12/02/2021

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

In the work we present to the 14th EBF Open Symposium, we show a 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

The method development focused on the challenges of balancing the detection of the 5 biomarkers; ensuring that each could be quantified within its own required range whilst maintaining a method that was fit for purpose for all 5 biomarkers. To enable this balance, the context of use for each biomarker was evaluated and the method adapted as required.

A fit for purpose method was established that met the requirements for the need and context of use. The method was used successfully in the quantification of human samples from a clinical study.

You may find below the poster for your ready reference:

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02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

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14/06/2021 15:46

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

READ MORE
04/06/2021 11:22

Qualification of a method for the measurement of a potentially therapeutic protein

A recent project completed by Anapharm involved qualifying an analytical method for the measurement of a therapeutic protein in human skin tissue. The protein of interest is one that is expressed ubiquitously in a number of tissues and cell types.

READ MORE

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

In the work we present to the 14th EBF Open Symposium, we show several factors that can affect analyte concentration and that might lead to erroneous conclusions regarding its stability.

These misleading phenomena are described in the poster and focus on three main points:
• Non-specific adsorption.
• Equilibrium time between erythrocytes and plasma.
• Plasma age.

You may find below the poster for your ready reference:

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02/12/2021 14:33

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

A 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

READ MORE
14/06/2021 15:46

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

READ MORE
04/06/2021 11:22

Qualification of a method for the measurement of a potentially therapeutic protein

A recent project completed by Anapharm involved qualifying an analytical method for the measurement of a therapeutic protein in human skin tissue. The protein of interest is one that is expressed ubiquitously in a number of tissues and cell types.

READ MORE

11/04/2021

Cholecalciferol (Vitamin D3)

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02/12/2021 14:33

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

A 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

READ MORE
02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

READ MORE
14/06/2021 15:46

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

READ MORE

Mesalazine

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02/12/2021 14:33

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

A 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

READ MORE
02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

READ MORE
14/06/2021 15:46

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

READ MORE

Tamoxifen Isomers E and Z

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02/12/2021 14:33

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

A 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

READ MORE
02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

READ MORE
14/06/2021 15:46

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

READ MORE

Aprepitant

MORE NEWS

02/12/2021 14:33

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

A 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

READ MORE
02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

READ MORE
14/06/2021 15:46

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

READ MORE

06/14/2021

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

In the work we presented during the 7th Young Scientist Symposium, we aim to provide ideas and strategies to help tackle and solve these problems, based on real issues encountered during method development.

The advice given in the poster when working with low mass compounds focuses on three main points:
• Evaluate alternative ionization sources in order to potentially reduce background noise and increase overall sensitivity.
• Carefully evaluate and investigate the evaporation step in order to reduce cross-contamination, even avoid it altogether when possible.
• Higher presence of endogenous isobaric compounds, that can have the same MRM transition, means that the chromatographic separation has to be extensively optimized. Good chromatographic separation of all isobaric compounds is paramount to avoid erroneous conclusions.

You may find the poster below for your ready reference:

MORE NEWS

02/12/2021 14:33

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

A 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

READ MORE
02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

READ MORE
04/06/2021 11:22

Qualification of a method for the measurement of a potentially therapeutic protein

A recent project completed by Anapharm involved qualifying an analytical method for the measurement of a therapeutic protein in human skin tissue. The protein of interest is one that is expressed ubiquitously in a number of tissues and cell types.

READ MORE

06/04/2021

Qualification of a method for the measurement of a potentially therapeutic protein

A recent project completed by Anapharm involved qualifying an analytical method for the measurement of a therapeutic protein in human skin tissue. The protein of interest is one that is expressed ubiquitously in a number of tissues and cell types.

While there are commercially available immunoassay kits to detect and measure this particular protein, the aim here was to develop a fit-for-purpose method, which produced reproducible results within a challenging matrix.

There were a number of factors that had a direct impact on the reproducibility of this assay and were addressed in the following ways:
• With limitations on the availability of the skin sample matrix and the presence of endogenous protein, a surrogate matrix of RIPA buffer was used.
• To maintain sample stability, samples were sliced on dry ice and removed only briefly for weighing.
• Cut samples were homogenised using the Precylls Evolution Homogeniser®, which allowed for the creation of repeatable programs at sub-zero temperatures.
• Additionally, during homogenisation, RIPA buffer, spiked with a protease inhibitor was added to prevent any enzyme degradation.
• To ensure that a similar amount of drug per mg of total protein was generated regardless of the size of sample section taken, a fixed ratio of RIPA buffer was added per mg of skin tissue.

In addition to method parameters normally assessed for a PK assay, such as precision and accuracy, selectivity, parallelism and stability, the qualification of this method also involved the evaluation and optimisation of the homogenization procedure. The use of an automated tissue homogeniser also helped in the generation of a highly reproducible homogenisation process, which then facilitated the completion of the qualification of a fit-for-purpose method.

Please find below the poster for your ready reference:

MORE NEWS

02/12/2021 14:33

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

A 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

READ MORE
02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

READ MORE
14/06/2021 15:46

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

READ MORE

02/25/2021

Silodosin Glucuronide

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02/12/2021 14:33

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

A 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

READ MORE
02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

READ MORE
14/06/2021 15:46

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

READ MORE

02/23/2021

Silodosin

MORE NEWS

02/12/2021 14:33

Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

A 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

READ MORE
02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

READ MORE
14/06/2021 15:46

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

READ MORE