Solid Phase Extraction

HybridSPE-Phospholipid

HybridSPE Phospholipid Technology
Ion Suppression & Phospholipid Contamination
Phospholipid Enrichment
Features & Benefits of HybridSPE-Phospholipid
Products
Technical Resources
SelectScience Award

 Enrichment of Phospholipids in Biological Samples Using HybridSPE (176Kb PDF) 

HybridSPE-PPT Brochure
HybridSPE-PPT Brochure


HybridSPE-Phospholipid Technology back to top

HybridSPE combines the simplicity of protein precipitation with the selectivity of Solid Phase Extraction (SPE) for the targeted removal of phospholipids and proteins in biological plasma/serum (Figure 1). The technology utilizes a zirconia-coated particle, and exhibits selective affinity towards phospholipids while remaining non-selective towards a range of basic, neutral and acidic compounds. The phospholipid retention mechanism is based on highly selective Lewis acid-base interaction between the proprietary zirconia ions (functionally bonded to the HybridSPE stationary phase) and the phosphate moiety consistent with all phospholipids (Figure 2).

Figure 1. HybridSPE “In-Well” Precipitation Method and Phospholipid Removal
hybrid spe ppt

View the details for each step in this process

  1. Precipitate Proteins
  2. Mix
  3. Apply vacuum
  4. Results
 
Figure 2. Lewis Acid-Base Interaction Between Hybrid SPE Zirconia Ions and Phospholipids
Proprietary HybridSPE
Zirconia Coated Silica
zirconium-phosphate interaction The phosphate moiety of phospholipids
Is a strong Lewis base
(electron donor) that interacts with Zr
atoms coated on the silica surface.


Ion-Suppression & Phospholipid Contamination back to top

In pharmaceutical bioanalysis, researchers develop and run various assays to quantitate drugs, pharmaceutical candidates, and their metabolites in biological fluids such as serum and plasma. As analysts strive for faster analyses, shorter run times, and lower limits of detection, ion-suppression due to inadequate removal endogenous sample interferences has become of great concern. It is well documented that one of the principal causes of ion-suppression is phospholipid contamination. Not only is signal suppression often evident in the positive ion electrospray mode (+ESI), phospholipids can often remain on the analytical column after sample analysis, and elute uncontrollably in a given LC run sequence.


Phospholipid Enrichment back to top

HybridSPE-Phospholipid can also be used to selectively capture and elute phospholipids for analysis and phospholipid profiling. The same Lewis-acid-base interactions that selectively remove phospholipids as interferences can also be used to recover phospholipids for analysis. A strong Lewis-base (5% ammonium hydroxide in acetonitrile) can be used to effectively and efficiently elute retained phospholipids from the HybridSPE sorbent.


Features & Benefits of HybridSPE: back to top
  • Merges both Protein Precipitation (PPT) & Solid Phase Extraction (SPE)
    • Offers simplicity & generic nature of protein precipitation
    • Selectivity approaches SPE via the targeted removal of phospholipids
  • 2-3 step generic procedure
  • 100% removal of phospholipids & precipitated proteins
  • Minimal to no method development
  • Available in 96-well and 1 mL cartridge dimensions
  • Can be used to enrich phospholipids for analysis 

 

hybrid spe ppt well plate

Products back to top
Description Qty Cat. No.
HybridSPE Products
Precipitation 96-well Plate, 50 mg/well 1 575656-U
 Precipitation 96-well Plate, 15 mg/well 1 52794-U
Precipitation Cartridge, 30 mg/1 mL 100 55261-U
Precipitation Cartridge, 500 mg/6 mL 30 55267-U
Related Products
96-well Protein Precipitation Filter Plate 1 55263-U
Supelco PlatePrep Vacuum Manifold 1 57192-U
96 Square/Deep Well Collection Plates, 0.35 mL, PP 50 575651-U
96 Square/Deep Well Collection Plates, 1 mL, PP 50 575652-U
96 Square/Deep Well Collection Plates, 2 mL, PP 50 575653-U
96 Square Well Pierceable Cap Mats 50 575655-U

Technical Resources back to top
Category / Title Type Size (kb) No. Pages I.D.
HybridSPE-Precipitation Technology Product Info. 483  T408102
Instructions/Data Sheet & Troubleshooting Guide for HybridSPE Data Sheet 76  T708008
Enrichment of Phospholipids in Biological Samples Using HybridSPE (Reporter 28.3) Article 176 T210003
Minimizing Phospholipid Matrix Effects in HILIC LC-MS using HybridSPE-Precipitation Small Volume Plates (Reporter 28.2) Article 338 T210002
Increased Bioanalytical Throughput and Recovery Utilizing HybridSPE-PPT Small Volume Plates (Reporter 28.1) Article 526 T210001
Troubleshooting Analyte Recovery when Using HybridSPE-Precipitation Technology (Reporter 27.2) Article 697 T209002
Increase Bioanalytical Assay Speed via Phospholipid Removal using HybridSPE™-Precipitation Technology (Reporter 26.5) Article 596 T208005
Introducing HybridSPE™-Precipitation Technology for Pharmaceutical Bioanalytical Sample Preparation (Reporter 26.3) Article 389 T208003
High Recovery Method of HybridSPE®-Phospholipid for Cleanup of Biological Samples Prior to LC-MS Analysis Presentation 465 22 T411056H
High Throughput Bioanalysis Utilizing Fused-Core™ Particle and HybridSPE®-PPT Technology  (HPLC 2010) Presentation 494 19 T410086H
Alternative Method of Hybrid SPE Sample Preparation for High Recovery LC-MS Analysis of Pharmaceutical Compounds in Plasma Presentation 712  17  T409228H
Comparison of Sample Preparation Techniques for Reduction of Matrix Interference in Biological Analysis by LC-MS-MS Presentation 216  22  T408163H
Depletion of Phospholipid Matrix Interference when Dealing with Small Volume Plasma Samples Presentation 587  20  T410128H
High Throughput Removal of Both Phospholipids and Proteins in Bioanalytical Sample Preparation Presentation 257  20  T409038H
Impact of Ion-Suppression Due to the Presence of Phospholipids on the Enantiomeric LC-MS Analysis of Clenbuterol Presentation 185  17  T409108H
Increased Bioanalytical Throughput Utilizing Fused-Core™ Particles with Selective Phospholipid Depletion Presentation 532  21  T409189H
Recovery of Pharmaceutical Drugs From Small Volume Biological Sample Using HybridSPE-PPT 96-well Plate Presentation 646  22  T410048H
Selective Depletion of Phospholipids Interference Utilizing HybridSPE™-Precipitation Technology Presentation 658  49  T408181H

SelectScience Award back to top