HPLC Column for Deproteination / Direct Injection of Biological Samples
When analyzing a target compound in a biological sample containing a large amount of protein such as serum by HPLC, deproteinization that requires time and labor is required. Wakopak® Wakosil® GP-N6 is a HPLC column that allows direct injection of protein-containing samples.
In HPLC analysis by Wakopak® Wakosil® GP-N6, macromolecules, such as proteins, pass through the column due to the hydrophilic structure of the modifier, while relatively small molecules, such as drugs, are retained by reverse-phase distribution, etc.
Features
- Capable of direct injection of biological samples
- Excellent durability
- Capable of analysis with high recovery rate and high reproducibility
Comparison with Conventional Pretreatment Procedures
Evaluation of Column Durability
Analysis of blood uric acid
Injection and analysis of 5 μL human serum were repeated. There is almost no change in the peak shape and retention value of uric acid even after 500 analyses, and stable results are delivered.
| First: R.T.-3.96 min., Press.23 kg/cm2 300 analysis: R.T.-3.97 min., Press.27 kg/cm2 500 analysis: R.T.-3.94 min., Press.33 kg/cm2 |
|
| Column Size Eluent Flow rate Detection Sample Injection Volume Peak |
: 4.6 mm x 150 mm : CH3CN/0.2 M Na2HPO4 (pH 6.0)=2/98 (v/v) : 0.5 mL/min. at R.T. : UV 284 nm : Human Serum : 5 μL : 1. Uric Acid |
Example of Direct Analysis
| Column Eluent Flow rate Detection Sample |
：Wakopak® Wakosil GP-N6 (4.6 mm x 150 mm) ：CH3OH/0.1 M Na2HPO4 (pH 6.0)=10/90 (v/v) ：1.0 mL/min. at 40℃ ：UV 254 nm ：1．Acetaminophen 5 μg/mL 2．Sulfamethoxazole 5 μg/mL 3．Ethenzamide 20 μg/mL 4．Salicylamide 25 μg/mL |
Column Switching Method
Wakopak® Wakosil® GP-N6 can be used as a pretreatment column for "column switching analysis", which can be used in pretreatment of biological samples.
Example of analysis of drug contained in serum
| Column Eluent Flow rate |
：Wakopak® Wakosil GP-N6 (4.6 mm x 50 mm) ：CH3CN/0.2 M NaH2PO4, Na2HPO4 (pH 6.0)=2/98 (v/v) ：1.0 mL/min. at 30℃ |
【Analytical conditions】
| Column Eluent Flow rate Detection Column switching time Sample |
：Wakopak® Wakosil-Ⅱ 5C18 RS (4.6 mm x 150 mm) ：CH3CN/0.1 M NaH2PO4, Na2HPO4 (pH 6.0)=30/70 (v/v) ：1.0 mL/min. at 30℃ ：UV 254 nm 0.032AUFS ：4.8-6 min. ：Human Serum 20 μL 1．Phenobarbital 20 μg/mL 2．Carbamazepine 5 μg/mL 3．Phenytoin 40 μg/mL |
Drug recovery rate
| Name of drug | Blood concentration (μg/mL) | Recovery rate (%) | CV (%) |
|---|---|---|---|
| 1. Phenobarbital | 20 | 99.3 | 1.06 |
| 2. Carbamazepine | 5 | 96.5 | 1.12 |
| 3. Phenytoin | 40 | 102.3 | 0.78 |
Precautions on handling
- Please use the mobile phase with the pH range of 2.5 to 7.5.
- Use the organic solvent concentration in the mobile phase at a level that does not denature the proteins (for acetonitrile, we recommend 20% or lower). Do not use solvents that denature the proteins.
- Pass the sample through a 0.2 μm filter.
- To prevent clogging of the column filter, it is recommended that a line filter of approximately 2 μm should be connected between the autosampler and column.
- Please handle the column in the same manner as the reversed-phase columns.
Product List
- Open All
- Close All
For research use or further manufacturing use only. Not for use in diagnostic procedures.
Product content may differ from the actual image due to minor specification changes etc.
If the revision of product standards and packaging standards has been made, there is a case where the actual product specifications and images are different.
The prices are list prices in Japan.Please contact your local distributor for your retail price in your region.