GUT MALABSORPTION ASSAY

Gut permeability is an issue when there are changes and the absorption of nutrients is altered.

The Theory:

A solution of 3 sugars are given to patients at a set concentration, then either urine or blood samples are taken at set time points for analysis. The ratio of the Lactulose (disaccharide) to either Rhamnose or Mannitol (monosaccharides) indicates if and where there is a problem in the gut. These 3 sugars are not metabolised by the body, they just permeate across the gut wall.

How we are involved:

We have developed two analysis methods, based on published work, to measure these sugars in either urine or plasma. We have a lower limit of quantitation of 15µg/ml in Urine and 7.5µg/ml in Plasma, however we would advise that the actual lower levels should be 25µg/ml for urine and 15µg/ml for plasma to ensure that the lowest levels do not fall outside the calibration range. Figure 1 shows the timing and peaks for the sugars being sent round the HPLC.

Figure 1: Graph from the HPLC showing the 3 sugars separation and their associated retention times.

We are working with Mike Hewetson from the Royal Veterinary College to adapt the plasma method for use with equine serum. At present, there is no easy way of determining where a problem is in the equine gut and it is hoped that this test will offer an alternative to submitting the horse for a surgical procedure.

WHAT CAN WE DO FOR YOU?

Here at the Bio Analysis Centre we have the capability to run samples through our UHPLC (Ultra High Performance Liquid Chromatograph) or LCMS (Liquid Chromatograph Mass Spectrometer). Shimadzu (1), the manufacture of the machinery, ran a Multiple Residue Analysis of 210 Pesticides in Food Samples by Triple Quadrupole UHPLC – MS/MS.

 

Pesticide residue on food can severely affect human health, so governments and food manufacturers established regulations which set the limits in a range of 0.01 – 10 mg / kg depending on the toxicity of the pesticide. For food used in the production of baby foods, the detectable level of pesticide must be less than 0.01 mg / kg along with prohibiting the use of certain toxic pesticides.

 

Linearity was investigated over a nine point calibration, with samples ranging from 0.5 μg / kg – 0.2mg / kg or 0.5 – 200 parts per billion (ppb) analysed in duplicate; calibration samples were injected once in increasing order and once in decreasing order. From that the Mass Spectrometer calculated from the area of the peak the concentration by correlating to the calibration curve associated with the specific pesticide.

 

This exact assay can be conducted here at the Bio Analysis Centre, if you have a similar project or any enquiries for any other assay, please contact us.

 

References

  1. Multi-Residue Analysis of 210 Pesticides in Food Samples by Triple Quadrupole UHPLC-MS/MS [Internet]. Shimadzu; [cited 29 January 2020]. Available from: https://www.shimadzu.com/an/literature/lcms/jpo113017.html?fbclid=IwAR0vWv-SXKE4BKlRmm_RgKE7JkAFXR9i-JJn3KsZRGb2sYxMwxiyaWKkNWk
Bio-Analysis at the One Nucleus Bio Wednesday 2019

Bio-Wednesday One Nucleus hosted by LBIC at the RVC

We were delighted to be asked to join the panel last night at the One Nucleus event hosted by LBIC at the RVC.

 

The topic, ‘Staffing the 21st Century Life Sciences R&D Industry.’ bio-analysis HPLC and LCMS

As an employer of several graduates, undergraduates and summer placements in our 5 years of business, we were interested to be part of the discussion and hear other people’s point of view.

We have found the quality of students coming through to be excellent and a real asset to us. In a business like ours they receive a very hands on and varied approach to the commercial world of bio-science.  (A hugely different experience from starting out in a Big Pharma Organisation!)

Discussion topics resonated around what the future looks like for graduates and post graduates; what skills employers and recruitment professionals are looking for: risk takers, lateral thinkers, commercial acumen; what could be taught, what should be taught and the changing foot print of career paths.

For a business like ours, people skills are essential, along with self motivation and common sense.

 

We pride ourselves in offering an excellent technical and customer service.  Our HPLC and LC-MS training can be combined with method development and bespoke consultancy to help advance your Research and Development.  Once trained, clients can come back and use the equipment to continue their research projects.

As a business and an employer, we have a lot to offer so please get in touch.

 

Bio-Analysis at the On Nucleus Bio-Weds - HPLC and LCMS

 

 

 

GDPR Update

Bio-Analysis Centre is reviewing and updating our privacy policies to ensure that your stored data is used securely and transparently. Please view our updated Privacy Policy here.

 

deltaDOT’s High Performance Capillary Electrophoresis

The Bio-Analysis Centre offers High Performance (label free) Capillary Electrophoresis (HPCE) system from deltaDOT to its clients.

Whilst electrophoresis is the process during which ions undergo movement in a fluid or gel under the influence of an electric field, capillary electrophoresis is a technique that separates these ions based on their electrophoretic mobility with the use of an applied voltage. This mobility is dependent on the atom’s radius, the charge of the molecule, and the molecule’s viscosity. The rate at which the charged particle moves is directly proportional to the applied electric field – as the field strength increases the mobility increases also.

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method development

Chromatographic analysis – An Intern’s Perspective

Chromatographic analysis is often an indispensable technique for a life scientist.

The gadgetry and the skills required, for the successful application of such techniques, are less so common. Having spent the summer months of 2017, working for the Bio Analysis Centre, I have been a very keen observer of the company’s modus operandi. I have been most impressed by the assembly of systems, developed and maintained by the laboratory manager, Dr. Hyde. This framework for the BAC ensures the smooth running of the services it provides.

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Triple-Quad MS Used to Confirm Food Safety

As the global population continues to grow exponentially the demand for food becomes ever more pressing. In order to supply this mounting demand, the world’s crop production must increase through optimised methods, fertilisers, agrochemicals and pesticides. Nonetheless, with strict regulations regarding contaminants including pesticides, mycotoxins and heavy metals the manufacturing of food and drinks must be monitored vigilantly. Analytical instrumentation technologies and methods on a mass spectrometer alongside gas chromatography allow this food safety promise to be guaranteed.

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An Introduction to Solid Phase Extraction (SPE)

Analytical scientists utilise a range of sample preparation tools to aid their method development, how they approach this challenge can significantly impact their success.

Solid Phase Extraction (SPE) is a type of sample preparation technology that uses solid particle and chromatographic packing material to chemically separate the different components of a sample.

Samples are usually in a liquid state and are run through stationary phase particles in a cartridge. The chromatographic bed can be used to selectively remove interferences to ensure subsequent analytical testing is more successful.

 

Liquid-Solid Phase Extraction carries the same basic principles of liquid chromatography used in HPLC, but for different reasons. In SPE, chromatography is used to prepare a sample prior to analytical testing.

 

Samples used in SPE can originate from a wide range of sources. They can be biological fluids (eg. Plasma, saliva, urine), food products (eg. Grain and meat), environmental samples (eg. Water, air, soil), pharmaceuticals, beverages or industrial products.

 

There are numerous benefits to using SPE:

 

  • The procedure can simplify a complex sample matrix and aid in purifying the compound. If there are a large number of interfering constituents or substances in the sample matrix, it makes analysis extremely difficult.
  • SPE can reduce ion suppression or enhancement in MS applications. With an appropriate method this effect will be minimised by cleaning the interferences from the compound, resulting in a more accurate reported value.
  • It has the ability to fractionate a sample matrix to allow analysis of compounds by class. If a sample contains many compounds, separating them by class can be useful so that further analysis can be carried out much more efficiently.
  • SPE allows better analysis of trace concentration of very low level compounds. The chromatographic packing material has retention capabilities which allow the ability to trace concentrate. This would be very difficult with other sample preparation techniques.

Comparative Oncology: The Next Frontier