Application Of Mass Spectrometry Research Articles

Molecular insight into algae-derived dissolved organic matters via Fourier-transform ion cyclotron resonance mass spectrometry: Effects of pretreatment methods and electrospray ionization modes

The release of algae-derived dissolved organic matter (ADOM) significantly increased in serious eutrophication waters, posing great threats to drinking water safety. Thus, the molecular composition decipherment is urgently in need. However, due to unsatisfactory pretreatment and ionization effects, the application of Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) on ADOM was limited. Therefore, the effects of pretreatment methods (cartridge type and loading) during solid-phase extraction (SPE) and electrospray ionization (ESI) modes with FT-ICR-MS on the molecular composition of ADOM were evaluated. The results showed compared with silica-based octadecyl (C18) cartridge, styrene-divinylbenzene polymer (PPL) cartridge exhibited higher recovery efficiency and retained more saturated and oxygenated compounds, such as carbohydrate-like and tannin-like. Furthermore, the recovery efficiency decreased with increasing loading, and hydrophilic and high-oxygenated carbohydrate-like and tannin-like were continuously replaced by hydrophobic and low-oxygenated aliphatic and aromatic compounds. Moreover, compared to negative ESI mode, the addition of positive ESI mode increased the molecular chemodiversity, especially more lipid-like and protein-like compounds. Thus, we proposed < 1:500 DOC/PPL mass ratio during SPE and dual ESI modes coupled with FT-ICR-MS could identify ADOM molecules more comprehensively. This work contributes to more comprehensive understanding of the molecular composition of ADOM and provides more references for pretreatment and characterization strategies of severely eutrophic waters.

Evaluation of photocathode materials in combination with commercial fiber-coupled UV photon sources as a compact electron source in mass spectrometry

Abstract The design of a compact electron source for mass spectrometry applications based on commercially available plug-and-play light sources and sturdy materials is presented, and its performance of photo-electron production is evaluated. We report quantum yields ranging between 1 – 2.5 x10-5 for aluminum, 9 x10-8 – 3 x10-6 for stainless steel, 1 – 2.2 x10-6 for molybdenum, 6.3 x10-7 – 5.1 x10-6 for tantalum, and 1.4 – 2.1 x10-6 for titanium, depending on the wavelength range of the irradiation source. In terms of quantum yield, the deuterium lamp provided better results compared to the two UV LEDs for all metals. However, due to their higher optical output power, the LEDs nevertheless yield a higher photoelectron current for all metals except for stainless steel. The measured photo-currents ranged from 1 – 2 nA for aluminum to 0.01 nA for Stainless Steel samples.

Comparative Investigation of Aroma-Active Volatiles in ("Ruixue", "Liangzhi", "Crystal Fuji," and "Guifei") Apples by Application of Gas Chromatography-Mass Spectrometry-Olfactometry (GC-MS-O) and Two-Dimensional Gas Chromatography-Quadrupole Mass Spectrometry (GC × GC-qMS) Coupled with Sensory Molecular Science.

Aroma dramatically impacts the overall flavor profiles and consumer acceptance; therefore, it is necessary to conduct a comprehensive analysis of the aroma characteristics of apples. In this study, the aroma differences among four popular apple varieties ("Ruixue", "Liangzhi", "Crystal Fuji," and "Guifei") were compared using two extraction methods (headspace-solid phase microextraction, and solvent-assisted flavor evaporation) coupled with gas chromatography-mass spectrometry-olfactometry (GC-MS-O) and two-dimensional gas chromatography-quadrupole mass spectrometry (GC × GC-qMS). A total of 82 odorants were identified via GC-MS-O, and 143 volatiles were identified by GC× GC-qMS. Among them, 41 key aroma-active compounds (butanal, ethyl acetate, 3-methylbutanal, methyl butanoate, 2-methylpropyl acetate, ethyl 2-methylbutanoate, ethyl 3-methylbutanoate, butyl acetate, hexanal, 2-methylbutyl acetate, 1-butanol, 2-methylpropyl butanoate, 3-methylbutyl acetate, (E)-2-hexenal, butyl butanoate, butyl 2-methylbutanoate, hexyl acetate, hexyl butanoate, hexyl, 2-methylbutanoate, 1-octen-3-ol, 3-methylthiopropanol, 1,3-octanediol, linalyl acetate, and so on) with high odor activity values (OAVs) and AI value (odor activity values ≥1 or aroma intensity ≥3) were identified. Partial least-squares-discriminant analysis showed that Ruixue exhibited a high "fruity" note, Guifei and Crystal Fuji had the greatest "wood," "floral," and "sweet" notes, while Liangzhi presented a significant "green" note. This study provided flavor chemistry support for the apple quality control and production.

Spatially heterogeneous lipid dysregulation in tuberculous meningitis

Tuberculous (TB) meningitis is the deadliest form of extrapulmonary TB which disproportionately affects children and immunocompromised individuals. Studies in pulmonary TB have shown that Mycobacterium tuberculosis can alter host lipid metabolism to evade the immune system. Cholesterol lowering drugs (i.e., statins) reduce the risk of infection, making them a promising host-directed therapy in pulmonary TB. However, the effect of M. tuberculosis infection on the young or adult brain lipidome has not been studied. The brain is the second-most lipid-rich organ, after adipose tissue, with a temporally and spatially heterogeneous lipidome that changes from infancy to adulthood. The young, developing brain in children may be uniquely vulnerable to alterations in lipid composition and homeostasis, as perturbations in cholesterol metabolism can cause developmental disorders leading to intellectual disabilities. To begin to understand the alterations to the brain lipidome in pediatric TB meningitis, we utilized our previously published young rabbit model of TB meningitis and applied mass spectrometry (MS) techniques to elucidate spatial differences. We used matrix assisted laser desorption/ionization-MS imaging (MALDI-MSI) and complemented it with region-specific liquid chromatography (LC)-MS/MS developed to identify and quantify sterols and oxysterols difficult to identify by MALDI-MSI. MALDI-MSI revealed several sphingolipids, glycerolipids and glycerophospholipids that were downregulated in brain lesions. LC-MS/MS revealed the downregulation of cholesterol, several sterol intermediates along the cholesterol biosynthesis pathway and enzymatically produced oxysterols as a direct result of M. tuberculosis infection. However, oxysterols produced by oxidative stress were increased in brain lesions. Together, these results demonstrate significant spatially regulated brain lipidome dysregulation in pediatric TB meningitis.

Validation and application of quick polar pesticide (QuPPe) extraction and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to analyze perchlorate in vegetables from Mexico

Perchlorate is a contaminant and endocrine disruptor present in vegetables. To address the lack of information on this problem in Mexico, we developed an analytical method to quantify perchlorate in commonly consumed vegetables based on quick polar pesticide (QuPPe) extraction and ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). The method was validated through the parameters of selectivity, accuracy, precision, linearity, limit of detection (LOD), and limit of quantification (LOQ) according to SANTE guidelines. The developed method exhibited a linear range of 0.005–0.100 mg/kg with LOD and LOQ values of 0.001 and 0.003 mg/kg, respectively. Recovery in a fortified tomato matrix was 96.85 ± 12.90% with a coefficient of variation (CV) of 11.55 ± 0.84%. The method was applied to analyze 31 vegetable samples collected from different regions in Mexico. Perchlorate concentrations ranged from not detected (ND) to 0.016 mg/kg, fresh weight. This method generates robust scientific evidence to implement and strengthen perchlorate monitoring programs for food matrices, supporting efforts to regulate this analyte in Mexico.

Advances in the application of liquid chromatography tandem mass spectrometry for therapeutic drug monitoring

Therapeutic drug monitoring is a good interpretation of personalized medicine, which helps to improve the safety and effectiveness of patient medication. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has gradually been widely accepted by clinical laboratories due to its characteristics of specificity, sensitivity, and flexibility in method development. This article introduces the importance and analytical techniques for therapeutic drug monitoring, describes the application and development of liquid chromatography tandem mass spectrometry for therapeutic drug monitoring, new advances in the detection of different drug types and sample types, and challenges in automation, convenience, and standardization.

Potential and application of tandem mass spectrometry (MS/MS) in the analysis and identification of novel bacteriocins: a review

SummaryBacteriocins are antimicrobial peptides synthesised ribosomally by Gram‐positive or Gram‐negative bacteria to gain a competitive advantage. The majority of bacteriocins are derived from Gram‐positive bacteria, with lactic acid bacteria being the most common source. Because they are considered ‘natural’, there is currently significant development of bacteriocins for application as food preservative agents. As a preservative agent, bacteriocin activity is highly dependent on purity, down to the amino acid profile and sequence. Therefore, bacteriocin identification is important. Currently, MS is a cutting‐edge tool in bacteriocin identification. This method has high selectivity, sensitivity and resolution. To the best of our knowledge, systematic reviews focusing on the application of MS for bacteriocin identification are currently limited. In light of this, the objective of this study is to provide a comprehensive review and summary of MS technologies in bacteriocin research, with a particular focus on the discovery and characterisation of novel sources of bacteriocin. Additionally, studies related to the discovery of bacteriocins from various sources, their role as antimicrobial agents, and their synthesis are emphasised. Thus, this study presents a comprehensive analysis of the advantages, limitations, and future perspectives of the methods employed.

Application of mass spectrometry in single-cell metabolomics

Application of mass spectrometry in single-cell metabolomics

Design, Synthesis, Biological Evaluation and Molecular Docking Studies of a New Series of Maleimide Derivatives.

A series of novel maleimide derivatives were synthesized, with various heterocyclic compounds serving as side chains in the synthesis process. The structural characteristics of these compounds were elucidated through the application of 1H-NMR spectroscopy, 13C-NMR (APT) spectroscopy, and high-resolution mass spectrometry (HRMS). The anti-cancer potential of these compounds was subsequently assessed in vitro, utilizing two distinct breast cancer cell lines, namely MDA-MB-231 and MCF-7, via MTT assay. Among the 12 newly synthesized compounds, 4 a, 4 b, 4 c, 4 d, 5 a, 5 b, 5 c and 5 d were determined to show the most promising anti-cancer activity against both breast cancer cell lines. Moreover, the morphological changes induced in the cells following a 24-hour incubation period with these compounds were observed using light microscopy. Additionally, molecular dynamics simulations were conducted to assess the stability of the bound conformations of the compounds to the target protein GSK-3β as obtained through molecular docking calculations.

Application of Mass Spectrometry for Analysis of Nucleobases, Nucleosides and Nucleotides in Tea and Selected Herbs: A Critical Review of the Mass Spectrometric Data.

The main and most commonly known biological function of nucleobases, nucleosides, and nucleotides is usually associated with the fact that they are the building blocks of nucleic acids. However, these compounds also belong to plant secondary metabolites, although in that role they have attracted less attention than the others, e.g., terpenes, phenolics, or alkaloids. The former compounds are also important constituents of the human diet, e.g., as ingredients of tea and herbs, endowing them with specific taste qualities and pharmacological activities. Liquid chromatography-mass spectrometry seems to be the most important analytical method that permits the identification and determination of nucleobases, nucleosides, and nucleotides, along with the other metabolites. The main goal of this review is to discuss in detail the aspects of mass spectrometric detection of nucleobases, nucleosides, and nucleotides in tea and selected herbs. An important conclusion is that the identification of the compounds of interest should be performed not only on the basis of [M + H]+/[M - H]- ions but should also be confirmed by the respective product ions; however, as discussed in detail in this review, it may sometimes be problematic. It also clear that all difficulties that may be encountered when analyzing plant material are caused by the complexity of the analyzed samples and the need to analyze different classes of compounds, and this review absolutely does not debase any of the mentioned papers.

Application of electrospray triple quadrupole mass spectrometry in assessing thalassemia in a population.

Thalassemia is widely prevalent in Asian countries and the Middle East region of the world. While the prevalence of α thal is around 5-20% in the population, the incidence of β thal is around 3-4%. The available data on thalassemia in Indians is based on chromatographic or electrophoretic techniques. The present study demonstrates the utility of mass spectrometry in detecting thalassemia in a small, healthy population from Tiptur, Karnataka. The study included a population of 315 individuals, spanning both children and adults, with ages ranging from 4 to 76 years. EDTA whole blood was collected after obtaining their written consent. The intensities of various globin chains were measured using a triple quadrupole mass spectrometer. The ratio of the peak areas of globin chains (α/β, δ/β (%)) served as biomarkers to detect thalassemia. Thirty random samples of α thalassemia were analyzed using GapPCR. The prevalence of a thal is 21.65% (68/314) based on α/β (< =0.8) and the prevalence of β thal is 3.8% (12/314) based on δ/β (%) (> =11.0). GapPCR detected three heteromutants of α deletion, one mutation in the HBA1 gene, and one a triplication (16.7%, 5/30). One sample was excluded from the analysis due to the presence of a variant. The utility of mass spectrometry for screening thalassemia in a population is demonstrated. The higher occurrence of α-thalassemia in the studied population underscores the importance of screening for thalassemia in regions with reportedly high prevalence rates.

Mass Spectrometry Advancements and Applications for Biomarker Discovery, Diagnostic Innovations, and Personalized Medicine.

Mass spectrometry (MS) has revolutionized clinical chemistry, offering unparalleled capabilities for biomolecule analysis. This review explores the growing significance of mass spectrometry (MS), particularly when coupled with liquid chromatography (LC), in identifying disease biomarkers and quantifying biomolecules for diagnostic and prognostic purposes. The unique advantages of MS in accurately identifying and quantifying diverse molecules have positioned it as a cornerstone in personalized-medicine advancement. MS-based technologies have transformed precision medicine, enabling a comprehensive understanding of disease mechanisms and patient-specific treatment responses. LC-MS has shown exceptional utility in analyzing complex biological matrices, while high-resolution MS has expanded analytical capabilities, allowing the detection of low-abundance molecules and the elucidation of complex biological pathways. The integration of MS with other techniques, such as ion mobility spectrometry, has opened new avenues for biomarker discovery and validation. As we progress toward precision medicine, MS-based technologies will be crucial in addressing the challenges of individualized patient care, driving innovations in disease diagnosis, prognosis, and treatment strategies.

Applications of MALDI mass spectrometry in forensic science.

Forensic chemistry literature has grown exponentially, with many analytical techniques being used to provide valuable information to help solve criminal cases. Among them, matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS), particularly MALDI MS imaging (MALDI MSI), has shown much potential in forensic applications. Due to its high specificity, MALDI MSI can analyze a wide variety of compounds in complex samples without extensive sample preparation, providing chemical profiles and spatial distributions of given analyte(s). This review introduces MALDI MS(I) to forensic scientists with a focus on its basic principles and the applications of MALDI MS(I) to the analysis of fingerprints, drugs of abuse, and their metabolites in hair, medicine samples, animal tissues, and inks in documents.

Advances in LDI-MS Analysis: The Role of Chemical Vapor Deposition-Synthesized Silver Nanoparticles in Enhancing Detection of Low-Molecular-Weight Biomolecules.

In this investigation, we detail the synthesis of silver nanoparticles (AgNPs) via a precise chemical vacuum deposition (CVD) methodology, aimed at augmenting the analytical performance of laser desorption/ionization mass spectrometry (LDI-MS) for the detection of low-molecular-weight analytes. Employing a precursor supply rate of 0.0014 mg/s facilitated the formation of uniformly dispersed AgNPs, characterized by SEM and AFM to have an average diameter of 33.5 ± 1.5 nm and a surface roughness (Ra) of 11.8 nm, indicative of their homogeneous coverage and spherical morphology. XPS and SEM-EDX analyses confirmed the metallic silver composition of the nanoparticles with Ag peak splitting, reflecting the successful synthesis of metallic Ag. Comparative analytical evaluation with traditional MALDI matrices revealed that AgNPs significantly reduce signal suppression, thereby enhancing the sensitivity and specificity of LDI-MS for low-molecular-weight compounds such as triglycerides, saccharides, amino acids, and carboxylic acids. Notably, the application of AgNPs demonstrated a superior linear response for triglyceride signals with regression coefficients surpassing 0.99, markedly outperforming conventional matrices. The study further extends into quantitative analysis through nanoparticle-based laser desorption/ionization (NALDI), where AgNPs exhibited enhanced ionization efficiency, characterized by substantially lower limits of detection (LOD) and quantification (LOQ) for tested standards. Particular attention was paid to lipids with a detailed examination of their fragmentation pathways. These results highlight the significant potential of AgNPs synthesized via CVD to transform the analytical detection and quantification of low-molecular-weight compounds using NALDI. This approach offers a promising avenue for expanding the scope of analytical applications in mass spectrometry and introducing innovative methodologies for enhanced precision and sensitivity.

LEAD-LEAD DATING REVEALS PERMIAN REMOBILIZATION OF NIOBIUM MINERALIZATION AT BAYAN OBO

Abstract The Bayan Obo deposit hosted by the H8 unit is a world-class rare earth element (REE) deposit with considerable niobium (Nb) and iron (Fe). Permian granites are widely exposed in the mining area and have a close spatial association with the Nb mineralization. Whether the granites contributed Nb or only remobilized existing mineralization is important for understanding the controls of ore formation. Previous studies have mostly focused on the REEs, whereas research on Nb has been limited. This is due mainly to the difficulty of accurately determining the age of the Nb mineralization because of the fine-grained and texturally complex nature of the Nb-bearing minerals and their exceptionally low U content. Although microbeam techniques show promise in tackling the aforementioned challenges, their application is hampered by matrix effects caused by the diverse composition of Nb-bearing minerals. Here we report the application of a high-precision secondary ion mass spectrometry (SIMS) Pb-Pb isochron approach that enables young samples (i.e., &amp;lt;500 Ma) to be dated without matrix-matched reference materials. A variety of Nb-bearing minerals from eastern Bayan Obo were analyzed, yielding Pb-Pb isochron ages of 276 ± 10 Ma (pyrochlore, 394–6,864 ppm U in the rim and 6,563–19,858 ppm in the core), 277 ± 36 Ma (fersmite, 18–61 ppm U; fergusonite-Ce, 45–95 ppm U), and 257 ± 46 Ma (aeschynite, 342–1,006 ppm U). In combination with the deposit geology and petrographic observations, these ages link the Nb mineralization to ~270 Ma granites. As these granites are not particularly rich in Nb, skarn formation during granite emplacement is interpreted to have remobilized the existing Nb mineralization, which increased the grain size of the Nb-bearing minerals—a key factor facilitating their extraction. Our study shows that high-precision SIMS Pb-Pb analysis holds promise for directly dating mineralization without matrix-matched reference materials. It also emphasizes the need to consider the role of the Nb remobilization at Bayan Obo and elsewhere.

Forensic application of sandpaper spray ionization mass spectrometry (SPS-MS): Direct analysis of solid pharmaceutical formulations and edible cannabis products.

In this work, we employed a new ambient ionization mass spectrometry technique, sandpaper spray mass spectrometry (SPS-MS), as an efficient tool to analyze pills and tablets of pharmaceutical formulations. The following samples were analyzed: regulators of blood pressure, cholesterol, and diabetes, and drugs for the treatment of erectile dysfunction (ED). Additionally, a hard candy of Cannabis sativa containing Δ9-tetrahydrocannabinol (Δ9-THC) and its related isomer cannabidiol (CBD) was also evaluated. The surfaces of the samples, without any prior treatment, were sanded onto triangular-cut sandpaper, and full MS scans (and MS/MS) were acquired in both positive and negative ionization modes. SPS-MS (and MS/MS) allowed for prompt detection of the active pharmaceutical ingredients (APIs) in each formulation. Other components of the formulations, added as excipients, were also tentatively identified. The results described herein indicate that the SPS-MS technique can be applied to fast screening of pills and tablets being potentially used as an efficient tool to detect counterfeit pharmaceutical and illicit products, a current issue of increasing concern.

Automated preparation of plasma lipids, metabolites, and proteins for LC/MS-based analysis of a high-fat diet in mice

Blood plasma is one of the most commonly analyzed and easily accessible biological samples. Here, we describe an automated liquid-liquid extraction platform that generates accurate, precise, and reproducible samples for metabolomic, lipidomic, and proteomic analyses from a single aliquot of plasma while minimizing hands-on time and avoiding contamination from plasticware. We applied mass spectrometry to examine the metabolome, lipidome, and proteome of 90 plasma samples to determine the effects of age, time of day, and a high-fat diet in mice. From 25 μl of mouse plasma, we identified 907 lipid species from 16 different lipid classes and subclasses, 233 polar metabolites, and 344 proteins. We found that the high-fat diet induced only mild changes in the polar metabolome, upregulated apolipoproteins, and induced substantial shifts in the lipidome, including a significant increase in arachidonic acid and a decrease in eicosapentaenoic acid content across all lipid classes.

Structural characterization of wax esters using ultraviolet photodissociation mass spectrometry

Wax esters play critical roles in biological systems, serving functions from energy storage to chemical signaling. Their diversity is attributed to variations in alcohol and acyl chains, including their length, branching, and the stereochemistry of double bonds. Traditional analysis by mass spectrometry with collisional activations (CID, HCD) offers insights into acyl chain lengths and unsaturation level. Still, it falls short in pinpointing more nuanced structural features like the position of double bonds. As a solution, this study explores the application of 213-nm ultraviolet photodissociation (UVPD) for the detailed structural analysis of wax esters. It is shown that lithium adducts provide unique fragments as a result of Norrish and Norrish-Yang reactions at the ester moieties and photoinduced cleavages of double bonds. The product ions are useful for determining chain lengths and localizing double bonds. UVPD spectra of various wax esters are presented systematically, and the effect of activation time is discussed. The applicability of tandem mass spectrometry with UVPD is demonstrated for wax esters from natural sources. The UHPLC analysis of jojoba oil proves the compatibility of MS2 UVPD with the chromatography time scale, and a direct infusion is used to analyze wax esters from vernix caseosa. Data shows the potential of UVPD and its combination with CID or HCD in advancing our understanding of wax ester structures.Graphical

Applications of native mass spectrometry and ultraviolet photodissociation in protein structure and interaction analysis

Dynamic changes in the structures and interactions of proteins are closely correlated with their biological functions. However, the precise detection and analysis of these molecules are challenging. Native mass spectrometry (nMS) introduces proteins or protein complexes into the gas phase by electrospray ionization, and then performs MS analysis under near-physiological conditions that preserve the folded state of proteins and their complexes in solution. nMS can provide information on stoichiometry, assembly, and dissociation constants by directly determining the relative molecular masses of protein complexes through high-resolution MS. It can also integrate various MS dissociation technologies, such as collision-induced dissociation (CID), surface-induced dissociation (SID), and ultraviolet photodissociation (UVPD), to analyze the conformational changes, binding interfaces, and active sites of protein complexes, thereby revealing the relationship between their interactions and biological functions. UVPD, especially 193 nm excimer laser UVPD, is a rapidly evolving MS dissociation method that can directly dissociate the covalent bonds of protein backbones with a single pulse. It can generate different types of fragment ions, while preserving noncovalent interactions such as hydrogen bonds within these ions, thereby enabling the MS analysis of protein structures with single-amino-acid-site resolution. This review outlines the applications and recent progress of nMS and UVPD in protein dynamic structure and interaction analyses. It covers the nMS techniques used to analyze protein-small-molecule ligand interactions, the structures of membrane proteins and their complexes, and protein-protein interactions. The discussion on UVPD includes the analysis of gas-phase protein structures and interactions, as well as alterations in protein dynamic structures, and interactions resulting from mutations and ligand binding. Finally, this review describes the future development prospects for protein analysis by nMS and new-generation advanced extreme UV light sources with higher brightness and shorter pulses.

Hyphenation of microflow chromatography with electrospray ionization mass spectrometry for bioanalytical applications focusing on low molecular weight compounds: A tutorial review.

Benefits of miniaturized chromatography with various detection modes, such as increased sensitivity, chromatographic efficiency, and speed, were recognized nearly 50 years ago. Over the past two decades, this approach has experienced rapid growth, driven by the emergence of mass spectrometry applications serving -omics sciences and the need for analyzing minute volumes of precious samples with ever higher sensitivity. While nanoscale liquid chromatography (flow rates <1 μL/min) has gained widespread recognition in proteomics, the adoption of microscale setups (flow rates ranging from 1 to 100 μL/min) for low molecular weight compound applications, including metabolomics, has been surprisingly slow, despite the inherent advantages of the approach. Highly heterogeneous matrices and chemical structures accompanied by a relative lack of options for both selective sample preparation and user-friendly equipment are usually reported as major hindrances. To facilitate the wider implementation of microscale analyses, we present here a comprehensive tutorial encompassing important theoretical and practical considerations. We provide fundamental principles in micro-chromatography and guide the reader through the main elements of a microflow workflow, from LC pumps to ionization devices. Finally, based on both our literature overview and experience, illustrated by some in-house data, we highlight the critical importance of the ionization source design and its careful optimization to achieve significant sensitivity improvement.