Preparative Chromatography Redefined
7
Preparative Chromatography Redefined
Transcript of Preparative Chromatography Redefined
Preparative Chromatography Redefined
Conventional Slurry Packing
Axia™ PackedHydraulic Piston
Compression Packing
Conventional Slurry Packing
Axia™ PackedHydraulic Piston
Compression Packing
Ef�
cien
cy (N
)
Pea
k A
sym
met
ry
1
1.1
1.2
1.3
1.4
40,000
60,000
80,000
100,000
120,000
53
54
55
56
57
58
59
60
61
62
Den
sity
(%)
Conventional Slurry Packing
Axia™ PackedHydraulic Piston
Compression Packing
Reproducible Column-to-Column EfficiencyAverage Efficiency (N) with Synergi™ 4 µm Hydro-RP 100 x 21.2 mm
Reproducible Column-to-Column Peak AsymmetryAverage Peak Asymmetry with Gemini® 5 µm C18 50 x 21.2 mm
Density Comparison of Packed Beds
27 % Improved Avg. Efficiency
13 % Improved Avg. Peak Shape
Expect an Excellent Axia™ Column. Every Time.A completely automated packing system offers feedback control and infinite tuning of packing density to specific media characteristics such as mechanical strength and porosity. An optimum higher bed density can be consistently reproduced column-to-column. This directly trans-
lates into consistent efficiency and peak asymmetry measurements and decreases the column variability seen in traditionally packed preparative columns.
Consistent Quality. Column-to-Column. Batch-to-Batch.
% RSD % RSD
11.8
2.5
9.2
5.2
Phenomenex Web: www.phenomenex.com
Worldwide accepted and trusted phases •such as C18(2), C8(2), and Phenyl-Hexyl
High surface area (100 Å; 400 m• 2/g) for increased sample loading
Available in bulk media for process scale •purifications
Strong hydrophobic selectivity in both •volatile and non-volatile buffers
Extremely high loading capacity from a •large surface area particle (110 Å; 375 m2/g)
pH stable 1-12 for maximum durability•
Four unique selectivities resolve and •separate mixtures of polar and non-polar compounds
High surface area (80 Å; 475 m• 2/g) for increased sample loading
Available in bulk media for process scale •purifications
Small Molecules
Selectivity Options. You Decide.
Chiral Molecules Proteins & Peptides Synthetic DNA/RNA
Engineered to provide a straightforward •approach to enantiomeric recognition and separation through HPLC and SFC
Lux Cellulose-2 incorporates an •advanced halogenated derivative, leading to unique enantioselectivity compared to previously commercialized cellulose phases
Lux Cellulose-1 features a classical •cellulose derivative used industry-wide for many enantiomeric separations and is a cost-effective replacement for CHIRALCEL® OD-H®
300 Å C18 and C4 columns designed to •analyze and purify intact proteins
Provides excellent resolution between •proteins with similar properties
90 Å C12 (Proteo) columns engineered •for increased peak capacity and resolution of peptide separations
Direct scale-up to preparative and bulk •materials
Easily separate N-1 failure sequences •from target oligo with > 90 % purities
Baseline separation/collection of desired •peak at preparative scale (5 µmole and greater)
Long column lifetime due to extended pH •stability and mechanical strength
An advanced column packing and hardware design, Axia™ incorporates patent-pending Hydraulic Piston Compression technology to eliminate bed collapse as a source of failure in short preparative columns. Ideal bed density is custom calculated and automated for each support, chemistry, and column size. Computer control of the
entire process assures both proper bed density and uniformity. Using a single, controlled hydraulic compression, the piston assembly is locked in place without allowing the media to decompress or “relax,” thus maintaining media and column bed integrity. Recompression of the bed is not required, as it is for other packing methodologies.
Axia Packing Process
Packing Piston
Media packed under ideal pressure
The packing piston head is integrated into the column and locked by the piston
retainers, so the pressure is never released.
Media is never allowed to relax, eliminating
voids and dramatically improving reproducibility,
column to column.
Extended column lifetime•
Reproducibility: column-to-column •
Higher efficiency•
Improved peak shape•
Increased loadability•
Stability under high flow rates•
The result is an improved, repeatable packing process, giving preparative chromatographers:
Hydraulic Piston Compression
See the technology in action! Please visit www.AxiaPrep.com
Phenomenex Web: www.phenomenex.com
0 2 4 6 8 min 0 10 20 min
0 1 2 3 min
0 1 2 3 min
0 1 2 3 min
0 1 2 3 min
Ap
p ID
180
62A
pp
ID 1
8077
Ap
p ID
180
78A
pp
ID 1
8064
Ap
p ID
180
63
Ap
p ID
180
65
Phenomenex Web: www.phenomenex.com
Option 2: Increase Column IDFor maximizing load without increasing the run time, consider scaling up to a larger column ID. Axia packed columns provide the three important benefits you need.
Reproducible performance across all column •diameters
Increased throughput without sacrificing purity•
High efficiency from analytical to preparative•
Option 1: Increase Column LengthIncrease sample load without increasing your flow rate by using a longer column. With Axia technology, each preparative column is optimized for:
Analytical-like efficiency•
Long column lifetime•
High sample load with high-surface area media such •as Gemini®-NX, Luna® or Synergi™
As a result, load generally increases as a direct proportion to column length. In this example the sample load tripled by increasing column length.
Column: Luna 5 μm C18(2)Gradient: 5 min, 5 % to 95 % Water/Acetonitrile, 0.5 % TFA
Flow Rate: 1.5 mL/min on 4.6 mm30 mL/min on 21.2 mm60 mL/min on 30 mm 150 mL/min on 50 mm
Detection: UV @ 254 nmSample: 1. Propranolol
2. Diphenhydramine
Axia™ ScalabilityTwo Options to Increase Sample Load
Gradient: 10 min on 100 mm length column 25 min on 250 mm length column 5 % to 95 % Acetonitrile, 0.5 % TFA
Flow Rate: 30 mL/min
50 x 4.6 mm64 µg SF=1Plates/Meter 87120Symmetry 1.16
50 x 21.2 mm 1.2 mg SF=20Plates/Meter 84138Symmetry 1.17
50 x 50 mm 7.3 mg SF=118Plates/Meter 88280Symmetry 1.01
50 x 30 mm 2.4 mg SF=40Plates/Meter 83235Symmetry 0.97
96 mg on 250 x 21.2 mmAxia Packed Column
32 mg on 100 x 21.2 mmAxia Packed Column
SF = Scaling Factor
Trad
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D-44
42-P
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—00
G-44
42-P
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—00
D-44
42-U
0-AX
—00
G-44
42-U
0-AX
——
——
AJ0-
8210
AJ0-
8310
Clar
ity 1
0 µm
Olig
o-RP
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00F-
4445
-P0-
AX00
G-44
45-P
0-AX
——
00F-
4445
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——
00F-
4445
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G-44
45-V
0-AX
AJ0-
8210
AJ0-
8310
Gem
ini 5
µm
C18
00
B-44
35-P
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00D-
4435
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AX00
F-44
35-P
0-AX
00G-
4435
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B-44
35-U
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00D-
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F-44
35-U
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00G-
4435
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B-44
35-V
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——
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0-78
46AJ
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08
Gem
ini 1
0 µm
C18
00
B-44
36-P
0-AX
00D-
4436
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F-44
36-P
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00G-
4436
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B-44
36-U
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F-44
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00G-
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B-44
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F-44
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00G-
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08
Gem
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C6-
Phen
yl 00
B-44
44-P
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F-44
44-P
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00G-
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B-44
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F-44
44-U
0-AX
00G-
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——
—AJ
0-78
41AJ
0-83
03
Gem
ini-N
X 5
µm C
18
00B-
4454
-P0-
AX00
D-44
54-P
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00F-
4454
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AX00
G-44
54-P
0-AX
00B-
4454
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D-44
54-U
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00F-
4454
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G-44
54-U
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4454
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AX—
00F-
4454
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AX—
AJ0-
8370
AJ0-
8371
Gem
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µm
C18
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B-44
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00D-
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F-44
55-P
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00G-
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F-44
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B-44
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F-44
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00G-
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AXAJ
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71
Jupit
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C18
—00
D-40
55-P
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——
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D-40
55-U
0-AX
—00
G-40
55-U
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——
——
AJ0-
7230
AJ0-
8313
Jupit
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0 µm
C4
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D-41
68-P
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00F-
4168
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AX00
G-41
68-P
0-AX
——
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G-41
68-U
0-AX
——
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AJ0-
7231
AJ0-
8314
Jupit
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µm
Pro
teo
——
—00
G-43
96-P
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——
00F-
4396
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AX00
G-43
96-U
0-AX
——
——
AJ0-
7842
AJ0-
8304
Jupit
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Pro
teo
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D-43
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00F-
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97-P
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97-U
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G-43
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G-43
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7842
AJ0-
8304
Luna
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B-42
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00D-
4252
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F-42
52-P
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00G-
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B-42
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00D-
4252
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F-42
52-U
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4252
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B-42
52-V
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——
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0-78
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Luna
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B-42
53-P
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00D-
4253
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F-42
53-P
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00G-
4253
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AX00
B-42
53-U
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00D-
4253
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F-42
53-U
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00G-
4253
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B-42
53-V
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4253
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4253
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Luna
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4249
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F-42
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4249
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AX00
B-42
49-U
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4249
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——
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Luna
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00
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Luna
5 µ
m C
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AX—
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4255
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4255
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11
Luna
10
µm C
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11
Luna
5 µ
m N
H 2—
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4378
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AX00
F-43
78-P
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4378
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4378
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78-U
0-AX
00G-
4378
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AX—
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0-81
62AJ
0-83
09
Luna
10
µm N
H 2—
——
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4379
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AX—
—00
F-43
79-U
0-AX
00G-
4379
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AX—
——
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4379
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AXAJ
0-81
62AJ
0-83
09
Luna
5 µ
m P
FP(2
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B-44
48-P
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00D-
4448
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AX00
F-44
48-P
0-AX
00G-
4448
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AX00
B-44
48-U
0-AX
00D-
4448
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AX00
F-44
48-U
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00G-
4448
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——
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0-83
77AJ
0-83
78
Luna
5 µ
m P
heny
l-Hex
yl 00
B-42
57-P
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00D-
4257
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AX00
F-42
57-P
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00G-
4257
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AX00
B-42
57-U
0-AX
00D-
4257
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AX00
F-42
57-U
0-AX
00G-
4257
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AX—
——
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0-78
41AJ
0-83
03
Luna
10
µm P
heny
l-Hex
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F-42
85-P
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00G-
4285
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00D-
4285
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F-42
85-U
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00G-
4285
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B-42
85-V
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00D-
4285
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AX—
00G-
4285
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AXAJ
0-78
41AJ
0-83
03
Luna
5 µ
m S
CX—
00D-
4398
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AX—
00G-
4398
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AX—
——
——
——
——
—
Luna
5 µ
m S
ilica(
2)
00B-
4274
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AX00
D-42
74-P
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00F-
4274
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AX00
G-42
74-P
0-AX
00B-
4274
-U0-
AX00
D-42
74-U
0-AX
00F-
4274
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AX00
G-42
74-U
0-AX
——
——
AJ0-
7229
AJ0-
8312
Luna
10
µm S
ilica(
2)
—00
D-40
91-P
0-AX
—00
G-40
91-P
0-AX
——
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4091
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AX00
G-40
91-U
0-AX
——
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G-40
91-V
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AJ0-
7229
AJ0-
8312
Lux 5
µm
Cell
ulose
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00D-
4459
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AX00
F-44
59-P
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4459
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AX—
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4459
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AX00
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59-U
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4459
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4459
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0-84
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Lux 5
µm
Cell
ulose
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00D-
4457
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AX00
F-44
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4457
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4457
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AX00
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57-U
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4457
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AX—
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00G-
4457
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0-84
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µm
Max
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00B-
4337
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AX00
D-43
37-P
0-AX
00F-
4337
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AX00
G-43
37-P
0-AX
00B-
4337
-U0-
AX00
D-43
37-U
0-AX
00F-
4337
-U0-
AX00
G-43
37-U
0-AX
——
——
AJ0-
7842
AJ0-
8304
Syne
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Max
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—00
D-43
50-P
0-AX
00F-
4350
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AX00
G-43
50-P
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00B-
4350
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D-43
50-U
0-AX
00F-
4350
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AX00
G-43
50-U
0-AX
——
00F-
4350
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AX00
G-43
50-V
0-AX
AJ0-
7842
AJ0-
8304
Syne
rgi 4
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B-43
75-P
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4375
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F-43
75-P
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00G-
4375
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B-43
75-U
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F-43
75-U
0-AX
00G-
4375
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AX—
——
—AJ
0-78
43AJ
0-83
05
Syne
rgi 1
0 µm
Hyd
ro-R
P —
—00
F-43
76-P
0-AX
00G-
4376
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AX00
B-43
76-U
0-AX
——
00G-
4376
-U0-
AX—
——
00G-
4376
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AXAJ
0-78
43AJ
0-83
05
Syne
rgi 4
µm
Pola
r-RP
00B-
4336
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AX00
D-43
36-P
0-AX
00F-
4336
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AX00
G-43
36-P
0-AX
00B-
4336
-U0-
AX00
D-43
36-U
0-AX
00F-
4336
-U0-
AX00
G-43
36-U
0-AX
——
——
AJ0-
7845
AJ0-
8407
Syne
rgi 1
0 µm
Pola
r-RP
——
—00
G-43
51-P
0-AX
00B-
4351
-U0-
AX00
D-43
51-U
0-AX
—00
G-43
51-U
0-AX
——
—00
G-43
51-V
0-AX
AJ0-
7845
AJ0-
8407
Syne
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µm
Fus
ion-R
P 00
B-44
24-P
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00D-
4424
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Aw
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Rec
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t: f:
Australia02-9428-6444 02-9428-6445 [email protected]
t: f:
Austria 01-319-1301 01-319-1300 [email protected]
t: f:
Belgium+31 (0)30-2418700 +31 (0)30-2383749 [email protected]
t: f:
Canada(800) 543-3681 (310) 328-7768 [email protected]
t: f:
Denmark4824 8048 4810 6265 [email protected]
t: f:
France01 30 09 21 10 01 30 09 21 11 [email protected]
t: f:
Germany 06021-58830-0 06021-58830-11 [email protected]
t: f:
Ireland 01 247 5405 +44 1625-501796 [email protected]
t: f:
Italy 051 6327511 051 6327555 [email protected]
t: f:
Luxembourg+31 (0)30-2418700 +31 (0)30-2383749 [email protected]
t: f:
Netherlands030-2418700 030-2383749 [email protected]
t: f:
New Zealand09-4780951 09-4780952 [email protected]
t: f:
Puerto Rico (800) 541-HPLC (310) 328-7768 [email protected]
t: f:
United Kingdom 01625-501367 01625-501796 [email protected]
t: f:
All other countries: Corporate Office USA (310) 212-0555 (310) 328-7768 [email protected]
www.phenomenex.comPhenomenex products are available worldwide. For the distributor in your country, contact Phenomenex USA, International Department at [email protected]
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