Process Media


TOYOPEARL Butyl-650M is a hydrophobic interaction chromatography (HIC) resin for biomolecule purification. The resin is composed of a base material of hydroxylated methacrylic polymer beads that have been functionalized with a Butyl ligand group.  TOYOPEARL Butyl-650M resin has a high degree of chemical and mechanical stability, allowing cleaning-in-place (CIP) and very high flow rates. TOYOPEARL Butyl-650M resin has a 100 nm pore size and a 65 µm bead size and is the second most hydrophobic of the TOYOPEARL HIC resins.

TOYOPEARL Butyl-650M resin can be used for high throughput capture, intermediate purification, and polishing process steps. The primary applications of this resin are for the separation of proteins, their isoforms, and aggregate removal. If greater resolution is needed, the smaller particle TOYOPEARL Butyl-650S resin may be used. If lower resolution is desired, the TOYOPEARL Butyl-650C can be used.

TOYOPEARL Butyl-650M resin has a dynamic binding capacity for lysozyme of 30 to 50 g/L. TOYOPEARL Butyl-650M resin maintains capacity at increased linear velocities, has good pressure-flow characteristics, and will withstand back pressures up to 0.3 MPa.


P/NDescriptionResin VolumeParticle Size (µm)   Price
0019802TOYOPEARL Butyl-650M25 mL65  186
0007477TOYOPEARL Butyl-650M100 mL65  564
0014702TOYOPEARL Butyl-650M1 L65  -
0007976TOYOPEARL Butyl-650M5 L65  -
0018355TOYOPEARL Butyl-650M50 L65  -


Product Attributes

Base bead: HW-65
Pore size (mean): 100 nm
Particle size (mean): 65 µm 
Ligand type: Butyl
DBC: 30 - 50 g/L 
Shipping buffer: 20% ethanol
pH stability: 1-13
Stability data: 10 years




Ether < PPG < Phenyl < Butyl < Hexyl
(least to most)


Separation of Lipase Isozymes from Impurity


Resin: TOYOPEARL Butyl-650M
Column size: 34 mm ID × 29 cm
Mobile phase: linear gradient, 810 mL (NH4)2SO4, 25% to 0% saturation in 0.01 mol/L acetate buffer, pH 5.6
Flow rate: 2 cm/hr
Detection: UV @ 254 nm
Sample: lipase from Geotrichum candidum
Sample load: 375 mg

For additional applications, please see our applications database.


Influence of Salt-type on Resolution


Chromatography on a TOYOPEARL Butyl-substituted support.

Resin: TOYOPEARL Butyl-650M
Mobile phase: 4.1 mm ID × 4 cm 
Mobile phase: Buffer A: 20 mmol/L phosphate buffer in 1.0 mol/L indicated salt, pH 7.0
Buffer B: buffer A with 1.0 mol/L indicated salt
Flow rate: 484 cm/hr (1 mL/min)
Detection: UV @ 280 nm

Fausnaugh, J.; Kennedy, L.; Regnier, F. J. Chromatography, 1984, 141, 317.

Benzyl Alcohol Removal


A 2% solution of benzyl alcohol in water has been identified as a suitable alternative to 20% ethanol as a preservative in resin storage solutions. A sample of TOYOPEARL Butyl-650M resin was prepared by adding 100 mL of aqueous 2% benzyl alcohol to 100 mL of suction filtered resin. A 100 mL aliquot of DI water was added to the filtered resin and stirred to make a slurry. This resin/ DI water slurry was allowed to stand for 5 minutes and was then suction filtered to remove the supernatant. This procedure was repeated 14 more times, for a total of 15 washes.

Samples of the filtered supernatant from the TOYOPEARL Butyl-650M resin was taken after the 5th, 10th, and 15th washes and analyzed for benzyl alcohol concentration. As demonstrated in the figure, a 2% benzyl alcohol solution can be removed from the TOYOPEARL Butyl-650M resin by thorough washing with DI water.

1.6 cm ID × 10 cm (20.1 mL)
1.6 cm ID × 10 cm (20.1 mL)
Code #DescriptionLiterature Type
AN056 Clearance of 2% Benzyl Alcohol from TOYOPEARL ResinsApplication Note
AN117 Simple and Effective Method for Purification of DMT-on Oligonucleotides using Hydrophobic Interaction Chromatography ResinsApplication Note
IM02 Instruction Manual for TOYOPEARL and TSKgel PW-type ResinsInstruction Manual
IM14 Instruction Manual -- Principles of Hydrophobic Interaction ChromatographyInstruction Manual
PCAT03 TOYOPEARL, TSKgel and Ca⁺⁺Pure-HA Bulk Resin 2019 Product Guide -- Hydrophobic InteractionProduct Catalog
RIS0012 TOYOPEARL Butyl-650Resin Information Sheet
TP231 An Effective One-step Purification of 5’-DMT-On Oligonucleotides Using Hydrophobic Interaction ChromatographyTechnical Presentation
P/NDescriptionPrice ($) 
OC41MDHIC-96Resin Seeker TOYOPEARL HIC 96-well plate kit-
0045235SkillPak HIC 1 mL column library-
0045233SkillPak HIC 1 mL column library284
0045215SkillPak TOYOPEARL Butyl-650M 1 mL columns (qty. 5)284
0045251SkillPak TOYOPEARL Butyl-650M 5 mL column235
0021376ToyoScreen Butyl-650M, 1 mL (qty. 6)-
0021377ToyoScreen Butyl-650M, 5 mL (qty. 6)-
0021398ToyoScreen HIC Mix Pack, 1 mL (qty. 5)-
0021399ToyoScreen HIC Mix Pack, 5 mL (qty. 5)-