Investigation into the design and application of solid core stationary phases has led to a better understanding of how the phases work and has resulted in their design aligned to the structure of the analytes being separated. The current range of columns available is discussed both in terms of selectivities, and also morphologies, allowing informed decisions to be made by the chromatographer. Using real life examples, coupled with advanced modeling, the effects of the particle size and morphology will be given for both small and large molecules, offering an insight into what the future holds for solid core products.
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.
Over the past decade, the number of mAb candidates entering the clinical pipeline has grown significantly. In addition, the number of ADCs that use mAb specificity to carry drug payloads to target sites has increased. As a result, analytical characterization is in high demand.
This webinar discusses new innovations in sample preparation, column technology, UHPLC, and high resolution mass spectroscopy (HRMS) that allow the development of analytical methods with run times of less than 5 minutes for all routine methods.
A recent study conducted by an independent environmental laboratory proves that TIGG’s 5DC 12 x 30 activated carbon is effective at removing TTHMs from municipal drinking water. The test was sponsored by a municipality that needed to come into compliance with the EPA’s DPB Stage 2 regulations.
The study sponsor historically had high levels of TTHM in its water that ranged between 105 and 171 µg/L. The municipality talked to TIGG about a water purification adsorption system. While some in the industry insist that coal-based activated carbon is the only effective means of removing disinfection byproducts from municipal drinking water, TIGG experts recommended their 5DC 12 x 30 virgin coconut activated carbon.
The municipality had an independent laboratory run a pilot study using a rapid small-scale column test. They used TIGG’s 5DC 12 x 30 NSF activated carbon, which is designed to remove low concentrations of contaminants from potable water. The laboratory found that initial TTHM concentrations of 138 µg/L were reduced to < 1.0 µg/L following treatment with virgin coconut activated carbon. Based on the scale-up of the testing, it was determined that a full scale carbon adsorber would effectively treat TTHMs for 325 days before the bed needed a changeout.
Not only is coconut-based activated carbon effective, it is typically more economical than coal-based activated carbon. The following report shows the laboratory’s results over a 24-day period.
Multi-Element Determination of Cu, Mn, and Se using Electrothermal Atomic Abs...IOSR Journals
Simultaneous multi-element graphite furnace atomic absorption spectrometer (SIMAA 6000) is used to get a new multi-element determinations methodology for Cu, Mn, and Se. Firstly, the optimum conditions for single-element mode are determined (which include: pyrolysis and atomization temperatures). Secondly, the optimum conditions for multi-element mode are also determined. The conditions in the two modes have been compared in terms of the characteristic masses, detection limits and pyrolysis and atomization temperatures. The effect of the matrix on the determination has been studied using urine standard sample from Seronorm (LOT 0511545). The accuracy of the developing methods has been confirmed by analysis different biological reference materials. Simultaneous multi-element GF-AAS offers a rapid, low cost and sensitive method for the analysis of trace elements
Investigation into the design and application of solid core stationary phases has led to a better understanding of how the phases work and has resulted in their design aligned to the structure of the analytes being separated. The current range of columns available is discussed both in terms of selectivities, and also morphologies, allowing informed decisions to be made by the chromatographer. Using real life examples, coupled with advanced modeling, the effects of the particle size and morphology will be given for both small and large molecules, offering an insight into what the future holds for solid core products.
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.
Over the past decade, the number of mAb candidates entering the clinical pipeline has grown significantly. In addition, the number of ADCs that use mAb specificity to carry drug payloads to target sites has increased. As a result, analytical characterization is in high demand.
This webinar discusses new innovations in sample preparation, column technology, UHPLC, and high resolution mass spectroscopy (HRMS) that allow the development of analytical methods with run times of less than 5 minutes for all routine methods.
A recent study conducted by an independent environmental laboratory proves that TIGG’s 5DC 12 x 30 activated carbon is effective at removing TTHMs from municipal drinking water. The test was sponsored by a municipality that needed to come into compliance with the EPA’s DPB Stage 2 regulations.
The study sponsor historically had high levels of TTHM in its water that ranged between 105 and 171 µg/L. The municipality talked to TIGG about a water purification adsorption system. While some in the industry insist that coal-based activated carbon is the only effective means of removing disinfection byproducts from municipal drinking water, TIGG experts recommended their 5DC 12 x 30 virgin coconut activated carbon.
The municipality had an independent laboratory run a pilot study using a rapid small-scale column test. They used TIGG’s 5DC 12 x 30 NSF activated carbon, which is designed to remove low concentrations of contaminants from potable water. The laboratory found that initial TTHM concentrations of 138 µg/L were reduced to < 1.0 µg/L following treatment with virgin coconut activated carbon. Based on the scale-up of the testing, it was determined that a full scale carbon adsorber would effectively treat TTHMs for 325 days before the bed needed a changeout.
Not only is coconut-based activated carbon effective, it is typically more economical than coal-based activated carbon. The following report shows the laboratory’s results over a 24-day period.
Multi-Element Determination of Cu, Mn, and Se using Electrothermal Atomic Abs...IOSR Journals
Simultaneous multi-element graphite furnace atomic absorption spectrometer (SIMAA 6000) is used to get a new multi-element determinations methodology for Cu, Mn, and Se. Firstly, the optimum conditions for single-element mode are determined (which include: pyrolysis and atomization temperatures). Secondly, the optimum conditions for multi-element mode are also determined. The conditions in the two modes have been compared in terms of the characteristic masses, detection limits and pyrolysis and atomization temperatures. The effect of the matrix on the determination has been studied using urine standard sample from Seronorm (LOT 0511545). The accuracy of the developing methods has been confirmed by analysis different biological reference materials. Simultaneous multi-element GF-AAS offers a rapid, low cost and sensitive method for the analysis of trace elements
In the pharmaceutical arena there is great interest in solid core technology, where there is a broad range of sample types as well as requirements throughout the process of developing new chemical entities. The presentation looks at how solid core technology can be readily adapted to cope with the challenges associated with the pharmaceutical sector, looking at various sample matrices and molecular entities, from small molecules to large biomolecules. The presentation gives an insight into how varying the solid core to porous layer allows the user to optimize separation performance by reducing extra band broadening. Data presented demonstrates how this technology is more robust than fully porous systems when analyzing biological extracts, routinely used in DMPK departments, resulting in longer column lifetimes.
The present work was carried out to evaluate the removal of p-nitrophenol by adsorption onto olive cake based activated carbon having a BET surface area of 672 m²/g. The batch adsorption experimental results indicated that the equilibrium time for nitrophenol adsorption by olive cake-based activated carbon was 120min. The adsorption data was modeled by equilibrium and kinetic models. The pseudo- first and second order as well as the Elovichkinetic models were applied to fit the experimental data and the intraparticle diffusion model was assessed for describing the mechanism of adsorption. The data were found to be best fitted to the pseudo-second order model with a correlation coefficient (R2=0.986). The intraparticle diffusion mechanism also showed a good fit to the experimental data, showing two distinct linear parts assuming that more than one step could be involved in the adsorption of nitrophenol by the activated carbon. The equilibrium study was performed using three models including Langmuir, Freundlich and Temkin. The results revealed that the Temkin equilibrium model is the best model fitting the experimental data (R2=0.944). The results of the present study proved the efficiency of using olive cake based activated carbon as a novel adsorbent for the removal of nitrophenol from aqueous solution.
26th John L Waters Symposium - Hyphenated Techniques in Thermal AnalysisKevin Menard, Ph.D. MBA
Hyphenated thermal analysis system allow a greater understanding of materials and processes. This talk from the John L Waters Symposium at PittCon 2015 discusses the history and current state of the techniques
Application of ICP-MS and LC-ICP-MS in Drug DevelopmentQPS Holdings, LLC
Inductively coupled mass spectroscopy plasma (ICP-MS) has big potential in preclinical and clinical studies of new drug candidates. One particular area is metallodrugs.
Short-chain alcohols are a group of volatile organic compounds (VOCs) that are often found in workplaces and laboratories, as well as medical, pharmaceutical, and food industries. Realtime monitoring of alcohol vapors is essential because exposure to alcohol vapors with concentrations of 0.15–0.30 mg·L−1 may be harmful to human health. This study aims to improve the detection capabilities of quartz crystal microbalance (QCM)-based sensors for the analysis of alcohol vapors. The active layer of chitosan was immobilized onto the QCM substrate through a selfassembled monolayer of L-cysteine using glutaraldehyde as a cross-linking agent. Before alcohol analysis, the QCM sensing chip was exposed to humidity because water vapor significantly interferes with QCM gas sensing. The prepared QCM sensor chip was tested for the detection of four different alcohols: n-propanol, ethanol, isoamyl alcohol, and n-amyl alcohol. For comparison, a non-alcohol of acetone was also tested. The prepared QCM sensing chip is selective to alcohols because of hydrogen bond formation between the hydroxyl groups of chitosan and the analyte. The highest response was achieved when the QCM sensing chip was exposed to n-amyl alcohol vapor, with a sensitivity of about 4.4 Hz·mg−1·L. Generally, the sensitivity of the QCM sensing chip is dependent on the molecular weight of alcohol. Moreover, the developed QCM sensing chips are stable after 10 days of repeated measurements, with a rapid response time of only 26 s. The QCM sensing chip provides an alternative method to established analytical methods such as gas chromatography for the detection of short-chain alcohol vapors.
Analysis of Herbicide Atrazine and Its Degradation Products in Agricultural S...Agriculture Journal IJOEAR
Abstract— A novel ultra-performance liquid chromatography‒mass spectrometry (UPLC‒MS) method was developed for the determination of herbicide atrazine (ATR) and its principal metabolites namely deisopropylatrazine (DIA), deethylatrazine (DEA) and hydroxyatrazine (HA) in soils. The limit of detection ranged from 0.06 μg kg‒1 (DEA) to 0.25 μg kg‒1 (HA). Recoveries for the four target analytes at three spiked levels ranged from 73.2 to 110% with relative standard deviation of 5.1‒8.1%. In the cases of the three control soil samples spiked with ATR were treated for 60d, the sum content of the three degraded products is 3, 6.4, and 6.8 times greater than ATR residue, respectively. Analyzing 80 soil samples from four counties evaluated this method. ATR of 1.1‒125 μg kg‒1 in 80 of 80 samples, ATR of 0.5‒7.8 μg kg‒1 in 39 of 80 samples, and DIA of 0.5 and 0.6 μg kg‒1 in 2 of 80 samples were found. The proposed method can ensure the rapid and highly sensitive analysis of atrazine and its degradation products in soil, and can provide a direction for proper application of atrazine and a base for evaluating their hazards to the environment.
Using Tunisian Phosphate Rock and Her Converted Hydroxyapatite for Lead Remov...IJERA Editor
Natural and synthesis apatites represent a cost effective soil amendment, which can be used for in situ reduction of lead bioavailability and mobility. In our previous work, we selected Tunisian Phosphate Rock (TPR) and Hydroxyapatite (CaHAp) as promising minerals for the removal of lead from aqueous solutions. X-ray powder diffraction patterns (DRX), Infra Red (IR), Thermogravimetric analysis (TGA) and Scanning Electron Microscopy (SEM) were used to characterize TPR and CaHAp. CaHAp was prepared from TPR and employed for the removal of Pb2+ ions at different concentrations from aqueous solution to determine the adsorption properties of CaHAp and compare them with those of a TPR. The kinetic data obtained indicated that the adsorption performances of the adsorbents depended both on their specific surface area and crystallinity. Complexation of lead ion on the adsorbent surface favoured the dissolution of hydroxyapatites characterized by a Ca/Pb molar ratio of 1.69. The maximum adsorption capacity of CaHAp for Pb2+ ions at 25 °C was 1.806 mmol /g relative to 1.035 mmol /g for TPR at the same temperature. The higher capacity of CaHAp was explained in terms of its porosity and crystallinity. The Pb2+ ions sorption results could be modelled by the Langmuir and Freundlich isotherms. The simulations of adsorption isotherms of Pb2+ on CaHAp allow us to conclude that there is a good correlation between the experimental data and the Langmuir model. On TPR, we show a good correlation between the experimental data and the Langmuir and Freundlich model.
Change of Peptides and Free -Amino Acids Contents during Nanjing Dry-Cured Du...Agriculture Journal IJOEAR
— In order to explore the relationship between the change of peptides and free-amino acid (FAA) and its unique flavour, Dry-cured duck samples of different processing phases were used to study the change of free-amino acid by High Performance Liquid Chromatography (HPLC) in this paper, meanwhile the trichloroacetic acid precipitation method for modeling use to establish the quantitative predicated peptides. The changes of small peptides and free amino acids in the process were studied. The results showed that the level and amount of proteolysis increased with the processing time at traditional technology, meanwhile the amount of peptides were positively correlated with FAA contents (R 2 =0.86).
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
Noter til oplægget i DM-regi, "LinkedIn som karriereredskab". Præsentationen er ikke et slideshow, men en service for kursusdeltageren, som blot skal være til aktivt til stede under oplægget, og ikke behøver at skrive pointer ned undervejs.
In the pharmaceutical arena there is great interest in solid core technology, where there is a broad range of sample types as well as requirements throughout the process of developing new chemical entities. The presentation looks at how solid core technology can be readily adapted to cope with the challenges associated with the pharmaceutical sector, looking at various sample matrices and molecular entities, from small molecules to large biomolecules. The presentation gives an insight into how varying the solid core to porous layer allows the user to optimize separation performance by reducing extra band broadening. Data presented demonstrates how this technology is more robust than fully porous systems when analyzing biological extracts, routinely used in DMPK departments, resulting in longer column lifetimes.
The present work was carried out to evaluate the removal of p-nitrophenol by adsorption onto olive cake based activated carbon having a BET surface area of 672 m²/g. The batch adsorption experimental results indicated that the equilibrium time for nitrophenol adsorption by olive cake-based activated carbon was 120min. The adsorption data was modeled by equilibrium and kinetic models. The pseudo- first and second order as well as the Elovichkinetic models were applied to fit the experimental data and the intraparticle diffusion model was assessed for describing the mechanism of adsorption. The data were found to be best fitted to the pseudo-second order model with a correlation coefficient (R2=0.986). The intraparticle diffusion mechanism also showed a good fit to the experimental data, showing two distinct linear parts assuming that more than one step could be involved in the adsorption of nitrophenol by the activated carbon. The equilibrium study was performed using three models including Langmuir, Freundlich and Temkin. The results revealed that the Temkin equilibrium model is the best model fitting the experimental data (R2=0.944). The results of the present study proved the efficiency of using olive cake based activated carbon as a novel adsorbent for the removal of nitrophenol from aqueous solution.
26th John L Waters Symposium - Hyphenated Techniques in Thermal AnalysisKevin Menard, Ph.D. MBA
Hyphenated thermal analysis system allow a greater understanding of materials and processes. This talk from the John L Waters Symposium at PittCon 2015 discusses the history and current state of the techniques
Application of ICP-MS and LC-ICP-MS in Drug DevelopmentQPS Holdings, LLC
Inductively coupled mass spectroscopy plasma (ICP-MS) has big potential in preclinical and clinical studies of new drug candidates. One particular area is metallodrugs.
Short-chain alcohols are a group of volatile organic compounds (VOCs) that are often found in workplaces and laboratories, as well as medical, pharmaceutical, and food industries. Realtime monitoring of alcohol vapors is essential because exposure to alcohol vapors with concentrations of 0.15–0.30 mg·L−1 may be harmful to human health. This study aims to improve the detection capabilities of quartz crystal microbalance (QCM)-based sensors for the analysis of alcohol vapors. The active layer of chitosan was immobilized onto the QCM substrate through a selfassembled monolayer of L-cysteine using glutaraldehyde as a cross-linking agent. Before alcohol analysis, the QCM sensing chip was exposed to humidity because water vapor significantly interferes with QCM gas sensing. The prepared QCM sensor chip was tested for the detection of four different alcohols: n-propanol, ethanol, isoamyl alcohol, and n-amyl alcohol. For comparison, a non-alcohol of acetone was also tested. The prepared QCM sensing chip is selective to alcohols because of hydrogen bond formation between the hydroxyl groups of chitosan and the analyte. The highest response was achieved when the QCM sensing chip was exposed to n-amyl alcohol vapor, with a sensitivity of about 4.4 Hz·mg−1·L. Generally, the sensitivity of the QCM sensing chip is dependent on the molecular weight of alcohol. Moreover, the developed QCM sensing chips are stable after 10 days of repeated measurements, with a rapid response time of only 26 s. The QCM sensing chip provides an alternative method to established analytical methods such as gas chromatography for the detection of short-chain alcohol vapors.
Analysis of Herbicide Atrazine and Its Degradation Products in Agricultural S...Agriculture Journal IJOEAR
Abstract— A novel ultra-performance liquid chromatography‒mass spectrometry (UPLC‒MS) method was developed for the determination of herbicide atrazine (ATR) and its principal metabolites namely deisopropylatrazine (DIA), deethylatrazine (DEA) and hydroxyatrazine (HA) in soils. The limit of detection ranged from 0.06 μg kg‒1 (DEA) to 0.25 μg kg‒1 (HA). Recoveries for the four target analytes at three spiked levels ranged from 73.2 to 110% with relative standard deviation of 5.1‒8.1%. In the cases of the three control soil samples spiked with ATR were treated for 60d, the sum content of the three degraded products is 3, 6.4, and 6.8 times greater than ATR residue, respectively. Analyzing 80 soil samples from four counties evaluated this method. ATR of 1.1‒125 μg kg‒1 in 80 of 80 samples, ATR of 0.5‒7.8 μg kg‒1 in 39 of 80 samples, and DIA of 0.5 and 0.6 μg kg‒1 in 2 of 80 samples were found. The proposed method can ensure the rapid and highly sensitive analysis of atrazine and its degradation products in soil, and can provide a direction for proper application of atrazine and a base for evaluating their hazards to the environment.
Using Tunisian Phosphate Rock and Her Converted Hydroxyapatite for Lead Remov...IJERA Editor
Natural and synthesis apatites represent a cost effective soil amendment, which can be used for in situ reduction of lead bioavailability and mobility. In our previous work, we selected Tunisian Phosphate Rock (TPR) and Hydroxyapatite (CaHAp) as promising minerals for the removal of lead from aqueous solutions. X-ray powder diffraction patterns (DRX), Infra Red (IR), Thermogravimetric analysis (TGA) and Scanning Electron Microscopy (SEM) were used to characterize TPR and CaHAp. CaHAp was prepared from TPR and employed for the removal of Pb2+ ions at different concentrations from aqueous solution to determine the adsorption properties of CaHAp and compare them with those of a TPR. The kinetic data obtained indicated that the adsorption performances of the adsorbents depended both on their specific surface area and crystallinity. Complexation of lead ion on the adsorbent surface favoured the dissolution of hydroxyapatites characterized by a Ca/Pb molar ratio of 1.69. The maximum adsorption capacity of CaHAp for Pb2+ ions at 25 °C was 1.806 mmol /g relative to 1.035 mmol /g for TPR at the same temperature. The higher capacity of CaHAp was explained in terms of its porosity and crystallinity. The Pb2+ ions sorption results could be modelled by the Langmuir and Freundlich isotherms. The simulations of adsorption isotherms of Pb2+ on CaHAp allow us to conclude that there is a good correlation between the experimental data and the Langmuir model. On TPR, we show a good correlation between the experimental data and the Langmuir and Freundlich model.
Change of Peptides and Free -Amino Acids Contents during Nanjing Dry-Cured Du...Agriculture Journal IJOEAR
— In order to explore the relationship between the change of peptides and free-amino acid (FAA) and its unique flavour, Dry-cured duck samples of different processing phases were used to study the change of free-amino acid by High Performance Liquid Chromatography (HPLC) in this paper, meanwhile the trichloroacetic acid precipitation method for modeling use to establish the quantitative predicated peptides. The changes of small peptides and free amino acids in the process were studied. The results showed that the level and amount of proteolysis increased with the processing time at traditional technology, meanwhile the amount of peptides were positively correlated with FAA contents (R 2 =0.86).
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
Noter til oplægget i DM-regi, "LinkedIn som karriereredskab". Præsentationen er ikke et slideshow, men en service for kursusdeltageren, som blot skal være til aktivt til stede under oplægget, og ikke behøver at skrive pointer ned undervejs.
Este curso de 5 meses visa abordar áreas fundamentais na comunicação de negócios:
Módulo I = Foundation ( Opinion, Suggestion, Job and Company)
Módulo II = Telephoning ( Answering, Taking a Message, Appointment, Checking an order)
Módulo III = Meetings ( Clarifycation, Interruptions, Action Points, Presentations, Summarizing)
Assessing Entrepreneurial Learning in Arts Entrepreneurship ProgramsJason C. White
Slides utilized at the 2015 Society for Arts Entrepreneurship Education Conference at The Ohio State University. For questions, please contact white.2374@osu.edu.
Learn about Waters technologies for analyzing oligonucleotides with LC-MS. We offer solutions for both oligo characterization and QC monitoring. Learn more: http://www.waters.com/oligos
Measuring pKas, logP and Solubility by Automated titrationJon Mole
Presentation by Sirius Analytical covering measurement of pKa, LogP, LogD, Solubility, Supersaturation and precipitation kinetics.
For more details visit www.sirius-analytical.com
Jane Cooper, Senior Applications Scientist
Separation by UPC2 is an ideal alternative to both HPLC and GC analysis
- Ability to run LC and GC amenable compounds in single analysis
-Fast 7 minute analysis of the 24 regulated allergens and 6 additional compounds containing:
– Different classes of compounds
– Different polarities
- UC2 with MS detection offers an orthogonal technique, which enables greater selectivity and specificity compared to either HPLC or GC analysis alon
- The developed 7 minute UPC2 method, is greater than 6 times faster than existing HPLC and GC
TOYOPEARL MX-Trp-650M is designed for efficient mixed-mode steps in protein purification:
Multimodal cation exchanger
High salt tolerance
High binding capacity for IgG
Novel composite electrodes:Preparation and application to the electroanalytic...Université de Dschang
M. Tchieno Melataguia Francis Merlin a soutenu une thèse de Doctorat/Phd en Chimie Inorganique ce 06 juin 2016 dans la salle des conférences de l'Université de Dschang. A l'issue de cette soutenance devant le jury présidé par le Prof. Emmanuel Ngameni lui a décerné la mention très honorable à l'unanimité de ses membres.
31015 Waeghe 1045 am Systematic Method Development with Novel Inert Solid-Core
1. Systematic Method Development with
Novel, Inert Solid-Core Bonded Phases
Thomas J. Waeghe, Ph.D.,
Geoffrey Faden, Carl L. Zimmerman, Alan P. McKeown*
MAC-MOD Analytical, Inc.
103 Commons Court
Chadds Ford, PA 19317
*Advanced Chromatography Technologies
1 Berry Street, Aberdeen, AB25 1HF, Scotland
2. Systematic Method Development with
Novel, Inert Solid-Core Bonded Phases
Encapsulated bonding technology (EBT) is a new approach to
stationary phase endcapping that provides exceptional inertness
and excellent phase stability across a broad pH range from 1.5 to
11.0. This technology has previously been successfully applied to
2, 3 and 5 micron totally porous (non-core) packing materials, but
has more recently been implemented for 2.5 and 5 micron solid-core
packings.
The ability to use stationary phase chemistry (C18, phenyl-hexyl),
organic modifier choice, and pH as variables in a systematic
approach is a significant advantage for UHPLC and HPLC method
development. The usefulness of such a method development
strategy will be described and demonstrated with an appropriate
example.
23/10/2015 Pittcon 2015 New Orleans
3. 1.00 1.05 1.10 1.15 1.20 1.25
0.0
0.5
1.0
1.5
2.0
2.5
3.0
0 5000 10000 15000 20000 25000
0 5 10 15 20 25
N
k
N
k
Resolution(Rs)
Zhao, J.H. and P.W. Carr. Analytical Chemistry, (1999) 71, 2623-2632
Selectivity: the most powerful
variable for increasing resolution
Efficiency
Selectivity
Retention
Rs =
k
1+k
4
√ N
3
0
0
t
tt
k R
1
2
k
k
)(16
base
r
w
t
N
2
Selectivity has
greatest impact on Rs
Rs changes
most rapidly
with
3/10/2015 Pittcon 2015 New Orleans
4. 4
Which LC Parameters Affect
Selectivity Most? 1,2
Isocratic Separations
1. Mobile phase pH
(for ionised analytes only)
2. Column Stationary Phase
3. Organic modifier
4. % Organic modifier
5. Column temperature
• Buffer choice
• Buffer concentration
• Additive concentration
Gradient Separations
• All parameters for isocratic
separations and,
• Gradient steepness
• k* (that is tG, F, , VM, MW)
• Delay volume
• Column dimensions
1 Adapted from ‘Introduction to Modern Liquid Chromatography”,
3rd Edition, Snyder, Kirkland, and Dolan, 2010, p.29, Wiley & Sons
SV
Ft
k
m
G
85*
MOST
Influence
LEAST
Influence
Relative Impact of Different Changes in
RPLC Parameters on Selectivity2
Parameter
Change in
Parameter
Maximum
| |
pH 5 pH units 0.70
Organic Modifier
CH3CNC
H3OH
0.20
Gradient Time (tG) 10-fold 0.20
Orthogonal Column Fs ≥ ~65 0.19
% Organic Modifier 10% (v/v) 0.08
Column Temperature 20 C 0.07
Buffer Concentration 2-fold 0.02
3/10/2015 Pittcon 2015 New Orleans
2 Journal of Chromatography A, 1101 (2006) 122–135, “Orthogonal”
separations for reversed-phase liquid chromatography, L.R. Snyder et al.
5. ACE UltraCore Solid-Core Columns
ACE UltraCore 2.5 m:
Total particle diameter = 2.5 µm
Shell thickness = 0.45 µm
ACE UltraCore 5 m:
Total particle diameter = 5 µm
Shell thickness = 0.7 µm
3/10/2015 Pittcon 2015 New Orleans 5
UltraCore SuperC18 and
SuperPhenylHexyl Columns
• Alternate selectivities: hydrophobic
and - interactions
• stable to 1000 bar (14,500 psi)
• 2 m frits for improved ruggedness
and uptime
• 20,000 column volume lifetime
minimum (≤ 40 C, pH 811.0)
6. Advantages of Encapsulated Bonding
Technology (EBT™)
Encapsulated Bonding Technology
• Uniquely developed for ACE
UltraCore SuperC18 and UltraCore
SuperPhenylHexyl
• EBT bonding and endcapping
– dramatically higher ligand coverage
– effectively eliminates the negative
effects of unbonded silanol groups
Benefits of EBT
• Inertness—superb peak shape
– for bases, acids, and neutrals
(pH 1.5–11.0)
• Stability
– silica protected from eluent at
mid and high pH
– use with volatile buffers for max.
stability, ideal for LC and LC-MS
(NH4OH, NH4OAc, etc.)
• Versatility
– no memory effects from switching
among eluents at different pHs
3/10/2015 Pittcon 2015 New Orleans 6
7. 73/10/2015 Pittcon 2015 New Orleans
x
x
x
x
trcolumn 1
tr column 2
R2 = 0.9987
Uwe Neue Selectivity Descriptor, S, as
measure of orthogonality
√
For collection of representative, diverse analytes,
plot analyte gradient retention times (obtained under
the same conditions) for different stationary phases,
or you can plot gradient RTs for different
combinations of analysis conditions and columns)
versus
R2 is coefficient of determination (measure of how close
the data are to the fitted regression line.
Neue Selectivity Factor
8. 3/10/2015 Pittcon 2015 New Orleans 8
UltraCore Phases: Orthogonality at low pH
UltraCore SuperPhenylHexyl at
low pH vs. SuperC18 at low pH
CH3OH
Low pH
S = 19
S ~10 or higher is effective for adjusting
9. SuperPhenylHexyl Orthogonality:
High pH vs. Low pH
3/10/2015 Pittcon 2015 New Orleans 9
UltraCore SuperPhenylHexyl (high pH)
vs. SuperPhenylHexyl (low pH)
SuperC18
SuperC18
pH 10.7
SuperPhHexyl
S = 12
SuperPhHexyl
S = 11
S=25
S=22
MeOH
MeCN
Nice pattern...
CH3OH
High pH
S = 83
S values between phases
and organic modifiers
at high pH
10. Objective: Explore performance of
ACE UltraCore columns for basic analytes
Parameters
– Stationary phase
• ACE UltraCore SuperC18 and
SuperPhenylHexyl
– Organic modifier
• CH3CN, CH3OH
– pH
• 2.8, 3.8, 8.2, 9.2, 10.2; 9.7 and 10.7
Samples
– 9 Appetite Suppressants
– 16 Drugs of Abuse
103/10/2015 Pittcon 2015 New Orleans
12. More Complex Appetite Suppressant
Mixture: optimum pH from DryLab®
0 10 20
Time (min)
0.00E+002.00E+084.00E+086.00E+08
Intensity
caf
ephed
phent unk
phenolphthalein
chlordiazepoxide
lorcaserin
fenfluramine
fluoxetine
unk3
DEP
sertraline
DidesSib
Rimonabant
NDesSib
sibutramine
orlistat
unk4
12
predicted optimum pH
based on DryLab using
only pH not gradient
time or temperature
2.1 x 100 mm, 2 m ACE Excel SuperC18
0.5 mL/min, 25C, 10–90% CH3CN/20 mM pH 9.3 buffer in 20 min.
MS Detection Data courtesy of Phyllis Wilson, U.S. FDA
3/10/2015 Pittcon 2015 New Orleans
14. UltraCore SuperC18: Examples with CH3CN
and CH3OH at pH 3.8 and pH 10.2
3/10/2015 Pittcon 2015 New Orleans 14
0 2 4 6 8 10
Time (min)
0.0000.0020.0040.0060.0080.010
Volts
1.150
1.849
2.366
2.782
3.154
3.296
6.350
7.354
9.800
0 2 4 6 8 10
Time (min)
0.000.02
Volts
1.981 2.053
3.047 4.029
4.672
4.762
5.692
7.306
8.365
UltraCore SuperC18
CH3CN/pH 3.8
2-50% in 13.7 min.
k* = 14.3
0.6 mL/min, 30 C for all 4
UltraCore SuperC18
CH3CN/pH 10.2
5-70% in 10 min.
k* = 15.5
0 2 4 6 8 10 12 14 16
Time (min)
0.0000.0020.0040.0060.008
Volts
1.283
2.173
2.656
3.516
4.215
5.008
9.145
14.456
15.258
2 4 6 8 10 12
Time (min)
0.0000.0100.020
Volts
2.266
2.345
3.517
4.959
6.309
6.455
9.610
10.337
11.785
UltraCore SuperPhenHex
CH3OH/pH 3.8
2-50% in 13.7 min.
k* = 14.3
UltraCore SuperPhenHex
CH3OH/pH 10.2
5-70% in 10 min.
k* = 15.5
2.2 2.4
Time (min)
Volts
2.266 2.345
1.8 2.0
Time (min)
Volts
1.981
2.053
15. Sample: 16 Drugs of Abuse
3/10/2015 Pittcon 2015 New Orleans 15
MW Compound Name pKa log P pI
RT SPH
2‐50%
pH 2.8
271.31 Normorphine 10.12 0.26 10.12 0.707
285.34 Morphine 9.66 0.90 9.66 1.051
301.34 Oxymorphone 9.14 0.78 9.14 1.405
285.34 Hydromorphone 9.34 1.62 9.34 1.789
301.38 Dihydrocodeine 9.33 1.55 11.74 2.432
299.36 Codeine 9.19 1.34 11.49 2.519
301.34 Noroxycodone 9.14 0.78 9.14 2.663
315.36 Oxycodone 8.21 1.03 10.89 2.807
327.37 6‐Acetylmorphine 8.42 1.09 9.25 2.874
299.36 Hydrocodone 8.61 1.96 13.30 2.989
289.33 Benzoylecgonine 9.54 ‐0.59 6.49 3.268
193.24 MDMA 10.14 1.86 NA 3.518
369.41 Heroin 9.10 1.55 NA 3.966
303.35 Cocaine 8.85 2.28 NA 4.051
243.39 Phencyclidine (PCP) 10.64 4.49 NA 4.616
271.40 Dextromethorphan 9.85 3.49 NA 4.793
Isobaric compounds highlighted in same color
RTs for SuperPhenylHexyl 2-50% gradient
16. UltraCore SuperC18, 2.1 x 50 mm, 2.5 m
4 pHs with CH3CN
0 2 4 6 8
Time (min)
0.0000.0020.004
Volts
0.649
0.881
1.187
1.697
2.613
2.729
3.023
3.118
3.227
3.402
4.021
6.547
6.958
7.188
7.399
2 4 6 8
Time (min)
0.0000.0020.0040.0060.0080.0100.0120.014
Volts
1.972
3.305
3.476
3.731
3.997
4.147
4.274
4.505
4.994
5.180 5.622
6.211
2 4 6 8 10
Time (min)
0.0000.0020.0040.0060.0080.0100.0120.014
Volts
1.948
3.154
3.537
3.804
3.9104.012
4.215
4.581
4.732
5.102
5.635
6.350
6.653
7.072
8.931
CH3CN/pH 3.8
250% in 13.7 min.
k* = 14.3
0.6 mL/min, 30 C for all 4
CH3CN/pH 8.2
270% in 11.9 min.
k* = 14.3
CH3CN/ pH 9.2
270% in 11.9 min.
k* = 14.3
3.0 4.0
Time (min)
Volts
3.154
3.537
3.8043.910 4.012
4.215
4.581
4.732
2 4 6 8 10
Time (min)
0.0000.0020.0040.0060.008
Volts
1.930
3.315
3.638
3.926
4.155
4.719
4.779
4.894
5.178
5.299
5.462
5.865
6.058
6.150
6.661
7.315 Gradient artifact
7.922
9.219
CH3CN/ pH 10.2
270% in 11.9 min.
k* = 14.3
4.0 5.0 6.0
Time (min)
Volts
3.305
3.476
3.731
3.997 4.147
4.274
4.505
4.994
5.180 5.622
3/26/2015 16
17. 0 2 4 6 8 10 12 14
Time (min)
0.0000.0020.0040.006
Volts
3.824
5.303
5.764
6.564
6.941
7.454
8.226
8.849
9.230
9.545
9.887
10.370
11.273
12.040
0 2 4 6 8 10 12
Time (min)
0.0000.002
Volts
0.874
1.533
1.982
2.496
4.132
4.354
4.536
4.938
5.225 6.727
10.829 12.065
0 2 4 6 8 10 12 14
Time (min)
UltraCore SuperPhenylHexyl, 2.1 x 50 mm, 2.5 m:
4 pHs with CH3OH
3/10/2015 Pittcon 2015 New Orleans 17
CH3OH/pH 3.8
250% in 13.7 min.
k* = 14.3
0.6 mL/min, 30 C for all 4
CH3OH/pH 8.2
270% in 11.9 min.
k* = 14.3
CH3OH/pH 9.2
270% in 11.9 min.
k* = 14.3
0 2 4 6 8 10 12 14
Time (min)
0.0000.0020.004
Volts
CH3OH/pH 10.2
270% in 11.9 min.
k* = 14.3
18. 0 10 20
Time (min)
0.0000.0020.0040.0060.0080.0100.012
Volts
2.775
4.987
6.431
7.540
10.166
10.483
10.895
11.340
11.527
11.705
13.944
18.774
19.514
20.190
0 2 4 6 8 10 12
Time (min)
0.0000.0020.0040.0060.008
Volts
0.857
1.478
1.960
2.466
4.091
4.287
4.455
4.878
5.194
6.018
6.712
10.673
11.830
Scaling pH 3.8 and 10.2 Separations to
3 x 100 mm UltraCore SuperPhenylHexyl
18
2 4 6 8 10 12 14 16 18
Time (min)
0.0000.0020.0040.0060.0080.010
Volts
3.481
5.736
6.477 6.951
7.468
13.959
14.597
16.927
17.472
0 10 20
Time (min)
0.0000.0100.020
Volts 4.940
8.565
9.681
10.317
10.804
11.418
12.867
13.354
14.060
14.287
14.526
15.254
15.594
15.989
18.006 20.229
21.322
2.1 x 50 mm, 2.5 m
250% in 13.7 min., 0.6 mL/min, 30 C
CH3OH/pH 3.8 NH4OAc (10 mM), 2 L
k* = 14.35
3 x 100 mm, 2.5 m
1.0 mL/min
270% in 23.8 min.
k* = 14.35
2.1 x 50 mm, 2.5 m
295% in 16.3 min., 0.49 mL/min, 30 C
k* = 14.3
CH3OH/pH 10.2 NH4OAc, 2 L
CH3OH
3 x 100 mm, 2.5 m
1.0 mL/min, 30C
295% in 32.5 min.
CH3OH/pH 10.2 NH4OAc (10 mM), 8 L
pH
3.8
CH3OH
pH
10.2
3/10/2015 Pittcon 2015 New Orleans
14.0 15.0 16.0
Time (min)
Volts
13.354
14.060
14.287 14.526
15.254
15.594
15.766
11.0 12.0
Time (min)
Volts
10.166
10.483
10.895 11.340
11.527
11.705
19. Separation on 3 x 100 mm, 2.5 m
SuperPhenylHexyl: All 16 separated
0 2 4 6 8 10 12
Time (min)
0.0000.0100.020
Volts
1.711
2.5863.025
3.516 4.641
4.941
5.279
5.465
5.755
5.832
6.132 6.576
9.159
9.490
11.21811.886
5.0 6.0
Time (min)
Volts
4.641
4.941
5.279
5.465
5.755
5.832
6.132 6.576
0 2 4 6 8 10
Time (min)
0.0000.0100.020
Volts
2.022
2.653
2.931
3.212
3.805
3.971
4.169
4.3554.417
4.621
4.853
7.307 7.569
3.8 4.0 4.2 4.4 4.6 4.8
Time (min)
Volts
3.805
3.971
4.169
4.355
4.417
4.621
4.853
3/10/2015 Pittcon 2015 New Orleans 19
2–70% ACN/pH 3.8 in 30 min. slope2–70% ACN/pH 3.8 in 15 min. slope
Additional
optimization with
DryLab
20. Summary
• ACE UltraCore SuperC18 and SuperPhenylHexyl columns were
evaluated at several pHs for two mixtures of basic analytes.
• Peak shape and selectivity were best at pH 3.8 and pH 10.2 with
ammonium formate and ammonium acetate/NH4OH mobile phases,
respectively.
• Use of 2.1 x 50 mm, 2.5 m column geometries allows for rapid, yet
thorough examination of various combinations of organic modifier,
stationary phase and pH.
• Method transfer to larger column geometries went well, although
inability to use injection delays did not permit completely accurate
transfer.
• Useable conditions were found for both of the best conditions for
each of the columns at low and high pH.
• For the drugs of abuse sample, pH 10.7 did not afford as good
resolution as pH 10.2.
203/10/2015 Pittcon 2015 New Orleans
