Use Of HPLC-MS Research Articles

Progress in the analysis of phytocannabinoids by HPLC and UPLC (or UHPLC) during 2020-2023.

Organic molecules that bind to cannabinoid receptors are known as cannabinoids. These molecules possess pharmacological properties similar to those produced by Cannabis sativa L. High-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography (UPLC, also known as ultra-high-performance liquid chromatography, UHPLC) have become the most widely used analytical tools for detection and quantification of phytocannabinoids in various matrices. HPLC and UPLC (or UHPLC) are usually coupled to an ultraviolet (UV), photodiode array (PDA), or mass spectrometric (MS) detector. To critically appraise the literature on the application of HPLC and UPLC (or UHPLC) methods for the analysis of phytocannabinoids published from January 2020 to December 2023. An extensive literature search was conducted using Web of Science, PubMed, and Google Scholar and published materials including relevant books. In various combinations, using cannabinoid in all combinations, cannabis, hemp, hashish, C. sativa, marijuana, analysis, HPLC, UHPLC, UPLC, and quantitative, qualitative, and quality control were used as the keywords for the literature search. Several HPLC- and UPLC (or UHPLC)-based methods for the analysis of phytocannabinoids were reported. While simple HPLC-UV or HPLC-PDA-based methods were common, the use of HPLC-MS, HPLC-MS/MS, UPLC (or UHPLC)-PDA, UPLC (or UHPLC)-MS, and UPLC (or UHPLC)-MS/MS was also reported. Applications of mathematical and computational models for optimization of protocols were noted. Pre-analyses included various environmentally friendly extraction protocols. During the last 4 years, HPLC and UPLC (or UHPLC) remained the main analytical tools for phytocannabinoid analysis in different matrices.

Use of HPLC-MS to Determine the Loss of Metabolites in Apple Juices under Different Storage Conditions.

The focus of this experiment was to compare the color and metabolic profile of apple juice from the red-fleshed cultivar 'Baya Marisa' with the white-fleshed cultivar 'Golden Delicious'. The changes in the phenolic compounds, organic acids, and sugar content during high-temperature short-time pasteurization and after one year of storage under different storage conditions were analyzed. A total of 26 individual phenolic compounds were identified and quantified. The total analyzed phenolics content (TAPC) decreased after pasteurization of the juices of both cultivars. The TAPC of fresh 'Baya Marisa' juice after pasteurization increased or remained the same compared to one-year stored 'Baya Marisa' juice, depending on the storage method. The sucrose content of the apple juice of both cultivars remained the same after pasteurization; interestingly, it decreased significantly after one year of storage, while the fructose and glucose content remained the same after pasteurization and increased significantly after one year of storage for both cultivars.

Disease resistance induction by K. marxianus against green mold in citrus via transcriptome analysis and verification based on metabolites

This study examined the transcriptional composition and changes in gene expression patterns in citrus fruit tissue treated with Kluyveromyces marxianus and its effects on key defense-related enzyme activities and metabolites. This analysis was carried out by the technique of RNA sequencing, the use of HPLC-MS, and GC–MS equipment. The sequencing data revealed that the induction of K. marxianus in the citrus fruit peel activated the expression of genes in 90 biological pathways. A total of 3785 genes were transcribed between the yeast-treated and control fruits. 1287 genes were expressed significantly, 682 genes were up-regulated representing 52.99 %, and 605 genes were down-regulated representing 47.00 %. Gene expressions in some of the pathways in the treated citrus stimulated an increase in the production of some defense-related enzymes, the synthesis of secondary metabolites such as phenolic compounds, flavonoids, phenolic acids, and an enhanced lignin production. The accumulation of these compounds contributed to the improvement of the antifungal activity and defense responses in the citrus. 8 phenolic acids, 12 sugars, 6 organic acids, and 17 amino acids were detected at varying concentrations. The investigations revealed that the production of the metabolites was significantly enhanced in the K. marxianus-treated fruits as compared to the control fruits. The study provides some fundamental understanding of the induced resistance mechanism in citrus by K. marxianus.

Quantitative Analysis and Differential Evaluation of Radix Bupleuri Cultivated in Different Regions Based on HPLC-MS and GC-MS Combined with Multivariate Statistical Analysis.

The quality of Radix Bupleuri is greatly affected by its growing environment. In this study, Radix Bupleuri samples that were harvested from seven different regions across northwest China were examined by high-performance liquid chromatography (HPLC) and gas chromatography (GC) coupled with mass spectrometry (MS) to reveal significant differences in quality contributed by the cultivation region. An HPLC-MS method was firstly established and used in the multiple reaction monitoring mode for the quantitative analysis of five saikosaponins in Radix Bupleuri so as to evaluate the difference in the absolute content of saikosaponins attributable to the cultivation region. The effect on the components of Radix Bupleuri was further investigated based on the profiles of the representative saponins and volatile compounds, which were extracted from the Radix Bupleuri samples and analyzed by HPLC-MS and GC-MS. Multivariate statistical analysis was employed to differentiate the Radix Bupleuri samples cultivated in different regions and to discover the differential compositions. The developed quantitative method was validated to be accurate, stable, sensitive, and repeatable for the determination of five saikosaponins. Further statistical tests revealed that the collected Radix Bupleuri samples were distinctly different from each other in terms of both saponins and volatile compounds, based on the provinces where they were grown. In addition, twenty-eight saponins and fifty-eight volatile compounds were identified as the differentially accumulated compositions that contributed to the discrimination of the Radix Bupleuri samples. The Radix Bupleuri samples grown in Shouyang county showed the highest content of saikosaponins. All of the results indicated that the cultivation region significantly affected the accumulation and diversity of the main chemical components of Radix Bupleuri. The findings of this research provide insights into the effect of the cultivation region on the quality of Radix Bupleuri and the differentiation of Radix Bupleuri cultivated in different regions based on the use of HPLC-MS and GC-MS combined with multivariate statistical analysis.

Bioavailability, bioactivity and impact on health of dietary flavonoids and related compounds: an update.

There is substantial interest in the role of plant secondary metabolites as protective dietary agents. In particular, the involvement of flavonoids and related compounds has become a major topic in human nutrition research. Evidence from epidemiological and human intervention studies is emerging regarding the protective effects of various (poly)phenol-rich foods against several chronic diseases, including neurodegeneration, cancer and cardiovascular diseases. In recent years, the use of HPLC-MS for the analysis of flavonoids and related compounds in foods and biological samples has significantly enhanced our understanding of (poly)phenol bioavailability. These advancements have also led to improvements in the available food composition and metabolomic databases, and consequently in the development of biomarkers of (poly)phenol intake to use in epidemiological studies. Efforts to create adequate standardised materials and well-matched controls to use in randomised controlled trials have also improved the quality of the available data. In vitro investigations using physiologically achievable concentrations of (poly)phenol metabolites and catabolites with appropriate model test systems have provided new and interesting insights on potential mechanisms of actions. This article will summarise recent findings on the bioavailability and biological activity of (poly)phenols, focusing on the epidemiological and clinical evidence of beneficial effects of flavonoids and related compounds on urinary tract infections, cognitive function and age-related cognitive decline, cancer and cardiovascular disease.

DEVELOPMENT OF DIRECT SERIAL COUPLING OF HYDROPHILIC INTERACTION LIQUID CHROMATOGRAPHY AND REVERSED-PHASE COLUMNS FOR THE SIMULTANEOUS ANALYSIS OF ANTI-TUBERCULOSIS DRUGS

The use of HPLC-MS and -UV systems are preferred methods for the detection of pharmaceutical drugs. In this study, direct serial coupling of HILIC to a reversed-phase column via a zero-volume union was applied to resolve a mixture of first-line anti-tuberculosis drugs, including rifampicin (RIF), isoniazid (INH), pyrazinamide (PZA), ethambutol (EMB), and streptomycin (STP). These analytes, detected by either UV or MS, were chosen for their wide range of polarities and their analysis was achieved in a single HPLC injection with a 16 min runtime per sample and a flow rate of 0.6 mL/min. In addition, optimization of the chromatographic conditions required to separate the antibiotics in the direct serial coupling setup are also discussed.

Production of Metabolites from the Penicillium roqueforti Complex

Penicillium roqueforti comprises three accepted species: P. carneum, which is associated with meat, cheese, and bread; P. paneum, associated primarily with bread and silage; and P. roqueforti, which is associated with various processed foods and silage. This paper reports the use of HPLC-MS and HPLC-NMR to investigate the metabolites of silage-derived strains from two areas where silage toxicoses are regularly observed (Scandinavia and eastern Canada). Only modest differences were seen between the metabolites produced by strains from Canada and Scandinavia; however, silage strains of P. paneum isolated from Quebec were poor producers of patulin. This paper reports for the first time the production of festuclavine from P. paneum. This may be important as a possible explanation for the ill thrift observed when this species is dominant in poorly ensiled materials fed to dairy cows.

The Turnover Kinetics of Major Histocompatibility Complex Peptides of Human Cancer Cells

Peptides presented by the major histocompatibility complex (MHC) are derived from the degradation of cellular proteins. Thus, the repertoire of these peptides (the MHC peptidome) should correlate better with the cellular protein degradation scheme (the degradome) than with the cellular proteome. To test the validity of this statement and to determine whether the majority of MHC peptides are derived from short lived proteins, from defective ribosome products, or from regular long lived cellular proteins we analyzed in parallel the turnover kinetics of both MHC peptides and cellular proteins in the same cancer cells. The analysis was performed by pulse-chase experiments based on stable isotope labeling in tissue culture followed by capillary chromatography and tandem mass spectrometry. Indeed only a limited correlation was observed between the proteome and the MHC peptidome observed in the same cells. Moreover a detailed analysis of the turnover kinetics of the MHC peptides helped to assign their origin to normal, to short lived or long lived proteins, or to the defective ribosome products. Furthermore the analysis of the MHC peptides turnover kinetics helped to direct attention to abnormalities in the degradation schemes of their source proteins. These observations can be extended to search for cancer-related abnormalities in protein degradation, including those that lead to loss of tumor suppressors and cell cycle regulatory proteins.

Analysis of neem oils by LC-MS and degradation kinetics of azadirachtin-A in a controlled environment. Characterization of degradation products by HPLC-MS-MS.

Since it was first isolated, the oil extracted from seeds of neem (Azadirachtin indica A juss) has been extensively studied in terms of its efficacy as an insecticide. Several industrial formulations are produced as emulsifiable solutions containing a stated titer of the active ingredient azadirachtin-A (AZ-A). The work reported here is the characterization of a formulation of this insecticide marketed under the name of Neem-azal T/S and kinetic studies of the major active ingredient of this formulation. We initially performed liquid-liquid extraction to isolate the neem oil from other ingredients in the commercial mixture. This was followed by a purification using flash chromatography and semi-preparative chromatography, leading to (13)C NMR identification of structures such as azadirachtin-A, azadirachtin-B, and azadirachtin-H. The neem extract was also characterized by HPLC-MS using two ionization sources, APCI (atmospheric pressure chemical ionization) and ESI (electrospray ionization) in positive and negative ion modes of detection. This led to the identification of other compounds present in the extract-azadirachtin-D, azadirachtin-I, deacetylnimbin, deacetylsalannin, nimbin, and salannin. The comparative study of data gathered by use of the two ionization sources is discussed and shows that the ESI source enables the largest number of structures to be identified. In a second part, kinetic changes in the main product (AZ-A) were studied under precise conditions of pH (2, 4, 6, and 8), temperature (40 to 70 degrees C), and light (UV, dark room and in daylight). This enabled us to determine the degradation kinetics of the product (AZ-A) over time. The activation energy of the molecule (75+/-9 kJ mol(-1)) was determined by examining thermal stability in the range 40 to 70 degrees C. The degradation products of this compound were identified by use of HPLC-MS and HPLC-MS-MS. The results enabled proposal of a chemical degradation reaction route for AZ-A under different conditions of pH and temperature. The data show that at room temperature and pH between 4 and 5 the product degrades into two preferential forms that are hydrolyzed to a single product over time and as a function of pH change.

Measurement of Trace Elements in Marine Environmental Samples Using Solution ICPMS. Current and Future Applications

AbstractFor Abstract see ChemInform Abstract in Full Text.

The use of HPLC-MS in T-cell epitope identification

The hunt for T-cell epitopes is going on because hopes are set on such peptide sequences for diagnosis and vaccine development in the fight against infectious and tumor diseases. In addition to a variety of other techniques used in T-cell epitope identification, mass spectrometers coupled to microcapillary liquid chromatography have now become an important and sensitive tool in separation, detection, and sequence analysis of highly complex natural major histocompatibility complex (MHC) ligand mixtures. In this article, we review the basics of mass spectrometric techniques and their on-line coupling to microcapillary liquid chromatography (μcap-LC). Furthermore, we introduce current strategies for the identification of new T-cell epitopes using microcapillary liquid chromatography–mass spectrometry (μcap-LC-MS).

Measurement of Trace Elements in Marine Environmental Samples using Solution ICPMS. Current and Future Applications

The strengths and weaknesses of using inductively coupled plasma mass spectrometer (ICPMS) measurements of samples in solution for marine environmental analyses using real world examples is discussed. ICPMS can detect nanogram per litre concentrations of trace elements but suffers from polyatomic interferences generated from the sample matrix. Most of the routine trace elements measured in marine biological tissue and sedimentdigests, with the notable exceptions of iron, chromium, vanadium, and selenium, are not subject to severe interferences. Low recoveries of trace elements from sediments are due to the inability of extraction acids to remove trace elements such as chromium and nickel from sediment matrices. The use of ICPMS offers the advantage that elements such as phosphorus, which previously required elaborate digestion procedures and a colorimetric determination, can be rapidly determined using nitric acid digestion alone. The use of flow injection coupled with ICPMS allows on-line preconcentration of trace metals and metalloids using chelation by ion-exchange resins or hydride generation and trapping as well as separation from matrix elements. Thus, the routine determination of trace elements and inorganic and methylated arsenic, antimony, mercury, and germanium species in open-ocean waters is possible. The coupling of HPLC and GC to ICPMS allows the measurement of metal and metalloid species in biological and sediment extracts. However, extraction of unaltered species from matrices presents a challenge. Many of the species found in the environmental samples are not known and analytical standards are not available. The concurrent use of HPLC-MS is needed to confirm these species.