Timmerman P, McDougall S, Adcock N
… +17 more, Arfvidsson C, Barfield M, Blech S, Cowan KJ, Ferrari L, Greco A, Golob M, Goodwin L, Hughes R, Ivanova T, Laurén A, Nelson R, Neitzel S, Verhaeghe T, van de Merbel N, Wright M, White S
Bioanalysis
· 2025 Sep · PMID 40916553
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The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use M10 guideline provides a global framework for bioanalytical method validation in studies intended for regulatory sub...The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use M10 guideline provides a global framework for bioanalytical method validation in studies intended for regulatory submission. While its structure ensures consistency and data reliability, the guideline also acknowledges that not all studies require the same level of validation. This paper examines where full compliance is essential and where scientific judgment allows for leaner, context-driven validation, such as in early-stage development, for additional matrices, metabolites, nonstandard biological matrices or studies intended for internal decision-making. Drawing on recent recommendations from the European Bioanalysis Forum, we introduce a decision-making flowchart and parameter table to support consistent application of a Context-of-Use approach to validation. These tools help guide when flexibility is appropriate while ensuring transparency and robustness in the data. The paper advocates continued dialogue both with end users of the data and the regulatory authorities to support a modernized, risk-based validation framework that remains aligned with patient needs and scientific integrity. We believe the recommendations in this paper are fully in alignment with the intent and core principles of ICH M10, while encouraging their application in a way that remains scientifically driven and proportionate to the purpose of the assay.
Bioanalysis
· 2025 Sep · PMID 40910749
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BACKGROUND: High-throughput solid-phase extraction coupled with tandem mass spectrometry (HT-SPE-MS/MS) is an automated sample delivery system to mass spectrometry that operates without chromatographic separation. The ty...BACKGROUND: High-throughput solid-phase extraction coupled with tandem mass spectrometry (HT-SPE-MS/MS) is an automated sample delivery system to mass spectrometry that operates without chromatographic separation. The typical analysis time per sample using this platform is 10-30 s. While the HT-SPE-MS/MS system has demonstrated efficacy for in vitro assays, its application to the analysis of biological samples from in vivo bioavailability and bioequivalence studies presents challenges due to the complexity of the sample matrix. Three critical issues - matrix effect, specificity, and carryover - have not been thoroughly evaluated in complex biological matrices such as plasma. RESEARCH DESIGN AND METHODS: This study assessed the feasibility of utilizing HT-SPE-MS/MS for the analysis of three metabolically related compounds (bupropion, hydroxybupropion, and threobupropion) in human plasma samples from a clinical bioequivalence study. Critical bioanalytical parameters, including matrix effect, specificity, accuracy, precision, and carryover, were systematically investigated. RESULTS: These methods were subsequently applied to a bioequivalence study of bupropion. The HT-SPE-MS/MS approach achieved comparable accuracy, precision, linearity, and sensitivity to conventional ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) methods, while offering 20- to 30-fold higher analysis speeds. CONCLUSION: The results of this study indicate that the HT-SPE-MS/MS system shows potential for high-throughput in vivo bioanalysis, particularly in bioavailability and bioequivalence studies.
Bioanalysis
· 2025 Sep · PMID 40904267
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AIMS: Risdiplam is a small molecule approved for the treatment of spinal muscular atrophy (SMA). The drug and its major metabolite had to be measured in plasma and tissue from several animal species and in human plasma a...AIMS: Risdiplam is a small molecule approved for the treatment of spinal muscular atrophy (SMA). The drug and its major metabolite had to be measured in plasma and tissue from several animal species and in human plasma and urine. Bioanalytical challenges including light sensitivity, instability, carryover, nonspecific binding, and complex tissue analysis, had to be overcome. MATERIALS & METHODS: Liquid chromatography tandem mass spectrometry with reversed-phase separation after protein precipitation/dilution was applied. Ascorbic acid was used as a stabilizer to mitigate degradation of the metabolite, and a surfactant additive prevented nonspecific binding in urine. Tissues were efficiently homogenized by bead beating and matrix-matched with plasma. RESULTS AND CONCLUSIONS: The above challenges were successfully addressed with bioanalytical methods tailored to study needs. Validations and regulatory analyses met requirements of current guidelines, including successful incurred sample reanalysis (ISR) in GLP and clinical studies. The 3R principles (Replacement, Reduction, Refinement) were applied in animal studies to minimize the use of real matrices. Pediatric studies were supported with rapid analysis and microsampling. Bioanalysis supported patient-centric approaches in dose finding and sampling and was key in answering important questions to enable risdiplam to the market.
Wessels U, Pöhler A, Ros F
… +1 more, Stubenrauch KG
Bioanalysis
· 2025 Aug · PMID 40899655
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AIM: To confirm the observation of IgM interference in the anti-adeno-associated virus (AAV) IgG immune complex (IC) assay format and to verify that IgM-specific digestion can improve anti-AAV IgG detection in IC assays....AIM: To confirm the observation of IgM interference in the anti-adeno-associated virus (AAV) IgG immune complex (IC) assay format and to verify that IgM-specific digestion can improve anti-AAV IgG detection in IC assays. METHODS: Treatment-emergent anti-AAV2 and anti-AAV9 IgG signals were measured in IC assays with and without IgM-specific digestion. Anti-AAV2 and anti-AAV9 IgM signals were measured in parallel. RESULTS: IgM digestion increased anti-AAV2 and anti-AAV9 IgG signals when anti-AAV2 or anti-AAV9 IgM were present in the matrix. CONCLUSIONS: Co-existing anti-AAV IgM cause interference in the anti-AAV IC assay format. Selective IgM digestion improves the detection of anti-AAV IgG in the IC assay.
Ye S, Gambardella J, Zaslavskaia L
… +7 more, Kim D, Konovalov A, Kostuk S, Samareh Afsari H, Place C, Coble K, Johnson AJ
Bioanalysis
· 2025 Aug · PMID 40891520
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BACKGROUND: Drug bridging immunoassays are widely employed as the standard approach for detecting anti-drug antibodies (ADAs) in the development of new biological entities. A major challenge in these assays is mitigating...BACKGROUND: Drug bridging immunoassays are widely employed as the standard approach for detecting anti-drug antibodies (ADAs) in the development of new biological entities. A major challenge in these assays is mitigating target interference, particularly when the soluble target exists in dimeric forms, which can result in false positive signals and compromise assay specificity. RESEARCH DESIGN AND METHODS: We developed sensitive and robust ADA assays capable of overcoming target interference to detect antibodies against BI X in both cynomolgus monkey (cyno) plasma and human serum matrices. This was achieved through the implementation of simple sample treatment techniques, specifically, acidification using a panel of different acids, to disrupt dimeric target interactions and minimize the interference. RESULTS: Optimization of the acid dissociation and subsequent neutralization steps significantly reduced target interference in both cyno and human matrices. These improvements were achieved without the need for additional assay development or complex depletion strategies. CONCLUSIONS: Compared to previously reported methods for mitigating target interference, the acid panel treatment approach is simpler, more time-efficient, and cost-effective. This user-friendly strategy can be readily applied to eliminate soluble dimeric targets during ADA method development, particularly in cases where alternative methodologies are not feasible or applicable.
Yu Y, Liu Y, Zhang J
… +7 more, Dai S, Wu R, Wan F, Yao C, Yao Y, Nan F, Li Y
Bioanalysis
· 2025 Sep · PMID 40884750
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AIM: We aim to establish a rapid and sensitive UPLC-MS/MS method to analyze the pharmacokinetics of oral anisodine hydrobromide (AH) tablets. METHODS: Comparison of pharmacokinetic differences between AH (present in vivo...AIM: We aim to establish a rapid and sensitive UPLC-MS/MS method to analyze the pharmacokinetics of oral anisodine hydrobromide (AH) tablets. METHODS: Comparison of pharmacokinetic differences between AH (present in vivo as anisodine) and its metabolite NOAT3 in two groups of rats after administration of AH at doses of 5, 10, and 20 mg/kg. In addition, plasma concentrations of AH were used as a pharmacokinetic parameter, and interleukin-1β (IL-1β) and lactic dehydrogenase (LDH) were selected as pharmacodynamic indicators to establish a pharmacokinetics-pharmacodynamics (PK-PD) combined model in physiological and CIRI model rats, to analyze the protective effect of AH against CIRI. RESULTS & CONCLUSION: The study demonstrated that AH could be rapidly and extensively distributed to various tissues in rats. AH is a promising drug for the treatment of rat models of CIRI.
Bioanalysis
· 2025 Aug · PMID 40851466
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Hybridization-based LC-MS is rapidly emerging as a bioanalytical platform for oligonucleotides, particularly when both high sensitivity and high specificity are needed. When used to analyze single-stranded antisense olig...Hybridization-based LC-MS is rapidly emerging as a bioanalytical platform for oligonucleotides, particularly when both high sensitivity and high specificity are needed. When used to analyze single-stranded antisense oligonucleotide (ASO) therapeutics, the workflows are relatively well established, but the analysis of double-stranded small interfering RNA (siRNA) therapeutics presents additional challenges due to competition for binding from the sense strand. In the last two years, the authors have independently published extensively on hybridization-based LC-MS bioanalysis of siRNA therapeutics, and now we take a step back to evaluate the progress we have made and offer our thoughts on the future of this platform. We touch upon aspects of the sample preparation and analytical process that can either be improved upon, made more efficient, or expanded to maximize the information that can be gained from a single sample. Additionally, we discuss how hybridization-based LC-MS compares to other common oligonucleotide bioanalytical workflows, and its potential to become a frontline assay platform for use in supporting regulatory submissions. Overall, we are excited about the potential hybridization-based LC-MS has demonstrated as a bioanalytical platform and are eager to begin the conversation on where this workflow goes next.
Gong Y, Gong W, Chen J
… +3 more, Man Z, Li W, Shi W
Bioanalysis
· 2025 Aug · PMID 40851465
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AIMS: To develop a UPLC-MS/MS method for quantifying tigecycline (TIG), meropenem (MER), polymyxin B1 (PB1), and PB2 in human plasma, and analyze their unbound fractions and influencing factors in HSCT patients. METHODS:...AIMS: To develop a UPLC-MS/MS method for quantifying tigecycline (TIG), meropenem (MER), polymyxin B1 (PB1), and PB2 in human plasma, and analyze their unbound fractions and influencing factors in HSCT patients. METHODS: Plasma samples from HSCT patients were analyzed using the developed UPLC-MS/MS technique. RESULTS: The method showed high accuracy, precision, and stability. Protein binding rates were 82.4% for TIG, 83.7%-79.3% for PB1, and 86.7%-82.6% for PB2, with lower binding at trough vs. peak concentrations. CONCLUSIONS: Preliminary analysis suggests that BMI and serum albumin may influence PB1/PB2 binding in CRE-infected HSCT patients though larger cohorts are needed for confirmation. (clinical trial registration # 2021-0938-01).
Protti M, Mandrioli R, Santos HM
… +3 more, Lodeiro C, Capelo-Martínez JL, Mercolini L
Bioanalysis
· 2025 Aug · PMID 40851463
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Microsampling, using minute amounts of biological specimen, is uniquely suited for carrying out human and animal pharmacokinetic and toxicokinetic studies. Indeed, it provides important advantages over common blood-drawi...Microsampling, using minute amounts of biological specimen, is uniquely suited for carrying out human and animal pharmacokinetic and toxicokinetic studies. Indeed, it provides important advantages over common blood-drawing procedures (e.g. increased analyte stability, simpler and cheaper storage and shipping, self-sampling possibility, reduced consumption of materials and animals) while also maintaining similar performance and result reliability. In the microsampling space, volumetric absorptive microsampling (VAMS) is currently considered one of the most important and widespread technologies, thanks to its volume accuracy capabilities, making it natively suitable for quantitative analysis. In this review, an exhaustive treatment of pharmacokinetic and toxicokinetic studies using VAMS is presented, as well as several examples of analytical methods prospectively enabling the employment of VAMS for the same purpose.
Bioanalysis
· 2025 Aug · PMID 40851441
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INTRODUCTION: Oral metronomic chemotherapy employs a low-dose combination of chemotherapeutics administered regularly to minimize toxicity while enhancing anticancer efficacy. The clinical utility of Doxorubicin (DOX) is...INTRODUCTION: Oral metronomic chemotherapy employs a low-dose combination of chemotherapeutics administered regularly to minimize toxicity while enhancing anticancer efficacy. The clinical utility of Doxorubicin (DOX) is limited due to severe cardiotoxicity. Interestingly, Piperine (PIP) has been explored to mitigate DOX-induced toxicity while enhancing its therapeutic efficacy. Solid lipid nanoparticles (SLNs) offer an efficient drug delivery approach to improve oral bioavailability and controlled release of DOX and PIP. AREAS COVERED: LC-MS/MS method was developed and validated per US-FDA bioanalytical guidelines to quantify DOX and PIP in plasma simultaneously. SLNs were developed and optimized using design expert software and exhibited particle size of 151.56 ± 0.32 nm, polydispersity index (PDI) of 0.172 ± 0.02, and surface charge of -22.83 ± 0.66 mV. Pharmacokinetic evaluation in female Sprague-Dawley rats showed enhanced AUC₀-∞ for DOX (31911.78 ± 226.92 ng/mL) and PIP (7377.66 ± 655.78 ng/mL), indicating improved systemic exposure. EXPERT OPINION: The findings highlight the potential of SLN-based co-delivery of DOX and PIP for oral metronomic chemotherapy. The validated LC-MS/MS method ensures precise pharmacokinetic assessment, which is crucial for future clinical translation. This study provides a promising strategy for enhancing oral chemotherapy efficacy.
Ogunesan O, Zaborske CD, Newsom C
… +3 more, Shafer MM, Zierold KM, Wickliffe JK
Bioanalysis
· 2025 Sep · PMID 40844138
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BACKGROUND: Volumetric Absorptive Microsampling (VAMS) is a blood sample collection method proposed as an alternative to venipuncture for metals/elements biomonitoring. However, the microsampler background concentration...BACKGROUND: Volumetric Absorptive Microsampling (VAMS) is a blood sample collection method proposed as an alternative to venipuncture for metals/elements biomonitoring. However, the microsampler background concentration of metals and small blood volume remains critical limitations, particularly for metals or trace element analysis in environmental health and epidemiological research. MATERIALS & METHODS: Trace element analysis was performed to measure metal concentration in blood samples collected via VAMS and venipuncture using Inductively Coupled Plasma Mass Spectrometry (ICP-MS). VAMS blanks showed elevated background concentration, and cleaning was attempted using a multi-step cleaning protocol. Detection limits and efficacy of background concentration reduction were evaluated. RESULTS: Initial analysis showed elevated background metal concentrations in the VAMS blank samplers, at or exceeding levels found in venous blood samples. VAMS blank with background concentration post-cleaning result indicated reductions in concentration for some metals; however, the concentration for most detected metals remained persistent. CONCLUSIONS: The efficacy of VAMS in environmental health and epidemiological biomarker research demonstrates both the promising potential and limitations, and the effectiveness of a rigorous cleaning protocol in reducing or eliminating background metal concentrations on microsampler tips.
Bioanalysis
· 2025 Aug · PMID 40829890
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Antibody-drug conjugates (ADCs) represent a rapidly advancing class of biotherapeutics for oncology and immunological indications. Comprehensive pharmacokinetic (PK) characterization is critical for assessing ADCs effica...Antibody-drug conjugates (ADCs) represent a rapidly advancing class of biotherapeutics for oncology and immunological indications. Comprehensive pharmacokinetic (PK) characterization is critical for assessing ADCs efficacy, safety, and overall therapeutic performance. Ligand binding assays (LBAs) are widely employed in both academic and industrial settings for the quantitative and semi-quantitative analysis of biologics. These assays rely on specific molecular interactions - commonly between antigens and antibodies or ligands and receptors - and offer high sensitivity, robustness, and cost-efficiency. In ADC bioanalysis, LBAs are utilized to quantify multiple types of analytes, including total antibody and antibody-drug conjugate. However, the development of LBA methods for ADCs is challenged by the structural heterogeneity of these molecules, analyte instability, and the need for high selectivity and sensitivity. This review summarizes the application of LBAs in ADC PK studies, outlines common methodological challenges, and discusses strategic considerations for assay development to ensure accurate and reliable bioanalytical measurements.
Qiu L, Lin J, Su Y
… +4 more, Kong Y, Zhou Y, Chen Y, Yu Y
Bioanalysis
· 2025 Aug · PMID 40827693
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AIMS: To establish a rapid, sensitive HPLC-MS/MS method for quantifying isavuconazonium in human plasma and characterize its pharmacokinetics in healthy Chinese subjects under fasting and postprandial conditions. MATERIA...AIMS: To establish a rapid, sensitive HPLC-MS/MS method for quantifying isavuconazonium in human plasma and characterize its pharmacokinetics in healthy Chinese subjects under fasting and postprandial conditions. MATERIALS & METHODS: Plasma samples were processed via acetonitrile protein precipitation. Separation was performed on an LC-20ADXR Plus C18 column with gradient elution (0.01% formic acid/acetonitrile). Detection used a Triple Quad 4500 mass spectrometer with MRM; isavuconazole-d4 was the internal standard. The method was validated, then applied to a crossover study in 32 healthy subjects (fasting vs. postprandial). RESULTS: The method showed good linearity (4-4000 ng/mL, R ≥0.9801) with LLOQ 4 ng/mL. Stability was confirmed under various conditions (e.g. 53 days at -20°C, 66 days at -80°C). Pharmacokinetic results revealed food delayed Tmax (2.5 vs. 5.0 h) and reduced Cmax (1929.68 vs. 1300.17 ng/mL) but did not affect AUC. CONCLUSIONS: The validated HPLC-MS/MS method is rapid and reliable for therapeutic drug monitoring. Food affects absorption but not total exposure, guiding clinical dosing in Chinese populations.
Riccardi Sirtori F, Di Ianni A, Crosasso C
… +5 more, Bertotti E, Molinaro F, De Salve I, Barbero L, Cowan KJ
Bioanalysis
· 2025 Aug · PMID 40827507
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The development of biotherapeutics, particularly multi-domain biotherapeutics (MDBs) and antibody drug conjugates (ADCs), presents unique challenges due to their complexity and increased immunogenicity risks. Bioanalytic...The development of biotherapeutics, particularly multi-domain biotherapeutics (MDBs) and antibody drug conjugates (ADCs), presents unique challenges due to their complexity and increased immunogenicity risks. Bioanalytical approaches play a pivotal role in addressing these challenges. Robust bioanalytical methods are essential for effectively assessing the pharmacokinetics (PK), pharmacodynamics (PD), and immunogenic responses of novel biologics. Advanced techniques, such as multiplexed assays using ligand binding assays (LBAs) and Liquid Chromatography-Mass Spectrometry (LC-MS) methods, are employed to handle the complexity of MDBs and ADCs effectively. Additionally, high-resolution mass spectrometry can investigate the potential biotransformation of biologics directly in in vivo studies. Furthermore, the Context of Use (CoU) is critical for defining assay purposes across nonclinical and clinical settings, ensuring their relevance and efficiency. Regulatory compliance is paramount to ensure assays meet standards for drug approval. Practical considerations include assay sensitivity, specificity, and the method's ability to investigate the structural integrity of MDBs and ADCs in studies. This paper aims to review recent publications on the application of bioanalytical techniques, specifically LBA and LC-MS, in supporting the development of pioneering biotherapeutics. Additionally, it will discuss optimal strategies for bioanalytical methods and immunogenicity assessment.
Liu K, Huang Y, Wang T
… +2 more, Mu R, Rosenbaum AI
Bioanalysis
· 2025 Jul · PMID 40827388
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AIM: Oral peptide therapeutics typically have short half-lives due to rapid degradation by digestive enzymes. Systematic peptide engineering and formulation optimization led to the development of a clinical candidate MED...AIM: Oral peptide therapeutics typically have short half-lives due to rapid degradation by digestive enzymes. Systematic peptide engineering and formulation optimization led to the development of a clinical candidate MEDI7219, an orally bioavailable glucagon-like peptide 1 (GLP-1) peptide, with greater stability than wild-type GLP-1 or semaglutide:~60% of MEDI7219 remained intact after 2 h in vitro incubation with simulated intestinal fluid. This study further investigates proteolytic stability by elucidating biotransformation products of MEDI7219 using liquid chromatography-mass spectrometry (LC-MS) methods. METHOD: Peptide metabolism was assessed using in vitro pancreatin assay followed by analysis utilizing liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID) and electron-activated dissociation (EAD) approaches. RESULTS: We have confidently identified 13 metabolites. Time course profiles of parent and metabolite peaks are consistent with sequential enzymatic cleavage pattern. The 13 metabolites mapped to 8 cleavage sites. Most of these cleavage sites can be explained by the specificity of digestive enzymes, trypsin, pepsin, and elastase. However, α-methyl-L-phenylalanine appeared to be well protected from chymotrypsin and pepsin digestion since no cleavage peptides ending with α-methyl-L-phenylalanine were observed. CONCLUSION: These study results provide further structural details explaining previously published stability data and provide new insights into potential GLP1 proteolytic liabilities for future engineering.
Zimmer J, O'dell M, Johnson D
… +44 more, Hays A, Gorityala S, Tary-Lehmann M, Malone K, Diaz M, Xu T, Dwivedi V, Dufield D, Iles T, Sikkema D, Majumdar R, Riccitelli N, Yuan M, Lowes S, Liu A, Matys K, Turksma A, Pirro J, Bergeron J, Karnik S, Voelker T, Salha D, Reynolds G, Lamy C, Yu M, Garofolo W, Nadarajah S, Leskovar A, Pellerin M, Veeramachaneni R, Kane C, Xu A, Hyer E, Carlsson A, Sanghvi M, Dompkowski E, Rundlett S, Bower J, Spytko A, Liu J, Gu W, Rocha A, O'Brien K, Fang X
Bioanalysis
· 2025 Jun · PMID 40823769
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The 17th GCC Closed Forum was held in San Antonio, TX, USA, on 10 May 2024. Representatives from international bioanalytical Contract Research Organizations were in attendance in order to discuss scientific and regulator...The 17th GCC Closed Forum was held in San Antonio, TX, USA, on 10 May 2024. Representatives from international bioanalytical Contract Research Organizations were in attendance in order to discuss scientific and regulatory issues specific to bioanalysis. The topics discussed at the meeting included integrative bioanalysis, patient-centric sampling, emerging technologies, data integrity, sample reconciliation efforts, discrepancies of ELISpot data, cross-validation harmonization, ultrasensitive platforms for immunogenicity assays, remote regulatory assessments, shedding assays by qPCR/dPCR, and biomarker assays. Conclusions and consensus from discussions of these topics are included in this article.
Jordan G, Pöhler A, Jany C
… +2 more, Bach T, Staack RF
Bioanalysis
· 2025 Aug · PMID 40817673
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AIMS: Immunogenicity testing is a key component of protein drug development, with ADA bridging assays recognized as the gold standard method. These assays employ labeled therapeutic drugs for capture and detection, with...AIMS: Immunogenicity testing is a key component of protein drug development, with ADA bridging assays recognized as the gold standard method. These assays employ labeled therapeutic drugs for capture and detection, with the substrate playing a critical role in generating a detectable signal to differentiate the presence of anti-drug antibodies (ADAs) from nonspecific binding. This study investigates the impact of substrate choice on the assay's ability to capture individual sample variability, focusing on screening and confirmatory cut points (SCP and CCP). METHODS: We compared the colorimetric substrate 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) with the fluorescent substrate 3-(4-hydroxyphenyl)propionic acid (HPPA) in ELISA-based ADA bridging assays for two monoclonal antibodies. RESULTS: Switching the substrate from ABTS to HPPA improved the differentiation of individual signals from baseline noise, enabling a more robust ADA identification. Additionally, the use of HPPA resulted in higher SCP and CCP values. Further investigation revealed that technical reader noise affected a normally distributed sample set when using ABTS. CONCLUSION: The substrate switch, while maintaining all other assay parameters, proved to be a convenient approach to increase SCP and CCP while improving the ability to capture individual sample variability.
Bioanalysis
· 2025 Aug · PMID 40810980
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With the ongoing advancements in the field of ambient ionization for mass spectrometry, systems with a high-throughput capability on the order of 1 sample/second are readily available. This has led to the adoption of mas...With the ongoing advancements in the field of ambient ionization for mass spectrometry, systems with a high-throughput capability on the order of 1 sample/second are readily available. This has led to the adoption of mass spectrometry for a wide variety of applications including those in the drug discovery process. Mass spectrometers have traditionally relied on pre-separation technologies such as high-pressure liquid chromatography for sample clean-up and isobaric separations, but such techniques are not high-throughput compatible. Differential mobility spectrometry is a high-speed atmospheric separation device with separations orthogonal to m/z that can be coupled with the high-throughput sample introduction devices such as acoustic ejection mass spectrometer to address this gap. In this article we highlight the significance of the recent reports on this topic and provide some insights into expanding the use of this technique for new applications. We believe this is a promising new development and will help propel the high-throughput mass spectrometry beyond isobaric interferences.