Time is Money: Utilizing Modernization, Automation and Innovation to Reduce Costs and Keep Your Lab Running Efficiently
Thursday, April 28, 2022 ◉ 1:00 pm EST
Organizer: Rachel Lieberman, Shimadzu Scientific Instruments
This symposium will focus on new strategies for laboratories to reduce costs and keep the lab running efficiently with presentations focused on automation, non-destructive techniques and innovative designs to adapt to the ever-changing analytical landscape. Particular focus will be on speeding up analysis in toxicology laboratories, explosive analysis and utilizing automation to reduce the ability for human error.
Multi-Target Screening and Quantitation of 24 Drugs in Blood, Serum and Urine Using Automated Sample Preparation Coupled Directly to LC-MS/MS
Xiaomeng Kate Xia, Shimadzu Scientific Instruments
Determining what analytes are present and the concentration of those compounds in a variety of biological matrices can be complex. Sample preparations in most forensic laboratories commonly use solid-phase extraction (SPE) or liquid-liquid extraction (LLE), which are not only time-consuming, but also introduce human errors during the multi-step protocols. We describe an automated alternative platform using a sample preparation module connected to a LC-MS/MS for multi-target screening and quantitation of 24 commonly abused drugs in blood, serum and urine. A human serum free of the target analytes was used as the blank and matrix to prepare spiked samples. A mixture of 24 drugs and four internal standards (IS) were spiked at varying concentrations using certified reference standards. Samples were prepared entirely inside the CLAM-2030 module, then transferred directly into the autosampler in the LCMS-8060 system. A ready-to-use method package Forensic Toxicology Database was used to set up the LC-MS/MS method for the analytes. Retention times were updated using certified reference standards. Two or more MRM events were used to identify compounds. Following the method LC conditions, the chromatographic separation of the drugs was achieved by a gradient elution in 17 minutes. Additionally, screening results were compared to the forensic toxicology database library for identification and confirmation using similarity scores. Validation of this quantitative method was completed following ANSI/ASB Standard 036. Limit of quantitation (LOQ) were as low as 1 ng/mL for all 24 analytes. Six points linear calibration curves in the range of 10-1000 ng/mL were established for the 24 analytes with R2 ≥ 0.990. Bias and precision were evaluated at three concentrations (8 ppb, 40 ppb, 80 ppb) and none exceeded the 20% acceptance range. No significant carryover was observed, even after the highest calibrator. No interferences from matrices and internal standards were observed.
A Step Ahead: Postmortem Forensic Toxicology in the 21st Century
Joseph Kahl, Miami-Dade Medical Examiner Department
Operating a forensic toxicology laboratory in the 21st century is an arduous task, one that requires securing additional funding for big-ticket items and routinely looking for mechanisms to improve workflow to reduce turnaround time for client satisfaction. The ability to acquire new instrumentation to improve analytical methodologies and expand the scope of testing, as well as efficiency in the development and validation of new analytical methods, are key. Since 2011, an influx of constantly evolving synthetic compounds has been observed in Miami, FL. The ability to identify and quantify these novel psychoactive substances (NPS) proved challenging in the early years for the Miami-Dade Medical Examiner (MDME) Toxicology Laboratory due to their apparent potency and low concentrations in postmortem specimens. After securing funding through a grant from the National Institute of Justice, the MDME Toxicology Laboratory purchased a Shimadzu LCMS-8060 triple quadrupole mass spectrometer to quantify these emerging NPS in postmortem specimens, in addition to improving the laboratory’s workflow of quantifying typical drugs of abuse. Utilizing the LCMS-8060’s sensitivity, specificity, fast scan rate, and ability for high throughput, the laboratory was able to quickly develop and validate multiple analytical methods to quantify select synthetic cathinones, NBOMe hallucinogens, synthetic opioids, and fentanyl analogs that were prevalent in casework. Additionally, the laboratory developed and validated a single method to quantify seven drugs of abuse that were previously quantified using four different methods on three different types of instrumentation. Once these analytical methods were validated and brought online, the backlog of cases requiring a quantitation of one or more NPS was eliminated altogether, and the turnaround time of all cases was reduced from 60 days to approximately 45 days.
Probe Electrospray Ionization Mass spectrometry (PESI) and Isotopically Dilution Approach for Ultrafast Measurement of Drugs: The Example of Metformin
Franck Saint-Marcoux, Service de Pharmacologie
Background: Metformin (MtF) is a first-line treatment used for type 2 diabetes. Lactic acidosis is a serious complication induced by or associated with elevated MtF plasma concentrations (therapeutic range: 0.5 to 2 mg/L; toxicity: > 5mg/L). When coupled with a mass spectrometry system (MS), the Probe Electrospray Ionization (PESI) method allows a direct analysis of liquid samples without pre-treatment. The objective was to develop a PESI-MS method for an ultra-rapid determination of MTF in plasma by isotopically dilution. Material and methods: We used a tandem mass spectrometer (LCMS-8060; Shimadzu®) equipped with a PESI source and using a selected reaction monitoring mode for the quantitation of MtF conducted in positive mode. Metformine-d5 (Mtf-d5) was chosen as an internal standard for the isotopically dilution approach. A 6-concentration calibration range (0.5 to 50 mg/L) was chosen and a complete validation of the method was performed. The results of this method were then compared to those of 2 classical methods (LC-MS/MS and LC-DAD) method in 60 real patient samples. Results: The sample preparation was a dilution of 10µL plasma in 1000µL ethanol/ ammonium formate buffer including Mtf -d5 at a fixed concentration of 5 mg/L. The measurement time was 0.31 minute. The limit of detection and quantitation was 0,5 mg/L. The accuracy (bias) and precision (RSD%) of the method were within acceptable ranges (min-max: -12.1-15.8% and 1.0-17.1%, respectively). Matrix effect at 10 mg/L was 7.7%. The agreement for diagnosis of a lactic acidosis with classical methods was 100% in 60 real sample patients. Conclusion: By combining the PESI-MS approach with no sample preparation and the isotopically dilution approach needing no calibration curve building, we propose a sensitive, accurate and ultra-fast solution for the measurement of Mtf. This could be helpful in a core Lab when rapid diagnosis of lactic acidosis is needed.
Leading the Way to the Future of Increased Productivity and Performance with High Throughput LCMS Multiplexing System and Integrated Analytical Intelligence
Theresa Meli, Quest Diagnostics
With laboratory demands increasing and high throughput LCMS capabilities becoming the standard testing technique, evolving everyday operations and robustness remains important in the industry. The Shimadzu Nexera QX Multiplexing LCMS instrument is the next generation LCMS platform created with advanced Analytical Intelligence (AI) features that offer high concentration sample detection with subsequent rinsing methods, software made for simplistic and customizable user interfaces, and a coupled 8060NX mass spectrometer with enhanced sensitivity. This system allows for quicker run times as a multichannel unit while the AI offers automatic carryover correction in place of technologist reinjections. The durability and consistency of the instrument makes for less frequent required maintenance and therefore reducing downtime from sample analysis. Using established procedures of solid-phase extraction, urine samples were extracted following two different opioid drug class panel protocols. Methods of analysis were optimized for each assay and dedicated protocols for both the needle and stream rinsing were designed to eliminate carryover. These assays were used to test the robustness of the system over a 30-day period, as well as the abilities of the incorporated AI technology. Over the 30 days, sensitivity remained constant, and the instrument required no repairs. For example, the retention time %RSD for norfentanyl/fentanyl was 1.7/0.5%, respectively, and the %RSD of calibration standards’ peak area was 9.7/8.9%, respectively. The AI also proved successful in its study showing that carryover could decrease from over 10% calibrator Cutoff level peak area (1.1x Cutoff) to less than 5% of Cutoff (0.05x Cutoff). With the newly designed hardware and integrated technology of this instrumentation, increased productivity was achieved for these methods. The system provides maximum uptime and lowered maintenance as well as a novel approach to combating carryover.
UT Southwestern Metabolic Phenotyping Core Mass Spectrometry Resources: Sample Preparation and Data Processing for Metabolite Analysis
Ruth Gordillo, UT Southwestern Medical Center
The Metabolic Phenotyping Core (MPC) provides comprehensive state-of-the-art analytical and phenotypical measures to the scientific community both inside and outside the UT Southwestern Medical Center. Our goal is to expand the scope of techniques available to investigators, standardize key methodologies, and expedite the completion of research on metabolic disorders (obesity and diabetes), cancer, aging, neurological disorders, etc. The MPC is associated with the Touchstone Diabetes Center. The center is devoted to the study of the cells and tissues that are affected by obesity and contribute to the development of diabetes and other obesity-associated diseases. Research in the Touchstone Diabetes Center focuses on both basic and clinical aspects of type 1 and type 2 diabetes, and questions related to the impact of obesity and diabetes on cardiovascular disease, renal disease, and cancer progression. Our services portfolio includes rodent metabolic chambers, specialized surgeries, glucose-insulin clamp studies, clinical analyzer metabolic panels, ELISA, and multiplexed immunoassays as well as a large selection of liquid chromatography tandem mass spectrometry (LC-MS/MS)-based metabolic analytical panels. This presentation focuses on the MPC’s mass spectrometry platforms specifically, including topics such as automated sample preparation, instrument provider pre-developed solution system metabolite analytical packages, as well as software packages for data analysis and bioinformatics tools. The seminar illustrates the direct application of these resources in studies of animal models of disease.