Chrom

Report
New Generic Approach for HPLC analysis:
The Modern Silica-Based HPLC Columns; SiliaChrom® Series
Charles Lévesque, M.Sc.
Product Manager-Analytical Chemistry
Vincent Bédard, B.Sc.
HPLC Specialist
François Béland, Ph.D.
Vice-President of R&D
Outlines
HPLC Listing of all C18
SiliaChrom Strategy
SiliaChrom—Chemistry Review
 New Techologies-SB,AQ,XT
Evaluation of SiliaChrom
Recent SiliaChrom Applications
Generic Approach for HPLC applications
SiliaChrom Focus
HPLC C18 column listing (L1-USP)
ACE 300 C18
ACE AQ
ACE C18
ACE C18-AR
AccQTag Ultra
ACQUITY UPLC BEH130
ACQUITY UPLC BEH 300
ACQUITY UPLC BEH C18
ACQUITY UPLC BEH Shield RP18
ACQUITY UPLC HSS C18
ACQUITY UPLC HSS C18 SB
ACQUITY UPLC HSS T3
ACQUITY UPLC OST C18
Adsorbosil C18
Adsorbosphere C18
Adsorbosphere HS C18
Adsorbosphere UHS C18
Adsorbosphere XL 300 C18
Adsorbosphere XL C18
Adsorbosphere XL C18-B
Allsphere ODS-1
Allsphere ODS-2
Allure C18
Allure Aqueous C18
Alltima C18
Alltima C18 LL
Alltima HP C18
Alltima HP C18 EPS
Alltima HP C18 Hi-Load
Alltima HP C18 AQ
Alltima HP C18 Amide
Alltima WP C18
AlphaBond C18
Amethyst C18
AminoQuant
APEX ODS
Apollo C18
AQUA C18
Aquapore OD-300 C18
AQUASIL C18
Ascentis C18
Ascentis Express C18
ASI C18
ASI microC18
Atlantis dC18
Atlantis T3
Axxi-Chrom C18, ODS
Bakerbond C18
BetaBasic 18
BetaMax Neutral
BETASIL C18
Bio-C18
BioBasic 18
Bio-Sil ODS
Bio-Sil ODS-10
Bondclone C18
BR-C18
Burdick & Jackson C18
Cadenza CD-C18
Cadenza 5CD-C18
Cadenza CL-C18
Cadenza 5CL-C18
Cadenza CW-C18
Cadenza 5CW-C18
Cadenza HS-C18
Capcell C18 AG 120
Capcell C18 SG 120, 300
Capcell C18 UG 80, 120, 300
Capcell C18 MG
Capcell C18 MGII
Capcell C18 MGIII
Capcell C18 ACR
Capcell C18 AQ
Capcell C18 IF
Chemcosorb 5-ODS-H
Chromegabond C18
Chromegabond WR-C18
Chromolith RP-18e
Clarity Oligo-RP
Clipeus C18
Cogent HPS C18
Cogent hQ C18
Cogent Simulare C18
Cogent MicroBee C18
Cogent Bidentate C18
Cogent e-Column C18
Cosmosil C18 MS-II
Cosmosil C18 AR-II
Cosmosil C18 AR-300
Cosmosil C18 PAQ
Delta-Pak C18
Denali
Develosil ODS
Diazem 2060 C18 60A
Diazem 300A C18 4-222
Diazem 4-206 C18
Diazem 4-230 C18
Diazem 4-246 C18
Diazem 4-247 C18EC
Diazem C18
Diazem C18 1000
Diazem C18 1120
Diazem C18 120A
Diazem C18 120A series
43,44,45 and 46
Diazem C18 series 200A,
300A
Diazem C18 series 2000
Diazem C18 series 3000
Diazem C18 series 4000
Diazem C18 60A
HPLC C18 column listing (L1-USP)
HyPURITY AQUASTAR
HyPURITY C18
IBM Octadecyl
IB-SIL C18
IB-SIL C18 BD
Inertsil ODS
Inertsil ODS series 1,2,3
ISCO C-18
J’Sphere ODS-H80
J’Sphere ODS-L80
J’Sphere ODS-M80
Jupiter C18
Kinetex C18
Kromasil C18
LiChrospher PAH
LiChrosorb RP-18
LiChrospher 100 RP-18
LiChrospher RP18e
Luna C18
Luna C18(2)
Diazem C18 60A series 1,2,3
Diazem C18 series EC
Diazem C18 Dual-Zone
Discovery C18
Discovery HS C18
Discovery Bo Wide Pore C18
Econosil C18
Econosphere C18
Everest C18
Fortis C18
Fortis H2o
Gammabond C18
Gemini C18
Gemini-NX
Genesis C18
GP-C18
GraceSmart C18
Haisil 100, 300 C18
Haisil HL C18
HALO C18
Hi Chrom Reversible ODS
Hibar II C18
Hibar ODS
HP-C18
HxSil C18
HydroBond PS-C18
High Strength Silica (HSS) HPLC
C18
High Strength Silica (HSS) HPLC
C18 SB
High Strength Silica (HSS) HPLC T3
Hyperprep HS C18
Hypersil 100 C18
Hypersil BDS C18
Hypersil GOLD
Hypersil GOLD aQ
Hypersil-ODS
Hypersil ODS-2
Macrosphere 300 C18
MetaSil AQ C18
MetaSil ODS
MetroSep C18H
MetroSep C18AQ
MetroSep C18 AQ+
MetroSep C18SH
MetroSep C18EPS
MPLC RP-18
µBondapak C18
µBondapak C18 Radial-Pak
Microsorb C18
MicroSpher C18
MonoChrom C18
nanoAcquity BEH130
nanoAcquity BEH300
Nova-Pak C18
NUCLEODUR C18 Gravity
NUCLEODUR C18 Isis
NUCLEODUR C18 Pyramid
NUCLEODUR 100 C18 ec
NUCLEODUR Sphinx RP
Nucleosil 100 C18
Nucleosil C18
Nucleosil C18 AB
Nucleosil C18 HD
Nucleosil C18 Nautilus
Nucleosil C18 MPN
Nucleosil C18 PAH
Nucleosil C18 PPN
ODS Ultrapak
OmniSpher C18
Onyx C18
Partisil ODS
Partisil ODS2
Partisil ODS3
Partisphere C18
Peco HCODS
Pecosil C18
Pecosphere 3x8C
Pecosphere 3x8CR C18
Pecosphere 5x15CRT C18
Pecosphere-3C C18
Pecosphere-3CR C18
Pecosphere-3x3 C18
Peptica C18 serresA,B,C
Pfarma C18 series A,B,C
Pinnacle II C18
Pinnacle DB C18
Pinnacle DB Aqueous C18
Platinum C18
Platinum EPS C18
Platinum 300 C18
Polaris C18-A
Polaris C18-Ether
Polygosil 60-7 C18
Polygosil C18
HPLC C18 column listing (L1-USP)
Shodex ODSpak F-411
Shodex Silica C18M
Shodex Silica C18P
Spectraphysics C18
Spheri-(5 and 10) RP-18
Spheri-(5 and 10) ODS C18
Spherisorb ODS1
Spherisorb ODS2
Spherisorb ODS-B
STR ODS-II
STR ODS-M
Sunfire C18
Supelcosil LC-18
Supelcosil LC-18-DB
Supelcosil LC-318
Superiorex
Superspher 100 RP-18
Symmetry C18
SymmetryPrep C18
Symmetry300 C18
SymmetryShield RP18
SynChropak RPP
Polygosil C18 60ª
Premier C18
Prep Nova-Pak HR C18
Prevail C18
Prevail OA
Prevail Select C18
PrimeSphere C18-HC
Prodigy ODS(3)
ProntoSIL C18 H
ProntoSIL C18 SH
ProntoSIL C18 AQ
ProntoSIL C18 AQ Plus
ProntoSIL C18 ace-EPS
ProSphere 300 C18
ProSphere 300 PAH
ProSphere HP C18
ProSphere HP C18 AQ
ProSphere HP C18 ZAP!
ProteCol-G C18 HQ203
ProteCol-P C18 HQ105
ProteCol-P C18 HPH125
ProteCol-P C18 GP125
Purospher RP-18
Purospher RP18e
Purospher STAR RP18e
Pursuit C18
Pursuit XRs C18
Radial-Pak C18
Reduced Activity C18
Regis ODS
Resolve C18
Resolvex C18
Sarasep P C18
Scherzo SM-C18
SelectaPore 90M, 300P, 300M
SynChropak RPP-1000
SynChropak RPP-4000
SynChropak RPP-100
SynChropak RPS-100
Synergi Fusion-RP
Synergi Hydro-RP
Targa C18
TSK-GEL ODS-100V
TSK-GEL ODS-100Z
TSK-GEL ODS-100S
TSK-GEL Super-ODS
TSK-GEL ODS-80Tm
TSK-GEL ODS-80Ts
TSK-GEL ODS-120A
TSK-GEL ODS-120T
TSK-GEL ODS-140HTP
TSK-GEL OligoDNA-RP
Ultra C18
Ultra Aqueous C18
Ultra II C18
Ultra II Aqueous C18
Ultracarb ODS
Ultrasphere ODS
Ultrasphere C-18
Ultrasphere C-18-IP
Ultron VX-ODS
Unison UK-C18
Unison US-C18
Velosep RP-18
VisionHT C18
VisionHT C18-B
VisionHT C18HL
VisionHT C18-P
Viva C18
Vydac 201TP
Vydac 202TP
Vydac 218MS
Vydac 218TP
Vydac 238MS
Vydac C18
Wakosil C18 RS 3 and 5
Wakosil 5C18 HG and AR
Wakopak Wakosil-II 3C18 HG
Wakopak Wakosil-II 3C18 AR
Wakopak Wakosil-II 3C18 RS
Wakopak Wakosil-II 5C18
Wakopak Wakosil-II 5C18-100
Wakopak Wakosil-II 5C18 HG
Wakopak Wakosil-II 5C18 AR
Wakopak Wakosil-II 5C18 RS
Wakopak Wakosil-II 5C18 HG Prep
Wakopak Wakosil-II 5C18 AR Prep
Wakopak Wakosil-II 5C18 RS Prep
Wakopak Handy ODS
Wakopak Wakosil 5C18
Wakopak Wakosil 5C18T
Wakopak Wakosil 5C18N
HPLC C18 column listing (L1-USP)
YMC-Pack Hydrosphere C18
YMC-Pack ODS-A
YMC-Pack ODS-AL
YMC-Pack ODS-AM
YMC-Pack ODS-AQ
YMC-Pack Pro C18
YMC-Pack Pro C18 RS
Zodiac 120 C18H
Zodiac 120 C18AQ
Zodiac 120 C18 Eurobond
Zodiac 120 AC EPS
Zodiac 120 C18SH
Zodiac ODS
Zodiac ODS H
ZORBAX ODS C18
ZORBAX ODS Classic
ZORBAX Eclipse Plus C18
ZORBAX Eclipse PAH
ZORBAX Eclipse XDB-C18
ZORBAX Extend-C18
ZORBAX Rx C18
ZORBAX StableBond C18
ZORBAX 300 StableBond C18
Wakopak Wakosil 10C18
Wakopak Wakosil 5C18 AR
Wakopak Wakosil 5C18-200
Wakopak Wakosil 5C18-200T
Wakopak Wakosil 5C18-200N
Wakopak Wakosil 10C18-200
Wakopak MS-5C18 GT
Wakopak Navi C18-5
Wakopak Combi ODS
Wakopak Combi ODS Fast
XBridge BEH130
XBridge BEH300
XBridge C18
XBridge OST C18
XBridge Shield RP18
XTerra MS C18
XTerra RP18
378 HPLC column as C18 in 2010
 Which is the best one!
 Do I need to buy all!
 What is my C18 strategy?
...etc...
$$$
A new modern strategy is absolutely necessary
in 2010 for HPLC end-users
New Generic Approach to Find Appropriate HPLC column
(Total Solution for HPLC end-users)
Better matrix (silica-gel)
-Stationary Phase Specification (manufacturers)
Phase according to Polarity (acidic and basic analytes)
Phase according to Hydrophobicity (% carbon loading)
Phase according to Low Metal Content and Silianol Activity
Phase compatible with new LC-MS technology
Phase for very aqueous mobile phase (100% buffer or water)
Etc...
Note: Every Pittcon and Analytica Events present new C18,
more and more HPLC C18/year
New Modern HPLC Strategy
 How to improve today silica-based HPLC columns?
 Better bonding chemistry/ improved control for better reproducibility.
 What about the matrix? (support—silica)










High Quality Level of the Support (Silica)
narrowest particule size distribution
perfectly spherical
high level of purity
High surface area (m2/g)
Efficiency pore size (Å)
No fines
SiliaSphere (SiliCycle)
Low metal content
High mechanical stability
Reproducible loading (critical in reversed-phase)
X
X
Supplier A (top)
Supplier B (bottom)
(both are real HPLC supports)
Generic Approach/Most popular applications
Now better support (Silica)
Similar chemistries than other HPLC suppliers
 Improved quality and efficiency
HPLC column for low pH (not hybrid, pure silica based)
HPLC column for high pH (pseudo-hybrid)
HPLC column for 100% aqueous mobile phase (or buffer)
HPLC column for LC-MS (no bleeding)
HPLC column for QC/QA
validation and method development (easy method)
No tailling, better sensitivity and peak shape for quaternary ammonium compounds)
HPLC column for HILIC technology (extremely polar compounds)
Portofolio of SiliaChrom
Columns
A column for every need: ONLY 7 Chemistries!
SiliaChrom SB C18 and SB C8
SiliaChrom XDB1 C18 ,C8, C4, CN, AMINO, PHENYL, DIOL, Silica…
SiliaChrom XDB and XDB2 (C8&C18)
SiliaChrom AQ C18 (dtC18)
SiliaChrom HILIC
SiliaChrom XT C18 (XT Fidelity C18)
NO MORE CHEMISTRIES
New Technology
SiliaChrom-Chemistries Review
Silica-Based HPLC columns for
Extremely Acidic Compounds
H 3C
H 3C
R=C8 or C18, pH<2.0
O
O
O
C
H 3C
Si
O
OH
H 3C
CH3
R
C
Si
Si
O
C CH3
CH3
CH3
CH3
Steric Effect
CH3
C
H3O +
CH3
CH3
Si
O
O
H 3C
Si
C
R
C CH3 CH3
CH3
Note: Not Endcapped silica gel
H 3O +
New Technology
SiliaChrom-Chemistries Review
Silica-Based HPLC columns for
all type of compounds (pH=2-9)
TMS endcapped
Free silianol groups (OH)
R=C18
Polar Surface
Ratio of TMS-OH-C18 is well-balanced
Note: Partially End capped silica gel
SiliaChrom-Chemistries Review
Silica-Based HPLC column for
EXTREMELY POLAR compounds
O
HN
SiliaChrom HILIC
H3C
Si
Si O
Normal-Reversed
Phases Conditions:
CH 2
CH 2
CH 3
O
Si OH
Only SiliaChrom HILIC
Note: Not Endcapped silica gel
CH 2
NH 2
SiliaChrom-Chemistries Review
H3C
Silica-based HPLC columns for low
to high hydrophobic compounds
pH=1.5-10.0
SiliaChrom XDB C8 (8%) and C18 (15%)
SiliaChrom XDB1 C8 (14%) and C18 (22%)
SiliaChrom XDB2 C18 (18%)
1) Different carbon loadings (8-22%C)
2) XDB2=Luna C18(2)
3) XDB1= Phenyl, NH2, CN, 300A, etc…
Si
Si O
(CH 2 ) 7 CH 3
CH 3
O
Si O
Si(CH 3 ) 3
H3C
Si
Si O
(CH 2 ) 17 CH 3
CH 3
O
Si O
Si(CH 3 ) 3
Note: Double Endcapped silica gel
New Technology
SiliaChrom-Chemistries Review
Silica-Based HPLC columns for
EXTREMELY Basic compounds
pH=1.5-12
Replace OH by CH3
SiliaChrom XT C18 Series (Pseudo-Hybrid tech.)
H 3C
Si
O
CH3
O
Si
O
Si
O
CH3
CH3
O
O
Si
O
Si
H 3C
O
Si
CH3
O
Si
O
No bridge
O
CH3
Si
R
H 3C
O
CH3
Si
CH3
CH3
Si
(No Backpressure issue as XBridge (Waters))
SiliaChrom-Chemistries Review
SiliaChrom Series
HPLC Classification-USP/NIST Approaches
All SiliaChrom HPLC columns are evaluated by USP and NIST approaches:
 Uracil: void volume marker
 Toluene: hydrophobic retention
 Ethylbenzene: methylene selectivity and plate count determination
 Amitriptyline: activity towards bases (silianol activity evaluation) (A)
 Quinizarin: activity towards chelating reagents (metal content evaluation) (B)
(Pharmacopeial
Forum, Vol. 31(2) March-Apr. 2005, p.637)
(NIST, SRM 870, Certificat Issue Date:December 19th 2002, Valid until September 30th 2010)
(A)
(B)
SiliaChrom Evaluation
HPLC System: Thermo Surveyor with PDA
(ChromQuest-Data handling version 5.0)
Mobile Phase: MeOH/buffer (80/20, v/v)
Buffer: 20mM of phosphate buffer adjusted at pH=7.0
Flow rate: 1.0 mL/min
Detector: UV scan (PDA), Total scan 200-600nm
Temperature: 23C
All HPLC columns: 4.6x150mm, 5µm
Phenomenex Luna C18(2)
150000
140000
130000
120000
110000
1.55
100000
Low Plate Count (see Results Table)
90000
uAU
7.34
80000
9.73
70000
60000
3.88
50000
4.86
40000
30000
20000
10000
0
-10000
0
1
2
3
4
5
6
7
8
9
10
11
12
13
Time (min)
Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline
14
15
Agilent Zorbax SB C18
180000
170000
Important Tailling
160000
150000
140000
1.38
130000
120000
110000
uAU
100000
90000
6.85
80000
70000
3.29
60000
4.16
50000
12.99
40000
30000
20000
10000
4.73
0
-10000
0
1
2
3
4
5
6
7
8
9
10
11
12
Time (min)
Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline
13
14
15
Waters Sunfire C18
Asymetry
150000
Metal Contamination
140000
130000
120000
1.49
110000
100000
uAU
90000
80000
70000
10.18
60000
4.05
50000
40000
8.71
5.14
30000
20000
10000
0
-10000
0
1
2
3
4
5
6
7
8
9
10
11
12
13
Time (min)
Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline
14
15
SiliaChrom XDB2 C18
190000
180000
170000
160000
150000
Very good peak shapes
1.61
uAU
140000
130000
120000
110000
100000
90000
80000
70000
60000
50000
40000
8.75
4.15
12.02
5.19
30000
20000
10000
0
-10000
0
1
2
3
4
5
6
7
8
9
10
11
12
13
Time (min)
Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline
14
15
Summary Results-Case Study
K’

N
Ethylbenzene
Ethyl/Tol
Ethylbenzene
SiliaChrom XDB2 C18
2.22
1.41
Luna C18(2)
2.13
Zorbax SB C18
R
AS
(Quinizarine)
Tf
(Amitriptyline)
28000
3.30
1.10
1.18
1.42
22000
2.90
1.23
1.20
2.01
1.45
25000
3.20
1.10
1.55
Sunfire C18
2.45
1.43
35500
3.90
1.80
1.12
SiliaChrom XDB1 C18
3.09
1.44
45000
4.73
1.09
1.65
HPLC Column
Basic analyte: SiliaChrom XDB2
Less polar analyte: SiliaChrom XDB1
Recent SiliaChrom Applications
Acidic and Basic Compounds (8) with one SiliaChrom HPLC column (AQ C18)
Fluoroquinolones for Veterinary use by HPLC-Marbofloxacin and Sarafloxacin
Halogenated Herbicides by HPLC-Dicamba, Atrazine and 2,4-D
Catecholamines by HPLC- Polar Compounds
Faster HPLC with SiliaChrom-Polar and mid-polar Compounds
Isomer separation for Chlorophenol series by simple HPLC method
HPLC study for Naloxone-pH=2.5, 7.0 and 10.5
HPLC Chromatogram of Ganciclovir– Polar Compound (diol-amine)
HPLC Chromtogram of Cefotaxime—Extremely Polar Compound (acid-amine-sulfur)
And more….
Recent SiliaChrom Applications
Acidic and Basic Compounds (8) with same SiliaChrom HPLC column
1-Maleic Acid
4-Chlorpheniramine
7-Dichlorphen
2-Acetaminophen
5-Benzoic Acid
3-Pseudoephedrine
8-Ibuprofen
6-Salicylic Acid
Recent SiliaChrom Applications
Acidic and Basic Compounds (8) with same SiliaChrom HPLC column
Phenomenex Luna C18(2) 5µm 4.6x150mm
210 nm
1
2
2
5
2
230
230nm
nm
4
1
1
3
6
7
8
Recent SiliaChrom Applications
Acidic and Basic Compounds (8) with same SiliaChrom HPLC column (same HPLC conditions than Luna C18(2))
SiliaChrom AQ C18 5µm 4.6x150mm
210 nm
2
5
230 nm
4
1
3
6
7
8
Recent SiliaChrom Applications
Fluoroquinolones for Veterinary use by HPLC-Marbofloxacin and Sarafloxacin
SiliaChrom AQ C18 5µm 4.6x150mm
Zwitterion
Molecules
Waters Atlantis dC18 5µm 4.6x150mm
Asymmetry
SiliaChrom AQ C18
1.12
1.09
Atlantis dC18
1.29
1.14
Note: SiliCycle has the perfect equivalent (better) with SiliaChrom AQ C18 (or dtC18)
than Atlantis dC18 (Waters)
Recent SiliaChrom Applications
Halogenated Herbicides by HPLC-Dicamba, Atrazine and 2,4-D
SiliaChrom dtC18 5µm 4.6x150mm
Recent SiliaChrom Applications
Halogenated Herbicides by HPLC-Dicamba, Atrazine and 2,4-D
Supelco Ascentis C18 5µm 4.6x150mm
Asymmetry
SiliaChrom dtC18
1.01
1.00
1.02
Ascentis C18
1.53
0.97
1.77
Recent SiliaChrom Applications
Catecholamines by HPLC-Polar Compounds
SiliaChrom AQ C18 5µm 4.6x150mm
Mobile Phase: Water 1.0% AcOH only!
Epinephrine Norepinephrine
Dopamine
Tyrosine
Levodopa
Recent SiliaChrom Applications
Faster HPLC with SiliaChrom-Polar and mid-polar Compounds
SiliaChrom AQ C18 5µm 3.0x50mm
95/5 0.1%TFA in H2O/ACN – 5/95 0.1% TFA in H2O/ACN in 2.25 min. 1.75
mL/min, 23°C, 254 nm (LC-MS conditions from AstraZeneca Canada)
Test Mix to evaluate HPLC column from AstraZeneca
Caffeine Chlorthalidone
Albendazole
(Canada)
Warfarin Tolnaftate
Dioctyl Phatalate
Recent SiliaChrom Applications
SiliaChrom XDB1 PHENYL 5µm 4.6x150mm
60/40, H2O/MeOH, 1.00 mL/min, 23°C, 254nm : Easy
Method!
OH
OH
OH
Cl
Cl
Cl
Recent SiliaChrom Applications
HPLC study for Naloxone-pH=2.5, 7.0 and 10.5
SiliaChrom XT C18, 5µm, 4.6x150mm
80/20, Phosphate Buffer/MeOH, 1.00 mL/min, 23°C, 254nm
Fronting ?
Assymetry = 0.85
pH=2.5
50/50, Phosphate Buffer/MeOH, 1.00 mL/min, 23°C, 254nm
Assymetry = 0.89
pH=7.0
Recent SiliaChrom Applications
HPLC study for Naloxone-pH=2.5, 7.0 and 10.5
SiliaChrom XT C18, 5µm, 4.6x150mm
50/50, H2O/MeOH 0.1% NH4,OH, 1.00 mL/min, 23°C, 254nm
pH=10.5
Assymetry = 0.99
Remember, pH range for XT is 1.5-12—Good Chromatography Tool
Recent SiliaChrom Applications
HPLC Chromatogram Ganciclovir – Polar Compounds (diol-amine)
SiliaChrom AQ C18 (4.6x150mm, 5µm)
MeOH:Water, 5:95
UV 252nm
1mL/min
Recent SiliaChrom Applications
HPLC Chromatogram of Cefotaxime—Extremely Polar compound
(acid-amine-sulfur)
SiliaChrom AQ C18 (4.6x150mm, 5µm)
0.4 mM KH2PO4 + 8mM K2HPO4: MeOH, 89:11
UV 254nm
1mL/min
Recent SiliaChrom Applications
HPLC for mixture of Fenitrothion-Pyriproxyfen—Different Level of Hydrophobicity
SiliaChrom XDB1 C18 (4.6x150mm, 5µm)
ACN:Water, 70:30
UV 254nm
1mL/min
Recent SiliaChrom Applications
HPLC Determination for Ephedrine (from Ephedra Sample)
Zorbax SB C18
(4.6x150mm, 5µm)
SiliaChrom XDB2 C18
(4.6x150mm, 5µm)
Recent SiliaChrom Applications
HPLC chromatogram of Berberine-Metal Content Effect
Sunfire C18
(4.6x250mm 5µm)
SiliaChrom dtC18
(4.6x250mm, 5µm)
ACN: 1%TFA, 35:65
UV 346nm
1 mL/min
Durability Testing at pH-1.00
SiliaChrom SB C18
Time/h
0
24
48
72
96
120
SD
Mean
RSD (%)
Theoritical
Retention time Tailing Factor
Plates #
(Ethylbenzene) (Ethylbenzene)
(Ethylbenzene)
(min)
(USP)
(USP)
5.91
1.01
14014
5.89
1.02
14085
5.77
1.02
14023
5.83
1.02
14076
5.85
1.01
14087
5.84
1.02
14050
0.05
5.85
0.84
0.01
1.02
0.51
32
14056
0.23
0 hr
120 hrs
SiliaChrom SB C18 (4.6x150mm, 5µm)
59:39:2 ACN:Water:TFA (pH = 1.00)
UV 270 nm, 1 mL/min
(Uracil, Phenol, Toluene and Ethylbenzene)
•Poster presented at Pittcon 2010, Orlando FL, USA
•LC-GC Application Note—September 2009
Durability Testing at pH-11.5
SiliaChrom XT C18
Naphthalene
Time/h
Tailing Factor
Retention time
(Naphthalene)
(min)
0
1.10
4.857
260
1.14
4.732
300
1.14
4.832
400
1.14
4.873
SiliaChrom XT C18 (4.6x150mm, 5µm)
MeOH: Water/TEA, pH=11.5, 40:60
UV 215nm, 1 mL/min
2.25 2.75
1.91
2.23
1.84
1 hr
2.77
400 hrs
Uracil, Phenol, Nitrobenzene and Naphtalene
Durability Testing at pH=11.50, 90C
SiliaChrom XT C18 Fidelity
0 hr
SiliaChrom XT C18 (4.6x150mm, 5µm)
54.9-44.9-0.2 (ACN-Water-TEA)
90C, UV 270 nm
1 mL/min
120 hrs
(Uracil, Phenol, Toluene and Ethylbenzene)
SiliaChrom HILIC, Vitamins
SiliaChrom HILIC (4.6x150mm, 5µm)
ACN 0.1% TFA: Water 0.1%TFA, 10:90
UV 280nm
1 mL/min
Waters HILIC Silica HPLC column
(4.6x150mm, 5µm)
B6
C
B1
B2
Vitamin B1, Vitamin B6, Vitamin C and Vitamin B2
Generic Approach with SiliaChrom Series
Acidic, Basic
or Neutral
Compounds
Resolution
Polarity
Final Step
87% of
HPLC success
in my Lab!
Not enough
retention
Acidic
SiliaChrom SB
pH=1-2 or HILIC
Basic
SiliaChrom XT
SiliaChrom AQ
(MeOH/H2O) or
(ACN/H2O), pH=2-9
Neutral
Good Retention, but
not enough
resolution
SiliaChrom SB,
pH=9-12 Series
or HILIC
SiliaChrom XDB,
XDB1 or XDB2
XT or XT Fidelity
(low and/or high pH)
Note: SiliaChrom AQ is an universal HPLC column
(see how particle is designed : TMS-OH-C18)
Conclusion
Low pH High pH
(0.5-2.0) (9-12)
SiliaChrom
SB
XT
High level of Water
Mobile Phase
Hydrophobicity
(different Level)
Low Metal
Content
AQ
XDB, XDB1
XDB2
dt
Polar
Extemely
Analyte Polar Analyte
AQ
HILIC
ONLY 8 Chemistries to cover more than
85% of Regular HPLC Application
using Silica-Based as support
Be aware for your HPLC column strategy, Marketing is not real SCIENCE!
www.silicycle.com
Thank you!

similar documents