Industrial Laboratories around the world are trying to find ways to minimize sample preparation and enhance productivity. The adaptation of modern mass spectrometry instrumentation is desired due to the high sensitivity and selectivity they provide. This presentation will describe how different sample preparation techniques can be simplified and automated for LC/MS/MS analyses.
Cleaning validation is necessary to establish the quality and safety of pharmaceutical drug products. In cleaning validation protocols, direct sampling is performed with swabs, which are sticks with textiles at one end. The sample on the swab after swabbing the surface of equipment is analyzed with a TOC analyzer and HPLC. Recently, HPLC has been more preferable because of the growing need for the individual analysis of products. Before the HPLC analysis, manual processes such as a sample extraction and a sample condensation are required. Such manual processes may affect to the quality of results. Thus, we evaluated the application of a novel on-line supercritical fluid extraction/chromatography system for the cleaning validation.
For more information, go to ssi.shimadzu.com and follow us on Twitter @ShimdzuSSI
The presentation describes the automated process of the system and present a number of applications from sample matrices such as food, polymers, and pharmaceuticals to show the utility of the system.
This webinar will provide pesticides residue analysts with valuable information on the development and optimization of chromatographic separations and mass spectrometry methods for the analysis of pesticide residues in food. The expert speakers will share their knowledge in understanding the critical aspects of the method, assisting analysts in optimizing their methods for the most challenging analyses.
Key Learning Objectives
- Learn how the use of automated software can make SRM development faster and more highly optimized.
- Learn how the use of a compound data store can further simplify method creation.
- Learn how the use of retention time-based SRM acquisition can increase MS/MS sensitivity and make method maintenance easier.
Event Overview:
In recent years, Gas Chromatography-triple quadrupole mass spectrometry has increased in popularity due to its ability to offer lower detection limits in complex matrices, simplified sample prep requirements, and faster analysis times. Of course, new instrument technology presents the need for the acquiring of new skills to harness the advantages offered by its adoption into current workflows.
In this webinar, a strategy for addressing both of these challenges is discussed in the context of new software designed to automate common method development and method maintenance tasks. Also, in addition to making the triple quadrupole easier to use, this strategy can increase sensitivity of the analysis, which will be demonstrated using a complex SRM pesticide method as an example.
For more information: www.thermoscientific.com/tsq8000
Industrial Laboratories around the world are trying to find ways to minimize sample preparation and enhance productivity. The adaptation of modern mass spectrometry instrumentation is desired due to the high sensitivity and selectivity they provide. This presentation will describe how different sample preparation techniques can be simplified and automated for LC/MS/MS analyses.
Cleaning validation is necessary to establish the quality and safety of pharmaceutical drug products. In cleaning validation protocols, direct sampling is performed with swabs, which are sticks with textiles at one end. The sample on the swab after swabbing the surface of equipment is analyzed with a TOC analyzer and HPLC. Recently, HPLC has been more preferable because of the growing need for the individual analysis of products. Before the HPLC analysis, manual processes such as a sample extraction and a sample condensation are required. Such manual processes may affect to the quality of results. Thus, we evaluated the application of a novel on-line supercritical fluid extraction/chromatography system for the cleaning validation.
For more information, go to ssi.shimadzu.com and follow us on Twitter @ShimdzuSSI
The presentation describes the automated process of the system and present a number of applications from sample matrices such as food, polymers, and pharmaceuticals to show the utility of the system.
This webinar will provide pesticides residue analysts with valuable information on the development and optimization of chromatographic separations and mass spectrometry methods for the analysis of pesticide residues in food. The expert speakers will share their knowledge in understanding the critical aspects of the method, assisting analysts in optimizing their methods for the most challenging analyses.
Key Learning Objectives
- Learn how the use of automated software can make SRM development faster and more highly optimized.
- Learn how the use of a compound data store can further simplify method creation.
- Learn how the use of retention time-based SRM acquisition can increase MS/MS sensitivity and make method maintenance easier.
Event Overview:
In recent years, Gas Chromatography-triple quadrupole mass spectrometry has increased in popularity due to its ability to offer lower detection limits in complex matrices, simplified sample prep requirements, and faster analysis times. Of course, new instrument technology presents the need for the acquiring of new skills to harness the advantages offered by its adoption into current workflows.
In this webinar, a strategy for addressing both of these challenges is discussed in the context of new software designed to automate common method development and method maintenance tasks. Also, in addition to making the triple quadrupole easier to use, this strategy can increase sensitivity of the analysis, which will be demonstrated using a complex SRM pesticide method as an example.
For more information: www.thermoscientific.com/tsq8000
This webinar will provide pesticides residue analysts with valuable information on software method development and data processing for the analysis of pesticide residues in food for both LC–MS and GC–MS. Technical experts will review the latest in software advances to help with data interpretation and reporting.
The feature of the new general chapter 621 is that a column packed with small particles can be used if column length and particle ratio (L/dp) is kept constant between the designated and modified column. This enables high speed analysis of USP methods more than ever.
In this study, a USP method was successfully transferred to an ultra-high speed method with the system suitability requirements met.
For more information, go to SSI.Shimadzu.com. Thanks for viewing.
The presence of Per- and Polyfluorinated Alkyl Substances (PFAS) in drinking water is being thoroughly studied due to the persistence of these compounds in the environment and their potential health effects. However, there is limited knowledge about the occurrence of these chemicals in bottled water, despite the increasing concerns about PFAS in the food supply. This poster shows results from a fast and simple direct injection method similar to draft EPA method 8237, using the Shimadzu triple quad LCMS-8050 to analyze seven commercially available samples of bottled water for 24 PFAS.
Many people pursue ideas of “efficiency” as an ideal for daily life; the same can be true in the HPLC laboratory. In this work, we demonstrate the efficiency, throughput, and reliability of a dual injection system for finished pharmaceutical products and in-process active pharmaceutical ingredients
The automation of sample preparation has become an increasingly important component for reproducible and operator-independent experiments. This work outlines novel strategies that are being utilized for automated online and offline sample preparation to achieve specific goals, such as a host of applications including targeting post-translationally modified proteins, non-tryptic peptides, and intact proteins.
Glycans in antibody drugs may play roles in the antigenicity, pharmacokinetics and high-order structural stability of drugs, which could adversely affect drug safety and effectiveness. It is therefore necessary to investigate which glycans are present in antibody pharmaceuticals. This presentation introduces an example of the analysis of glycans. For more information, go to ssi.shimadzu.com.
The growth of, and the confidence in, hemp products will require applicable testing to ensure product quality and safety. Chromatography technology will play a large role in this as the technique is used for potency testing. This study optimizes a quantitative chromatographic determination of 15 cannabinoids using the Shimadzu Hemp Analyzer.
A single chromatographic method was developed for the separation and quantitation of ten common seized drugs. The single quadrupole mass spectrometer, LCMS 2020, demonstrated its capability for simultaneous detection and confirmation using in source fragmentation of all analytes. Linear calibration curves were acquired for each analyte.
Fast, selective, and sensitive methods can be developed for the analysis of impurities
Offering many business benefits using UPLC and UPC2
Increase in sample throughput
Reduction in toxic solvent usage
Using mass spectral detection over UV detection provides
Improvement in sensitivity and selectivity
Reduced matrix effects
PDA and mass detection provide complementary information for peak assignment and structural confirmation of impurities
Total workflow solutions that cater every budget, performance or throughput requirement for confirmatory dioxin analysis were discussed in the Thermo Scientific Lunch Seminar at the Dioxin 2014 conference. D. Hope, CEO & Owner Pacific Rim Laboratoris, presented about the economies of POPs analysis from the point of view of a leading laboratory using the very latest dioxin method kits. C. Cojocariu, Thermo Fisher Scientific, discussed recent changes in EU regulations which bring new opportunities for more labs to participate in dioxin analysis and about validating methods using Gas Chromatography triple quadrupole for PCDD/Fs with reference to the new EU Commission Regulation No. 709/2014.
This webinar will provide pesticides residue analysts with valuable information on software method development and data processing for the analysis of pesticide residues in food for both LC–MS and GC–MS. Technical experts will review the latest in software advances to help with data interpretation and reporting.
The feature of the new general chapter 621 is that a column packed with small particles can be used if column length and particle ratio (L/dp) is kept constant between the designated and modified column. This enables high speed analysis of USP methods more than ever.
In this study, a USP method was successfully transferred to an ultra-high speed method with the system suitability requirements met.
For more information, go to SSI.Shimadzu.com. Thanks for viewing.
The presence of Per- and Polyfluorinated Alkyl Substances (PFAS) in drinking water is being thoroughly studied due to the persistence of these compounds in the environment and their potential health effects. However, there is limited knowledge about the occurrence of these chemicals in bottled water, despite the increasing concerns about PFAS in the food supply. This poster shows results from a fast and simple direct injection method similar to draft EPA method 8237, using the Shimadzu triple quad LCMS-8050 to analyze seven commercially available samples of bottled water for 24 PFAS.
Many people pursue ideas of “efficiency” as an ideal for daily life; the same can be true in the HPLC laboratory. In this work, we demonstrate the efficiency, throughput, and reliability of a dual injection system for finished pharmaceutical products and in-process active pharmaceutical ingredients
The automation of sample preparation has become an increasingly important component for reproducible and operator-independent experiments. This work outlines novel strategies that are being utilized for automated online and offline sample preparation to achieve specific goals, such as a host of applications including targeting post-translationally modified proteins, non-tryptic peptides, and intact proteins.
Glycans in antibody drugs may play roles in the antigenicity, pharmacokinetics and high-order structural stability of drugs, which could adversely affect drug safety and effectiveness. It is therefore necessary to investigate which glycans are present in antibody pharmaceuticals. This presentation introduces an example of the analysis of glycans. For more information, go to ssi.shimadzu.com.
The growth of, and the confidence in, hemp products will require applicable testing to ensure product quality and safety. Chromatography technology will play a large role in this as the technique is used for potency testing. This study optimizes a quantitative chromatographic determination of 15 cannabinoids using the Shimadzu Hemp Analyzer.
A single chromatographic method was developed for the separation and quantitation of ten common seized drugs. The single quadrupole mass spectrometer, LCMS 2020, demonstrated its capability for simultaneous detection and confirmation using in source fragmentation of all analytes. Linear calibration curves were acquired for each analyte.
Fast, selective, and sensitive methods can be developed for the analysis of impurities
Offering many business benefits using UPLC and UPC2
Increase in sample throughput
Reduction in toxic solvent usage
Using mass spectral detection over UV detection provides
Improvement in sensitivity and selectivity
Reduced matrix effects
PDA and mass detection provide complementary information for peak assignment and structural confirmation of impurities
Total workflow solutions that cater every budget, performance or throughput requirement for confirmatory dioxin analysis were discussed in the Thermo Scientific Lunch Seminar at the Dioxin 2014 conference. D. Hope, CEO & Owner Pacific Rim Laboratoris, presented about the economies of POPs analysis from the point of view of a leading laboratory using the very latest dioxin method kits. C. Cojocariu, Thermo Fisher Scientific, discussed recent changes in EU regulations which bring new opportunities for more labs to participate in dioxin analysis and about validating methods using Gas Chromatography triple quadrupole for PCDD/Fs with reference to the new EU Commission Regulation No. 709/2014.
Automated sample hydrolysis for a forensic toxicology urine screening LC-MS/M...SCIEX
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Automated sample preparation using the GERSTEL MPS Series and MAESTRO softwar...GERSTEL
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Specific applications that use automated sample preparation prior to injection into an LCMSMS system.
The wide variety of automated sample preparation options offered by GERSTEL.
Automated Dried Blood Spot extraction and analysis using LC/MS/MS.
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Automated SPE of drugs of abuse from oral fluid samples and analysis by LC/MS/MS.
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Get an exclusive demo of the new family of UiPath LLMs – GenAI models specialized for processing different types of documents and messages
This is a hands-on session specifically designed for automation developers and AI enthusiasts seeking to enhance their knowledge in leveraging the latest intelligent document processing capabilities offered by UiPath.
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However, this ease of use means that the subject of security in Kubernetes is often left for later, or even neglected. This exposes companies to significant risks.
In this talk, I'll show you step-by-step how to secure your Kubernetes cluster for greater peace of mind and reliability.
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Do you want to learn how to model and simulate an electrical network from scratch in under an hour?
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During the webinar, you will discover the PowSyBl ecosystem as well as handle and study an electrical network through an interactive Python notebook.
PowSyBl is an open source project hosted by LF Energy, which offers a comprehensive set of features for electrical grid modelling and simulation. Among other advanced features, PowSyBl provides:
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GDG Cloud Southlake #33: Boule & Rebala: Effective AppSec in SDLC using Deplo...James Anderson
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Automated SPE-LC/MS/MS method development using ITSP
1. Step 3. Sample Breakthrough - The macro ITSP SPE MD
Sample Loading Capacity simply loads the ITSP device with
sample of increasing volumes in increments of 10µL until
breakthrough occurs. In this pictorial representation, the blue food
coloring begins to breakthrough at between 30 and 40µL. If the
sample is relatively clean, then direct injection without a wash can
offer specific data for sample breakthrough.
However, if the sample is dirty it could clog the orifice or damage
the analytical column, so the device would require regular SPE
wash and elution steps, as in Step 4 below.
An Automated Procedure for Determining ITSP SPE Analytical Parameters for the Analysis of Cortisol and Cortisone in Urine
Rick Youngblood1, Kim Gamble1, Thurman Allsup2, and Ken Lewis 2
1 MicroLiter Analytical Supplies, Inc., Suwanee GA, 2OpAns LLC, Durham NC
Overview
•A CTC Analytics HTS sample handler was used to extract and
clean-up urine samples using ITSP SPE devices and inject the
extracts onto LC/MS/MS.
•This method uses only standard laboratory equipment.
•A fully automated method has been developed for the
optimization of the ITSP SPE extraction.
•This procedure eliminates most of the manual sample
manipulation found in the traditional assay development.
For Further Information
www.MicroLiter.com
Contact Rick Youngblood at RYoungblood@MicroLiter.com
MicroLiter Analytical Supplies, Inc. PO Box 808 Suwanee, GA 30024
Phone (770) 932-6565
www.OpAns.com
Contact Thurman Allsup at TAllsup@OpAns.com
OpAns, LLC, 4134 S. Alston Ave. Durham, NC 27713
Phone (919) 323-4300
Introduction
References
1. McCurdy D, Lin Z, Inn KGW, Bell R, Wagner S, Efurd, DW,
Steiner R, et al. Second interlaboratory comparison study for the
analysis of 239PU in synthetic urine at the µBq (~100 aCi) level by
mass spectrometry. Journal of Radioanalytical and Nuclear
Chemistry. 2005;263/2:447-445.
Results
Using ITSP SPE method development macros for automating
sample preparation, in less than an hour it has been shown
that the optimum wash solvent is 30-40% methanol in water
and the optimum elution solvent is 70-80% methanol in water.
The experiment determining retention times for the test
compounds while changing mobile phases confirmed that the
test compounds just started to elute at 30-40% methanol and
were eluted quickly at 80-90% methanol. Using the sample
load macro, in less than an hour it was determined that the
sample capacity was above 45 µg in 900 µL urine.
Conclusion
One of the advantages of ITSP SPE is the ability to perform
automated sample extraction, cleanup, and extract injection in-
line. A simplified method for the optimization of an ITSP SPE
method has been demonstrated here.
Step Solvent Volume (µL) Flow Rate
(µL/sec)
Condition B 100 15
Condition A 100 15
Load Sample 100 10
Flush Air 100 15
Elute C 100 5
Flush Air 100 40
Solvent A: Water
Solvent B: Methanol
Solvent C: Methanol:Water (A gradient of mixtures from100%
Methanol to 100% water in 10% increments)
Analysis Method
All analysis was on a HPLC/MS/MS using a Finnigan TSQ
Quantum Ultra AM triple quadrupole with a Positive Ion (HESI)
Heated Electrospray Ionization Probe and an Agilent 1200
Rapid Resolution HPLC. Area ratios of compound area to
internal standard area were calculated.
.
Extraction Hardware
ITSP Cartridges: C8 10 mg (MicroLiter Product No.: 07-C810-20A)
A CTC Analytics PAL HTS sample handler was used to prepare the
samples. The PAL was configured with a 100 µl syringe and two
tray holders. Each tray holder held 2 microplates, one of which was
designed to hold the ITSP hardware kit (Product No.: 07-ITSP-HW).
ITSP SPE Find Solvent Strength Method
Methods
Sample Preparation Method
A standard of Cortisol and Cortisone was prepared in synthetic
urine1 at a nominal concentration 50 ng/mL for the solvent
strength and C8 HPLC elution experiments and at 50 µg/mL for
the sample loading experiment. Cortisol stable label internal
standard (50 ng) was added to the eluant wells. The sample
preparation and method development macros written by
MicroLiter were independently tested and verified by Thurman
Allsup at OpAns LLC.
Step Solvent Volume (µL) Flow Rate
(µL/sec)
Condition B 100 15
Condition A 100 15
Load Sample 100-900 10
Flush Air 100 15
Elute C 100 5
Flush Air 100 40
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
0 20 40 60 80 100 120
InverseRT
% MeOH
C8 HPLC Elution Conditions
Cortisone RT
Cortisol RT
0
500
1000
1500
2000
2500
3000
0 10 20 30 40 50
AreaRatio
µg Loaded
Sample Loading µg
Cortisone
Cortisol
ITSP (Instrument Top Sample Prep) is a consumable device
that allows for a common autosampler used on most
chromatographs to prepare client samples using Solid Phase
Extraction (SPE) or filtration.
ITSP’s septum seals and grips the needle creating a closed
environment above the bedmass. By removing the volume
above the column bedmass and replacing it with the analytical
instrument’s syringe, solutions can be passed across the
bedmass using the hydraulic pressure of the syringe plunger.
The gripping action of the septum also allows for the ITSP
device to be transported to any location on the autosampler.
By defining waste and elution locations the device can be
prepped in one location and moved to elute the collected
compounds in another. Once eluted into a clean receptacle
the instrument can discard the device and inject the eluate
onto the analytical instrument.
The following diagrams illustrate the ITSP device and the
typical set-up on an LC/MS/MS autosampler.
ITSP Device Components
8mm Crimp Seal
SPE or Filter Media Funnel Cup
Autosampler Needle Guide
Autosampler Syringe Needle
Needle Centering AttachmentNeedle Guide
Septum
ITSP Autosampler Setup
Autosampler
Sample Tray Elution Tray
Solvent Reservoirs ITSP Tray
Injection Valve
Wash Station
Analytical Syringe
ITSP (Instrument Top Sample Prep) is patented under the United States Patent and Trademark Office under the following patents: US Patents
6,859,615 and 7,001,774; Canadian Patent 2,316,648; and European Patent 1.171.701. Other patents pending in all of the above mentioned offices.
Step 1. Blend Solvents – The macro ITSP SPE MD Blend
Solvents blends two solvents beginning with 100% in
position one decreasing to 0% in position 11. Likewise
Solvent two starts with 0% in position 1 and ends with 100%
in position 11.
Step 2. Solvent Strength – The macro ITSP SPE MD
Solvent Strength simply loads the device with sample and
elutes with the blended solvents. As illustrated, the sample
begins to elute at approximately 37% Methanol, which yields
the optimum wash solvent strength.
At approximately 70% Methanol the sample elution is
complete, which yields the optimum elution solvent strength,
and both parameters are set. One other note: One of the
chromatograms was produced using an ITSP device twice
proving that ,in situations where chain-of-custody is not a
factor, the device can be reused.
MicroLiter has
developed a 96 well
microplate and a set
of macros that
automates method
development based
on application notes
provided by most
sorbent
manufacturers. It is
also an excellent tool
for scaling down
methods you believe
can increase the
Method Development Tray Setup
Solvent Strength
Eluate
Sample
Solvent Blend
ITSP
productivity of your lab while at the same time reducing the
costs of solvents, raw sample requirements and hazardous
waste disposal.
Automated Method Development
Step 4 - This chart illustrates the 10mg C8 sorbent had no
issues loading the sample within the requirements of the assay
using the macro ITSP SPE MD Full Load Wash Elute Capacity.
C8 HPLC Elution Conditions
A Zorbax Eclipse XDB-C8 HPLC column with a flow of 0.6 mL/min
was used to determine retention times for cortisol and cortisone
while varying isocratic mobile phases in 10% increments from 30-
100% methanol and water.
ITSP SPE Find Sample Loading Method
Solvent A: Water
Solvent B: Methanol
Solvent C: Methanol:Water (A gradient of mixtures from100%
methanol to 100% water in 10% increments)
Verification – An experiment to determine retention times
for the test compounds on a C8 HPLC column was
performed to verify the data from the solvent strength
experiment.
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0 20 40 60 80 100
AreaRatio
% MeOH
Solvent Strength Method
Cortisone
Cortisol