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Bioanalytical Services

​SingHealth Clinical Pharmacology provides method development, method validation and sample analysis of small and large molecules in blood, saliva, synovial fluid, urine, culture supernatant and tissue specimens to support clients in advancing their research programs. We offer methodologies based on LC-MS/MS, HPLC, ELISA and AlphaLISA. Biologics are one of the fastest growing classes of drugs in the past decade and we offer to develop ELISA based method for level monitoring to identify individuals with loss of responses.

In order to provide the flexibility for researchers and commercial partners to meet their budget and diverse project's needs, we offer customised bioanalytical services either in non-GLP setting or according to GLP requirements. We will also help to advise on alternative assay scenarios, if feasible, to help manage cost and minimise time.

For non-GLP services, it allows for shorter development and analysis time. Once the analyte(s) has been proven to be useful in clinical setting or an important lead in drug discovery process, these research grade assays can be transferred to method development for GLP bioanalytical testing. This will be valuable in preliminary pharmacokinetics studies.


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Small Molecules Bioanalysis

Small active molecules (usually < 1 kDa) make up over 90% of the drugs available on the market today and with our experience in analytical methodologies, we provide analysis of

•Small molecule drugs and its metabolites

•New chemical entities

•Small biomarkers

The available platforms to support these analyses are LC-MS/MS and HPLC.


Large Molecules Bioanalysis.pngLarge Molecules Bioanalysis

Large molecule drugs, which are typically therapeutic proteins (biologics) have in recent years becoming increasingly important. Due to the high cost of biologics, monitoring of therapeutic antibody levels may be a cost effective option especially in view of possible anti-drug responses.

We provide analysis of biologics and large biomarkers using ELISA, AlphaLISA and LC-MS/MS (limited).





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Method Development

To support in vitro and in vivo studies, quantitative method development is provided for

• Single compound

• Cassette doses

• Compound series

This comprises of optimisation of the sample extraction and selected analytical techniques as well as establishing the calibration ranges. The time required is dependent on

                                                                        • Complexity of the studied molecules

                                                                        • Availability of established methodologies

                                                                        • Detection sensitivity required for the study


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Method Validation

Validation of method provides documented evidence that the developed analytical procedure is suitable for its intended use in term of quality, reliability and consistency of results. It is performed in compliance with GLP and is patterned after the current FDA "Guidance for Industry - Bioanalytical Method Validation" and/or EMA "Guideline on Bioanalytical Method Validation. Clients can also tailor the validation details according to their needs and budget. Some of the typical validations are:

• Precision and accuracy

• Selectivity

                                                                        • Recovery efficiency

                                                                        • Dilution integrity

                                                                        • Matrix effect

                                                                        • Re-injectability

                                                                        • Temperature stability

                                                                        • Repeated freeze-thaw stability

                                                                        • Stock solution stability

                                                                        • Working solution stability


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Sample Analysis

Analytical methods are employed in the analysis of in vitro and in vivo samples. Sample matrix and calibration ranges are flexible and are tailored to meet individual study requirements. In general, results are reported as test compound concentration per sample volume or weight in Excel format.

In addition to developing new analytical methods, we also provide analysis of biomarkers using commercially available ELISA, AlphaLISA® and AlphaScreen® kits for research studies. Let us know your requirement and we can assist to source for the appropriate kits.


 

Instruments for Bioanalytical Analyses

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  • Multiplex drug quantification

Able to measure two or more drugs1 in a single analytical run due to its ability to identify different ionised pairs even if they are in the same retention time. Saving time and sample.

  • Improved sensitivity and specificity

A combination of physical separation ability of HPLC and mass analysis ability of mass spectrometry (replacing the UV detection of HPLC) to provide increased sensitivity (up to femtomole2) and specificity.

  • Low sample volume

Due to its increased sensitivity, sample as low as 5 - 10 µL can be analysed but subjected to the concentration and properties of the target analytes, and sample extraction efficiency.

  • High throughput due to shorter analytical time

Depending on the properties of target analyte and methodology, each sample may be analysed in less than 1 minute.

  • Suitable for wide array of biological matrices besides blood and plasma

Also suitable for saliva, cerebrospinal fluid, urine, tissues etc.

1 Subjected to whether the analytes can be efficiently extracted in a single methodology
2 Subjected to analyte recovery efficiency during extraction as well as matrix interferences

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  • Broad spectrum drug screening

Allows for screening of a wide classes of drugs regardless of their structure and/or polarity.

  • Less complexity than LC-MS/MS

Fewer considerations in method development. Separation by physical and chemical properties such as molecular sizes, solubility and detection by UV absorbance. However, unlike LC-MS, it does not have resolving power to differentiate between two analytes with the same retention time.

  • Lower sensitivity than LC-MS/MS

Due to the limitation of UV detection, pico-level may be possible subjected to target analytes and extraction protocol. Usually higher sample volume (> 100 µL) is required.

  • Allows for multiplex drug quantification

Able to measure two or more drugs in a single analytical run, provided if they have different retention times.

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Absorbance & fluorogenic assays based eg. ELISA

• Chemiluminesence based assays eg. CLIA

Alpha technology based assays eg, AlphaLISA 

• Time-resolved fluorescence®, AlphaScreen®

Developed Bioanalytical Assays
We have developed the analytical methodologies for the below analytes. 

 

AnalytesPlatform​Validated Matrices​
ColchicineLC-MS/MS​Human plasma, synovial fluid​
5-MethyltetrahydrofolateLC-MS/MS​Human plasma​
5- FluorouracilHPLC​Mouse plasma​
Imatinib​HPLC​Human plasma
Uracil & DihydrouracilLC-MS/MS​Human plasma, mammalian cell lines​
Clozapine​HPLCHuman plasma, saliva​