Project Description

reversed phase polymer chromatography

EPRUI Biotech Co. Ltd. offers a new generation of reversed phase polymer HPLC packing material. Our polymer chromatography column packing material is made of highly crosslinked monodisperse polymer resins including highly crosslinked polystyrene divinylbenzen, polymethyl methacrylate, copolymers of PS/DVB and PMMA.

With precise control of particle size, particle size distribution, specific surface area, pore size structure and surface functional groups, our monodisperse polymer HPLC packing material has great properties including high resolution and loading capacity, strong rigidity, acid and base resistant, low back pressure, long lifetime and low non-specific adsorption.

Our polymer based reverse phase chromatography packings have been widely used in high performance liquid chromatography (HPLC) analysis, isolation and purification of organic compounds, proteins, polypeptides, nucleic acids, natural products and synthetic pharmaceuticals.

EPRUI supplies 2um nonporous polystyrene microspheres which is for UPLC application; 5um and 10um porous polystyrene microspheres for HPLC analysis use; 10um, 15um, 30um, 40um, 50um porous porous PS and PMMA microspheres for preparative chromatography use. Pore sizes can be chosen from 100A, 300A, 500A, 1000A which can satisfy various biological purification requirements.

  1. Highly uniform particle size
  2. Perfect spherical morphology
  3. Optimized pore structure
  4. Good chemical and PH stability
  5. High mechanical strength
  6. Scale production

1. Highly uniform particle size and perfect spherical morphology

It is very important for chromatographic application by controlling particle size and monodispersity of microspheres because column efficiency is usually decided by particle size and its distribution. During chromatographic separation, the longitudinal diffusion of solutes is the main reason for broadening of chromatographic bands and peaks. Columns packed with uniform microspheres can effectively narrow the bands and peaks which provide the lowest back pressure and make the columns with high efficiency and high resolution.

Compared with other commercial chromatography media, our monodisperse microspheres have demonstrated great advantages of strong rigidity, easy column packing, low back pressure, high column efficiency, good resolution, stable column bed and lower risk of frit-plugging.

figue1-monodisperse-polymer-microspheres-with-various-particle-size

Figue1: Monodisperse Polymer Microspheres with various particle size

figure2-particle-size-distribution-of-30um-polystyrene-microspheres

Figure2: Particle Size Distribution of 30um Polystyrene Microspheres

2. Optimized pore structure

Pore size and specific surface area have a strong influence on the separation performance of chromatography media. The choice of a chromatography media with correct pore structure would improve the loading capacity, the column efficiency and the final purity of target molecule. We offer media with pore sizes typically from 50Å up to 1000Å.

figure3-polymer-microspheres-with-different-pore-size

Figure3: Polymer microspheres with different pore size

figure4-comparison-of-separation-result-with-different-pore-size

Figure4: Comparison of separation result with different pore size

Samples: Ribonuclease B, Recombinant human insulin, Cytochrome C, Lysozyme,  BSA
Mobile Phase A: 0.1% TFA in water
Mobile Phase B: 0.1%TFA in acetonitrile
Gradient: 20%B to 40% B,20 min; 40% B,10 min

 

3. Superior chemical and pH stability

Reverse Phase PS/DVB V.S.  Silica Gel C18

The biggest advantage of highly cross-linked PS/DVB chromatography over silica gel chromatography is its excellent pH stability over a wide range of pH 1-14. Polymer chromatography media maintain their outstanding physical and chemical stability even in extreme acid or alkaline solutions (for e.g. 1 M NaOH/HCL) and organic solvents (such as methanol, ethanol, acetone, n-propanol, isopropanol, DMSO, THF, acetonitrile, 6 M guanidine hydrochloride, etc.). Besides, EPRUI provides the customers a wider choice of conditions for process development and optimization. The outstanding chemical stability, have been proven to meet the requirements of Cleaning-in-Place (CIP) in cGMP operations. For example, the loading capacity and separation efficiency remain unchanged, after 10um 300A PS/DVB column was soaked in 1 M NaOH at 60°C for 40 days (Figure 1, used for Insulin purification).

Insulin-purification

Figure 1:Insulin loading capacity when EPRUI-PSD10-300 column was soaked in 1M NaOH (at 60℃) for 40 days.

4. Mass production and Great batch reproducibility

Our advanced manufacturing facilities and  a precise quality control system guarantee consistent high quality and batch-to-batch reproducibility of the bulk production of media. Figures below show the results of the performance test of more than twenty different lots of the chromatography media, indicating the good batch-to-batch reproducibility in our production.

figure5-consistent-particle-size-distribution-30um

Figue5:Consistent Particle Size Distribution—30um

figure6-consistent-particle-size-distribution-40um

Figue6:Consistent Particle Size Distribution—40um

figure7-stable-pore-volume-and-specific-surface-area

Figure7: Stable pore volume and specific surface area

figure8-reproducible-isolation-of-four-proteins-by-18-lots
Figure8: Reproducible isolation of four proteins by 18 lots

chromatographic column: 4.6 mm I.D. x 250 mm, EPRUI- 40PS-300
Mobile Phase A: 0.1% TFA in water
Mobile Phase B: 0.1%TFA in acetonitrile
Gradient: 20%B to 40% B,20 min; 40% B,10 min
Flow Velocity: 1 ml/min;80%B to 95%B,10min
Wavelength: UV @ 280nm

Our monodisperse polymer microspheres have been successfully applied in the isolation and purification of plant extracts, antibiotics, polypeptides and others.

Plant Extracts Antibiotics Polypeptides Others
Taxol Vancomycin Insulin Iohexol
Docetaxel Derivatives of vancomycin Thymalfasin Iopamidol
Huperzine Daptomycin Octreotide Acarbose
Ascutellarin Telavancin Oxytocin
Stevioside Caspofungin
Neomangiferin Pneumocandin B0
Myricetin Teicoplanin
Diosgenin Dalbavancin
Vinorelbine
Ecdyson
Arbutin

Purification Case of Insulin

EPRUI develops a whole set of insulin purification technology which can successfully improve the purity of insulin from 45% to 99%

purification-case-of-insulin

Condition:

Instrument: High pressure equipment

Loading Amount: 100mg/ml

Test: 214nm

Item Particle Size Pore Size Matrix
EPRUI-PSD3 3um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSD5 5um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSD10 10um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSD15 15um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSD20 20um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSD25 15um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSD30 30um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSD40 40um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSD50 50um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSD100 100um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSD200 200um Nonporous,100,300,500,1000A PS/DVB
EPRUI-PSM10 10um Nonporous,100,300,500,1000A Poly DVB/acrylate
EPRUI-PSM15 15um Nonporous,100,300,500,1000A Poly DVB/acrylate
EPRUI-PSM20 20um Nonporous,100,300,500,1000A Poly DVB/acrylate
EPRUI-PSM30 30um Nonporous,100,300,500,1000A Poly DVB/acrylate
EPRUI-PSM40 40um Nonporous,100,300,500,1000A Poly DVB/acrylate
EPRUI-PSM50 50um Nonporous,100,300,500,1000A Poly DVB/acrylate
EPRUI-PSM100 100um Nonporous,100,300,500,1000A Poly DVB/acrylate
EPRUI-PSM200 200um Nonporous,100,300,500,1000A Poly DVB/acrylate
EPRUI-PMM10 10um Nonporous,100,300,500,1000A Polyacrylate
EPRUI-PMM15 15um Nonporous,100,300,500,1000A Polyacrylate
EPRUI-PMM20 20um Nonporous,100,300,500,1000A Polyacrylate
EPRUI-PMM30 30um Nonporous,100,300,500,1000A Polyacrylate
EPRUI-PMM40 40um Nonporous,100,300,500,1000A Polyacrylate
EPRUI-PMM50 50um Nonporous,100,300,500,1000A Polyacrylate
EPRUI-PMM100 100um Nonporous,100,300,500,1000A Polyacrylate
EPRUI-PMM200 200um Nonporous,100,300,500,1000A Polyacrylate