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Applications of High-Performance Liquid Chromatography (HPLC) in Biological Sample Analysis
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Applications of High-Performance Liquid Chromatography (HPLC) in Biological Sample Analysis

A special issue of Molecules (ISSN 1420-3049). This special issue belongs to the section "Analytical Chemistry".

Deadline for manuscript submissions: closed (31 December 2023) | Viewed by 12501

Special Issue Editor

Dr. Ritin Sharma

E-Mail Website
Guest Editor
Translational Genomics Research Institute, Phoenix, AZ, USA
Interests: biological Mass Spectrometry; proteomics; phosphoproteomics; targeted proteomics; metaproteomics; microbial proteomics; cancer biology; kinases; signal transduction; PTMs; neurological disorders
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Liquid chromatography for biological sample analysis has greatly advanced in the last decade. With the improvements in design and engineering of HPLC instrumentation, nanoscale to high-flow separation is now routinely performed in analytical labs throughout the world. However, the complexity associated with various biological matrices such as high dynamic range of blood plasma or cellular heterogeneity in tissues or tumors present additional challenges for successful analyte measurement. Researchers are constantly striving to increase sensitivity and precision of biological assays by improving separation methods, developing novel column chemistries or enhancing detection methods.

The focus of this special issue is to highlight novel approaches for analysis of biomolecules (proteins, peptides, post-translational modifications, metabolites, lipids, nucleic acids...) in various biological matrices using HPLC/UHPLC coupled to different detection methods such as mass spectrometry, UV and NMR. The issue welcomes original research articles, technical notes or review articles describing detection of targeted panel of analytes, application of liquid chromatography in proteomics, lipidomics or metabolomics, methods showcasing advancement in sample preparation for liquid chromatography and methods reporting novel chromatographic techniques. The issue also welcome submission from researchers involved in development of new software or tools for LC analysis.

Dr. Ritin Sharma
Guest Editor

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Keywords

  • liquid chromatography
  • mass spectrometry
  • small molecules
  • proteomics
  • peptides
  • lipidomics
  • metabolomics
  • HPLC

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Published Papers (5 papers)

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Research

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11 pages, 2957 KiB  
Article
Quantitative Analysis of Flavonoids in Fruiting Bodies of Sanghuangporus Using Ultra-High-Performance Liquid Chromatography Coupled with Triple Quadrupole Mass Spectrometry
by Zhongjing Zhou, Zhiping Deng, Shuang Liang, Xiaowei Zou, Yi Teng, Weike Wang and Lizhong Fu
Molecules 2023, 28(13), 5166; https://doi.org/10.3390/molecules28135166 - 2 Jul 2023
Cited by 3 | Viewed by 1864
Abstract
A rapid, precise, and dependable method for quantifying flavonoids in the fruiting bodies of Sanghuangporus was established using ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS). Separation was achieved using a ZORBAX Eclipse Plus C18 column (1.8 μm, 3.0 mm × [...] Read more.
A rapid, precise, and dependable method for quantifying flavonoids in the fruiting bodies of Sanghuangporus was established using ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS). Separation was achieved using a ZORBAX Eclipse Plus C18 column (1.8 μm, 3.0 mm × 100 mm) with a 15 min gradient of a mobile phase consisting of 0.01% aqueous formic acid and 2 mm/L ammonium formate (mobile phase A), and 0.01% formic acid and 2 mm/L ammonium formate in methanol (mobile phase B). A mass spectrometry analysis was performed using the multiple reaction monitoring (MRM) mode with an electrospray ion source. This method enabled the simultaneous detection of 10 flavonoids (sakuranetin, quercitrin, myricitrin, kaempferol, luteolin, rutin, hyperoside, kaempferol-3-O-rutinoside, catechin, and catechin gallate) in the fruiting bodies of Sanghuangporus. Additionally, we applied this method to analyze the flavonoid content in fruiting bodies of various Sanghuangporus species. The results revealed substantial variations in flavonoid content, up to a 100-fold difference, among different species, with myricitrin, hyperoside, and rutin identified as the most abundant flavonoids. This protocol serves as a valuable tool for quantifying flavonoid compounds in different Sanghuangporus species or under diverse cultivation conditions, particularly for identifying species with high levels of specific flavonoid compounds. Full article
(This article belongs to the Special Issue Applications of High-Performance Liquid Chromatography (HPLC) in Biological Sample Analysis)
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11 pages, 1211 KiB  
Article
Dissipation Dynamic, Residue Distribution and Risk Assessment of Emamectin Benzoate in Longan by High-Performance Liquid Chromatography with Fluorescence Detection
by Yanping Liu, Haibin Sun, Xiaonan Wang, Hong Chang and Siwei Wang
Molecules 2023, 28(8), 3346; https://doi.org/10.3390/molecules28083346 - 10 Apr 2023
Cited by 2 | Viewed by 1653
Abstract
A derivatization method combined with high-performance liquid chromatography–fluorescence detection (HPLC–FLD) was used to evaluate the dissipation, residue distribution and risk assessment of emamectin benzoate in whole longan and pulp. The average recoveries were 82–111% with relative standard deviation (RSD) less than 11%. The [...] Read more.
A derivatization method combined with high-performance liquid chromatography–fluorescence detection (HPLC–FLD) was used to evaluate the dissipation, residue distribution and risk assessment of emamectin benzoate in whole longan and pulp. The average recoveries were 82–111% with relative standard deviation (RSD) less than 11%. The limit of quantification (LOQ) was 0.001 mg/kg in longan and pulp. The half-lives were 3.3–4.2 days. The terminal residues in whole longan were <0.001–0.025 mg/kg applied two and three times at two levels of dosage with PHIs of 10, 14, and 21 days. The residues in whole longan had a higher quantity than those in the pulp, and the terminal residues of pulp were all lower than LOQ (0.001 mg/kg). The chronic risk of emamectin benzoate was not negligible to humans depending on ADI% value, which was higher than 1; and the acute risk was acceptable to the consumer. This study could provide guidance for the safe use of emamectin benzoate in longan and serve as a reference for the establishment of maximum residue limits (MRLs) in China. Full article
(This article belongs to the Special Issue Applications of High-Performance Liquid Chromatography (HPLC) in Biological Sample Analysis)
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12 pages, 1285 KiB  
Article
Simultaneous Determination of Orelabrutinib, Zanubrutinib, Ibrutinib and Its Active Metabolite in Human Plasma Using LC-MS/MS
by Lu-Ning Sun, Yang Zhao, Zhou-Yi Qian, Xiang-Long Chen, Hong Ma, Yu-Jiao Guo, Hao Shen and Yong-Qing Wang
Molecules 2023, 28(3), 1205; https://doi.org/10.3390/molecules28031205 - 26 Jan 2023
Cited by 3 | Viewed by 2313
Abstract
Ibrutinib, orelabrutinib, and zanubrutinib are all Bruton’s tyrosine kinase inhibitors, which have greatly improved the treatment of B-cell malignancies. In this study, an LC-MS/MS method was developed and validated for the determination of orelabrutinib, zanubrutinib, ibrutinib, and its active metabolite dihydrodiol ibrutinib in [...] Read more.
Ibrutinib, orelabrutinib, and zanubrutinib are all Bruton’s tyrosine kinase inhibitors, which have greatly improved the treatment of B-cell malignancies. In this study, an LC-MS/MS method was developed and validated for the determination of orelabrutinib, zanubrutinib, ibrutinib, and its active metabolite dihydrodiol ibrutinib in human plasma. The Ibrutinib-d5 was used as the internal standard. Pretreatment was performed using a simple protein precipitation step using acetonitrile. The ACQUITY UPLC HSS T3 column (2.1×50 mm, 1.8 μm) was used to separate the analytes, and the run time was 6.5 min. The mobile phase consisted of acetonitrile and 10 mM of ammonium formate, which contained 0.1% formic acid. The multiple reactions’ monitoring transitions were selected at m/z 428.1→411.2, 472.2→455.2, 441.1→304.2, 475.2→304.2 and 446.2→309.2 respectively for orelabrutinib, zanubrutinib, ibrutinib, dihydrodiol ibrutinib and ibrutinib-d5 using positive ion electrospray ionization. The standard curves were linear, from 0.400 to 200 ng/mL for ibrutinib and dihydrodiol ibrutinib, 1.00–500 ng/mL for orelabrutinib, and 2.00–1000 ng/mL for zanubrutinib. Selectivity, the lower limit of quantitation, precision, accuracy, matrix effect, recovery, stability, and dilution integrity all met the acceptance criteria of FDA guidance. This method was used to quantify the plasma levels of orelabrutinib, zanubrutinib, ibrutinib, and dihydrodiol ibrutinib in clinical patients. Full article
(This article belongs to the Special Issue Applications of High-Performance Liquid Chromatography (HPLC) in Biological Sample Analysis)
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13 pages, 2477 KiB  
Article
A Comparative Study on the Reduction Modes for Quinone to Determine Ubiquinone by HPLC with Luminol Chemiluminescence Detection Based on the Redox Reaction
by Naoya Kishikawa, Mahmoud El-Maghrabey, Miharu Tobo and Naotaka Kuroda
Molecules 2023, 28(1), 96; https://doi.org/10.3390/molecules28010096 - 22 Dec 2022
Cited by 1 | Viewed by 1892
Abstract
Ubiquinone (UQ) is considered one of the important biologically active molecules in the human body. Ubiquinone determination in human plasma is important for the investigation of its bioavailability, and also its plasma level is considered an indicator of many illnesses. We have previously [...] Read more.
Ubiquinone (UQ) is considered one of the important biologically active molecules in the human body. Ubiquinone determination in human plasma is important for the investigation of its bioavailability, and also its plasma level is considered an indicator of many illnesses. We have previously developed sensitive and selective chemiluminescence (CL) method for the determination of UQ in human plasma based on its redox cycle with dithiothreitol (DTT) and luminol. However, this method requires an additional pump to deliver DTT as a post-column reagent and has the problems of high DTT consumption and broadening of the UQ peak due to online mixing with DTT. Herein, an HPLC (high-performance liquid chromatography) system equipped with two types of online reduction systems (electrolytic flow cell or platinum catalyst-packed reduction column) that play the role of DTT was constructed to reduce reagent consumption and simplify the system. The newly proposed two methods were carefully optimized and validated, and the analytical performance for UQ determination was compared with that of the conventional DTT method. Among the tested systems, the electrolytic reduction system showed ten times higher sensitivity than the DTT method, with a limit of detection of 3.1 nM. In addition, it showed a better chromatographic performance and the best peak shape with a number of theoretical plates exceeding 6500. Consequently, it was applied to the determination of UQ in healthy human plasma, and it showed good recovery (≥97.9%) and reliable precision (≤6.8%) without any interference from plasma components. Full article
(This article belongs to the Special Issue Applications of High-Performance Liquid Chromatography (HPLC) in Biological Sample Analysis)
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Review

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15 pages, 3610 KiB  
Review
The High-Precision Liquid Chromatography with Electrochemical Detection (HPLC-ECD) for Monoamines Neurotransmitters and Their Metabolites: A Review
by Bruno P. Guiard and Guillaume Gotti
Molecules 2024, 29(2), 496; https://doi.org/10.3390/molecules29020496 - 19 Jan 2024
Cited by 5 | Viewed by 3295
Abstract
This review highlights the advantages of high-precision liquid chromatography with an electrochemical detector (HPLC-ECD) in detecting and quantifying biological samples obtained through intracerebral microdialysis, specifically the serotonergic and dopaminergic systems: Serotonin (5-HT), 5-hydroxyindolacetic acid (5-HIAA), 3,4-dihydroxyphenylacetic acid (DOPAC), dopamine (DA), 3-metoxytryptamin (3-MT) and [...] Read more.
This review highlights the advantages of high-precision liquid chromatography with an electrochemical detector (HPLC-ECD) in detecting and quantifying biological samples obtained through intracerebral microdialysis, specifically the serotonergic and dopaminergic systems: Serotonin (5-HT), 5-hydroxyindolacetic acid (5-HIAA), 3,4-dihydroxyphenylacetic acid (DOPAC), dopamine (DA), 3-metoxytryptamin (3-MT) and homovanillic acid (HVA). Recognized for its speed and selectivity, HPLC enables direct analysis of intracerebral microdialysis samples without complex derivatization. Various chromatographic methods, including reverse phase (RP), are explored for neurotransmitters (NTs) and metabolites separation. Electrochemical detector (ECD), particularly with glassy carbon (GC) electrodes, is emphasized for its simplicity and sensitivity, aimed at enhancing reproducibility through optimization strategies such as modified electrode materials. This paper underscores the determination of limits of detection (LOD) and quantification (LOQ) and the linear range (L.R.) showcasing the potential for real-time monitoring of compounds concentrations. A non-exhaustive compilation of literature values for LOD, LOQ, and L.R. from recent publications is included. Full article
(This article belongs to the Special Issue Applications of High-Performance Liquid Chromatography (HPLC) in Biological Sample Analysis)
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