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Analyzing potential biomarkers in sweat by ultra-high-performance supercritical fluid chromatography-mass spectrometry (CAT#: STEM-CT-0027-LJX)

Introduction

Liquid chromatography-mass spectrometry methods were required to afford the rapid separation and detection of purines and small organic acids. These compounds are found in sweat and sebum and are potential biomarkers for the early detection of pressures sores. Two ultra-high-performance supercritical fluid chromatography-mass spectrometry assays have been successfully developed for both classes of compounds. Separation for purines was achieved using a gradient of supercritical carbon dioxide and methanol with a 1-aminoanthracene sub 2 μm particle size column followed by positive ion electrospray ionization. Separation for organic acids was achieved using a gradient of supercritical carbon dioxide and methanol (50 mM ammonium acetate 2% water) with a Diol sub 2 μm particle size column followed by negative ion electrospray ionization. Calibration curves were created in the absence of internal standards and R2 values > 0.96 were achieved using single ion monitoring methods for the protonated purines and the deprotonated acids. The two new assays afford rapid analytical methods for the separation and detection of potential biomarkers in human sweat leading to the early detection and prevention of pressure sores.




Principle

Supercritical fluid is a substance that has both gaseous and liquid properties above the critical point. Supercritical fluids have the advantages of high diffusion coefficient, low viscosity, adjustable solubility and high vapor phase density, so they can provide efficient mass spectrometry ionization and separation results.
Supercritical fluid chromatography (SFC) is an efficient separation technique that uses supercritical fluid as a mobile phase. The samples are packed into short tubes or SPE columns, and the samples are compressed and regulated by supercritical fluid to obtain good solubility. The sample components are then separated by column interaction, thus achieving the separation of different compounds.

Applications

For efficient separation of substances
Widely used in biology, chemistry, environmental protection and other fields

Procedure

1. Sample injection
2. The high pressure pump increases the pressure of the sample and mobile phase
3. The sample and mobile phase enter the chromatographic column
4. Flow limiter assists sample separation

Materials

• Sample Type:
Sweat

Notes

1. In the process of separation, the control of pressure and temperature of supercritical fluid is very important, which affects the properties and separation efficiency of supercritical fluid.
2. In addition, the selection of the appropriate column, packing and moving equivalent factors will also affect the separation effect.
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