 11 Apr 2019

Cited by 20 | Viewed by 3992

Abstract

Trichosanthes kirilowii Maxim. is one of the original plants for traditional Chinese medicines Trichosanthis Fructus, Trichosanthis Semen, Trichosanthis Pericarpium and Trichosanthis Radix. Amino acids, nucleosides and carbohydrates are usually considered to have nutritional value and health-care efficacy. In this study, methods involving high-performance [...] Read more.

Trichosanthes kirilowii Maxim. is one of the original plants for traditional Chinese medicines Trichosanthis Fructus, Trichosanthis Semen, Trichosanthis Pericarpium and Trichosanthis Radix. Amino acids, nucleosides and carbohydrates are usually considered to have nutritional value and health-care efficacy. In this study, methods involving high-performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD), UV-visible spectrophotometry and ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) were established for quantifying carbohydrates (fructose, glucose, stachyose, raffinose and polysaccharide), fourteen nucleosides and twenty one amino acids. Moreover, sixty-three samples from nine different parts, including pericarp, seed, fruit pulp, stem, leaf, main root, main root bark, lateral root and lateral root bark of T. kirilowii from different cultivated varieties were examined. The established methods were validated with good linearity, precision, repeatability, stability, and recovery. The results showed that the average content of total amino acids in roots (15.39 mg/g) and root barks (16.38 mg/g) were relatively higher than for others. Contents of nucleosides in all parts of T. kirilowii were below 1.5 mg/g. For carbohydrates, fruit pulp has a higher content than others for glucose (22.91%), fructose (20.63%) and polysaccharides (27.29%). By using partial least-squared discriminate analysis (PLS-DA), Variable importance in the projection (VIP) plots and analysis of variance (ANOVA) analysis, the characteristic components of the different organs (fruit, stems and leaves, roots) were found. This analysis suggested there were potential medicinal and nutritive health care values in various parts of the T. kirilowii, which provided valuable information for the development and utilization of T. kirilowii. Full article

► Show Figures

Graphical abstract

15 pages, 2262 KiB

Open AccessArticle

A New Method for Simultaneous Determination of Phenolic Acids, Alkaloids and Limonoids in Phellodendri Amurensis Cortex

by Yao Chen, Zhao Zhang, Yang Zhang, Xiaomei Zhang, Zhipeng Zhang, Yonghong Liao and Bengang Zhang

Molecules 2019, 24(4), 709; https://doi.org/10.3390/molecules24040709 - 15 Feb 2019

Cited by 22 | Viewed by 3559

Abstract

Phellodendri Amurensis Cortex (PAC) is a well-known herbal medicine in China with complex components, but the previous research has mostly focused on its alkaloids analysis. For the first time, a simpler and more efficient method was proposed in this paper to simultaneously determine [...] Read more.

Phellodendri Amurensis Cortex (PAC) is a well-known herbal medicine in China with complex components, but the previous research has mostly focused on its alkaloids analysis. For the first time, a simpler and more efficient method was proposed in this paper to simultaneously determine the content of three different kinds of compounds—phenolic acids, alkaloids and limonoids—in PAC. The phenolic acids included 3-O-feruloylquinic acid, 4-O-feruloylquinic acid and syringin. The alkaloids include magnoflorine, phellodendrine, jatrorrhizine, palmatine and berberine, while the limonoids include obaculactone and obacunone. An approach combining multi-wavelength and HPLC-DAD was used in this study due to the great difference in maximum absorption wavelength of the various components. Four wavelengths at 215, 275, 280 and 310 nm, respectively, were chosen for monitoring. It has been indicated through appropriate tests that this approach is of high accuracy, good repeatability and stability and provides a scientific basis for the quality assessment of PAC and associated derivatives. In addition, the chromatographic fingerprints method combined with multivariate statistical analysis chosen in this study was proved to be effective and reasonable for an accurate classification of 33 batches of samples collected from different locations. Full article

► Show Figures

Figure 1

2018

Jump to: 2024, 2023, 2022, 2021, 2020, 2019

17 pages, 1629 KiB

Open AccessFeature PaperArticle

Profile of Phosphatidylserine Modifications under Nitroxidative Stress Conditions Using a Liquid Chromatography-Mass Spectrometry Based Approach

by Bruna Neves, Pedro Domingues, Maria Manuel Oliveira, Maria do Rosário Domingues and Tânia Melo

Molecules 2019, 24(1), 107; https://doi.org/10.3390/molecules24010107 - 29 Dec 2018

Cited by 9 | Viewed by 4354

Abstract

Nitrated lipids have been detected in vitro and in vivo, usually associated with a protective effect. While nitrated fatty acids have been widely studied, few studies reported the nitration and nitroxidation of the phospholipid classes phosphatidylcholine, and phosphatidylethanolamine. However, no information regarding nitrated [...] Read more.

Nitrated lipids have been detected in vitro and in vivo, usually associated with a protective effect. While nitrated fatty acids have been widely studied, few studies reported the nitration and nitroxidation of the phospholipid classes phosphatidylcholine, and phosphatidylethanolamine. However, no information regarding nitrated and nitroxidized phosphatidylserine can be found in the literature. This work aims to identify and characterize the nitrated and nitroxidized derivatives of 1-palmitoyl-2-oleoyl-sn-3-glycero-phosphoserine (POPS), obtained after incubation with nitronium tetrafluoroborate, by liquid chromatography (LC) coupled to mass spectrometry (MS) and tandem MS (MS/MS). Several nitrated and nitroxidized products were identified, namely, nitro, nitroso, nitronitroso, and dinitro derivatives, as well as some nitroxidized species such as nitrosohydroxy, nitrohydroxy, and nitrohydroperoxy. The fragmentation pathways identified were structure-dependent and included the loss of HNO and HNO₂ for nitroso and nitro derivatives, respectively. Combined losses of PS polar head group plus HNO or HNO₂ and carboxylate anions of modified fatty acyl chain were also observed. The nitrated POPS also showed antiradical potential, demonstrated by the ability to scavenge the ABTS^●+ and DPPH^● radicals. Overall, this in vitro model of nitration based on LC-MS/MS provided additional insights into the nitrated and nitroxidized derivatives of PS and their fragmentation fingerprinting. This information is a valuable tool for targeted analysis of these modified PS in complex biological samples, to further explore the new clues on the antioxidant potential of nitrated POPS. Full article

► Show Figures

Figure 1

9 pages, 1220 KiB

Open AccessArticle

Confirmatory Analysis of Nitroimidazoles and Hydroxy Metabolites in Honey by Dispersive-Solid Phase Extraction and Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

by Xiaowei Li, Yuebin Ke, Yingyu Wang, Chengfei Wang, Dongyang Ye, Xue Hu, Lan Zhou and Xi Xia

Molecules 2018, 23(12), 3350; https://doi.org/10.3390/molecules23123350 - 18 Dec 2018

Cited by 7 | Viewed by 3172

Abstract

An ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry (UHPLC-MS/MS) method was developed and validated for confirmatory analysis of four nitroimidazoles and three hydroxy metabolites in honey. Honey samples were dissolved in 2% formic acid solution and nitroimidazoles and metabolites were isolated and enriched [...] Read more.

An ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry (UHPLC-MS/MS) method was developed and validated for confirmatory analysis of four nitroimidazoles and three hydroxy metabolites in honey. Honey samples were dissolved in 2% formic acid solution and nitroimidazoles and metabolites were isolated and enriched by dispersive-solid phase extraction using mixed-mode strong cation-exchange sorbent. The determination involves separation of analytes on an UHPLC C₁₈ column and detection by multiple reaction monitoring in positive ionization mode. The recovery of the method was ranged from 90.2 to 105.6% with inter-day relative standard deviations of less than 11.2%. The limits of detection and limits of quantification were in the ranges of 0.02–0.07 µg/kg and 0.05–0.2 µg/kg, respectively. Honey samples from the market were analyzed to demonstrate the applicability of the proposed method. Full article

► Show Figures

Figure 1

9 pages, 1646 KiB

Open AccessArticle

A Comparative Study of Advanced Stationary Phases for Fast Liquid Chromatography Separation of Synthetic Food Colorants

by Ivona Lhotská, Petr Solich and Dalibor Šatínský

Molecules 2018, 23(12), 3335; https://doi.org/10.3390/molecules23123335 - 15 Dec 2018

Cited by 2 | Viewed by 3308

Abstract

Food analysis demands fast methods for routine control and high throughput of samples. Chromatographic separation enables simultaneous determination of numerous compounds in complex matrices, several approaches increasing separation efficiency and speed of analysis were involved. In this work, modern types of column with [...] Read more.

Food analysis demands fast methods for routine control and high throughput of samples. Chromatographic separation enables simultaneous determination of numerous compounds in complex matrices, several approaches increasing separation efficiency and speed of analysis were involved. In this work, modern types of column with monolithic rod or superficially porous particles were employed and compared for determination of eight synthetic food dyes, their chromatographic performance was evaluated. During method optimization, cyano stationary phase Chromolith Performance CN 100 × 4.6 mm and Ascentis Express ES-CN 100 × 4.6 mm, 5 µm were selected for the separation of polar colorants. The separation was performed by gradient elution of acetonitrile/methanol and 2% water solution of ammonium acetate at flow rate 2.0 mL min⁻¹. Mobile phase composition and the gradients were optimized in order to enable efficient separation on both columns. The method using fused-core particle column provided higher separation efficiency, narrow peaks of analytes resulted in increased peak capacity and shortening of analysis time. After the validation, the method was applied for analysis of coloured beers, soft drinks and candies. Full article

► Show Figures

Graphical abstract

17 pages, 958 KiB

Open AccessArticle

Extraction, Analysis, and Antioxidant Activity Evaluation of Phenolic Compounds in Different Italian Extra-Virgin Olive Oils

by Chiara Fanali, Susanna Della Posta, Alessandra Vilmercati, Laura Dugo, Marina Russo, Tommasangelo Petitti, Luigi Mondello and Laura De Gara

Molecules 2018, 23(12), 3249; https://doi.org/10.3390/molecules23123249 - 08 Dec 2018

Cited by 25 | Viewed by 5571

Abstract

The analysis of phenolic compounds in extra virgin olive oils was carried out by high-performance liquid chromatography utilizing photodiode array and mass spectrometry detectors. The chromatographic profile of thirty samples from four Italian Regions highlighted the presence of secoiridoids, phenolic alcohols, flavonoids, and [...] Read more.

The analysis of phenolic compounds in extra virgin olive oils was carried out by high-performance liquid chromatography utilizing photodiode array and mass spectrometry detectors. The chromatographic profile of thirty samples from four Italian Regions highlighted the presence of secoiridoids, phenolic alcohols, flavonoids, and phenolic acid classes. A similar qualitative profile was observed with some differences in peak area and fifteen compounds were tentatively identified. Quantitative analysis was performed by UV detection considering eight standard phenolic compounds. The chromatographic method, after optimization, was validated studying some parameters, e.g., intra-day and inter-day retention time precision, limit of detection, limit of quantification, and linearity. Recovery of the method was performed achieving good results (10 and 50 g·g⁻¹ with recovery of 72.9–92.1% (w/w) and 79.1–102.8% (w/w), respectively). In all samples secoiridoids were the main compounds ranging from 85 to more than 99% (w/w) of the total concentration of detected phenolic compounds while phenolic acids accounted for the lowest percentage (0.1–0.6%, w/w). Finally, total concentration of phenolic compounds and antioxidant activity were determined with different chemical assays. A good and significant correlation among total phenolic compound concentration and antioxidant activity was observed. A significant different phenolic compound concentration and antioxidant activity was determined between samples from Puglia and Sicily. This was studied performing statistical analysis by one-way analysis of variance (ANOVA) followed by Bonferroni post-hoc test. Full article

► Show Figures

Figure 1

18 pages, 266 KiB

Open AccessFeature PaperReview

Liquid Chromatographic Strategies for Separation of Bioactive Compounds in Food Matrices

by Chiara Cavaliere, Anna Laura Capriotti, Giorgia La Barbera, Carmela Maria Montone, Susy Piovesana and Aldo Laganà

Molecules 2018, 23(12), 3091; https://doi.org/10.3390/molecules23123091 - 27 Nov 2018

Cited by 18 | Viewed by 3511

Abstract

Nowadays, there is an increasing attention for nutraceuticals and, in general, bioactive compounds naturally present in food. Indeed, the possibility of preserving human health and preventing disease (e.g., cardiovascular diseases, cancer etc.) by the intake of healthy food is attractive for both consumers [...] Read more.

Nowadays, there is an increasing attention for nutraceuticals and, in general, bioactive compounds naturally present in food. Indeed, the possibility of preserving human health and preventing disease (e.g., cardiovascular diseases, cancer etc.) by the intake of healthy food is attractive for both consumers and food industries. In turn, research in this field was also prompted significantly, with the aim of characterizing these bioactive compounds and ascribe to them a specific activity. The bioactive compounds can belong to several chemical classes. However, their chemical diversity and presence in complex matrices, such as food, make it challenging both their isolation and characterization. To tackle this issue, efficient separation systems are needed, which are mainly based on chromatography. In this context, this mini-review aims to provide the reader with an overview of the most relevant and recent approaches for the separation of the most common bioactive compounds in food, in particular polyphenols, phenols, carotenoids, and peptides, by liquid chromatography approaches. Full article

12 pages, 1726 KiB

Open AccessArticle

A Novel Method for the Determination of Vancomycin in Serum by High-Performance Liquid Chromatography-Tandem Mass Spectrometry and Its Application in Patients with Diabetic Foot Infections

by Min Liu, Zhi-Hui Yang and Guo-Hui Li

Molecules 2018, 23(11), 2939; https://doi.org/10.3390/molecules23112939 - 10 Nov 2018

Cited by 26 | Viewed by 5041

Abstract

A novel, precise, and accurate high-performance liquid chromatography-tandem mass spectrometry (Q-trap-MS) method was developed, optimized, and validated for determination of vancomycin in human serum using norvancomycin as an internal standard. Effect of different parameters on the analysis was evaluated. ZORBAX SB-C₁₈ column [...] Read more.

A novel, precise, and accurate high-performance liquid chromatography-tandem mass spectrometry (Q-trap-MS) method was developed, optimized, and validated for determination of vancomycin in human serum using norvancomycin as an internal standard. Effect of different parameters on the analysis was evaluated. ZORBAX SB-C₁₈ column (150 × 4.6 mm, 5 μm) using water (containing 0.1% formic acid, v/v)–acetonitrile (containing 0.1% formic acid, v/v) as a mobile phase was chosen. The calibration curve was linear over the concentration ranges of 1 to 2000 ng/mL for vancomycin. The limit of detection (LOD) and limit of quantification (LOQ) for vancomycin were 0.3 and 1.0 ng/mL. Recoveries were between 87.2 and 102.3%, which gave satisfactory precision. A total of 100 serum samples (from 50 patients with diabetic foot proven Gram-positive infection and 50 nondiabetic patients with pneumonia requiring hospitalization and antibiotic therapy) were analyzed by this method. The trough vancomycin concentrations of diabetic foot infection (DFI) patients and nondiabetic patients were 8.20 ± 2.83 μg/mL (range: 4.80–14.2 μg/mL) and 15.80 ± 5.43 μg/mL (range: 8.60–19.5 μg/mL), respectively. The method is sensitive, precise, and reproducible, it could be applied for routine laboratory analysis of vancomycin in serum samples. Full article

► Show Figures

Figure 1

13 pages, 454 KiB

Open AccessArticle

HPLC-UV/Vis-APCI-MS/MS Determination of Major Carotenoids and Their Bioaccessibility from “Delica” (Cucurbita maxima) and “Violina” (Cucurbita moschata) Pumpkins as Food Traceability Markers

by Caterina Bergantin, Annalisa Maietti, Paola Tedeschi, Guillermina Font, Lara Manyes and Nicola Marchetti

Molecules 2018, 23(11), 2791; https://doi.org/10.3390/molecules23112791 - 27 Oct 2018

Cited by 36 | Viewed by 4565

Abstract

Carotenoids are a widespread group of fat-soluble pigments, and their major nutritional importance comes from their pro-vitamin A activity and their antioxidant capacity. In this study, two different pumpkin cultivars (Cucurbita maxima, also named ‘Delica’ and Cucurbita moschata, also known [...] Read more.

Carotenoids are a widespread group of fat-soluble pigments, and their major nutritional importance comes from their pro-vitamin A activity and their antioxidant capacity. In this study, two different pumpkin cultivars (Cucurbita maxima, also named ‘Delica’ and Cucurbita moschata, also known as ‘Violina’) from the southern Po Delta area were investigated in terms of carotenoid content and the influence of food processing on compositional changes and carotenoid bioaccessibility. Quali- and quantitative determination of carotenoids in sample extracts were performed on a C30 column by means of an online coupled HPLC-UV/Vis-APCI-MS/MS technique. The identification of separated compounds was tentatively achieved by merging (i) chromatographic data, (ii) UV-Vis spectra, and (iii) MS/MS fragmentation spectra. The chromatographic profiles for the two cultivars showed qualitative differences. Two major carotenoids were considered for quantification purposes and further investigations: lutein and

β

-carotene. Quantification of target carotenoids was performed with external calibration through analytical standards. The concentration of lutein and

β

-carotene was higher in C. maxima than in the other variety, C. moschata. Carotenoids are susceptible to degradation (isomerization and oxidation) during food processing (i.e., cooking), and the concentration of lutein and

β

-carotene were monitored in oven-cooked and steam-cooked pumpkins. The steam-cooking process was superior in terms of limiting carotenoid loss. A complete functional profile of pumpkins as a source of carotenoids was gained with the evaluation of their in vitro bioaccessibility and their bioavailability after intake during human digestion. Bioaccessibility of lutein and

β

-carotene were estimated by an in vitro static digestion model that involved salivary, gastric, and duodenal phases. Bioaccessibility values progressively increased from the salivary to the duodenal phase for both pumpkin varieties and cooking methods. Bioaccessibility of lutein was always lower than

β

-carotene for both cultivars and for both cooking methods. Bioaccessibility values for lutein and

β

-carotene changed from 1.93% to 2.34% vs. 4.94% and 8.83% in the salivary phase, from 2.7% to 4.63% vs. 7.83% and 15.60% in the gastric phase, and from 10.04% to 13.42% vs. 25.81% and 35.32% in the duodenal phase. For both target compounds, bioaccessibility in the duodenal phase was more than twice the gastric values, and it underlined that the type of cooking did not influence release from the initial matrix. Full article

► Show Figures

Figure 1

15 pages, 1743 KiB

Open AccessArticle

Preclinical Pharmacokinetics of Scoparone, Geniposide and Rhein in an Herbal Medicine Using a Validated LC-MS/MS Method

by Tun-Pin Hsueh and Tung-Hu Tsai

Molecules 2018, 23(10), 2716; https://doi.org/10.3390/molecules23102716 - 22 Oct 2018

Cited by 21 | Viewed by 4709

Abstract

The herbal formula Yin-Chen-Hao-Tang has been reported to have anti-fibrosis properties. The aim of this study was to reveal the pharmacokinetic characteristics of bioactive compounds in this herbal formula. A new high-performance liquid chromatography-tandem mass spectrometry method was developed and validated for simultaneous [...] Read more.

The herbal formula Yin-Chen-Hao-Tang has been reported to have anti-fibrosis properties. The aim of this study was to reveal the pharmacokinetic characteristics of bioactive compounds in this herbal formula. A new high-performance liquid chromatography-tandem mass spectrometry method was developed and validated for simultaneous determination of scoparone, geniposide and rhein in rat plasma. A pharmaceutical herbal powder was administered to rats at doses of 1 g/kg and 3 g/kg orally. The method showed excellent linearity (r² > 0.999) and validation was successfully conducted for the pharmacokinetic study. The results show that the C_max values and areas under the curve of scoparone, geniposide and rhein were higher and not proportional to the dose in rat plasma, while the T_max and half-life values were consistent in the group that received 1 g/kg. The clearance of the higher dose (3 g/kg) did not decrease proportionally to that of the low dose. The results showed the nonlinear pharmacokinetic properties of scoparone, geniposide and rhein in Yin-Chen-Hao-Tang that suggested possible accumulation of bioactive compounds through oral administration. This pharmacokinetic study reveals that an increased dose of this herbal formula would largely increase the maximum concentration and bioavailability of scoparone, geniposide and rhein. Full article

► Show Figures

Figure 1

12 pages, 2610 KiB

Open AccessFeature PaperArticle

The Way to Ultrafast, High-Throughput Enantioseparations of Bioactive Compounds in Liquid and Supercritical Fluid Chromatography

by Omar H. Ismail, Simona Felletti, Chiara De Luca, Luisa Pasti, Nicola Marchetti, Valentina Costa, Francesco Gasparrini, Alberto Cavazzini and Martina Catani

Molecules 2018, 23(10), 2709; https://doi.org/10.3390/molecules23102709 - 20 Oct 2018

Cited by 32 | Viewed by 3624

Abstract

Until less than 10 years ago, chiral separations were carried out with columns packed with 5 or 3

μ

m fully porous particles (FPPs). Times to resolve enantiomeric mixtures were easily larger than 30 min, or so. Pushed especially by stringent requirements from [...] Read more.

Until less than 10 years ago, chiral separations were carried out with columns packed with 5 or 3

μ

m fully porous particles (FPPs). Times to resolve enantiomeric mixtures were easily larger than 30 min, or so. Pushed especially by stringent requirements from medicinal and pharmaceutical industries, during the last years the field of chiral separations by liquid chromatography has undergone what can be defined a “true revolution”. With the purpose of developing ever faster and efficient method of separations, indeed, very efficient particle formats, such as superficially porous particles (SPPs) or Full article

► Show Figures

Figure 1

7 pages, 789 KiB

Open AccessArticle

Validation of UPLC-MS/MS Method for Determination of Urinary Lactulose/Mannitol

by Jacopo Gervasoni, Aniello Primiano, Cristina Graziani, Franco Scaldaferri, Antonio Gasbarrini, Andrea Urbani and Silvia Persichilli

Molecules 2018, 23(10), 2705; https://doi.org/10.3390/molecules23102705 - 20 Oct 2018

Cited by 14 | Viewed by 4974

Abstract

Determination of urinary lactulose/mannitol is one of the most used tests to evaluate intestinal barrier function. High-performance liquid chromatography (HPLC) separation with electrospray ionization tandem mass spectrometry guarantees high levels of selectivity and reproducibility. In this paper we report an upgrade of the [...] Read more.

Determination of urinary lactulose/mannitol is one of the most used tests to evaluate intestinal barrier function. High-performance liquid chromatography (HPLC) separation with electrospray ionization tandem mass spectrometry guarantees high levels of selectivity and reproducibility. In this paper we report an upgrade of the previous published liquid chromatography tandem mass spectrometry method, introducing more reliable internal standards and ultra-performance liquid chromatography with ethylene bridged hybrid amide columns. The ultra-performance liquid chromatography provided an efficient chromatographic separation of the two sugars in 5 min, compared to 15 min using the previous method. The limit of quantification was 10 µg/mL for mannitol and 2.5 µg/mL for lactulose, and the assay was linear up to 1000 µg/mL for mannitol and 1000 µg/mL for lactulose. The within-run precision and accuracy ranged from 0.7 to 2.9% and 97.2 to 101.2%, respectively. The between-run precision and accuracy ranged from 1.9 to 4.7% and 94.8 to 97.5%, respectively. Recovery was higher than 90.2% for both lactulose and mannitol, and the matrix effect for both lactulose and mannitol was lower than 15%. With this new method we have a real improvement in terms of accuracy and reproducibility, ensuring results in shorter time. The changes to the previous protocol make this method excellent for routine purposes. Full article

► Show Figures

Figure 1

12 pages, 2234 KiB

Open AccessArticle

Qualitative and Quantitative Analysis of the Major Constituents in WLJ Herbal Tea Using Multiple Chromatographic Techniques

by Chao-Zhan Lin, Run-Jing Zhang, Yu-Feng Yao, Xiao-Dan Huang, Rong-Bo Zheng, Bo-Jian Wu and Chen-Chen Zhu

Molecules 2018, 23(10), 2623; https://doi.org/10.3390/molecules23102623 - 12 Oct 2018

Cited by 9 | Viewed by 3280

Abstract

Quality control of Chinese herbal tea remains a challenge due to our poor knowledge of their complex chemical profile. This study aims to investigate the chemical composition of one of the best-selling and famous brand of beverage in China, Wanglaoji Herbal Tea (WLJHT), [...] Read more.

Quality control of Chinese herbal tea remains a challenge due to our poor knowledge of their complex chemical profile. This study aims to investigate the chemical composition of one of the best-selling and famous brand of beverage in China, Wanglaoji Herbal Tea (WLJHT), via a full component quantitative analysis. In this paper, a total of thirty-four representative constituents were identified or tentatively characterized using ultra-high performance liquid chromatography coupled with quadrupole tandem time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Moreover, the quantitative analyses of fourteen constituents were performed by high performance liquid chromatography with a triple quadruple tandem mass spectrometry (HPLC-MS/MS) method and saccharide compositions of WLJHT were also quantitatively determined by high performance liquid chromatography (HPLC) with evaporative light scattering detector (ELSD) on a Hilic column, separately. Using multiple chromatographic techniques presented a good precision, sensitivity, repeatability and stability, and was successfully applied to analyze 16 batches of WLJHT samples. Therefore, it would be a reliable and useful approach for the quality control of WLJHT. Full article

► Show Figures

Graphical abstract

16 pages, 3512 KiB

Open AccessArticle

Chemical Fingerprinting and Quantification of Chinese Cinnamomi Cortex by Ultra High Performance Liquid Chromatography Coupled with Chemometrics Methods

by Ninghui Ma, Yue Ding, Yong Zhang, Tong Zhang, Yaxiong Yi and Bing Wang

Molecules 2018, 23(9), 2214; https://doi.org/10.3390/molecules23092214 - 31 Aug 2018

Cited by 11 | Viewed by 3785

Abstract

To rapidly clarify and quantify the chemical profiling of Cinnamomi cortex a reliable and feasible strategy of chromatographic fingerprinting with a suite of chemometrics methods was developed and validated by ultra-high performance liquid chromatography coupled with diode array detection. Furthermore, to identify more [...] Read more.

To rapidly clarify and quantify the chemical profiling of Cinnamomi cortex a reliable and feasible strategy of chromatographic fingerprinting with a suite of chemometrics methods was developed and validated by ultra-high performance liquid chromatography coupled with diode array detection. Furthermore, to identify more meaningful chemical markers, the chemometrics methods including hierarchical cluster analysis (HCA), principal component analysis (PCA) and similarity, which all generate quality evaluations and correlation classifications of Cinnamomi cortex, were used to improve the Cinnamomi cortex quality control standards. A total of 12 characteristic peaks were confirmed, seven of which were identified by comparing their retention times, UV and MS spectra with authentic compounds. Moreover, 11 analytes were accurately determined, as a complementary quantification method of chromatographic fingerprinting. For quantitative analyses, selective detection was performed at 254, 280 and 340 nm. The tested samples were separated and determined using UPLC and a series of methodologies including linearity, precision, accuracy, limit of detection and quantification and extraction recoveries were validated. Meanwhile the method bias for all the analytes did not exceed 5%. A total of 42 samples were acquired in China and analyzed. The results demonstrated that chromatographic fingerprinting in combination with chemometrics methods provides a promising and practical method to more effectively and comprehensively control the quality of Cinnamomi cortex from various sources, which would be a useful reference for the development and further study of Cinnamomi cortex and related formulations. Full article

► Show Figures

Figure 1

16 pages, 1144 KiB

Open AccessArticle

Tracing the Thermal History of Seafood Products through Lysophospholipid Analysis by Hydrophilic Interaction Liquid Chromatography–Electrospray Ionization Fourier Transform Mass Spectrometry

by Ilario Losito, Laura Facchini, Rosa Catucci, Cosima Damiana Calvano, Tommaso R. I. Cataldi and Francesco Palmisano

Molecules 2018, 23(9), 2212; https://doi.org/10.3390/molecules23092212 - 31 Aug 2018

Cited by 7 | Viewed by 3245

Abstract

Low temperature treatments commonly applied to seafood products have been shown to influence their phospholipid (PL) profile through enzymatic hydrolysis. In the present study, the generation of lysophospholipids (LPL) resulting from this process was systematically investigated for selected, commercially relevant seafood products, namely [...] Read more.

Low temperature treatments commonly applied to seafood products have been shown to influence their phospholipid (PL) profile through enzymatic hydrolysis. In the present study, the generation of lysophospholipids (LPL) resulting from this process was systematically investigated for selected, commercially relevant seafood products, namely oysters, clams, octopuses, and shrimps. These products were subjected to thermal treatments like refrigeration or freezing after being purchased as fresh, defrozen, or frozen products depending on the case. The coupling between hydrophilic interaction liquid chromatography (HILIC) and electrospray ionization with high resolution/accuracy Fourier transform mass spectrometry (ESI-FTMS) was exploited to evaluate the PL profile of the cited products, especially the incidence of LPL related to the two main PL classes of seafood products—phosphatidylcholines (PC) and phosphatidylethanolamines (PE)—in the lipid extracts. The lyso forms of PE (LPE) were found to be generally more sensitive than those of PC (LPC) to thermal treatments, usually exhibiting a significant increase upon prolonged refrigeration at 4 °C in all types of investigated products except European flat oysters. Moreover, the distinction between fresh and frozen or defrozen products could be achieved in the case of octopuses and shrimps, respectively. Full article

► Show Figures

Graphical abstract

14 pages, 1325 KiB

Open AccessArticle

Changes in Nutrient Profile and Antioxidant Activities of Different Fish Soups, Before and After Simulated Gastrointestinal Digestion

by Gaonan Zhang, Shujian Zheng, Yuqi Feng, Guo Shen, Shanbai Xiong and Hongying Du

Molecules 2018, 23(8), 1965; https://doi.org/10.3390/molecules23081965 - 06 Aug 2018

Cited by 23 | Viewed by 4905

Abstract

Different kinds of freshwater fish soups show a diverse range of health functions, due to their different nutritional substances and corresponding bioactivities. In the current study, in order to learn the theoretical basis of the potential role fish soup plays in diet therapy [...] Read more.

Different kinds of freshwater fish soups show a diverse range of health functions, due to their different nutritional substances and corresponding bioactivities. In the current study, in order to learn the theoretical basis of the potential role fish soup plays in diet therapy functions, the changes of nutrient profiles and antioxidant activities in crucian carp soup and snakehead soup (before and after simulated gastrointestinal digestion) were investigated, such as chemical composition, free amino acids, mineral and fatty acid contents, DPPH radical scavenging activity, ferrous ion chelating activity, hydroxyl radical-scavenging activity and the reducing power effect. Results show that the content of mineral elements in snakehead fish soup was significantly higher than that of crucian carp soup, especially for the contents of Ca, Zn, Fe. The content of total amino acid (TAA) of crucian carp soup (82.51 mg/100 mL) was much higher than that of snakehead fish soup (47.54 mg/100 mL) (p < 0.05). Furthermore, the antioxidant capacity of crucian carp soup was stronger than that of snakehead soup. The intensive profiles of nutritional composition and antioxidant activities of these two kinds of fish soups were expected to partly provide the theoretical basis of therapeutic effects. Full article

► Show Figures

Graphical abstract

12 pages, 5337 KiB

Open AccessArticle

Quantitative and Chemical Fingerprint Analysis for the Quality Evaluation of Platycodi Radix Collected from Various Regions in China by HPLC Coupled with Chemometrics

by Haiyang Lu, Mengzhen Ju, Shanshan Chu, Tao Xu, Yuzhe Huang, Qingyun Chan, Huasheng Peng and Shuangying Gui

Molecules 2018, 23(7), 1823; https://doi.org/10.3390/molecules23071823 - 23 Jul 2018

Cited by 19 | Viewed by 4702

Abstract

Platycodi Radix (PR) is the root of Platycodon grandiflorum (Jacq.) A. DC., which has been used for a long time in China to treat pulmonary diseases. The present study aimed to evaluate the quality of PR samples collected from 23 regions of 11 [...] Read more.

Platycodi Radix (PR) is the root of Platycodon grandiflorum (Jacq.) A. DC., which has been used for a long time in China to treat pulmonary diseases. The present study aimed to evaluate the quality of PR samples collected from 23 regions of 11 provinces in China. Eight saponins were quantified using HPLC coupled with evaporative light scattering detection (HPLC-ELSD). The samples with the highest total contents of saponins were from southern China, such as Yunnan, Guangxi, Jiangxi, and Guangzhou. The fingerprint analysis of PR samples was conducted by HPLC-UV method. Nineteen common peaks were selected and the similarity values varied from 0.607 to 0.921. These findings indicated that the saponins contents of PR from different regions varied significantly, with PR samples from southern China having the highest contents of saponins. These comprehensive methods were successful in evaluating the quality of PR samples from northern and southern China, which will serve as a guide for the development of PR as a clinical medication. Full article

► Show Figures

Figure 1

14 pages, 904 KiB

Open AccessArticle

Development of Validated and Stability-Indicating LC-DAD and LC-MS/MS Methods for Determination of Avanafil in Pharmaceutical Preparations and Identification of a Novel Degradation Product by LCMS-IT-TOF

by Nafiz Öncü CAN

Molecules 2018, 23(7), 1771; https://doi.org/10.3390/molecules23071771 - 19 Jul 2018

Cited by 20 | Viewed by 5366

Abstract

Avanafil (AVA), one of the most effective drugs prescribed for erectile dysfunction, is a pyrimidine-derivative PDE5 inhibitor. In the current work, new LC methods were developed and validated for quantitative determination of avanafil and qualitative determination of its degradation products. The quantitative determination [...] Read more.

Avanafil (AVA), one of the most effective drugs prescribed for erectile dysfunction, is a pyrimidine-derivative PDE5 inhibitor. In the current work, new LC methods were developed and validated for quantitative determination of avanafil and qualitative determination of its degradation products. The quantitative determination of avanafil was carried out using liquid chromatography with photodiode array detection (LC-DAD) and liquid chromatography-tandem mass spectrometry LC-MS/MS methods, and fully validated according to the ICH Q2 (R1) guideline, while qualitative determination was performed using a liquid chromatography mass spectrometry-ion trap-time of flight (LCMS-IT-TOF) instrument. The separation of avanafil and its degradation products was carried out using the same reversed-phase chromatographic conditions, in which a second-generation C₁₈-bonded monolithic silica column (Chromolith^® High Resolution RP-18e, 100 × 4.6 mm, Merck KGaA) was used as stationary phase. Briefly, the methods enable quantitation of avanafil with high accuracy (recovery > 95%) and precision (RSD% < 2.0), within the ranges of 0.5–20 μg/mL for LC-DAD and 150–6000 ng/mL for LC-MS/MS. In the forced degradation studies, over and above currently existing data, a new oxidation-based degradation product, whose predicted m/z is 367.1168, was identified and its structure was confirmed by high-resolution mass spectrometric analysis. As the main advantage, either an LC-DAD or LC-MS/MS instrument can be chosen for interference-free quantitation of AVA, according to the facilities in quality-control laboratories. Full article

► Show Figures

Graphical abstract

12 pages, 2871 KiB

Open AccessArticle

Aflatoxins and A. flavus Reduction in Loaf Bread through the Use of Natural Ingredients

by Juan M. Quiles, Raquel Torrijos, Fernando B. Luciano, Jordi Mañes and Giuseppe Meca

Molecules 2018, 23(7), 1638; https://doi.org/10.3390/molecules23071638 - 04 Jul 2018

Cited by 9 | Viewed by 5133

Abstract

In this study, the antifungal activity of yellow mustard (YMF) and oriental mustard (OMF) meal extracts against 14 strains of fungi was tested on a solid medium. The results obtained with the YMF were next confirmed in liquid medium determining the minimum inhibitory [...] Read more.

In this study, the antifungal activity of yellow mustard (YMF) and oriental mustard (OMF) meal extracts against 14 strains of fungi was tested on a solid medium. The results obtained with the YMF were next confirmed in liquid medium determining the minimum inhibitory concentration (MIC) and the minimum fungicide concentration (MFC). Finally, the use of YMF as a natural preservative to extend the useful life of bread was evaluated. Breads with different concentrations of YMF (2, 4, 6 and 8 g/kg) were prepared and contaminated with Aspergillus flavus ISPA 8111 and Penicillium nordicum CECT 2320. For 10 days the formation of mycelium was observed, and after that the fungal growth and the mycotoxins production was determined. The results obtained with the YMF were compared with breads treated with the commercial additive sodium propionate (E-281). The results showed a significant reduction of the fungal population using 6 g/kg and 8 g/kg of YMF in bread contaminated with A. flavus and with P. nordicum and an extensions of the breads shelf life of 7 and 5 days, respectively, in comparison with the control experiment. A reduction of 78% of AFB₁ was observed using 6 g/kg of YMF while no AFB₁ production was detected employing 8 g/kg of YMF in bread preparation. Full article

► Show Figures

Figure 1

13 pages, 2102 KiB

Open AccessArticle

Upregulation of UDP-Glucuronosyltransferases 1a1 and 1a7 Are Involved in Altered Puerarin Pharmacokinetics in Type II Diabetic Rats

by Songtao Dong, Maofan Zhang, Huimin Niu, Kunyu Jiang, Jialei Jiang, Yinglin Ma, Xin Wang and Shengnan Meng

Molecules 2018, 23(6), 1487; https://doi.org/10.3390/molecules23061487 - 20 Jun 2018

Cited by 10 | Viewed by 3944

Abstract

Puerarin is an isoflavonoid extracted from Pueraria lobata roots, and displays a broad range of pharmacological activities, including antidiabetic activity. However, information about the pharmacokinetics of puerarin in diabetics is scarce. This study was conducted to investigate the difference in pharmacokinetic effects of [...] Read more.

Puerarin is an isoflavonoid extracted from Pueraria lobata roots, and displays a broad range of pharmacological activities, including antidiabetic activity. However, information about the pharmacokinetics of puerarin in diabetics is scarce. This study was conducted to investigate the difference in pharmacokinetic effects of puerarin in normal rats and rats with diabetes mellitus (DM), and the mechanism involved. DM was induced by a combined high-fat diet (HFD) and streptozotocin (STZ) injection. Plasma concentrations of puerarin in DM, HFD, and control rats were determined after intravenous (20 mg/kg) and oral administration (500 mg/kg) of puerarin, and pharmacokinetic parameters were estimated. The messenger RNA (mRNA) and protein expression levels of Ugt1a1 and Ugt1a7 in rat livers and intestines were measured using qRT-PCR and western blot, respectively. The area under the concentration–time curve and the clearance of puerarin in the DM rats statistically differed from those in the control rats (p <0.05) with both administration routes. The hepatic and intestinal gene and protein expressions of Ugt1a1 and Ugt1a7 were significantly increased in the DM rats (p <0.05). Therefore, the metabolic changes in diabetes could alter the pharmacokinetics of puerarin. This change could be caused by upregulated uridine diphosphate (UDP)-glucuronosyltransferase activity, which may enhance puerarin clearance, and alter its therapeutic effects. Full article

► Show Figures

Graphical abstract

15 pages, 4218 KiB

Open AccessArticle

Plasma Pharmacokinetic Determination of Canagliflozin and Its Metabolites in a Type 2 Diabetic Rat Model by UPLC-MS/MS

by Song-tao Dong, Hui-min Niu, Yin Wu, Jia-lei Jiang, Ying Li, Kun-yu Jiang, Xin Wang, Mao-fan Zhang, Ming-feng Han and Sheng-nan Meng

Molecules 2018, 23(5), 1229; https://doi.org/10.3390/molecules23051229 - 20 May 2018

Cited by 16 | Viewed by 5236

Abstract

Canagliflozin is a novel, orally selective inhibitor of sodium-dependent glucose co-transporter-2 (SGLT2) for the treatment of patients with type 2 diabetes mellitus. In this study, a sensitive and efficient UPLC-MS/MS method for the quantification of canagliflozin and its metabolites in rat plasma was [...] Read more.

Canagliflozin is a novel, orally selective inhibitor of sodium-dependent glucose co-transporter-2 (SGLT2) for the treatment of patients with type 2 diabetes mellitus. In this study, a sensitive and efficient UPLC-MS/MS method for the quantification of canagliflozin and its metabolites in rat plasma was established and applied to pharmacokinetics in a type 2 diabetic rat model. We firstly investigated the pharmacokinetic changes of canagliflozin and its metabolites in type 2 diabetic rats in order to use canagliflozin more safely, reasonably and effectively. We identified three types of O-glucuronide metabolites (M5, M7 and M17), two kinds of oxidation metabolites (M8 and M9) and one oxidation and glucuronide metabolite (M16) using API 5600 triple-TOF-MS/MS. Following liquid–liquid extraction by tert-butyl methyl ether, chromatographic separation of canagliflozin and its metabolites were performed on a Waters XBridge BEH C18 column (100 × 2.1 mm, 2.5 μm) using 0.1% acetonitrile–formic acid (75:15, v/v) as the mobile phase at a flow rate of 0.7 mL/min. Selected ion monitoring transitions of m/z 462.00→191.10, 451.20→153.10, 638.10→191.10 and 478.00→267.00 were chosen to quantify canagliflozin, empagliflozin (IS), O-glucuronide metabolites (M5, M7 and M17), and oxidation metabolites (M9) using an API 5500-triple-MS/MS in the positive electrospray ionization mode. The validation of the method was found to be of sufficient specificity, accuracy and precision. The pathological condition of diabetes could result in altered pharmacokinetic behaviors of canagliflozin and its metabolites. The pharmacokinetic parameters (AUC_0–t, AUC_0–∞, CL_z/F, and V_z/F) of canagliflozin were significantly different between the CTRL and DM group rats (p < 0.05 or p < 0.01), which may subsequently cause different therapeutic effects. Full article

► Show Figures

Graphical abstract

15 pages, 2186 KiB

Open AccessArticle

Antioxidant and ACE Inhibitory Activity of Enzymatic Hydrolysates from Ruditapes philippinarum

by Yue Yu, Fengjiao Fan, Di Wu, Cuiping Yu, Zhenyu Wang and Ming Du

Molecules 2018, 23(5), 1189; https://doi.org/10.3390/molecules23051189 - 16 May 2018

Cited by 32 | Viewed by 3855

Abstract

Ruditapes philippinarum proteins were hydrolyzed by trypsin, neutrase, and pepsin. The antioxidant activities and ACE inhibitory activity of hydrolysates were analyzed and the antioxidant activities were related to their molecular weight distribution and amino acid compositions. Results indicated the hydrolysis of proteins led [...] Read more.

Ruditapes philippinarum proteins were hydrolyzed by trypsin, neutrase, and pepsin. The antioxidant activities and ACE inhibitory activity of hydrolysates were analyzed and the antioxidant activities were related to their molecular weight distribution and amino acid compositions. Results indicated the hydrolysis of proteins led to an increase in small peptides and free amino acids. The antioxidant activities of Ruditapes philippinarum hydrolysates against DPPH radical scavenging, inhibition on linoleic acid peroxidation, and reducing power showed that the neutrase hydrolysate exhibited the strongest antioxidant activity. In addition, an ACE inhibition assay revealed that the pepsin hydrolysate had the highest ACE inhibitory ability. Ruditapes philippinarum protein hydrolysates could be a promising source of natural antioxidant and ACE inhibitory. Full article

► Show Figures

Figure 1

15 pages, 1417 KiB

Open AccessFeature PaperArticle

Fast Profiling of Natural Pigments in Different Spirulina (Arthrospira platensis) Dietary Supplements by DI-FT-ICR and Evaluation of their Antioxidant Potential by Pre-Column DPPH-UHPLC Assay

by Eduardo Sommella, Giulio Maria Conte, Emanuela Salviati, Giacomo Pepe, Alessia Bertamino, Carmine Ostacolo, Francesca Sansone, Francesco Del Prete, Rita Patrizia Aquino and Pietro Campiglia

Molecules 2018, 23(5), 1152; https://doi.org/10.3390/molecules23051152 - 11 May 2018

Cited by 38 | Viewed by 5375

Abstract

Arthrospira platensis, better known as Spirulina, is one of the most important microalgae species. This cyanobacterium possesses a rich metabolite pattern, including high amounts of natural pigments. In this study, we applied a combined strategy based on Fourier Transform Ion Cyclotron Resonance [...] Read more.

Arthrospira platensis, better known as Spirulina, is one of the most important microalgae species. This cyanobacterium possesses a rich metabolite pattern, including high amounts of natural pigments. In this study, we applied a combined strategy based on Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and Ultra High-Performance Liquid Chromatography (UHPLC) for the qualitative/quantitative characterization of Spirulina pigments in three different commercial dietary supplements. FT-ICR was employed to elucidate the qualitative profile of Spirulina pigments, in both direct infusion mode (DIMS) and coupled to UHPLC. DIMS showed to be a very fast (4 min) and accurate (mass accuracy ≤ 0.01 ppm) tool. 51 pigments were tentatively identified. The profile revealed different classes, such as carotenes, xanthophylls and chlorophylls. Moreover, the antioxidant evaluation of the major compounds was assessed by pre-column reaction with the DPPH radical followed by fast UHPLC-PDA separation, highlighting the contribution of single analytes to the antioxidant potential of the entire pigment fraction. β-carotene, diadinoxanthin and diatoxanthin showed the highest scavenging activity. The method took 40 min per sample, comprising reaction. This strategy could represent a valid tool for the fast and comprehensive characterization of Spirulina pigments in dietary supplements, as well as in other microalgae-based products. Full article

► Show Figures

Graphical abstract

16 pages, 1386 KiB

Open AccessArticle

Quantitative Analysis of Four Catechins from Green Tea Extract in Human Plasma Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry for Pharmacokinetic Studies

by Jeong-Eun Park, Tae-Eun Kim and Kwang-Hee Shin

Molecules 2018, 23(4), 984; https://doi.org/10.3390/molecules23040984 - 23 Apr 2018

Cited by 12 | Viewed by 6315

Abstract

Green tea is consumed as a beverage worldwide and has beneficial effects, such as a lower risk of cardiovascular disease and cancer. A quantitative analysis of the beneficial components in plasma is important for understanding the potential health benefits of green tea. Four [...] Read more.

Green tea is consumed as a beverage worldwide and has beneficial effects, such as a lower risk of cardiovascular disease and cancer. A quantitative analysis of the beneficial components in plasma is important for understanding the potential health benefits of green tea. Four catechins—epigallocatechin-3-gallate (EGCG), epicatechin-3-gallate (ECG), epigallocatechin (EGC), and epicatechin (EC)—which account for the majority of the components of green tea, were analyzed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). In this study, a validated method was optimized to obtain the blood concentrations after the one-time ingestion of 630 mg green tea extract with digoxin and then after the ingestion of 630 mg green tea repeatedly for 15 days. The calibration curve, including the LLOQ, was constructed over 1–500 ng/mL for EGCG, ECG, and EGC and 0.1–50 ng/mL for EC. The method for inter- and intra-validation was applied, acceptable for both accuracy and precision. We successfully developed an appropriate UPLC-MS/MS method for human plasma with good reproducibility and sensitivity. Thus, this method could be applied for future preclinical and clinical studies on EGCG, ECG, EGC, and EC. Full article

► Show Figures

Graphical abstract

18 pages, 12961 KiB

Open AccessArticle

Single Laboratory Validation of a Quantitative Core Shell-Based LC Separation for the Evaluation of Silymarin Variability and Associated Antioxidant Activity of Pakistani Ecotypes of Milk Thistle (Silybum Marianum L.)

by Samantha Drouet, Bilal Haider Abbasi, Annie Falguières, Waqar Ahmad, Sumaira, Clothilde Ferroud, Joël Doussot, Jean Raymond Vanier, Eric Lainé and Christophe Hano

Molecules 2018, 23(4), 904; https://doi.org/10.3390/molecules23040904 - 14 Apr 2018

Cited by 33 | Viewed by 5730

Abstract

Fruits of Silybum marianum (L.) Gaernt are the main source of taxifolin derived flavonolignans. Together, these molecules constitute a mixture called silymarin with many useful applications for cosmetic and pharmaceutic industries. Here, a validated method for the separation of the silymarin constituents has [...] Read more.

Fruits of Silybum marianum (L.) Gaernt are the main source of taxifolin derived flavonolignans. Together, these molecules constitute a mixture called silymarin with many useful applications for cosmetic and pharmaceutic industries. Here, a validated method for the separation of the silymarin constituents has been developed to ensure precision and accuracy in their quantification. Each compound was separated with a high reproducibility. Precision and repeatability of the quantification method were validated according to the AOAC recommendations. The method was then applied to study the natural variability of wild accessions of S. marianum. Analysis of the variation in the fruits composition of these 12 accessions from Pakistan evidenced a huge natural diversity. Correlation analysis suggested a synergistic action of the different flavonolignans to reach the maximal antioxidant activity, as determined by cupric ion reducing antioxidant capacity (CUPRAC) and ferric reducing antioxidant power (FRAP) assays. Principal component analysis (PCA) separated the 12 accessions into three distinct groups that were differing from their silymarin contents, whereas hierarchical clustering analysis (HCA) evidenced strong variations in their silymarin composition, leading to the identification of new silybin-rich chemotypes. These results proved that the present method allows for an efficient separation and quantification of the main flavonolignans with potent antioxidant activities. Full article

► Show Figures

Graphical abstract

18 pages, 2927 KiB

Open AccessArticle

New Findings in Prunus padus L. Fruits as a Source of Natural Compounds: Characterization of Metabolite Profiles and Preliminary Evaluation of Antioxidant Activity

by Dario Donno, Maria Gabriella Mellano, Marta De Biaggi, Isidoro Riondato, Ernest Naivonirina Rakotoniaina and Gabriele Loris Beccaro

Molecules 2018, 23(4), 725; https://doi.org/10.3390/molecules23040725 - 22 Mar 2018

Cited by 36 | Viewed by 4806

Abstract

European bird cherry (Prunus padus L.) has been known since the Middle Ages for its medical/food use and high health-promoting value. This study aimed to assess the potential of these fruits as a source of bioactive compounds through the characterization of its [...] Read more.

European bird cherry (Prunus padus L.) has been known since the Middle Ages for its medical/food use and high health-promoting value. This study aimed to assess the potential of these fruits as a source of bioactive compounds through the characterization of its physicochemical traits, nutraceutical properties, phytochemical composition via HPLC fingerprint, and antioxidant capacity. Fully ripened fruits of Prunus padus L. (Colorata cv) were collected in mid-July 2017 in Chieri, north-western Italy. The TPC (194.22 ± 32.83 mg_GAE/100 g_FW) and TAC (147.42 ± 0.58 mg_C3G/100 g_FW) values were obtained from the analyzed extracts. The most important phytochemical class was organic acids (48.62 ± 2.31%), followed by polyphenols (35.34 ± 1.80%), monoterpenes (9.36 ± 0.64%), and vitamin C (6.68 ± 0.22%). In this research the most important flavonols selected as marker were quercitrin (16.37 ± 3.51 mg/100 g_FW) and quercetin (11.86 ± 2.36 mg/100 g_FW). Data were reported based on fresh weight. Moreover, fresh fruits showed a mean antioxidant activity value of 17.78 ± 0.84 mmol Fe²⁺·kg⁻¹. Even though the seeds and leaves contain cyanogenic glycosides, this study showed that these fruits could be a natural source of bioactive compounds with high antioxidant properties, due to the contents of organic and phenolic acids, catechins, and a synergetic effect of vitamin C and flavonoids. Full article

► Show Figures

Figure 1

11 pages, 1185 KiB

Open AccessCommunication

Development and Validation of a HPLC-UV Method for Extraction Optimization and Biological Evaluation of Hot-Water and Ethanolic Extracts of Dendropanax morbifera Leaves

by Hyung-Jae Choi, Dae-Hun Park, Seung-Hui Song, In-Soo Yoon and Seung-Sik Cho

Molecules 2018, 23(3), 650; https://doi.org/10.3390/molecules23030650 - 13 Mar 2018

Cited by 19 | Viewed by 5282

Abstract

Dendropanax morbifera Leveille (Araliaceae) has been used in traditional oriental remedies for cancer, inflammation, diabetes, and thrombosis. However, a validated analytical method, standardization, and optimization of extraction conditions with respect to biological activity have not been reported. In this study, a simple and [...] Read more.

Dendropanax morbifera Leveille (Araliaceae) has been used in traditional oriental remedies for cancer, inflammation, diabetes, and thrombosis. However, a validated analytical method, standardization, and optimization of extraction conditions with respect to biological activity have not been reported. In this study, a simple and validated HPLC method for identifying and quantifying active substances in D. morbifera was developed. Hot water and ethanolic D. morbifera leaf extracts from different production regions were prepared and evaluated with regard to their chemical compositions and biological activities. The contents of active compounds such as rutin and chlorogenic acid were determined in four samples collected from different regions. The 80% ethanolic extract showed the best antioxidant activity, phenolic content, reducing power, and xanthine oxidase (XO) inhibitory activity. The validated HPLC method confirmed the presence of chlorogenic acid and rutin in D. morbifera leaf extracts. The antioxidant and XO inhibitory activity of D. morbifera extract could be attributed to the marker compounds. Collectively, these results suggest that D. morbifera leaves could be beneficial for the treatment or prevention of hyperuricemia-related disease, and the validated HPLC method could be a useful tool for the quality control of food or drug formulations containing D. morbifera. Full article

► Show Figures

Figure 1

11 pages, 1241 KiB

Open AccessArticle

Comparison of Multiple Bioactive Constituents in Different Parts of Eucommia ulmoides Based on UFLC-QTRAP-MS/MS Combined with PCA

by Ying Yan, Hui Zhao, Cuihua Chen, Lisi Zou, Xunhong Liu, Chuan Chai, Chengcheng Wang, Jingjing Shi and Shuyu Chen

Molecules 2018, 23(3), 643; https://doi.org/10.3390/molecules23030643 - 13 Mar 2018

Cited by 44 | Viewed by 4897

Abstract

Eucommia ulmoides Oilv. (EU), also called Du-zhong, is a classical traditional Chinese medicine. Its bark, leaf, and male flower are all used for medicinal purposes, called Eucommiae Cortex (EC), Eucommiae Folium (EF), and Eucommiae Flos Male (EFM). In order to study the difference [...] Read more.

Eucommia ulmoides Oilv. (EU), also called Du-zhong, is a classical traditional Chinese medicine. Its bark, leaf, and male flower are all used for medicinal purposes, called Eucommiae Cortex (EC), Eucommiae Folium (EF), and Eucommiae Flos Male (EFM). In order to study the difference in synthesis and the accumulation of metabolites in different parts of EU, a reliable method based on ultra-fast liquid chromatography tandem triple quadrupole mass spectrometry (UFLC-QTRAP-MS/MS) was developed for the simultaneous determination of a total of 21 constituents, including two lignans, 6 iridoids, 6 penylpropanoids, 6 flavonoids, and one phenol in the samples (EC, EF, and EFM). Furthermore, principal component analysis (PCA) was performed to evaluate and classify the samples according to the contents of these 21 constituents. All of the results demonstrated that the chemical compositions in EC, EF, and EFM were significantly different and the differential constituents (i.e., aucubin, geniposidic acid, chlorogenic acid, pinoresinol-di-O-β-d-glucopyranoside, geniposide, cryptochlorogenic acid, rutin, and quercetin) were remarkably associated with sample classifications. The research will provide the basic information for revealing the laws of metabolite accumulation in EC, EF, and EFM from the same origin. Full article

► Show Figures

Figure 1

14 pages, 629 KiB

Open AccessArticle

Quality Evaluation of Apocyni Veneti Folium from Different Habitats and Commercial Herbs Based on Simultaneous Determination of Multiple Bioactive Constituents Combined with Multivariate Statistical Analysis

by Cuihua Chen, Zixiu Liu, Lisi Zou, Xunhong Liu, Chuan Chai, Hui Zhao, Ying Yan and Chengcheng Wang

Molecules 2018, 23(3), 573; https://doi.org/10.3390/molecules23030573 - 03 Mar 2018

Cited by 19 | Viewed by 3861

Abstract

Apocyni Veneti Folium (AVF) is a kind of staple traditional Chinese medicine with vast clinical consumption because of its positive effects. However, due to the habitats and adulterants, its quality is uneven. To control the quality of this medicinal herb, in this study, [...] Read more.

Apocyni Veneti Folium (AVF) is a kind of staple traditional Chinese medicine with vast clinical consumption because of its positive effects. However, due to the habitats and adulterants, its quality is uneven. To control the quality of this medicinal herb, in this study, the quality of AVF was evaluated based on simultaneous determination of multiple bioactive constituents combined with multivariate statistical analysis. A reliable method based on ultra-fast liquid chromatography tandem triple quadrupole mass spectrometry (UFLC-QTRAP-MS/MS) was developed for the simultaneous determination of a total of 43 constituents, including 15 flavonoids, 6 organic acids, 13 amino acids, and 9 nucleosides in 41 Luobumaye samples from different habitats and commercial herbs. Furthermore, according to the contents of these 43 constituents, principal component analysis (PCA) was employed to classify and distinguish between AVF and its adulterants, leaves of Poacynum hendersonii (PHF), and gray relational analysis (GRA) was performed to evaluate the quality of the samples. The proposed method was successfully applied to the comprehensive quality evaluation of AVF, and all results demonstrated that the quality of AVF was higher than the PHF. This study will provide comprehensive information necessary for the quality control of AVF. Full article

► Show Figures

Figure 1

15 pages, 3676 KiB

Open AccessArticle

Molecular and Functional Properties of Protein Fractions and Isolate from Cashew Nut (Anacardium occidentale L.)

by Cheng-mei Liu, Qian Peng, Jun-zhen Zhong, Wei Liu, Ye-jun Zhong and Fang Wang

Molecules 2018, 23(2), 393; https://doi.org/10.3390/molecules23020393 - 12 Feb 2018

Cited by 65 | Viewed by 8118

Abstract

Some molecular and functional properties of albumin (83.6% protein), globulin (95.5% protein), glutelin (81.3% protein) as well as protein isolate (80.7% protein) from cashew nut were investigated. These proteins were subjected to molecular (circular dichroism, gel electrophoresis, scanning electron microscopy) and functional (solubility, [...] Read more.

Some molecular and functional properties of albumin (83.6% protein), globulin (95.5% protein), glutelin (81.3% protein) as well as protein isolate (80.7% protein) from cashew nut were investigated. These proteins were subjected to molecular (circular dichroism, gel electrophoresis, scanning electron microscopy) and functional (solubility, emulsification, foaming, water/oil holding capacity) tests. Cashew nut proteins represent an abundant nutrient with well-balanced amino acid composition and could meet the requirements recommended by FAO/WHO. SDS-PAGE pattern indicated cashew nut proteins were mainly composed of a polypeptide with molecular weight (MW) of 53 kDa, which presented two bands with MW of 32 and 21 kDa under reducing conditions. The far-UV CD spectra indicated that cashew proteins were rich in β-sheets. The surface hydrophobicity of the protein isolate was higher than that of the protein fractions. In pH 7.0, the solubility of protein fractions was above 70%, which was higher than protein isolate at any pH. Glutelin had the highest water/oil holding capacity and foaming properties. Protein isolate displayed better emulsifying properties than protein fractions. In summary, cashew nut kernel proteins have potential as valuable nutrition sources and could be used effectively in the food industry. Full article

► Show Figures

Graphical abstract

Journal Menu

Journal Browser

Advances in Liquid Separation Techniques for Food and Pharmaceutical Analysis (Closed)

Share This Topical Collection

Editors

Topical Collection Information

Keywords

Published Papers (46 papers)

2024

Jump to: 2023, 2022, 2021, 2020, 2019, 2018

2023

Jump to: 2024, 2022, 2021, 2020, 2019, 2018

2022

Jump to: 2024, 2023, 2021, 2020, 2019, 2018

2021

Jump to: 2024, 2023, 2022, 2020, 2019, 2018

2020

Jump to: 2024, 2023, 2022, 2021, 2019, 2018

2019

Jump to: 2024, 2023, 2022, 2021, 2020, 2018

2018

Jump to: 2024, 2023, 2022, 2021, 2020, 2019

Further Information

Guidelines

MDPI Initiatives

Follow MDPI