SPC: NMR

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The Singapore Phenome Centre (SPC) is the first in Singapore and South East Asia to be equipped with the In Vitro Diagnostics Research (IVDr) system. This 600 MHz NMR IVDr system comprises of the AVANCE III HD console, Ascend 600 magnet and a 5mm broadband inverse probe (Bruker, Z814601, PA BBI 600S3 H-BB- D-05 Z) that is a Z-gradient high resolution probe, targeted at screening complex mixtures by 1H NMR for rapid identification and quantification of metabolites.

The system also comes with an automated SampleJet sampler changer that can process up to 5 racks of 96 sample tubes and an additional 30 sample positions on its carousel. The SampleJet system has a cooling unit to cool samples to 4 °C prior to measurement. Typical sample preparation for NMR measurement requires a minimum of 0.9 mL of sample for urine while plasma or serum samples require a minimum of 0.4 mL.





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The IVDr system is a complete, proven and standardised nuclear magnetic resonance (NMR) spectroscopy platform aimed at enabling cost-effective, high-performance NMR based pre-clinical screening and diagnostic research of novel NMR assays of biofluids such as urine, plasma and serum. Based on Bruker’s validated standard operating procedures, high throughput NMR screening can be achieved in a highly reproducible and automated manner. This will assist the development of novel NMR methods for the diagnosis and prognosis of disease especially as the system is highly suited for the automated high throughput screening of large batch sizes of samples. This correlates well with the large sample size analysis required for “Epidemiology” and “Public Health” studies in development towards preventive healthcare and precision medicine. In addition, the IVDr system at SPC has unlimited access to Bruker’s in-house database library that can be used to quantify up to 750  compounds in urine, plasma and serum samples with high accuracy and speed.

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Typical NMR spectra for blood plasma or serum samples are recorded using a standard presat. 1D nuclear Overhauser effect spectroscopy (NOESY) pulse sequence as well as an additional Carr-Purcell- Meiboom-Gill (CPMG) pulse sequence. The CPMG experiment facilitates the detection of small molecule metabolites by reducing the large overlapped signals from biological macromolecules like protein and lipids.


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A 2D J-resolved spectroscopy experiment (JRES) is also done to resolve overlapping spectra peak signals and is capable of validating and quantifying up to 115 lipoproteins. Quantification is done via the “Electronic REference To access In vivo Concentrations” (ERETIC) method that allows accurate quantification without the need for the spiking in of internal standards. This is done by inducing an artificial electronic signal in the spectrum at a peak-free region. Target NMR peak signals specific to each individual compound have been identified for quantification and resolved using Bruker’s in-house algorithms, that rigorously fit the peak shapes to each individual compound’s specific NMR peak signal and are further validated against known authentic standards at various pH values. As a result, ERETIC both greatly simplifies the sample preparation process for NMR measurement and facilitates rapid and cost-effective metabolite quantitation.