Paper spray ionization: The faster path to mass spec analysis Save time and reduce costs through direct sample analysis of biological fluids BROUGHT TO YOU BY INDEPENDENT SCIENCE PUBLISHER IN PARTNERSHIP WITH Paper spray ionization Introduction Common concerns with using conventional chromatography-based MS include time- Contents consuming protocols, experiment backlogs, high costs per sample, equipment downtime • Detection of controlled substances for maintenance and the need for experienced in blood samples using the VeriSpray technicians. ion source with Thermo Scientific™ Using paper spray ionization such as the TSQ Altis™ MS for clinical research Thermo Scientific™ VeriSpray™ PaperSpray and forensic toxicology ion source, researchers in clinical and forensic toxicology labs with high throughput • Quantitative analysis of antifungal requirements can now perform direct Mass drugs using PaperSpray tandem Spectrometry (MS)-based analyses with mass spectrometry for clinical minimal solvent consumption and sample research preparation, thereby saving costs per test and • Robustness characterization of reducing the time involved. a quantitative whole blood assay The VeriSpray ion source solution involves using PaperSpray technology for fast, easy and direct introduction of samples clinical research to the mass spectrometer via electrospray ionization. By simply adding samples and • Combined PaperSpray and FAIMS solvent to paper cartridges and loading them technology for rapid quantification into the system, the VeriSpray ion source of immunosuppressants in whole technology minimizes time-consuming sample blood for clinical research preparation steps and offers a simplified workflow, unlike chromatography processes. • Improving limit of quantification In recent years, paper spray-MS has in clinical research by combining garnered interest in the fields of forensic PaperSpray with FAIMS technology toxicology and clinical research due to its • Analysis of Immunosuppressant robust quantitative analysis, high-throughput Drugs Directly from Whole Blood capabilities, short turnaround times and using PaperSpray Technology time-saving workflows. In this eBook, we provide case studies and proof-of-concept • Robust Quantitative Analysis of experiments that corroborate the varied EDDP by VeriSpray PaperSpray benefits of employing paper spray ionization Spectrometry methods. • Drugs of Abuse Screening and Quantification Using Minimal sample preparation PaperSpray Technology Paper spray ionization in mass spectrometry employs a “dilute and shoot” method, • Featured Products requiring minimal sample preparation • FAQ steps, such as derivatization and reducing/ removing simpler sample purification • Additional Resources processes. In a method optimized to analyze immunosuppressant drugs, 10 μl of human blood was directly spotted on sample method directly from the dried blood spots. plates, minimizing sample preparation that In another method developed for chromatography protocols would otherwise quantitative analysis of antifungal drugs, require. Cyclosporin A, Everolimus and 4 μl of serum samples spiked with internal Tacrolimus were analyzed using a 2-minute standards were spotted on the VeriSpray 2 Paper spray ionization cartridge and allowed to dry at ambient in EDDP quantitation, this time using urine temperature for 30 minutes. The drugs – samples. Upon analyzing 480 urine samples, voriconazole, itraconazole, and posaconazole researchers reported that the system runs for – were analyzed in a 2-minute protocol that extended periods of time without the need for included the extraction step, as well as the maintenance and yielded no significant loss MS analysis. in signal. Quick turnaround time for screening Improved selectivity and limits of detection and quantitation One way to circumvent the potential Researchers investigated the potential of low signal-to-noise ratio in paper spray paper spray ionization along with a triple applications is to couple it with high-field quadrupole mass spectrometer as a screening asymmetric ion mobility spectrometry (FAIMS). tool for commonly encountered drugs of A differential ion mobility technique, FAIMS abuse in a study at Indiana University–Purdue enables greater selectivity of ions entering the University Indianapolis. The sampling plate, mass spectrometer. Coupling PaperSpray with with 24 sample tips, each holding a separate FAIMS in a proof-of-concept method analyzing dried blood spot, was analyzed in minutes, the antidepressant drug, amitriptyline, offering quick turnaround times to screen reduced background interferences and drugs, an important consideration for forensic increased signal-to-noise ratio, resulting in and clinical labs. improved limits of detection. Another proof-of-concept research method, Another study quantifying immunosuppressants testing the rapid screening ability of the in human whole blood compared calibration PaperSpray technology on 19 drugs of abuse curves obtained using the PaperSpray demonstrated how the system analyzed technology with and without FAIMS. Without 480 injections with no loss in sensitivity or coupling to FAIMS, the lowest calibration reproducibility. level for two of the immunosuppressants, tacrolimus and everolimus, was excluded due High-throughput quantitative analysis: to being close to the background signal. With The short analysis times offered by paper the FAIMS interface coupled, however, the spray ionization serves as a high-throughput background signal was significantly reduced. quantitative assay, rapidly processing samples The data showed accuracy and linearity across and saving time for clinical researchers. In the measured range. a technical method for quantitative analysis A recent webinar on paper spray-MS of EDDP (2-ethylidene- 1,5-dimethyl-3,3- explores the background of paper spray diphenylpyrrolidine), an opioid metabolite, ionization in greater detail and digs deeper human whole blood samples were analyzed. into potential applications, along with Without the need for sample pre-treatment, workflows and additional examples. separation, sample clean-up or dilution, With simple workflows, faster analyses and the system maintained sensitivity, robustness reduced cost/sample, the PaperSpray ion and quantitative accuracy over all of the source technology is an excellent option for 480 samples. every clinical research and forensic toxicology Another method tested the high-throughput laboratory looking for faster, accurate, robust, capabilities of the PaperSpray-MS system and sensitive analyses. 3 Paper spray ionization TECHNICAL NOTE 65420 Detection of controlled substances in blood samples using the VeriSpray ion source with TSQ Altis MS for clinical research and forensic toxicology Authors Application benefits Greta Ren1, Nicholas Manicke1, • Quick turn-around time Cornelia Boeser2, • Reduced cost per sample, increased ease-of-use and robustness Neloni R. Wijeratne2 1Indiana University - • Six drugs of abuse analytes in single quantitative method Purdue University Indianapolis, Indianapolis, IN Goal To develop a robust, sensitive, reliable, and reproducible PaperSpray-mass 2Thermo Fisher Scientific, spectrometry workflow for detection of illicit drugs in blood for clinical San Jose, CA research and forensic toxicology using the Thermo Scientific™ TSQ Altis™ mass spectrometer connected with the Thermo Scientific™ VeriSpray™ Keywords PaperSpray system Illicit drugs, opiates, opioids, amphetamines, PS-MS/MS, Introduction TSQ Altis MS, VeriSpray PaperSpray The abuse of controlled substances is a serious ongoing problem worldwide, ion source, TraceFinder, forensic causing significant societal disruption and economic damage. One part of toxicology, benzodiazepines the overall strategy to mitigate the effects from the abuse of drugs requires high-performance methodologies for the screening and quantitation of these substances in biological matrices. Modern forensic toxicological and clinical research laboratories need simpler methods that provide higher throughput and faster analysis for the screening and quantitation of drugs of abuse. 4 Paper spray ionization PaperSpray technology combined with triple quadrupole ion source coupled to a triple quadrupole mass mass spectrometry is an ideal choice for rapid drug spectrometer as a drug screening tool for applications screening and quantitation in clinical research and in clinical research and forensic toxicology. Experiments forensic toxicology applications for two main reasons. were carried out using the VeriSpray PaperSpray ion First, studies have demonstrated that low ng/mL or source on a TSQ Altis triple-stage quadrupole mass lower detection limits are obtainable directly from blood, spectrometer. The VeriSpray system enables robust, which is sufficient for the detection of relevant drugs rapid, and automated PaperSpray analysis. Sample at target concentrations. Second, reduce the burden storage, extraction, and ionization all take place on on the laboratory by simplifying method development, VeriSpray sampling plates. Biofluids are spotted and reducing the amount of bench work and thus decreasing dried directly on the sampling plate. The plates contain time to result. Reports in the literature for screening by 24 individual PaperSpray tips, each of which analyzes PaperSpray MS include the detection of amphetamine- a separate sample (Figure 1A). Analysis of the plate is like designer drugs in oral fluid,1 agrichemicals in fruit,2 carried out automatically via the VeriSpray ion source targeted