Turn Imagination into Results
ProteCol™
HPLC Columns
t'MFYJCMFIBSEXBSFPQUJPOT
t)JHIRVBMJUZQIBTFT
t1FSGFDUNBUDIGPSBMM
ZPVSTFQBSBUJPOOFFET
AUSTRALIA & PACIFIC REGION
EUROPE
JAPAN
USA
ProteCol™ Column Range Introduction
5VSOJNBHJOBUJPOJOUPSFTVMUTXJUIUIF1SPUF$PM™SBOHF
PG)1-$DPMVNOT
8JUI 3FWFSTF 1IBTF /PSNBM 1IBTF BOE 4QFDJBMUZ
1IBTFT1SPUF$PMFOTVSFTZPVIBWFUIFSJHIUQIBTFGPS
ZPVSTFQBSBUJPOOFFET8JUIUIFPQUJPOPGDPNCJOJOH
JOFSU1&&,DPBUFEIBSEXBSFPSUSBEJUJPOBMTUBJOMFTTTUFFM
XJUI FBDI RVBMJUZ QIBTF UIF 1SPUF$PM )1-$ TPMVUJPO
EFMJWFSTUIFDPNCJOBUJPOZPVSFRVJSF
ProteCol Reverse Phase
tProteCol C18PGGFSTBGMFYJCMFSBOHFPG$CPOEFEQIBTFTJODMVEJOHQ)TUBCJMJUZBOEQPSFTJ[FPQUJPOT
tProteCol C8DPMVNOTIBWFQPSFTJ[FTUPTVJUZPVSBOBMZTJT
tProteCol C4DPMVNOTIBWFIJHIEVSBCJMJUZBOEFYUFOEFEBDJEJDBOEBMLBMJOFSFTJTUBODF
tProteCol Phenyl Hexyl DPMVNOTPGGFSVOJRVFTFMFDUJWJUZ
ProteCol Normal Phase
tProteCol AminoDPMVNOTFOBCMFTFQBSBUJPOJOCPUIOPSNBMBOESFWFSTFEQIBTF
tProteCol Cyano and SilicaFOBCMFPQUJPOTGPSOPSNBMQIBTFDISPNBUPHSBQIZ
ProteCol Specialty Phase
t ProteCol HILICSBOHFQSPWJEFTBQPMBSTUBUJPOBSZQIBTFBOEIJHIMZPSHBOJDNPCJMFQIBTFBMMPXJOHZPVUPSFUBJO
BOETFQBSBUFQPMBSBOBMZUFT
tProteCol ChiralDPMVNOTFOTVSFUIFJTPMBUJPOBOEBOBMZTJTPGQVSFFOBOUJPNFST
tProteCol PFPDPMVNOTBSFVTFGVMJOUIFTFQBSBUJPOPGFQJNFST
tProteCol SCXDPMVNOTIBWFBIJHIMPBEJOHDBQBDJUZBOEQSFTTVSFMJNJU
ProteCol Ultra Phase
t3BOHFPGQIBTFTGPS6)1-$VTF
Hardware Options
1SPUF$PM)1-$DPMVNOTBSFBWBJMBCMFJOUIFGPMMPXJOHGPSNBUT
t*OFSUIBSEXBSF1&&,DPBUFETUBJOMFTTTUFFMPS1&&,TJM™$BQJMMBSZIBSEXBSF
t$BQJMMBSZ)1-$
t4UBJOMFTTTUFFM
t6)1-$
t4FNJQSFQBSBUJWFBOEQSFQBSBUJWF
Inert Hardware
t1&&,DPBUFETUBJOMFTTTUFFM
t$BQJMMBSZ)1-$
t0QUJNJ[FEBOBMZUFSFDPWFSZ
t4VQFSJPSQFBLTIBQFBOESFQSPEVDJCJMJUZ
t-FTTBSUJGBDUTEVFUPSFEVDFEDBSSZPWFS
The Importance of Inert HPLC Column Design
/POTQFDJGJD JOUFSBDUJPOT CFUXFFO UIF UBSHFU BOBMZUF BOE UIF TJMJDB QBSUJDMFT JO UIF )1-$ DPMVNO BSF OPX
XFMM DPOUSPMMFE XJUI UIF BWBJMBCJMJUZ PG VMUSBQVSF TJMJDBT 5PEBZ DISPNBUPHSBQIFST FYQFDU TJMJDB TPVSDFE CZ
NBOVGBDUVSFSTUPCFPGUIFIJHIFTUQVSJUZ8IBUJTPGUFOOPUDPOTJEFSFEJTUIFSPMFDPMVNOIBSEXBSFNBZQMBZ
JOOPOTQFDJGJDJOUFSBDUJPOToUIFGSJUBOEJOUFSOBMDPMVNOIBSEXBSFDBOCPUIJOGMVFODFUIFCFIBWJPSPGBOBMZUFT
XJUILOPXONFUBMDIFMBUJOHBDUJWJUZ
1SPUF$PMPGGFSTUXPJOFSUIBSEXBSFPQUJPOT1&&,DPBUFETUBJOMFTTTUFFMPS1&&,TJM™DBQJMMBSZIBSEXBSF
1&&,-JOFE
$PMVNO
4BNQMF
$PMVNOT
#VGGFS
'MPX
1IBTF
.FUBMDIFMBUJOHQSPCF
¬HPGIZESPYZQZSJEJOF/PYJEF
NNYNN*%BU«$
N.&%5"JO8BUFS
NMNJO
1SPUF$PM™$¬N
ProteCol™
1&&, DPBUFE
F
4UBJOMFTT4UFFM
$PMVNO
E
D
A
B
C
"
#
$
%
&
'
A Colum
PEEK
Porou
Statio
PEEK
Stain
$PMVNOFOEDBQ B
1&&, GSJUIPVTJOH C
1PSPVT1&&, GSJU D
4UBUJPOBSZQIBTF E
1&&, MJOJOH
F
4UBJOMFTTTUFFM
DPMVNOCPEZ
.PTUQIBSNBDFVUJDBMMZBDUJWFDPNQPVOETBOEOBUVSBMQSPEVDUTIBWFUIFQPUFOUJBMUPJOUFSBDUXJUINFUBMT'PS
UIJTSFBTPONPMFDVMFTMJLFRVJOJ[BSJOUFUSBDZDMJOFPSDJDMPQJSPYGPSNUBJMJOHQFBLTJOUIFQSFTFODFPGNFUBMJO
UIFDPMVNOPSTZTUFN
Capillary HPLC
ProteCol Capillary HPLC Is Perfect For:
t4NBMM TBNQMFT CJPUFDIOPMPHZ NFEJDBM SFTFBSDI
QSPUFPNJDT
t&YPUJDTPMWFOUTEFVUFSBUFETPMWFOUTGPS-$/.3
t-PXDPODFOUSBUJPOTIJHIMZQPUFOUQIBSNBDFVUJDBMT
NFEJDBMSFTFBSDI
t*OTUSVNFOUBUJPOEJSFDUDPVQMJOHJOUP.4
Figure demonstrating the relationship of analysis sensitivity with low volume
use of solvents.
5IF1SPUF$PMDBQJMMBSZ)1-$EFTJHOJODMVEFTJOUFHSBUFE
DPOOFDUJPOUVCJOHXJUI1&&,TJM™GPSUIFDPMVNOCPEZ
BOE DPOOFDUJPO DBQJMMBSJFT EFMJWFSJOH SPCVTU EFTJHO
[FSPWPMVNFDPOOFDUJPOTBOEVOJOUFSSVQUFEGMPX
Stainless Steel Hardware
1SPUF$PM)1-$XJUITUBJOMFTTTUFFMIBSEXBSFIBTCPUIFYUFSOBMCPEZBOEFOEGJUUJOHTNBOVGBDUVSFEGSPN
IJHIRVBMJUZHSBEFTUBJOMFTTTUFFM
UHPLC
4UBJOMFTTTUFFMIBSEXBSFJTEFTJHOFETQFDJGJDBMMZGPS6)1-$VTFBOETUBOEBSEXJUIUIF1SPUF$PM6MUSBQIBTFT
5IFTFDPMVNOTBSFGPSVTFBUQTJ
Chemical Properties of ProteCol Range
Stability at pH 1:
$PMVNOTJOUIF1SPUF$PMSBOHFTIPXOPEFUFSJPSBUJPO
XIFOFYQPTFEUPQ)CVGGFST
Overlay of 40 chromatograms run at pH 1.0 spanning 1200 column volumes.
Long-term Reproducibility
$PMVNOT JO UIF 1SPUF$PM SBOHF TIPX B SFNBSLBCMF
SFQSPEVDJCJMJUZPGUIPVTBOETPGJOKFDUJPOTTVCKFDUUP
TBNQMFQVSJUZBOENPCJMFQIBTFDPOEJUJPOT
2500th Injection
2000th Injection
1500th Injection
900th Injection
500th Injection
1st Injection
Chromatograms of a test mix over a period of 2500 injections.
Analysis of Larger Molecules
8IFOBOBMZ[JOHTBNQMFTDPOUBJOJOHMBSHFNPMFDVMFTQFQUJEFTQSPUFJOTQPMZNFSTXJUI.8
UIFTJ[FPGUIF
NPMFDVMFBOEUIFTJ[FPGUIFQPSFTUSVDUVSFQMBZBOJNQPSUBOUSPMFJOUIFRVBMJUZPGUIFTFQBSBUJPO"TUIFBOBMZUF
JODSFBTFTJOTJ[FSFMBUJWFUPUIFQPSFTJ[F
UIFEJGGVTJPOSBUFJOTJEFUIFQPSFCFDPNFTTNBMMFSBOENBTTUSBOTGFSJO
BOEPVUPGUIFQPSFTZTUFNCFDPNFTTMPXMFBEJOHUPCBOECSPBEFOJOH0CWJPVTMZXIFOUIFBOBMZUFTJ[FJTFRVBM
UPPSCJHHFSUIBOUIFQPSFTJ[FUIFSFDBOCFOPQPSFEJGGVTJPO"NBUIFNBUJDBMEFTDSJQUJPOPGUIJTSFMBUJPOTIJQXBT
QVCMJTIFECZ3FOLJO&.3FOLJO+(FO1IZTJP
BOEIFMQTUPJMMVTUSBUFUIFQIFOPNFOPO
Relative Diffusion Coefficients
5kDa peptide in 1000Å pores
100kDa protein in 1000Å pores
5kDa peptide in 300Å pores
5IF3FOLJO.PEFMQSPWJEFTBOFRVBUJPOPGUIFFGGFDUPGQPSFBOEBOBMZUF
TJ[FPOUIFEJGGVTJPODPFGGJDJFOU"%Q%NWBMVFPGNFBOTUIFEJGGVTJPO
JOTJEFUIFQPSFJTBTGBTUBTJOUIFCVMLMJRVJE%Q%NNFBOTUIFNPMFDVMF
EPFTOPUNPWFJOTJEFUIFQPSFT
5kDa peptide in 200Å pores
5kDa peptide in 120Å pores
100kDa protein in 300Å pores
5kDa peptide in 100Å pores
100kDa protein in 200Å pores
100kDa protein in 120Å pores
100kDa protein in 100Å pores
An illustration of the relative diffusion rate of a 5kDa peptide and a 100kDa protein in a number of pore systems.
Reverse Phase ProteCol Range
tProteCol C18PGGFSTBGMFYJCMFSBOHFPG$CPOEFE
QIBTFTJODMVEJOHQ)TUBCJMJUZBOEQPSFTJ[FPQUJPOT
tProteCol C8 DPMVNOT IBWF QPSF TJ[FT UP TVJU ZPVS
BOBMZTJT
tProteCol C4 DPMVNOT IBWF IJHI EVSBCJMJUZ BOE
FYUFOEFEBDJEJDBOEBMLBMJOFSFTJTUBODF
tProteCol Phenyl Hexyl DPMVNOTPGGFSVOJRVFTFMFDUJWJUZ
Reverse Phase Column Selection Tree
Molecule
Type
Sample
Type
Size or pH
requirements
ProteCol
Selection
Proteins
> 15,000 Da
ProteCol – C8
Q 1000
< 5,000 Da
ProteCol – C18
G 200
< 15,000 Da
ProteCol – C18
G 300
Chelating
Compounds
pH 1-9
ProteCol – C18
Q 100
General Purpose
pH 1-12
ProteCol – C18
H 120
Hydrophobic
pH 1-12
ProteCol – C8
H 120
Hydrophilic
pH 1-9
ProteCol – C18
P 120
General Purpose
pH 1-9
ProteCol – C18
G 120
Large Molecules
> 1 KDa
Peptides
Small Molecules
> 1 KDa
General
ProteCol C18
t4VCTUBOUJBMMZSFEVDFETBNQMFBOEDPMVNOQSFQBSBUJPOUJNF
t*NQSPWFEQFBLTIBQFHJWJOHZPVJNQSPWFESFQSPEVDJCJMJUZBOETFOTJUJWJUZ
t'FXFSBSUJGBDUTEVFUPSFEVDFEDBSSZPWFS
t&OBCMFTVTFPG.4&-4%BOE$PSPOB$%"UFDIOJRVFT
Four Chemistries
Phase
Pore Size (Å)
Particle Size (¬m)
Pore Volume (ml)
Surface Area
Carbon Load %
C18 Q
C18 G
C18 G
C18 G
C18 H
C18 P
£
£
£
£
£
£
£
£
£
£
£
£
C18 Q
t6MUSBQVSFTJMJDB
t'VMMZ FOE DBQQFE PQUJNJ[FE
$QIBTFT
C18 H
t.PEJGJFE CPOEFE QIBTF
NBLJOH JU FBTJFS UP XPSL
GSPNMPXUPIJHIQ)VTJOH
UIFTBNFDPMVNO
t/PWFM DIFNJDBM CPOEJOH
FOTVSJOH TUBCJMJUZ VOEFS
FYUSFNF BMLBMJOF BOE BDJEJD
DPOEJUJPOT
tQ)TUBCJMJUZ
C18 G
t4UBCMFJOBRVFPVTDPOEJUJPOT
t3FEVDFTOPOTQFDJGJDBOBMZUF
JOUFSBDUJPOT
t4FQBSBUFTQPMBSDPNQPVOET
tQ)TUBCJMJUZ
C18 P
t1PMBSFNCFEFE$
tDPNQBUJCMFXJUIXBUFS
tQ)TUBCJMJUZ
$ISPNBUPHSBN PG UFUSBDZDMJOF BOUJCJPUJD
BOE JUT NBKPS
EFHSBEBUJPOQSPEVDU/PUFUIFQFBLCSPBEFOJOHPOUIF
CBTFPGUIFQFBLSVOUISPVHIUIFTUBJOMFTTTUFFMDPMVNO
*OTFU UIF UFUSBDZDMJOF NPMFDVMF EFQJDUJOH UIF UISFF
QPUFOUJBMDIFMBUJOHHSPVQT
$PMVNO
4BNQMF
.PCJMF1IBTF"
.PCJMF1IBTF#
(SBEJFOU
'MPX3BUF
5FNQFSBUVSF
%FUFDUJPO
YNN*%$)
$POUFOUPG7JUBNJO4VQQMFNFOU $BQTVMFT#MBDLNPSFT"VTUSBMJB
8BUFS
"DFUPOJUSJMF
UPNJOVUFTUP#
UPNJOVUFT#
N-NJO
«$
ON
1SPUF$PM™)QIBTFCPOEJOH
$PMVNO
YNN$(¬N
.PCJMF1IBTF N.,)PO Q))PO N-NJO
5FNQFSBUVSF «$
%FUFDUJPO
ON
4BNQMFT
J
5FTU.JY0SHBOJD"DJET
JJ
$SBOCFSSZ+VJDF
ProteCol C8
t±QPSFTJ[F
t*OUFSNFEJBUFQPMBSJUZ$QIBTF
t$POUJOVJUZ PG VTJOH )1-$ GPS BMM TFQBSBUJPO OFFET TJNQMJGJFT ZPVS
XPSLGMPX
t'BDJMJUBUFTUIFVTFPG.4
t&MJNJOBUF4%41"(&GSPNZPVSXPSLGMPX
Phase
Pore Size (Å)
$
$
Particle Size (µm) Pore Volume (ml)
£
£
Surface Area
Carbon Load %
£
£
$PMVNO
1SPUF$PM™$2¬N*%YNN
'MPXSBUF
¬-NJO
.PCJMFQIBTF" GPSNJDBDJE
.PCJMFQIBTF# BDFUPOJUSJMFGPSNJDBDJE
(SBEJFOU
NJO#NJO#NJO#
NJO#NJO#
Base peak chromatogram of ribosomal proteins.
Why choose 1000 Å pore size?
±QPSFTJ[FTJMJDBTFOBCMFMBSHFJSSFHVMBSTIBQFEQSPUFJOTUPCJOE
UPUIFCPOEFEQIBTFXJUIPVUSFTUSJDUJOHBDDFTTUPUIFQPSFDPNQBSFE
UP±TJMJDBTXIPTFQPSFTBSFFBTJMZCMPDLFECZMBSHFQSPUFJOT
L%BTQIFSJDBM
NPMFDVMFTJOB±QPSF
n
L%B SPETIBQFE
NPMFDVMFT JO B ±
QPSFBTQFDUSBUJP
L%BNPMFDVMFTBETPSCFEPO
UIFXBMMPGB±QPSF
ProteCol C4
Standard Proteins
$PMVNO
1SPUF$PM™$
YNN
.PCJMF1IBTF "
$)$/5'"
#
)5'"
"#
'MPXSBUF
N-NJO
%FUFDUJPO
67ON
5FNQ
«$
t ¬NNBUFSJBM
t1SPUFJOQVSJGJDBUJPO
t)JHIEVSBCJMJUZ
t&YUFOEFEBDJEJDBOEBMLBMJDSFTJTUBODF
t3FDPNNFOEFEGPSDPNQPVOETUPPTUSPOHMZSFUBJOFEPO$
BOE$QIBTFT
Phase
Pore Size (Å)
$
Particle Size (¬m) Particle Volume
Surface Area
% Carbon
Load
pH
4BNQMF
3JCPOVDMFBTF
$ZUPDISPNF
-ZTP[ZNF
#4"
.ZPHMPCJO
0WBMCVNJO
ProteCol Phenyl Hexyl
1SPUF$PM 1IFOZM )FYZM VTFT B IFYZMMJOLFE QIFOZM QIBTF XIFSF UIF
IFYZM BMLZM DIBJO EFMJWFST VOJRVF TFMFDUJWJUZ BOE JODSFBTFE IZESPMZUJD
TUBCJMJUZ XIFO DPNQBSFE UP QSPQZMMJOLFE DIFNJTUSZ 5IF FYBNQMF
QJDUVSFE IJHIMJHIUT UIF TFQBSBUJPO PG B NJYUVSF PG #FO[PEJB[BQJOFT
XIJDIJTEJGGJDVMUUPTFQBSBUFPOBTUBOEBSEQIFOZMUZQFDPMVNO
Phase
Pore Size (Å)
Particle Size (µm)
Surface Area
% Carbon Load
pH
1IFOZM)FYZM
$PMVNO
1SPUF$PM™1IFOZM)FYZM
.PCJMF1IBTF"DFUPOJUSJMF8BUFS
'MPXSBUF
N-NJO
%FUFDUJPO 67ON
4BNQMF
-PSNFUB[FQBN
%JB[FQBN
0YB[FQBN
Normal Phase ProteCol™ Range
tProteCol AminoDPMVNOBMMPXTCBTJDDPNQPVOETFQBSBUJPOJO
OPSNBMQIBTFBOEDBSCPIZESBUFBOBMZTJT
tProteCol Cyano DPMVNOT QSPWJEF DISPNBUPHSBQIJD SFUFOUJPO
CPUI JO OPSNBM BOE SFWFSTFE QIBTF TFQBSBUJPO EVF UP JUT
NPEFSBUFQPMBSJUZ
tProteCol Silica DPMVNOT IBWF IJHI EVSBCJMJUZ BOE FYUFOEFE
BDJEJDBOEBMLBMJOFSFTJTUBODF
ProteCol Amino
Specifications
t#POEFEXJUIBNJOPQSPQZMTJMBOF
Applications
t#BTJDDPNQPVOETFQBSBUJPOVOEFSOPSNBMQIBTF
DPOEJUJPOT
t4BDDIBSJEFTFQBSBUJPOVTJOHBDFUPOJUSJMFXBUFS
Phase
Pore Size (Å)
"NJOP
Particle Size
(¬m)
Malto-oligosaccharides
$PMVNO1SPUF$PM™"NJOP
YNN
&MVFOU$)$/)
'MPXSBUFN-NJO
%FUFDUJPO3*%
5FNQ«$
4BNQMF(MVDPTF
.BMUPTF
.BMUPUSJPTF
.BMUPUFUSBPTF
.BMUPQFOUBPTF
Tocopherol isomers
$PMVNO1SPUF$PM™"NJOP
YNN
&MVFOU)FYBOF&UIZMBDFUBUF
'MPXSBUFN-NJO
%FUFDUJPO67ON
5FNQ«$
4BNQMFĮTocopherol
ȕTocopherol
Ȗ Tocopherol
įTocopherol
Surface Area % Carbon Load
ProteCol Cyano
$PNNPOBQQMJDBUJPOTBSFGPSUIFTFQBSBUJPOPGGMBWPOPJET
FYUSBDUJPOPGQPMBSDPNQPVOETGSPNOPOQPMBSTBNQMFT
BTXFMMBTUIFBOBMZTJTPGTBNQMFTDPOUBJOJOHBOBMZUFTXJUI
BXJEFSBOHFPGIZESPQIPCJDJUZ
Phase
Pore Size (Å)
$ZBOP
Particle Size
(¬m)
Surface Area % Carbon Load
ProteCol Silica
)JHITVSGBDFBSFBBOENFDIBOJDBMTUSFOHUI
Phase
Pore Size (Å)
4JMJDB
Particle Size
(¬m)
Surface Area % Carbon Load
Specialty Phase ProteCol™ Range
tProteCol ChiralDPMVNOTFOTVSFUIFJTPMBUJPOBOEBOBMZTJTPGQVSF
FOBOUJPNFST
t ProteCol HILICSBOHFQSPWJEFTBQPMBSTUBUJPOBSZQIBTFFOBCMJOHUIF
SFUFOUJPOBOETFQBSBUJPOPGQPMBSBOBMZUFTVTJOHPSHBOJDNPCJMFQIBTFT
tProteCol SCXDPMVNOIBTBIJHIMPBEJOHDBQBDJUZBOEQSFTTVSFMJNJU
tProteCol PFPDPMVNOJTVTFGVMJOUIFTFQBSBUJPOPGFQJNFST
ProteCol Chiral
tProteCol Chiral CHMJTBNPEJGJFEDFMMVMPTFDPBUFEPOIJHIQVSJUZIJHIQFSGPSNBODFTQIFSJDBMTJMJDBQBSUJDMFT
5IFDIFNJDBMNPEJGJDBUJPOJODMVEFTUIFDIFNJDBMCPOEJOHPGDIMPSPNFUIZMQIFOZMDBSCBNBUFUPDFMMVMPTF
5IFVTFPGDFMMVMPTFNPEJGJFEXJUIDIMPSJOBUFEQIFOZMHSPVQTQSPWJEFTGPSUIFTFQBSBUJPOGPSNBOZQSFWJPVTMZ
VOSFTPMWFEPSQPPSMZSFTPMWFEDIJSBMNJYUVSFT
tProteCol Chiral CHCBSFQPMZTBDDIBSJEFDPBUFEDIJSBMDPMVNOTNBOVGBDUVSFEVTJOHBVOJRVFQSPEVDUJPOQSPDFTT
PGDPBUJOHUIFQSPWFODIJSBMTFMFDUPSUSJTEJNFUIZMQIFOZM
DBSCBNPZMDFMMVMPTFPOIJHIQVSJUZTJMJDBHFM
tProteCol Chiral CHAQPMZTBDDIBSJEFDPBUFEDIJSBMDPMVNOTBSFDSFBUFEVTJOHBVOJRVFQSPEVDUJPOQSPDFTTPG
DPBUJOHUIFQSPWFODIJSBMTFMFDUPSUSJTEJNFUIZMQIFOZM
DBSCBNPZMBNZMPTFPOIJHIQVSJUZTJMJDBHFM
tProteCol Chiral CH4VTFTBNPEJGJFEDFMMVMPTFDPBUFEPOIJHIQVSJUZIJHIQFSGPSNBODFTQIFSJDBMTJMJDBQBSUJDMFT
BOEDPOTJTUTPGUIFDIFNJDBMCPOEJOHPGDIMPSPNFUIZMQIFOZMDBSCBNBUFUPDFMMVMPTF5IFVTFPGDFMMVMPTF
NPEJGJFEXJUIDIMPSJOBUFEQIFOZMHSPVQTQSPWJEFTGPSUIFTFQBSBUJPOGPSNBOZQSFWJPVTMZVOSFTPMWFEPSQPPSMZ
SFTPMWFEDIJSBMNJYUVSFT
ProteCol™ CHC
Column Size:9NN
Particle Size:NJDSPO
Sample Name:"USPQJOF
Mobile Phase:)FYBOF&UIBOPM
Flow Rate:N-NJO
Injection Vol: ¬Pressure:CBS
ProteCol CHA
Column Size: 9NN
Particle Size: NJDSPO
Sample Name: )ZESPCFO[PJO
Mobile Phase: &UIBOPMJO)FYBOF
Flow Rate: N-NJO
Injection Vol: ¬Pressure: CBS
Phase
Sub Phase
$IJSBM
$)$
EJNFUIZMQIFOZMDBSCBNBUFDFMMVMPTF
Chemical Structure
Particle Size (¬m)
$IJSBM
$)"
EJNFUIZMQIFOZMDBSCBNBUFBNZMPTF
$IJSBM
$).
DIMPSPNFIZMQIFOZMDBSCBNBUFDFMMVMPTF
$IJSBM
$)
DIMPSPNFUIZMQIFOZMDBSCBNBUFBNZMPTF
$IJSBM
$)
DIMPSPNFUIZMQIFOZMDBSCBNBUFDFMMVMPTF
4FNJQSFQBSBUJWFBOEQSFQBSBUJWFGPSNBUTBSFBWBJMBCMF
ProteCol HILIC
)*-*$DISPNBUPHSBQIZVTFTNPCJMFQIBTFTDPOUBJOJOHCFUXFFOXBUFSGPSUIFSFUFOUJPOPGQPMBSDPNQPVOET
5IF1SPUF$PMSBOHFPG)*-*$DPMVNOTEFMJWFSTZPVTFQBSBUJPOTQFDJGJDGPSZPVSQPMBSBOBMZUFBOBMZTJT
ProteCol HILIC-RP
$PMVNO
1SPUF$PM™)*-*$31 YNN¬N
.PCJMF1IBTF.FUIBOPM8BUFS
%FUFDUJPO VW!ON
'MPX
N-NJO
4BNQMF
-PSNFUB[FQBN
%JB[FQBN
0YB[FQBO
1SPUF$PM)*-*$31DPMVNOTEFMJWFSBDPNCJOBUJPO
PG )*-*$ BOE SFWFSTFE QIBTF DISPNBUPHSBQIZ
o QFSGFDU GPS TBNQMFT DPOUBJOJOH QPMBS BOE
IZESPQIPCJD BOBMZUFT 5IF DPNQPTJUJPO PG CPUI
UIF QPMZIZESPYZMBUFE QPMZNFS BOE 0%4 HSPVQT
CPVOE UP TJMJDB QSPWJEFT IZESPYZM MFWFMT UIBU BSF
XFMM BCPWF DPOWFOUJPOBM IZESPYZM BOE EJPM UZQF
TUBUJPOBSZQIBTFT
$PMVNO
1SPUF$PM™)*-*$'
YNN¬N
.PCJMF1IBTFN.BNNPOJVN
BDFUBUFJOXBUFS
"DFUPOJUSJMF
'MPX
N-NJO
%FUFDUJPO
VW!ON
4BNQMF
.%"
"NQIFUBNJOF
.%."
.FUIBNQIFUBNJOF
$PMVNO
1SPUF$PM™)*-*$)$
YNN¬N
.PCJMF1IBTF"DFUPOJUSJMF
N.
"NNPOJVNBDFUBUF
%FUFDUJPO VW!ON
'MPX
N-NJO
*OKFDUJPO7PM ¬-
$PMVNO
1SPUF$PM™)*-*$31 YNN¬N
.PCJMF1IBTF.FUIBOPM8BUFS
%FUFDUJPO VW!ON
'MPX
N-NJO
4BNQMF
)ZESPDPSUJTPOF
$PSUJDPTUFSPOF
Į)ZESPYZQSPHFTUFSPOF
$PSUJTPOF"DFUBUF
,FUPQSPHFTUFSPOF
ProteCol HILIC-FL
1SPUF$PM)*-*$'-JTEFTJHOFEGPSSFUFOUJPOBOETFQBSBUJPO
PGQPMBSBOEOPOQPMBSDPNQPVOETUIBUBSFOPUSFUBJOFE
PSTFQBSBUFEPODPOWFOUJPOBMSFWFSTFEQIBTFDPMVNOT
*UDPOTJTUTPGBGMVPSJOBUFECBTFETUBUJPOBSZQIBTFCPVOE
UPTJMJDB5IJTDPNQPTJUJPOQSPWJEFTGPSFYDFMMFOUSFUFOUJPO
BOEQFBLTIBQFGPSQPMBSIBMPHFOBUFEQPMBSBNJOFTBOE
QPMBSBSPNBUJDDPNQPVOET
ProteCol HILIC-HC
1SPUF$PM )*-*$)$ IJHI DBQBDJUZ
JT DPNQPTFE PG B
QPMZIZESPYZMBUFE QPMZNFS DPBUFE BOE CPVOE UP TJMJDB
QSPWJEJOHIZESPYZMMFWFMTUIBUBSFXFMMBCPWFDPOWFOUJPOBM
IZESPYZMBOEEJPMUZQFTUBUJPOBSZQIBTFT
5IF DISPNBUPHSBN IJHIMJHIUT UIF VOJRVF DBQBCJMJUZ GPS
1SPUF$PM )*-*$)$ XIFSF UPMVFOF JT MFTT SFUBJOFE UIBO
VSBDJM6SBDJMIBTCFFOUSBEJUJPOBMMZVTFEBTBOVOSFUBJOFE
NBSLFS GPS UIF EFUFSNJOBUJPO PG WPJE WPMVNF IPXFWFS
XJUI1SPUF$PM)*-*$)$BOEBOBDFUPOJUSJMFNPCJMF
QIBTFVSBDJMDBOCFSFUBJOFE
$PMVNO
1SPUF$PM™)*-*$1*
YNN¬N
.PCJMF1IBTF"DFUPOJUSJMF8BUFS
%FUFDUJPO VW!ON
'MPX
N-NJO
/.FUIZM
$IMPSJEB[PO
$IMPSJEB[PO
ProteCol HILIC-PI
1SPUF$PM )*-*$1* DPOTJTUT PG BO BSPNBUJD BNJOF CBTFE
TUBUJPOBSZ QIBTF CPVOE UP TJMJDB 5IJT DPNQPTJUJPO
QSPWJEFTGPSFYDFMMFOUSFUFOUJPOBOEQFBLTIBQFGPSQPMBS
BNJOFDPNQPVOET
ProteCol SCX
1 SPUF$PM4$9JTBTJMJDBCBTFETUSPOHDBUJPOFYDIBOHFSTVJUBCMFGPSUIFBOBMZTJTPGTNBMMPSHBOJDCBTFT#BTFE
POBCPOEFEBSPNBUJDTVMGPOJDBDJEHSPVQBOEBWBJMBCMFJOBOE¬NQBSUJDMFTJ[F1SPUF$PM4$9XJMMEFMJWFS
TVQFSCQFSGPSNBODF
t)JHIMPBEJOHDBQBDJUZBOEQSFTTVSFMJNJU
t3PCVTUCPOEJOHUFDIOPMPHZ
Phase
Pore Size (Å)
Particle Size (¬m)
Surface Area
pH
t)JHIEFOTJUZCPOEJOH
4$9
t#POEJOHBSPNBUJDTVMGPOJDBDJEHSPVQT
ProteCol PFP
1'1 JT B USVMZ VOJRVF TUBUJPOBSZ QIBTF XJUI QSPQFSUJFT TJHOJGJDBOUMZ EJGGFSFOU GSPN 0%4 QIBTFT 5IJT VOJRVF
DIBSBDUFSSFTVMUTGSPNCPOEFEQFOUBGMVPSPQIFOZMHSPVQTJNQBSUJOHBQJQJFMFDUSPOJOUFSBDUJPOQSPEVDJOHBO
FOIBODFESFUFOUJPOGPSNBOZDPNQPVOETQBSUJDVMBSMZUIPTFUIBUDPOUBJOQPMBSJ[BCMFFMFDUSPOT.BOZDMBTTFT
PG DPNQPVOET BOE OBUVSBMMZ PDDVSSJOH DIFNJDBMT BMTP
DPOUBJOQPMBSJ[BCMFFMFDUSPOTBOEDBOCFTFQBSBUFEVTJOH
1'11'1IBTCFFOFYUSFNFMZVTFGVMJOUIFTFQBSBUJPOPG
FQJNFST &QJNFST BMTP FYJTU JO NBOZ OBUVSBM NJYUVSFT
TVDIBTQIBSNBDFVUJDBMMZBDUJWFOBUVSBMQBDMJUBYFM
3URWH&ROŒ3)3
Phase
Pore Size (Å)
Particle Size
(¬m)
Surface Area
% Carbon
Load
pH
PFP
Column Application Recommendations and USP Guide
Small Analyte
Large Analyte
Polar Analyte
Very
Hydrophobic Analyte
Low
Nonspecific Interaction
Fast Analysis
Extreme pH
Conditions
Chiral Analysis
Halogenated
Samples
Basic Samples
Range of
Hydrophobicities in
Analyte
HILIC
Applications
Aromatic
Samples
6TFUIF641DPEFUPTFMFDUUIF1SPUF$PMDPMVNOGPSZPVSOFFET
ProteCol C18 Q
Y
N
O
O
Y
N
N
N
Y
Y
Y
N
Y
ProteCol C18 H
Y
N
O
O
O
N
Y
N
Y
O
Y
N
Y
ProteCol C18 G
Y
N
O
O
O
N
N
N
Y
O
Y
N
Y
ProteCol C18 P
Y
N
Y
O
/"
N
N
N
Y
O
Y
N
Y
ProteCol C8 Q
Y
N
N
Y
O
N
N
N
Y
O
Y
N
Y
ProteCol C8 H
Y
NY
N
Y
O
N
N
N
Y
O
Y
N
Y
ProteCol C4 H
Y
NY
N
Y
O
N
N
N
Y
O
Y
N
Y
ProteCol Phenyl Hexyl
Y
N
O
O
O
N
N
N
Y
O
Y
N
Y
ProteCol Silica
Y
N
Y
N
/"
N
N
N
Y
Y
Y
Y
Y
ProteCol Amino
Y
N
Y
N
/"
N
N
N
Y
Y
Y
Y
Y
ProteCol Cyano
Y
N
Y
Y
/"
N
N
N
Y
O
Y
O
O
ProteCol HILIC RP
Y
N
Y
N
/"
N
N
N
Y
Y
Y
Y
Y
ProteCol HILIC PI
Y
N
Y
N
/"
N
N
N
Y
Y
Y
Y
Y
ProteCol HILIC FL
Y
N
Y
N
/"
N
N
N
Y
O
Y
Y
Y
ProteCol HILIC HC
Y
N
Y
N
/"
N
N
N
Y
Y
Y
Y
Y
ProteCol Chiral CHC
Y
N
/"
/"
/"
N
N
Y
/"
/"
/"
/"
/"
ProteCol Chiral CHM
Y
N
/"
/"
/"
N
N
Y
/"
/"
/"
/"
/"
ProteCol Chiral CHA
Y
N
/"
/"
/"
N
N
Y
/"
/"
/"
/"
/"
ProteCol Chiral CH5
Y
N
/"
/"
/"
N
N
Y
/"
/"
/"
/"
/"
ProteCol Chiral CH4
Y
N
/"
/"
/"
N
N
Y
/"
/"
/"
/"
/"
ProteCol SCX
Y
N
Y
N
/"
N
N
N
Y
Y
N
O
N
ProteCol PFP
Y
N
O
O
O
N
N
N
Y
O
Y
N
Y
ProteCol Ultra C18
Y
N
O
O
N
Y
N
N
Y
O
Y
N
Y
ProteCol Ultra C8
Y
N
N
Y
N
Y
N
N
Y
O
Y
N
Y
ProteCol Ultra Amino
Y
N
Y
N
/"
Y
N
N
Y
O
Y
Y
Y
ProteCol Ultra Cyano
Y
N
Y
Y
/"
Y
N
N
Y
O
Y
Y
O
ProteCol Ultra HILIC FL
Y
N
Y
N
/"
Y
N
N
Y
O
Y
N
O
ProteCol Ultra HILIC PI
Y
N
Y
N
/"
Y
N
N
Y
Y
Y
N
Y
ProteCol Ultra PFP
Y
N
O
O
N
Y
N
N
Y
O
Y
N
Y
ProteCol Ultra Phenyl
Y
N
N
Y
N
Y
N
N
Y
O
Y
N
Y
ProteCol Ultra Phenyl Hexyl
Y
N
O
O
N
Y
N
N
Y
O
Y
N
Y
ProteCol Ultra Polar
Y
N
Y
O
N
Y
N
N
Y
O
Y
N
Y
ProteCol Ultra SCX
Y
N
Y
N
/"
Y
N
N
Y
Y
N
O
N
1FOUBGMVPSPQIFOZMIBTBTQFDJBMTFMFDUJWJUZGPSIBMPHFOBUFETVCTUBODFTBOETIPVMECFVTFEXIFOTFQBSBUJPOPODPOWFOUJPOBM31QIBTFTJTEJGGJDVMU
1IFOZMBOE)FYBQIFOZMIBWFBTQFDJBMTFMFDUJWJUZGPSBSPNBUJDTVCTUBODFTBOETIPVMECFVTFEXIFOTFQBSBUJPOPODPOWFOUJPOBM31QIBTFTJTEJGGJDVMU
*OSFWFSTFEQIBTFNPEF
8IFOQBSUJDMFTXJUI±±PS±BSFDIPTFO
Y 3FDPNNFOEFE
N /PU3FDPNNFOEFE
O 0QUJPOBM
How to Order - Building your HPLC Column Part Number
5IF4(&1SPUF$PM™SBOHFPG)1-$DPMVNOTPGGFSTZPVNBOZDPNCJOBUJPOTXIFSFZPVDBOTFMFDUUIFQIBTFQBSUJDMF
TJ[FDPMVNOMFOHUIBOE*%BTXFMMBTDPMVNOIBSEXBSF
5PNBLFPSEFSJOHFBTJFSQMFBTFVTFUIFGPMMPXJOHHVJEFXIFOCVJMEJOHZPVSDPMVNOGPSZPVSBQQMJDBUJPO
t5IFQBSUOVNCFSTUBSUTXJUI1IBTF$PEF1BSUJDMF4J[F-FOHUI$PEF*%$PEF)BSEXBSF$PEFBOEXIFUIFSJUJTB
HVBSEDPMVNO
t*GZPVXBOUBHVBSEDPMVNOUPDPNQMFNFOUBOBOBMZUJDBMDPMVNOBEEi(wBTBTVGGJY
Phase
Particle
Size
Column
Length
Column
ID
Hardware
Part
Number
5 ¬N
250 mm
2.1 mm
PEEK coated
?
&YBNQMF
1SPUF$PM™$)
ID CODE
LENGTH CODE
PARTICLE SIZE
HARDWARE CODE
Particle Size Particle Size
Code
Length
Length
Code
ID
ID Code
Hardware
Hardware
Code
1.8
18
10
G
0.15
15
Stainless Steel
S
2.5
25
10
T
0.3
30
UHPLC
U
3
03
50
L
1
01
PEEKsil™
K
5
05
100
M
2.1
02
PEEK Coated
P
10
150
N
3
03
250
R
4.6
46
10
10
10
$
05
R
02
P
2C18205R02P
3FGFSUPUIFUBCMFPO
UIFOFYUQBHF
Build a column easily by downloading our HPLC Part Number Generator at www.sge.com/lc
PHASE
Phase
Length
ID
Pore Size (Å)
Phase Code
Particle Size (µm)
Hardware Code
1SPUF$PM™$2
-./3
$
4,1
1SPUF$PM™$)
-./3
$
4,1
1SPUF$PM™$(
-./3
$
4,1
1SPUF$PM™$1
-./3
10-
4,1
1SPUF$PM™$)
-./3
$
4,1
1SPUF$PM™$(
-./3
$
4,1
1SPUF$PM™$(
-./3
$
4,1
1SPUF$PM™$)
-./3
$
4,1
1SPUF$PM™$)
-./3
$
4,1
1SPUF$PM™1IFOZM)FYZM
-./3
/)
SP
1SPUF$PM™4JMJDB
-./3
4*-
SP
1SPUF$PM™"NJOP
-./3
".
SP
SP
1SPUF$PM™$ZBOP
-./3
$/
1SPUF$PM™)*-*$31
-./3
)-
SP
1SPUF$PM™)*-*$1*
-./3
)-
SP
1SPUF$PM™)*-*$'-
-./3
)-
SP
1SPUF$PM™)*-*$)$
-./3
)-
SP
1SPUF$PM™$IJSBM$)$
R
$)$
SP
1SPUF$PM™$IJSBM$).
R
$).
SP
1SPUF$PM™$IJSBM$)"
R
$)"
SP
1SPUF$PM™$IJSBM$)
R
$)
SP
1SPUF$PM™$IJSBM$)
R
$)
SP
1SPUF$PM™4$9
-./3
4$9
SP
1SPUF$PM™ PFP
-./3
1'1
SP
1SPUF$PM™6MUSB$
-./
6$
U
1SPUF$PM™6MUSB$
-./
6$
U
1SPUF$PM™6MUSB"NJOP
-./
6".
U
1SPUF$PM™6MUSB$ZBOP
-./
6$/
U
1SPUF$PM™6MUSB)*-*$'-
-./
6)-
U
1SPUF$PM™6MUSB)*-*$1*
-./
6)-
U
1SPUF$PM™6MUSB1'1
-./
61'1
U
1SPUF$PM™6MUSB1IFOZM
-./
61)
U
1SPUF$PM™6MUSB1IFOZM)FYZM
-./
6/)
U
1SPUF$PM™6MUSB1PMBS
-./
610-
U
1SPUF$PM™6MUSB4$9
-./
64$9
U
*Note no guard columns available.
Guard Columns (pack of three)
Trap Columns (single pack)
1SPUF$PM™$1±
NN*%
NN*%
NN*%
¬N*%
¬N*%
¬N*%
¬N*%
1SPUF$PM™$2±
NN*%
NN*%
NN*%
¬N*%
¬N*%
¬N*%
¬N*%
1SPUF$PM™$2±
NN*%
NN*%
NN*%
¬N*%
¬N*%
¬N*%
¬N*%
1SPUF$PM™$2±
NN*%
NN*%
NN*%
¬N*%
¬N*%
¬N*%
¬N*%
1SPUF$PM™$)±
NN*%
NN*%
NN*%
¬N*%
¬N*%
¬N*%
¬N*%
1SPUF$PM™$(±
NN*%
NN*%
NN*%
¬N*%
¬N*%
¬N*%
¬N*%
1SPUF$PM™$)±
NN*%
NN*%
NN*%
¬N*%
¬N*%
¬N*%
¬N*%
1SPUF$PM™$)±
NN*%
NN*%
NN*%
¬N*%
¬N*%
¬N*%
¬N*%
1SPUF$PM™$)±
NN*%
NN*%
NN*%
¬N*%
¬N*%
¬N*%
¬N*%
1SPUF$PM™4JMJDB±
NN*%
NN*%
NN*%
/"
/"
/"
/"
1SPUF$PM™"NJOP±
NN*%
NN*%
NN*%
/"
/"
/"
/"
1SPUF$PM™$ZBOP±
NN*%
NN*%
NN*%
/"
/"
/"
/"
Inner diameter of the guard columns provided. Particle size of the stationary phase is corresponding with the particle size of the main column (please specify when ordering).
HPLC Method Development
*GZPVBSFMPPLJOHUPEFWFMPQBNFUIPEBOEEFUFSNJOF
UIFNPTUTVJUBCMF)1-$DPMVNOGPSUIFBOBMZTJTGPMMPX
UIJT EFWFMPQNFOU GMPXDIBSU UP FTUBCMJTI UIF CFTU
NFUIPE
(BUIFS*OGPSNBUJPO
1VCMJTIFE.FUIPE
:FT
/P
1FSGPSN3VOVOEFS
$POEJUJPOTPGUIF
1VCMJTIFE.FUIPE
4UVEZ$IFNJTUSZPG
4VCTUBODF
*T4VCTUBODF
$IBSHFE
:FT
/P
"EKVTUQ)"XBZ
GSPNQ,B7BMVF
4UBSUXJUI$PO
QBSUJBMMZEFHSBEFEQSPEVDUPS
SFTPMVUJPONJYUVSFTUBOEBSE
3VO(SBEJFOUXJUI
"DFUPOJUSJMF
"MM1FBLT3FTPMWFE
:FT
/P
-PPLGPS$SJUJDBM1FBL1BJST
$IBOHF1BSBNFUFST
'MPX3BUF
5FNQFSBUVSF
Q)
(SBEJFOU1SPGJMFUP
3FTPMWF$MPTF1FBLT
"MM1FBLT3FTPMWFE
:FT
/P
$IBOHF0SHBOJD.PEJGJFS
UP.F0)PS5)'
$POTJEFS*POQBJSJOH
3FBHFOUT
"MM1FBLT3FTPMWFE
:FT
/P
%PFTUIF.FUIPE
8PSLPOUIF3FBM
4BNQMF
$IBOHFUP%JGGFSFOU#SBOE
PG$$PMVNO
/P
:FT
*TUIF$$PMVNO
4VJUBCMF
:FT
0QUJNJ[F.FUIPEGPS
4IPSU"OBMZTJT5JNF
/P
$IBOHFUP%JGGFSFOU31
$PMVNO1IFOZM$$/FUD
-BSHF(BQT#FUXFFO
BMM1FBLT
:FT
*ODSFBTF4MPQFPG(SBEJFOU
6TF4IPSUFS$PMVNO
/P
"MM1FBLT3FTPMWFE
:FT
/P
/P
$IFDLGPS.FUIPE
3PCVTUOFTTBOE$SJUJDBM
1BSBNFUFST
$IBOHFUP%JGGFSFOU-$.PEF
/PSNBM1IBTF*&$FUD
"MM1FBLT3FTPMWFE
1BSUT8JUI/P
1FBLT
:FT
Method Finished Documentation
/P
LC is Not Suitable for
the Analysis
Method Validation
:FT
1BSUJBMMZ*ODSFBTF(SBEJFOU
4MPQFUP4LJQ"SFBT
8JUI/P1FBL
HPLC Troubleshooting
Problem
System Related
-PXVOTUFBEZTZTUFN
QSFTTVSF
Reason
-FBL
"JSJOQVNQIFBE
%JSUJODIFDLWBMWFDIFDLXIFUIFSWBMWF
DBOOPUDMPTF
#MPDLBHFDPOUBNJOBUJPO
Resolution
$IFDLBMMDPOOFDUJPOTBOEUJHIUFODPOOFDUJPOTSFQMBDFTFBMT
%FHBTNPCJMFQIBTFBOEQVSHFTZTUFN
'JSTUMZUSZQVSHJOHTZTUFNBUIJHIGMPXSBUFUPEJTMPEHFDPOUBNJOBUJPO
4FDPOEMZEJTBTTFNCMFDIFDLWBMWFBOETPOJDBUF
)JHITZTUFNQSFTTVSF
0QFODPOOFDUJPOTTFRVFOUJBMMZGSPNUIFEFUFDUPSCBDLUPUIFQVNQ
UP MPDBUF CMPDLBHF 'MVTI DBQJMMBSJFT SFQMBDF JOMJOF GJMUFST PS HVBSE
DPMVNOTDMFBOJOKFDUPSWBMWFSFWFSTFDPMVNOGMPXXJUIPVUEFUFDUPS
JOMJOF
EFQFOEJOHPOXIFSFUIFCMPDLBHFXBTMPDBUFE
#MPDLBHF QSFDJQJUBUFE CVGGFS TBMUT
%JTDPOOFDU DPMVNO BOE GMVTI XJUI QVSF XBUFS BU MPX GMPX SBUF UP
DBO IBQQFO XIFO UIF TZTUFN PS EJTTPMWFCVGGFSTBMUTBHBJO
VTFS TVEEFOMZ DIBOHFT NPCJMF QIBTF
DPNQPTJUJPO GSPN IJHI PSHBOJD UP
BRVFPVTCVGGFSPSWJDFWFSTB
)JHIWJTDPTJUZNPCJMFQIBTF
*ODSFBTFUFNQFSBUVSFDIBOHFNPCJMFQIBTFPSEFDSFBTFGMPXSBUF
4NBMMTUBUJPOBSZQIBTFQBSUJDMFT
*ODSFBTFUFNQFSBUVSFSFEVDFGMPXSBUFVTFTIPSUFSDPMVNO
$SVTIFEQBSUJDMFTTVEEFOQSFTTVSFTQJLFT 3FQMBDFUIFDPMVNO
DBO DBVTF QPSPVT TJMJDB UP GSBDUVSF BOE
HFOFSBUFiGJOFTw
/PJTZGMVDUVBUJOHESJGUJOH 4ZTUFNDPOUBNJOBUJPO
%JTDPOOFDUDPMVNOBOESJOTFTZTUFNXJUIBDPNCJOBUJPOPGBDJE
CBTFMJOF
OJUSJDBDJEPSQIPTQIPSJDBDJEGPSBTIPSUQFSJPEPGUJNFGPMMPXFECZ
XBUFSBOEBPSHBOJDXBTIPGBDFUPOJUSJMF*1"PWFSOJHIU
%P
/05SVOUIFBDJEUISPVHIUIFDPMVNO
"HFPGUIF67MBNQ
3FQMBDFUIF67MBNQ
5FNQFSBUVSFGMVDUVBUJPOT
6TFDPMVNOPWFO
)JHIFS 67 BCTPSQUJPO PG FJUIFS NPCJMF 6TF)1-$HSBEFTPMWFOUTDIFDL67DVUPGGWBMVFTGPSNPCJMFQIBTF
QIBTF " PS # DBVTFT ESJGU JO HSBEJFOU DPNQPOFOUTDIBOHFUPIJHIFSXBWFMFOHUI
FMVUJPO
3FHVMBS QVMTJOH PG UIF "JSJOQVNQIFBEBMTPDBVTFTQVMTJOHPG %FHBTNPCJMFQIBTFBOEQVSHFTZTUFN
CBTFMJOF
UIFCBDLQSFTTVSF
%JSU JO DIFDL WBMWF BMTP DBVTFT QVMTJOH 'JSTUUSZQVSHJOHTZTUFNBUIJHIGMPXSBUFUPEJTMPEHFDPOUBNJOBUJPO
PGUIFCBDLQSFTTVSF
4FDPOEEJTBTTFNCMFDIFDLWBMWFBOETPOJDBUF
#VCCMF USBQQFE JO UIF GMPX DFMM o UIF %FHBTNPCJMFQIBTFBOEQVSHFTZTUFN
EFUFDUPSSFTQPOTFDIBOHFTESBNBUJDBMMZ
XIFOUIFEFUFDUPSPVUMFUJTUFNQPSBSJMZ
CMPDLFEXJUIBGJOHFS
The Chromatogram
5BJMJOHQFBLT
8SPOHQ)TPNFQFBLTBSFUBJMJOHXIJMF 5IFQ)PGUIFNPCJMFQIBTFTIPVMECFVOJUTPSNPSFBCPWFPSCFMPX
PUIFSTBSFTZNNFUSJDBM
UIFQ,BWBMVFPGUIFBOBMZUFUPIBWFBMMNPMFDVMFTFJUIFSJOUIFDIBSHFE
PSJOUIFOFVUSBMTUBUF
7PJEWPMVNFTBMMQFBLTBSFUBJMJOH
$IFDLDPOOFDUJPOTSFQMBDFHVBSEDPMVNOSFQMBDFDPMVNO
/POTQFDJGJD JOUFSBDUJPOT TPNFBMM 3FQMBDF DPMVNO XJUI BO JOFSU DPMVNO SFQMBDF NFUBM UVCJOH XJUI
TBNQMF DPNQPOFOUT DBO JOUFSBDU XJUI 1&&,TJM™ UVCJOH "EE BEEJUJWFT FH &%5"
JOUP NPCJMFQIBTF MPXFS
BDUJWF TJUFT JO UIF GMPXQBUI TJMBOPM Q)UPJOPSEFSUPQSPUPOBUFTJMBOPMHSPVQT
HSPVQT NFUBM TVSGBDFT PG UVCFT BOE
GSJUT
'SPOUJOHUBJMJOHQFBLT
$IBOOFMJOH
$IBOOFMJOH JOEJDBUFT B TFSJPVT QSPCMFN XJUI UIF DPMVNO BOE UIF
DPMVNO OFFET SFQMBDJOH 'PS UIF JOUFSJN ZPV DBO USZ UP SFWFSTF UIF
DPMVNOGMPXEJSFDUJPO
i7JTDPVT GJOHFSJOHw o IBQQFOT XIFO 5SZUPNBUDIUIFWJTDPTJUZPGUIFTBNQMFXJUIUIFNPCJMFQIBTF*EFBMMZ
UIFSFJTBMBSHFEJGGFSFODFCFUXFFOUIF BMXBZTVTFNPCJMFQIBTFBTUIFTBNQMFEJMVFOU
WJTDPTJUZPGUIFTBNQMFBOEUIFWJTDPTJUZ
PGUIFNPCJMFQIBTF
4UBUJPOBSZQIBTFEFHSBEBUJPO
-PTTPGMJHBOETXIFOUIFDPMVNOJTFYQPTFEUPFYUSFNFQ)PSXIFO
UIFDPMVNOJTWFSZPMEDBOMFBEUPQFBLGSPOUJOH3FQMBDFUIFDPMVNO
$PMVNOPWFSMPBEJOH
3FEVDFUIFBNPVOUPGTBNQMFJOKFDUFEPSVTFBDPMVNOXJUIBMBSHFS*%
More Than Just Packaging
HPLC Packaging
4(&IBTEFWFMPQFEJNQSPWFEQBDLBHJOHFOBCMJOHZPVUPSFDFJWFZPVSDPMVNOXJUIDPOGJEFODFBOETUPSFJUTFDVSFMZ
#ZEFTJHOJOHQBDLBHJOHUIBUDPNCJOFTTIJQQJOHBOETUPSBHFTPMVUJPOTDIBPUJDESBXFSTBOEFYQFOTJWF)1-$TQFDJGJD
TUPSBHFBSFFMJNJOBUFE
(VBSE$PMVNO
0QFOTGMBUUPTUPSF
DPMVNOTBOEBDDFTTPSJFT
JOZPVSESBXFS
-BCFMUPJEFOUJGZDPMVNO
5FTU3FQPSU
4UPSFVQUPUISFFGVMMTJ[FEBOBMZUJDBMDPMVNOT
Complementary HPLC Products and Supplies
MEPS™
.&14™ .JDSP &YUSBDUJPO CZ 1BDLFE 4PSCFOU
JT B NJDSP
41&TPMVUJPOUIBUJODPSQPSBUFTUIFTUBUJPOBSZQIBTFJOB
NJDSPDBSUSJEHFJOUFHSBUFEJOBIJHIRVBMJUZ4(&BOBMZUJDBM
TZSJOHF#BSSFM*OTFSUBOE/FFEMF#*/DPOGJHVSBUJPO
Conditioning
3FVTF.&14
NBOZUJNFT
EFQFOEFOU
PONBUSJY
.&14 JTUIFNJOJBUVSJ[BUJPOPGDPOWFOUJPOBM41&QBDLFE
CFEEFWJDFTGSPNN-UP¬-CFEWPMVNFT
Load
t3FQFBUMPBE
t4NBMMTBNQMF
WPMVNFT
t4NBMMWPMVNF
TPMWFOUSFRVJSFE
F7PM® .&14 TUBUJPOBSZ QIBTFT BWBJMBCMF $ $ $
"14%7#4%7#
PTFE
Sealing Ring
MEPS™
Packed bed
Needle
Elute
*OKFDUUIF
BOBMZUFEJSFDUMZ
JOUPUIFJOKFDUPS
To Barrel
Frits
End Plug
Schematic of the MEPS ™ stationary phase within the syringe needle
– SGE’s patented ‘Barrel Insert and Needle’ (BIN) configuration.
Wash
t3FNPWF
VOXBOUFE
BOBMZUF
t&OTVSFIJHI
SFDPWFSJFT
ProteCol™ Guard Columns and Filters
Analytical Protection
Capillary Protection
ProteCol Filter
5IF 1SPUF$PM GJMUFS JT EFTJHOFE UP GJMUFS UIF TBNQMF
QSJPS UP UIF QSFDPMVNO 1FBL CSPBEFOJOH IBT CFFO
FMJNJOBUFEEVFUPMPXEFBEWPMVNFJOFSUEFTJHO
t ;FSPEFBEWPMVNFGJMUFSEFTJHO
t ;FSPQSFTTVSFESPQBDSPTTGJMUFS
t ;FSPDPNQSPNJTFPOQFSGPSNBODF
ProteCol Guard Column
8FSFDPNNFOEUIFVTFPG1SPUF$PM(VBSEDPMVNOT
UP QSPUFDU UIF BOBMZUJDBM DPMVNO BOE FOTVSF
JU QFSGPSNT DPOTJTUFOUMZ 5IF HVBSE DPMVNO JT
EFTJHOFEUPGJUJOUPUIFCBDLPGB1&&,GJOHFSUJHIU
GJUUJOHQSPWJEFEXJUIUIFHVBSEDPMVNO
/PGVSUIFS
VOJPOTBSFSFRVJSFE
5IF1SPUF$PM *O-JOF'JMUFSJTBTJNQMFBOEFGGFDUJWF
XBZ UP QSPUFDU ZPVS DBQJMMBSZ DPMVNOT GSPN
QBSUJDVMBUFT*UQSPUFDUTZPVSTZTUFNGSPNCMPDLBHFT
BOE JODSFBTFE CBDL QSFTTVSFT XJUIPVU JOUSPEVDJOH
QFBL UBJMJOH PS MPTT PG SFTPMVUJPO 5IF GJMUFSJOH
FMFNFOU JT B NJDSPO QPSPTJUZ TDSFFO MPDBUFE
CFUXFFOUIFTRVBSFDVUBOEQPMJTIFEFOETPGUXP
MFOHUITPG1&&,TJM™ UVCJOHAUBJMT
Accessories
HPLC Tubing
1&&,TJM™ NBZ CF VTFE BT
B EJSFDU SFQMBDFNFOU GPS
DPOWFOUJPOBM TUBJOMFTT TUFFM
BT XFMM BT B SFQMBDFNFOU
GPS1&&,UVCJOHVTFEJO-$
TZTUFNT5IF1&&,QPMZNFS
FYUFSJPS DPBUJOH BOE UIF
GVTFETJMJDBDPNCJOBUJPONBLFT1&&,TJMWFSZSPCVTU
NBLJOH JU JEFBM GPS DBQJMMBSZ )1-$ BOE -$.4
BQQMJDBUJPOT
EasyLok
&BTZ-PL DPOOFDUJPOT DPNQSJTF
PG B LOVSMFE TUBJOMFTT TUFFM
OVU BOE B EPVCMF FOEFE 1&&,
GFSSVMF 5IF 1&&, GFSSVMF TMJEFT
PWFS BOZ 0% UVCJOH UP JUT SFRVJSFE QPTJUJPO
XIJMF UIF OVU JT GJOHFS UJHIUFOFE 6OMJLF TUBJOMFTT
TUFFMUIF1&&,GFSSVMFXJMMOPUDSVTIUIFUVCJOHBOE
DBO CF FBTJMZ SFBEKVTUFE GPS RVJDL DPMVNO DIBOHFT
5IFVOJRVFEPVCMFFOEFEGFSSVMFEFTJHOTFBMTBUUXP
QPJOUTUPQSFWFOUMFBLT
HPLC Connections
1SPUF$PM 6OJPOT TUBJOMFTT
TUFFMPS1&&,
BSFDPNCJOFE
XJUI SFVTBCMF 1&&, GFSSVMFT
GBDJMJUBUJOHDPOOFDUJOHBOZDPNCJOBUJPOPGNN
GVTFETJMJDBUVCJOHBOE1&&,TJM
t4UBJOMFTT TUFFM VOJPOT DBO CF GJOHFS UJHIUFOFE PS
UJHIUFOFE XJUI B XSFODI GPS IJHIQSFTTVSF
BQQMJDBUJPOT
t1&&, VOJPOT DBO CF GJOHFS UJHIUFOFE 5IFZ BSF
TMJHIUMZ MBSHFS UIBO TUBJOMFTT TUFFM VOJPOT CVU BMTP
MJHIUFSGPSMFTTTUSFTTPOZPVSUVCJOH
5IFGJUUJOHTBSFDPNQBUJCMFXJUIBOZTUBOEBSEGFNBMF
)1-$ GJUUJOH JODMVEJOH 4XBHFMPL® 1BSLFS™ 8BUFST®
7BMDP®BOE8IBUNBO®
Hexnut
*EFBM GPS BQQMJDBUJPOT XIFSF
DPSSPTJWF TPMWFOUT BSF CFJOH
VTFE )FYOVUT IBWF JOFSU
DPOUBDUTVSGBDFTNBLJOHUIFN
CJPDPNQBUJCMF 4UBJOMFTT TUFFM
UISFBE GJUUJOHT VTF B
OPOTXBHJOH ,FM'® PS 1&&,
SFQMBDFBCMFGFSSVMF
ProteCol™ Capillary HPLC Columns
Phase Type
ID
Length
(mm)
Pore Size
(Å)
Particle Size
(µm)
Column Type
Pack Size
Part No.
C8 HQ
150 µm
100
1000
3
PEEKsil™
1
250182
C8 HQ
150 µm
150
1000
3
PEEKsil™
1
250180
C8 HQ
300 µm
100
1000
3
PEEKsil™
1
250177
C8 HQ
300 µm
150
1000
3
PEEKsil™
1
250175
250162
ProteCol™ C8 HQ1003
ProteCol™ C18 HQ203 and HQ303
C18 HQ
150 µm
100
200
3
PEEKsil™
1
C18 HQ
150 µm
100
300
3
PEEKsil™
1
250142
C18 HQ
150 µm
150
200
3
PEEKsil™
1
250160
C18 HQ
150 µm
150
300
3
PEEKsil™
1
250140
C18 HQ
300 µm
100
200
3
PEEKsil™
1
250157
C18 HQ
300 µm
100
300
3
PEEKsil™
1
250137
C18 HQ
300 µm
150
200
3
PEEKsil™
1
250155
C18 HQ
300 µm
150
300
3
PEEKsil™
1
250135
Pack
Size
Part No.
2224058
ProteCol™ HPLC Capillary Trap Columns
Phase Type
ID
(µm)
Length
mm)
Pore Size
(Å)
Particle Size
(µm)
Tail ID
(µm)
Tail OD
C8
150
10
300
3
50
1/32"
1
C8
300
5
300
3
50
1/32"
1
2222158
C18
150
10
300
3
50
1/32"
1
2224054
C18
300
5
300
3
50
1/32"
1
2227054
C18
300
10
300
3
50
1/32"
1
2222054
ReproSil™ HPLC Capillary Trap Columns
ReproSil™ is the reference material for leading proteomics scientists. ReproSil™ captures
the broadest range of peptides from hydrophilic to hydrophobic.
Phase Type
ID
(µm)
Length
mm)
Pore Size
(Å)
Particle Size
(µm)
Tail ID
(µm)
Tail OD
C18-AQ
150
10
120
5
50
1/32"
3
2224072
C18-AQ
300
10
120
3
50
1/32"
3
2222066
C18-AQ
300
10
120
5
50
1/32"
3
2222072
ODS-3
300
10
120
5
50
1/32"
3
2222070
HPLC Columns
Pack
Size
Part No.
HPLC Columns and
Applications
HPLC Columns
ProteCol™ HPLC Guard Columns
Phase Type
ID
Length (mm)
Pore Size (Å)
Particle Size
(µm)
Column Type
Pack Size
Part No.
ProteCol™-P C18 HQ105
HPLC Columns and
Applications
C18 HQ
2.1 mm
10
100
5
Stainless Steel
(PEEK™ frit)
3
250007
C18 HQ
4 mm
10
100
5
Stainless Steel
(PEEK™ frit)
3
250009
ProteCol™-G C8 HQ125
C8 HQ
2.1 mm
10
120
5
Stainless Steel
(PEEK™ frit)
3
250003
C8 HQ
4 mm
10
120
5
Stainless Steel
(PEEK™ frit)
3
250005
ProteCol™-P C18 HPH125
C18 HPH
2.1 mm
10
120
5
Stainless Steel
3
250013
C18 HPH
4 mm
10
120
5
Stainless Steel
3
250015
Stainless Steel
(PEEK™ frit)
3
250021
ProteCol™-G C18 HQ203 and HQ303
C18 HQ
2.1 mm
10
200
3
C18 HQ
2.1 mm
10
300
3
Stainless Steel
(PEEK™ frit)
3
250029
C18 HQ
2.1 mm
10
300
5
Stainless Steel
(PEEK™ frit)
3
250025
C18 HQ
4 mm
10
200
3
Stainless Steel
(PEEK™ frit)
3
250023
C18 HQ
4 mm
10
300
3
Stainless Steel
(PEEK™ frit)
3
250031
C18 HQ
4 mm
10
300
5
Stainless Steel
(PEEK™ frit)
3
250027
ProteCol™-G HQ1003
C8 HQ
2.1 mm
10
1000
3
Stainless Steel
(PEEK™ frit)
3
250019
C8 HQ
4 mm
10
1000
3
Stainless Steel
(PEEK™ frit)
3
250017
4 mm
10
120
5
Stainless Steel
(PEEK™ frit)
3
250033
ProteCol™ GP125
C18 GP
ProteCol™ HPLC Capillary Guard Columns
Phase Type
ID
(µm)
Length
(mm)
Pore Size
(Å)
Particle Size
(µm)
Tail ID
(µm)
Tail OD
Pack Size
Part No.
C8
300
10
300
3
50
1/32"
3
222205
C18
150
10
300
3
50
1/32"
3
222404
C18
300
10
300
3
50
1/32"
3
222204
HPLC Columns
HPLC Application by Industry | Environmental
PAHs using EPA 610 on ProteCol™ C18 GP125
Column Part No.:
250212
Sample:
Polyaromatic Hydrocarbons
(EPA610 mix)
20 min 100% B
ProteCol™ C18 GP125
150 x 4.6 mm
50 min 100% B
Column:
Injection Volume:
1 µL
Mobile Phase A:
Water
Mobile Phase B:
100% Acetonitrile
Gradient:
0 min
70% B
51 min
Flow rate.:
1.0 ml/min
Temperature:
20 °C
Detection:
254 nm
45000
Components
6
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
40000
35000
30000
25000
1
20000
5
2
10
15000
8
10000
11
9
7
3 4
70% B
12
13 14
Naphthalene
Acenaphthylene
Acenaphthene
Fluorene
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benzo[a] Anthracene
Chrysene
Benzo[b] Fluoranthene
Benzo[k] Fluoranthene
Benzo[a] Pyrene
Dibenz[a,h] Anthracene
Benzo[g,h,i] Perylene
Indeno[1,2,3-cd] Pyrene
16
5000
15
0
0
5
10
15
20
25
30
35
40
45
HPLC Application by Industry | General Chemistry
Paraben Mixture on ProteCol™ C18 HPH125
1400000
1200000
1000000
800000
butylparaben
methylparaben
ethylparaben
propylparaben
1600000
Column Part No.:
250112
Sample:
Paraben Mix
Column:
O
ProteCol™ C18 HPH125
O-R150 x 4.6 mm
Injection Volume:
1 µL
Mobile Phase:
75% Methanol in Water
Flow rate.:
1.0 ml/min
OH
Temperature:
30 °C
Detection:
254 nm
heptylparaben
600000
400000
200000
0
0
5
10
HPLC Application by Industry
15
20
HPLC Columns and
Applications
HPLC Application by Industry | Food, Flavor, Fragrance
TP-0199-H | Measuring Fruit Juice Adulteration by Changes in Flavonoid
Content Using MEPS™ on ProteCol™ HQ105
Column Part No.:
250102
Column:
ProteCol™ HQ105
150 mm x 4.6 mm ID
Mobile Phase A:
0.1 % TFA in water
Mobile Phase B:
0.1 % TFA in 80 % methanol
LC System:
Shimadzu Prominance LC20.
Flow Rate:
1.0 ml/min
Gradient Profile:
0 min - 0 % B
20 min - 50 % B
30 min - 100 % B
40 min - 100 % B
41 min - 0 % B
60 min - 0 % B
Column Temperature:
40 °C
Detection:
254, 350 and 550 nm
HPLC Columns and
Applications
Chromatograms of cranberry juice prepared with different MEPS™ phases.
Flavonoid fingerprinting of various fruit juices on MEPS™ C18.
More chromatograms and application information can be found at
sge.com/documents/chromatogram-library
HPLC Application by Industry | Life Sciences
Enolase Trypsin Digest on ProteCol™ Trap Column C18 HQ303
Column Part No.:
250140
Sample:
Enolase Trypsin Digest
10 fmol/ µL
40 min 40% B
Column:
ProteCol™ C18 HQ303
150 mm x 150 µm ID
45 min 100% B
Injection Volume:
8 µL
Mobile Phase A:
0.1% Formic Acid in Water
Mobile Phase B:
0.1% Formic Acid in 90%
Acetonitrile
Gradient:
0 min
10% B
59 min 100% B
60 min
5% B
70 min
5% B
Flow rate.:
0.5 µL/min
Detection:
QSTAR Elite MS with
nanoLC source
HPLC Columns and
Applications
HPLC Applications by Industry
HPLC Application by Industry | Life Sciences
TA-0141-H | Ribosomal Proteins on ProteCol™ C8 HQ 1003
Column Part No.:
250170
Sample:
Ribosomal Proteins from
Rat Liver
Gradient:
0 min
™
Column:
ProteCol C8 HQ 1003
150 mm x 2 mm ID
Injection Volume:
10 µL
Solvent A:
Aqueous 0.1% (v/v) Formic Acid
Solvent B:
Acetonitrile/0.1% (v/v) Formic Acid
Agilent 1100 LC System
LC:
5% B
80 min
45% B
81 min
85% B
82 min
85% B
83 min
5% B
88 min
5% B
Flow rate.:
0.25 mL/min
Detection:
Agilent 6220 ESI-TOF LC/MS
HPLC Columns and
Applications
No.
1
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
16
17
17
18
19
20
21
21
22
214
RT
[min]
22.4
24.2
26.5
28.7
31.9
36.7
36.7
37.9
42
42.3
42.3
43.8
44.6
45.6
46.6
47.3
47.3
47.7
49.1
49.5
49.5
50
51
51.6
52.2
53.4
Mass
Protein
No.
10943
10943
6648
6276
12321
17279
9399
9270
18449
17623 17779
18448
15667
12122
15644
8087
12465
23922
23922
27908
27908
14164
18343
14421
15465
15466
15727
L37
L37
S30
L39
L36a(L44)
L26
S27a
S27a; cleaved C-term.
L21 NG to KR
L24 & L24 cleaved C-term.
L21
L27
L36
L28
L38
L35a
L13 (terminal KK)
L13 (terminal KK)
L8
L8
L31
S11
L35
S24
S24
L32
23
24
25
26
27
28
29
30
31
32
33
34
35
35
36
37
38
39
40
41
42
43
44
45
46
RT
[min]
53.9
54.5
55.3
55.3
56.2
56.7
56.7
57.8
60
60.8
61.4
63.2
63.2
63.6
64.5
64.5
65.8
66.8
67.4
69.5
71.6
72.5
75.3
77
78.5
Mass
Protein
28680
23191 23647
23345
16503
24015
9170
14776
15954
13284
21527
29464
29862
16314
16314
29466
15379
17091
17629
22169
22460
14708
21893
11772
30355
26585
S6
L14 (native & with mod)
L13a
L27a
L15
S21, N-acetylmethionine
L23 N-acetylserine
S19
S20
L18
S4
L7a
S16
S16
S4
S17
S13
S18 N-acetylserine
S7 N-acetylmethionine
S9
S15a
L9
P2
L7
S3 N-acetylalanine
This selection guide can be viewed as an electronic book at sge.com/selectionguide
BPI/EIC Chromatograms of BSA Tryptic Peptides on ProteCol™ C18 HQ303
Column Part No.:
250137
Gradient:
0 min
0% B (loading)
50% B
Sample:
BSA Tryptic Digest
8 min
Column:
ProteCol™ C18 HQ303
100 x 0.3 mm
26 min 80% B
LC:
Dionex Ultimate 3000
Solvent A:
0.1% Formic Acid
Solvent B:
0.1% Formic Acid in
Acetonitrile
27 min 80% B
37 min 80% B
38 min 0% B
45 min 0% B
Detection:
Bruker HCT ion trap
Mascot Search via Biotools:
Mascot Score: 1481
Sequence coverage:
71.5
2 pmol of sample injected,
(130 ng)
HPLC Columns and
Applications
Two hydrophilic marker peptides usually not reproducibly detected on different column materials are reproducibly retained and detected using
ProteCol™ C18 HQ303 column. Green and blue: Overlaid BPI chromatograms of 2 consecutive runs of 2 pmol of BSA (~130 ng) red and orange:
extracted ion chromatograms (EIC) of the two respective peptides.
HPLC Applications by Industry
215
HPLC Application by Industry | Life Sciences
BPI/EIC Chromatograms of Human α1-proteinase Inhibitor Tryptic Peptides on
ProteCol™ C18 HQ303
HPLC Columns and
Applications
Column Part No.:
250137
27 min 80% B
Sample:
Tryptic Digest of Human
α1-proteinase Inhibitor
37 min 80% B
Column:
ProteCol™ C18 HQ303
100 x 0.3 mm
LC:
Dionex Ultimate 3000
Solvent A:
0.1% Formic Acid
Solvent B:
0.1% Formic Acid in
Acetonitrile
Gradient:
0 min
0% B (loading)
8 min
50% B
38 min 0% B
45 min 0% B
Detection:
Bruker HCT ion trap
Mascot Search via Biotools:
Mascot Score: 1481
Sequence coverage:
64.9% (does not include the
glycopeptides that comprise
21% of the sequence).
10 pmol of sample injected,
(500 ng)
26 min 80% B
Tryptic peptides reflecting the protein termini are detected as well separated peaks, though the N-terminal peptide is isocratically
separated under loading conditions. The different glycopeptides carrying mainly disialylated, biantennary N-glycans as described
previously are well separated. Grey: BPI of A1PI tryptic peptides, other colors: Extracted ion chromatograms (EIC) of the respective
peptides as indicated.
216
More chromatograms and application information can be found at
sge.com/documents/chromatogram-library
HPLC Application by Industry | Pharmaceutical
TP-0190-H | NIST SRM 870 on ProteCol™ C18 HQ105
Column Part No.:
250100
Sample:
Uracil (28 µg/g),
Toluene (1400 µg/g),
Ethylbenzene (1700 µg/g),
Quinizarin (94 µg/g),
Amitriptyline (2800 µg/g)
in methanol
Column:
ProteCol™-P C18 HQ105
250 mm x 4.6 mm ID
Injection Volume:
1 µL
Mobile Phase:
4 mM phosphate pH7.0 in 80
% methanol
Flow rate.:
1.0 ml/min
Temperature:
23 °C
Detection:
254 nm
LC System
Shimadzu Prominence 20 AC
HPLC Columns and
Applications
ProteCol™-P C18 HQ105
Average competitor
Type I silica
This figure shows the chromatogram achieved under the described conditions in comparison with a competitor’s column and a column
packed with type I silica.
HPLC Applications by Industry
HPLC Application by Industry | Pharmaceutical
TA-0135-M | The Extraction and Analysis of Urinary Antitussive Metabolites
using MEPS™ and ESI-LCMSn on ProteCol™-P C18 HQ105
Abstract
HPLC Columns and
Applications
Urine specimens were first hydrolysed with
beta-glucuronidase and then the analytes
of interest were extracted using MEPS™
prior to analysis on a ProteCol™-P C18
HQ105 column using a 1 % v/v aqueous
acetic acid – methanol mobile phase.
Detection of the target analytes was by ESIMSMS with collision parameters selected
for specific analytes. The MEPS™ phases
were nominally 50 µm silicas modified with
C18, C8 or SCX chemistries. Speculative
structural elucidation of metabolites was
possible by mass fragmentography and
MS3 or MS4 as required.
Sample preparations
3 mL aliquots of urine from the samples
collected at 0 and 2 hours after oral
administration were diluted with 0.1 M
phosphate buffer (pH 6.0, 4.5 mL) and
the pH adjusted to 6.2-6.3. The samples
were then enzyme hydrolyzed with betaglucuronidase for 2 hours at 50 °C. Samples
were extracted by either a conventional
mixed mode SPE method or by a reversedphase MEPS™ method.
SPE extraction was performed on Bond-Elut
Certify™ columns using methods described
previously. (Wynne PM, Batty DC, Vine JH
and Simpson NKJ., Chromatographia, 59
(4/5), S50-S61, (2004)).
Experimental
Administration and Sample Collection
Oral administration of a single dose of:
10 mL Vicks® Cough Syrup (equivalent to
pentoxyverine (carbetapentane) citrate 15
mg), 10 mL Robitussin® DX Dry Cough Forte
syrup (equivalent to 30 mg dextromethorphan
hydrobromide) 2 Mersyndol® day strength
tablets (equivalent to paracetamol 1000 mg
and codeine phosphate 19.2 mg). Naturally
voided urine samples were collected at 0, 2,
3 and 4 hours following administration. Urine
samples were stored frozen at -20 °C until
required for analysis.
218
MEPS™ extraction was performed on C18
MEPS™ BINS fitted to a 100 µL MEPS™
syringe. MEPS™ BINS were conditioned
sequentially with 50 µL methanol and 100
µL water.
• 50 µL methanol conditioning.
• 100 µL water conditioning.
• 1 mL sample was drawn and expelled in
80 µL steps.
• 80 µL water wash.
• 50 µL sodium tetraborate pH adjustment.
• 80 µL water wash.
• 2 x 80 µL air drying.
• 2 x 20 µL methanol elution.
• 10 µL iso-propanol elution.
This selection guide can be viewed as an electronic book at sge.com/selectionguide
1.2E+07
Sample:
Uracil (28 µg/g),
Toluene (1400 µg/g),
Ethylbenzene (1700 µg/g),
Quinizarin (94 µg/g),
Amitriptyline (2800 µg/g)
in methanol
Column:
ProteCol™-P C18 HQ105
150 mm x 4.6 mm ID
Injection Volume:
10 µL
Mobile Phase A:
1 % aq. acetic acid in 10 %
methanol
1.0E+07
6E+06
8.0E+06
2.5E+06
240
250
260
270
280
290
288.00
250102
271.99
Column Part No.:
6.0E+06
4.0E+06
240
250
260
270
280
290
2.0E+06
1 % aq. acetic acid in 90 %
methanol
Flow rate.:
0.7 ml/min
Gradient:
20 min 0 to 100 % B
10 min at 100 % B
Temperature:
40 °C
0
5
10
*
*
*
15
*
20
25
30
1.8E+06
258.06
Mobile Phase B:
0E+00
5E+06
HPLC Columns and
Applications
H-N
O-CH3
1E+07
240
260
300
272.06
CH3
H-N
Cytochrome P450 2D6
Cytochrome P450 3A4
O-CH3
Dextromethorphan
(parent compound)
2E+07
240
3E+07
270
CH3
280
290
?
OH
H-N
OH
H-N
3E+07
260
H-N
OH
240
3E+07
250
260
270
240
274.00
244.09
OH
Dextrorphan
(active drug)
258.13
Thermo LCQ Classic positive
ion mode
250
1.8E+06
CH3
Detection:
280
3E+06
2E+07
250
260
270
280
290
Metabolites of Dextromethorphan
5E+06
4E+06
3E+06
5.0E+06
hydroxy metabolite (M+1=350)
hydroxy desethyl metabolite (M+1=322)
hydroxy metabolite (M+1=350)
dihydroxy metabolite (M+1=366)
dihydroxy desethyl metabolite (M+1=348)
dihydroxy metabolite (M+1=366)
hydroxy desethyl metabolite (M+1=348)
hydroxy metabolite (M+1=350)
hydroxy metabolite (M+1=350)
dihydroxy metabolite (M+1=366)
hydroxy metabolite (M+1=350)
didesethyl metabolite (M+1=278)
desethyl metabolite (M+1=306)
parent (M+1=334)
hydroxy metabolite or possible N-oxide (M+1=350)
O
C-OCH2CH2OCH2CH2-N
C2H5
C2H5
(#14: M+1=334)
1
3.5E+05
4
4.5E+06
HO
286.08
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
=
6E+06
III
N
O
CH3
H
HO
4.0E+06
14
Morphine (M=285.34)
270
3.5E+06
280
290
300
310
320
2
1.6E+06
H3C
3.0E+06
N
O
OH
OH
HO
2.5E+06
Hydroxycodeine (M=285.34)
270
280
290
300
310
320
2.0E+06
2E+06
1.5E+06
8
5
2
1E+06
1.0E+06
15
3
1
4,5
6
7
9 10
11
int.Std.
12
13
10
11
12
13
14
15
0.0E+00
16
1
5.0E+05
0E+00
17
18
19
0
2,3
5
10
15
20
25
3
1.6E+06
5.0E+05
H3C
1E+06
9
7
5
11
10
12
Hydroxycodeine (M=285.34)
1.5E+06
6
1
CH3
OH
H
HO
4
2E+06
N
O
1.0E+06
int.Std.
270
280
290
300
310
320
4
6E+06
15
3E+06
H3C
2.0E+06
N
O
3
2
30
20
13
8
CH3
H
HO
Codeine (M=285.34)
2.5E+06
270
4E+06
280
290
300
310
286.03
H3C
N
O
5E+06
3.5E+06
14
6E+06
Metabolites of Carbetapentane
4.0E+06
320
5
1.6E+06
3.0E+06
Signals are related to
carbetapentane which was
co-administrated
OH
H
HO
Norcodeine (M=285.34)
270
280
290
300
310
320
Metabolites of Codeine
HPLC Applications by Industry
219
HPLC Columns and
Applications
HPLC Supplies and Accessories
MEPS™ Micro SPE
ProteCol™ In-Line Filters
In-line Filter
PEEKsil™ Tubing Kits
PEEKsil™ Injection Loop
EasyLok™ Fittings
Hexnut™ Fittings
HPLC Column Replacement Frits
ProteCol™ Unions, Ferrules and Fittings
High Efficiency HPLC Column Couplers
Detector/Tubing Unions
ProteCol™ Accessory Kit
Guard Cartridge Systems
Solvent Filter
SilFlow™ - New Splitter Technology
ProteCol™ MicroFlow Meter
Microflow™ Meter
Using SGE fittings and connections to test ProteCol™ HPLC Columns
222-225
226
226
226
227
227
227
228
228
229
229
230
230
231
231
232
232
HPLC Supplies | MEPS™ Micro SPE
MEPS™ (Micro Extraction by Packed
Sorbent) has been created by SGE to
overcome the limitations of conventional
sample preparation methods. It has been
estimated that up to 75% of labor time
in a typical analytical laboratory workflow
is consumed in preparing and processing
samples prior to their analysis. Therefore,
any improvement in sample preparation
efficiency will increase sample throughput and deliver important time savings for
busy laboratories. Below is a comparison
of MEPS™ with other common sample
preparation techniques.
Comparison of Sample Preparation Methods
HPLC Supplies and
Accessories
Method Description
LLE
(Liquid-Liquid Extraction)
SPE
(Solid Phase Extraction)
MEPS™ (Micro Extraction by
Packed Sorbent)
• Relies on the difference
in solubility of analytes in
immiscible liquids.
• The organic phase is immobilized
on a stationary phase in a
cartridge.
• One
phase
is
aqueous
(hydrophilic) and the other is an
organic (hydrophobic) solvent.
• A suitable organic solvent mixture
is used to selectively elute the
compounds of interest.
• A miniaturized form of SPE with
a reduced stationary phase
(3mg) integrated into a high
quality SGE analytical syringe.
• Stationary phase does not move,
sample and solvents must be
moved to the cartridge.
• Stationary phase moves with
the syringe which aspirates and
dispenses the sample.
• Can be used to remove unwanted
interfering compounds.
• Can be used to remove unwanted
interfering compounds and to
increase sample concentration.
Sample Volume Required
• Generally large e.g. 10-100 mL.
• Small e.g. 3 mL
• Micro e.g. 50 µL.
Evaporation Step
• Solvent evaporation required to
increase sample concentration
to a level that can be analyzed.
• In the majority of cases, solvent
evaporation required to increase
sample concentration to a level
that can be analyzed.
• Evaporation
required.
Solvent Used
• Large solvent volumes required
e.g. 10-100 mL.
• Large solvent volumes required
e.g. 10 mL.
• Micro e.g. 500 µL.
Time
• Slow and labor intensive.
• Evaporation step can be time
consuming.
• Fast e.g. minutes, as one step
washes, loads and elutes.
Automation
• Entire process
automated.
• Can be automated.
• Can be fully automated.
Price
• High solvent
discard costs.
cannot
be
purchase
and
• High solvent
discard costs.
purchase
and
• Low solvent
discard costs.
generally
purchase
not
and
MEPS™ (Micro Extraction by Packed Sorbent)
is a micro SPE solution that incorporates
the stationary phase in a micro-cartridge
integrated in a high quality SGE analytical
syringe (Barrel Insert and Needle - BIN
configuration). MEPS™ is the miniarization
of conventional SPE packed bed devices
from mL to µL bed volumes.
MEPS™ stationary
C2, C8, C18, Silica
phases
available:
Schematic of the MEPS™ stationary phase within the syringe needle
– SGE’s patented ‘Barrel Insert and Needle’ (BIN) configuration.
To view a demonstration of MEPS™ visit sge.com/support/videos
How To Use MEPS™
• Step 1: Pump the sample
through the MEPS™ BIN (one
or more volumes may be
taken).
• Step 2: Wash the MEPS™
BIN once by pumping 20 µL
to 50 µL of wash solution
through the BIN to remove
interferences.
• Step 3: Elute the analyte by
drawing solvent through the
BIN into the syringe barrel.
• Step 4: Inject the analyte
directly into the injector.
• Pump 50 µL solvent followed
by 50 µL wash solution to
prepare BIN for the next
sample.
STEP
1
STEP
2
STEP
3
STEP
4
HPLC Supplies and
Accessories
SAMPLING
(1-N TIMES)
WASHING
ELUTION
SOLVENT
INJECTION
MEPS™ Is Reusable
Like conventional SPE, the number of times
the cartridge can be reused is dependent
on the sample matrix and the cleaning
regime between elutions. Since only 3mg of
stationary phase is used in MEPS™ it can be
washed effectively between each extraction
without the need for large solvent volumes.
For simple applications, MEPS™ devices have
been used successfully for up to 50 cycles
making it very cost effective.
MEPS™ Can Be Semi or Fully Automated
Semi-automation of MEPS™ can be achieved
by coupling MEPS™ syringes to SGE’s eVol®
automated analytical syringe to speed up
repetitive SPE and is ideal for rapid method
development. For more information on
eVol® see pages 22-24.
MEPS™ can also be fully automated on
autosamplers such as the CTC PAL for online SPE and injection.
MEPS™ Has Proven Accuracy and Precision Compared to
Other Sample Preparation Methods
Method
Ropivacaine
LOD (nM)
Accuracy (%)
MEPS™ / GC-MS
2
105
5.0
1 min
LLE / GC-MS
2
101
3.8
20 min
SPE / LC-UV
100
101
3.0
20 min
5
110
6.3
40 min
SPME / GC-MS
Precision (RSD%)
(Inter-assay)
Handling
Time
Comparison of accuracy and precision between MEPS™ and other methods for ropivacaine (local anesthetics).
HPLC Supplies | MEPS™ Micro SPE
223
HPLC Supplies | MEPS™ Micro SPE
MEPS™ Applications and Publications
MEPS™ micro SPE format is ideally suited to previously challenging applications. For an
example of MEPS™ used in a forensic application see page 12.
HPLC Supplies and
Accessories
Industry
Title
Author
Journal
Environmental
Determination of organic priority pollutants and emerging
compounds in wastewater and snow samples using multiresidue
protocols on the basis of microextraction by packed sorbents
coupled to large volume injection gas chromatography-mass
spectrometry analysis.
Prieto et al
J Chrom A,
2010,
1217:
6002-6011
Forensic and Pharmaceutical
Liquid chromatographic analysis of oxcarbazepine and its
metabolites in plasma and saliva after a novel microextraction by
packed sorbent procedure.
Saracino et al
Anal Chim Acta,
2010, 661: 222228
Environmental
At-line microextraction by packed sorbent-gas chromatography–
mass spectrometry for the determination of UV filter and
polycyclic musk compounds in water samples.
Moeder et al
J Chrom A, 2010,
1217:2925-2932
Forensic
Contribution of microextraction in packed sorbent for the
analysis of cotinine in human urine by GC–MS.
Lafay et al
Anal
Bioanal
Chem,
2010,
396: 937–941
General Chemistry and
Life Science
Recent advances in microextraction by packed sorbent for
bioanalysis.
Abdel-Rehim
J Chrom A,
2010,
1217:
2569-2580
Forensic
Rapid identification and quantification of methamphetamine and
amphetamine in hair by gas chromatography/mass spectrometry
coupled with micropulverized extraction, aqueous acetylation
and microextraction by packed sorbent.
Miyaguchi et al
J. Chrom A,
2009,
1216:
4063–4070
General Chemistry and
Life Science
Fully Automatic Sample Treatment by Integration of
Microextraction by Packed Sorbents into Commercial Capillary
Electrophoresis−Mass Spectrometry Equipment: Application to
the Determination of Fluoroquinolones in Urine.
Morales- Cid
et al
Anal.
Chem.,
2009, 81: 3188–
3193
Forensic
Screening of Cocaine and Its Metabolites in Human Urine
Samples by Direct Analysis in Real-Time Source Coupled to Timeof-Flight Mass Spectrometry After Online Preconcentration
Utilizing Microextraction by Packed Sorbent.
Jagerdeo
E,
Abdel-Rehim M
J Am Soc Mass
Spectrom. 2009
May;20(5):891899
Food and Flavour
Determinationof2,4,6-Trichloroanisoleand2,4,6-Tribromoanisole
in Wine using Microextraction in Packed Syringe and Gas
Chromatography−Mass Spectrometry.
Jönsson et al
J. Agric. Food
Chem., 2008,
56: 4962–4967
General Chemistry
Study of the factors affecting the performance of microextraction
by packed sorbent (MEPS) using liquid scintillation counter and
liquid chromatography-tandem mass spectrometry.
Altun
and
Abdel-Rehim
Anal Chim Acta.
2008, 630:116123
Pharmaceutical
MEPS™ as a rapid sample preparation method to handle unstable
compounds in a complex matrix: determination of AZD3409 in
plasma samples utilizing MEPS™-LC-MS-MS.
Abdel-Rehim M
et al
J
Chromatogr
Sci.
2008
46:518-523
Pharmaceutical and
Life Science
Rapid and Sensitive Method for Determination of
Cyclophosphamide in Patients Plasma Samples Utilizing
Microextraction by Packed Sorbent Online with Liquid
Chromatography-Tandem Mass Spectrometry (MEPS™-LC-MS/
MS).
Said et al
J. Liquid Chromatography &
Related
Technologies 2008,
31: 683–694
This selection guide can be viewed as an electronic book at sge.com/selectionguide
MEPS™ Syringe Options
All syringes may be used manually as well as with the listed autosamplers.
Description
100 µL Removable needle MEPS™ syringe for CTC Analytics, HTA 300A Plus & Varian 8400 systems.
Replacement plunger assembly for 005291.
250 µL Removable needle MEPS™ syringe for CTC Analytics, HTA 300A Plus & Varian 8400 systems.
Replacement plunger assembly for 006291.
250 µL Removable needle MEPS™ syringe for CTC Analytics systems.
Replacement plunger assembly for 006292.
100 µL Removable needle MEPS™ syringe for Agilent systems.
Replacement plunger assembly for 005292.
250 µL Removable needle MEPS™ syringe for Agilent systems.
Replacement plunger assembly for 006293.
100 µL Removable needle MEPS™ syringe for Shimadzu systems.
Replacement plunger assembly for 005293.
250 µL Removable needle MEPS™ syringe for Shimadzu systems.
Replacement plunger assembly for 006294.
# per Pack
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Part No.
005291
031826
006291
031831
006292
031831
005292
0318263
006293
0318303
005293
0318274
006294
0318305
HPLC Supplies and
Accessories
MEPS™ Barrel Insert and Needle (BIN) Assembly Options
For GC applications, needle is 23 gauge, 0.63 mm OD, Cone point style.
Description
MEPS™ BIN for CTC Analytics, HTA 300A Plus & Varian 8400 systems
C18
Silica
C8+SCX*
C2
C8
MEPS™ Development kit (contains 1 each of C18, C8, C2, SILICA and C8+SCX)
MEPS™ BIN for CTC Analytics systems using 250 µL syringes
C18
Silica
C8+SCX*
C2
C8
MEPS™ Development kit (contains 1 each of C18, C8, C2, SILICA and C8+SCX)
MEPS™ BIN for Agilent systems
C18
Silica
C8+SCX*
C2
C8
MEPS™ Development kit (contains 1 each of C18, C8, C2, SILICA and C8+SCX)
MEPS™ BIN for Shimadzu systems
C18
Silica
C8+SCX*
C2
C8
MEPS™ Development kit (contains 1 each of C18, C8, C2, SILICA and C8+SCX)
For Use with
MEPS™ Syringe P/N
# per Pack Part No.
005291 and 006291
005291 and 006291
005291 and 006291
005291 and 006291
005291 and 006291
005291 and 006291
5
5
5
5
5
1
2900101
2900102
2900103
2900104
2900106
2900105
006292
006292
006292
006292
006292
006292
5
5
5
5
5
1
2900301
2900302
2900303
2900304
2900306
2900305
005292 and 006293
005292 and 006293
005292 and 006293
005292 and 006293
005292 and 006293
005292 and 006293
5
5
5
5
5
1
2900601
2900602
2900603
2900604
2900606
2900605
005293 and 006294
005293 and 006294
005293 and 006294
005293 and 006294
005293 and 006294
005293 and 006294
5
5
5
5
5
1
2900601
2900602
2900603
2900604
2900606
2900605
For Use with
MEPS™ Syringe P/N
# per Pack Part No.
005291 and 006291
005291 and 006291
005291 and 006291
005291 and 006291
005291 and 006291
005291 and 006291
005291 and 006291
005291 and 006291
5
5
5
5
5
5
5
1
2900401
2900402
2900403
2900404
2900406
2900408
2900409
2900405
006292
006292
006292
006292
006292
006292
006292
006292
5
5
5
5
5
5
5
1
2900501
2900502
2900503
2900504
2900506
2900508
2900509
2900505
For LC Applications, needle is 22 gauge, 0.72 mm OD.
Description
MEPS™ BIN for CTC Analytics, HTA 300A Plus & Varian 8400 systems
C18
Silica
C8+SCX*
C2
C8
SCX
SAX
MEPS™ Development kit (contains 1 each of C18, C8, C2, SILICA and C8+SCX)
MEPS™ BIN for CTC Analytics systems using 250 µL syringe
C18
Silica
C8+SCX*
C2
C8
SCX
SAX
MEPS™ Development kit (contains 1 each of C18, C8, C2, SILICA and C8+SCX)
Base material is silica with mean particle size of 45 µm and pore size of 60 Å. *C8+SCX BINS are labelled as M1.
HPLC Supplies | MEPS™ Micro SPE
HPLC Supplies | ProteCol™ In-Line Filters
Particulate Protection for
Your Columns
SGE fittings, guard
cartridges and in-line
filters are completely
compatible with all
other manufacturers' HPLC columns
and systems.
HPLC Supplies and
Accessories
• Zero dead volume filter design.
• Zero pressure drop across filter.
• Zero compromise on performance.
The ProteCol™ In-Line Filter is a simple
and effective way to protect your capillary
columns from particulates. It protects your
system from blockages and increased back
pressures without introducing peak tailing
or loss of resolution. The filtering element
Description
is a 2 micron porosity screen, located
between the square-cut and polished ends
of two lengths of PEEKsil™ tubing (‘tails’). The
construction of the in-line filter minimizes
dead volume, providing optimum column
performance. In-line filters are available
with both 1/16" and 1/32" O.D. PEEKsil™
tails, providing versatility in connectivity and
flexibility. The ends of the tails are precisely
square cut and polished to achieve zero
dead volume connections using ProteCol™
Unions.
Inlet
Length
(mm)
Outlet
Length
(mm)
Tail ID
(µm)
Tail OD
Pack Size
Part No.
212005
ProteCol™ Filtering Connector 1/16"
50
50
50
1/16"
3
ProteCol™ Filtering Connector 1/32"
50
50
50
1/32"
3
222012
ProteCol™ 1/32" In-line Filter Kit Includes: 3 x 1/32"
In-Line Filters (P/N 222012), 2 x PEEK™ Fingertight
fittings for 6-40UNF port (P/N 222017), 2 x PEEK™
Fingertight fittings for 10/32 port (P/N 222002).
50
50
50
1/32"
See
description
222016
HPLC Supplies | In-line Filters
• Inert – biocompatible.
• Utilizes a replaceable 0.5 µm porosity frit
assembly.
• Simple, cost effective means of sample
filtration suitable for pre-column use.
• Low dead volume design eliminates
peak broadening and enhances column
resolving power.
• Supplied with Hexnuts™.
Description
Pack Size
Part No.
In-line Filter with a 0.5 µm Frit
(supplied with holder)
1
204002
Replacement In-line Filter Frit
Assembly
1
202009
If you can't pre-filter samples, make sure to use a low
dead volume in-line filter after the injector.
HPLC Supplies | PEEKsil™ Tubing Kits
Description
Pack
Size
Part No.
1/16" tubing kit (50 µm I.D. 100, 200,
and 300 mm lengths)
5
212012
1/32" tubing kit (50 µm I.D. 100, 200,
and 300 mm lengths)
5
222008
For other lengths of PEEKsil™ tubing see PEEKsil™ on page 239.
226
HPLC Supplies | ProteCol™ In-Line Filters, In-line Filters and PEEKsil™ Tubing Kits
HPLC Supplies | PEEKsil™ Injection Loop
• Smooth, non-reactive internal bore
reduces sample carry-over and sample
rinse times.
• Reduced sample loss due to shear or
adsorption.
• Maximum operating pressures are 3000 psi
for 10 µL, and 20 µL loops.
• Supplied with PEEKsil™ tubing and two
sets of stainless steel Hexnuts™ with
PEEK™ ferrule fittings.
• Compatible with most valves including
Rheodyne® and Valco®.
Volume
PEEKsil™ID
Length
Pack
Size
Part No.
10 µL
0.22 mm
(0.009")
26 cm
(10.24")
1
0650010
20 µL
0.22 mm
(0.009")
53 cm
(20.87")
1
0650020
For complete loop
fill, the syringe capacity should be
greater than twice
the loop volume.
The loop capacity
sets the injection
volume. For partial
loop fill, the injection volume should
be no greater than
half the loop capacity. The injection
size sets the injection volume.
HPLC Supplies and
Accessories
HPLC Supplies | EasyLok™ Fittings
EasyLok™ fittings are composed of a
knurled stainless steel nut and a double
ended PEEK™ ferrule. The PEEK™ ferrule
simply slides over any 1/16" OD tubing
to its required position, while the nut is
finger tightened. Unlike stainless steel, the
PEEK™ ferrule will not crush the tubing and
can be easily readjusted for quick column
changes.
The fittings are compatible with any
standard female HPLC fitting including
Swagelok®, Parker™, Waters®, Valco® and
Whatman®.
EasyLok™ fittings are recommended for use
with SGE GLT™ columns.
SPECIFICATIONS
Maximum Pressure Rating
5,000 psi
Thread Type
The unique double ended ferrule design
seals at two points to prevent leaks.
10-32
Description
Pack Size
Part No.
EasyLok™ Fitting
2 fittings,
2 ferrules
206102
10
206160
PEEK™ Double Ended Ferrules
For quick and
easy HPLC column
connections use
EasyLOK™ nuts.
HPLC Supplies | Hexnut™ Fittings
• Stainless steel 10-32 thread fittings use a
non-swaging Kel-F® or PEEK™ replaceable
ferrule.
• Inert contact surfaces make them
biocompatible.
• Ideal for applications where corrosive
solvents are being used.
• Hexnut™ with Kel-F® or PEEK™ ferrules are
recommended for use with SGE GLT™
columns.
Specifications
Maximum Pressure Rating
5,000 psi
Thread Type
10-32
Description
Pack Size
Part No.
Stainless Steel Hexnuts™ and PEEK™
Ferrules
2 hexnuts,
2 ferrules
1021003
Stainless Steel Hexnuts™ and PEEK™
Ferrules
10 hexnuts,
10 ferrules
1021011
Kel-F® Ferrules
10
0730010
Replacement PEEK™ Ferrules
2
0730004
Replacement PEEK™ Ferrules
10
0730014
HPLC Supplies | PEEKsil™ Injection Loop, EasyLok™ Fittings and Hexnut™ Fittings
227
HPLC Supplies | Column Replacement Frits
Accumulated impurities during a column's
life may block the column termination
frits, making a replacement necessary. Frit
porosity must be chosen to retain particles.
Select appropriate replacement frit for
column type and packing size.
Description
Pack Size
Part No.
1 mm ID micro column frit assembly
(3 µm packing)
2
202016
1 mm ID micro column frit assembly
PEEK®-PTFE frit
2
2020155
4 mm ID GLT ™ column frit assembly
(3 µm packing)
2
202002
2 mm ID GLT ™ column frit assembly
PEEK®-PTFE frit (5 µm packing)
2
2020035
Frit Removal Tool
1
200005
HPLC Supplies | ProteCol™ Unions, Ferrules and Fittings
HPLC Supplies and
Accessories
Perfect Connections,
Every Time
• Stainless steel unions can be inger
tightened or tightened with a 3/16"
wrench for high-pressure applications.
• PEEK™ unions can be finger tightened.
• Zero dead volume design for no peak
They are slightly larger than stainless
dispersion.
steel unions but also lighter - less stress
• Easytouse.
on your tubing.
• Completeversatility–allowsconnection
to the same or a range of different size Connectivity
tubing.
ProteCol™ Unions and reusable PEEK™
• AvailableinPEEK™ and stainless steel.
ferrules allow you to connect any
combinationof:
Stainless steel or PEEK™ unions
ProteCol™ Unions are available in either
stainless steel or PEEK™ (for all sizes). All use
the same range of reusable PEEK™ ferrules.
• 1/32"PEEKsil™.
• 1/16"PEEKsil™.
• 0.36mmfusedsilicatubing.
ProteCol™ Stainless Steel Unions
Description
Pack Size
Part No.
0.36 mm to 0.36 mm
2 unions and 4 PEEK™ Ferrules
222007
1/16" to 0.36 mm
2 unions and 4 PEEK™ Ferrules
212008
1/16" to 1/16"
2 unions and 4 PEEK™ Ferrules
212006
1/16" to 1/32"
2 unions and 4 PEEK™ Ferrules
212007
1/32" to 0.36 mm
2 unions and 4 PEEK™ Ferrules
222006
1/32" to 1/32"
2 unions and 4 PEEK™ Ferrules
222005
HPLC Supplies | Column Replacement Frits and ProteCol™ Unions, Ferrules and Fittings
ProteCol™ PEEK™ Unions
Description
Pack Size
Part No.
0.36 mm to 0.36 mm
2 unions and 4 PEEK™ Ferrules
222011
1/16" to 0.36 mm
2 unions and 4 PEEK™ Ferrules
212011
1/16" to 1/16"
2 unions and 4 PEEK™ Ferrules
212009
1/16" to 1/32"
2 unions and 4 PEEK™ Ferrules
212010
1/32" to 0.36 mm
2 unions and 4 PEEK™ Ferrules
222010
1/32" to 1/32"
2 unions and 4 PEEK™ Ferrules
222009
ProteCol™ PEEK™ Ferrules
Description
Pack Size
Part No.
0.36 mm to 0.36 mm
5
223007
1/16" to 0.36 mm
5
213008
1/16" to 1/16"
5
213006
1/16" to 1/32"
5
213007
1/32" to 0.36 mm
5
223006
1/32" to 1/32"
5
223005
PEEK™ Fingertight Fittings
Description
Pack Size
Part No.
5
2062752
1/16" to 10-32UNF
1/32" to 10-32UNF
5
222002
1/32" to 10-32UNF (long)
5
2062753
1/32" to 6-40UNF
5
222017
HPLC Supplies | High Efficiency HPLC Column Couplers
• Inert – biocompatible.
• Provide a rigid low dead volume,
metal free connection between two
components of an HPLC system.
• Ideal for connecting two HPLC columns, a
HPLC column to a guard cartridge or inline filter.
Specifications
Maximum Pressure Rating
5,000 psi
Thread Type
Description
Stainless Steel Hexnut™ Coupling Kit
10-32
Pack Size
Part No.
1
200009
5 x Replacement PEEKsil™ tubing
(1/16" x 175 µm ID x 50 mm)
Part No. 0624351
2 x Replacement PEEK™ Ferrules
Part No. 0730004
10 x Replacement PEEK™ Ferrules
Part No. 0730014
HPLC Supplies | Detector/Tubing Unions
• True zero dead volume unions enable
connection of existing detector and other
metal male fittings to the finger-tight
compatible SGE column and cartridge
systems.
• Available for Waters®, Swagelok®, Parker™,
Valco® and Rheodyne® male fittings.
Description
Pack Size
Part No.
Union for Waters and Rheodyne
tubing
1
2062746
Union for Swagelok®, Parker and
Valco tubing
1
2062747
HPLC Supplies | High Effi ciency Column Couplers and Detector/Tubing Unions
HPLC Supplies and
Accessories
HPLC Supplies | ProteCol™ Accessory Kits
Description
ProteCol™ 1/32" accessory kit includes:
Pack
Size
Part No.
1
222000
2 x 1/32" – 1/32" stainless steel union
3 x 1/32" – 1/32" double-end
PEEK™ ferrules
1 x in-line filter with 1/32" 50 µm x 50
mm PEEKsil™ tubing
1 x 1/32" 50 µm x 100 mm
PEEKsil™ tubing
1 x 1/32" 50 µm x 150 mm
PEEKsil™ tubing
Positioning toolkit (acrylic clamp,
wrench and depth gauge)
HPLC Supplies and
Accessories
HPLC Supplies | Guard Cartridge Systems
• SGE recommends that 4.0 mm ID Guards
be used to protect all 4.6 mm and 4.0 mm
ID HPLC columns.
• Vital for HPLC column protection.
• Inert, metal free - biocompatible flow
path.
• Improved biological and ion chromatography.
• Pre-packed in a range of packing
materials.
These inert, biocompatible cartridge
systems offer maximum efficiency to
protect the analytical column and begin the
separation process.
Always use a guard
cartridge to prolong
column life.
230
Each of these guard cartridges has an
effective length of 10 mm.
Guard Cartridge Holder
Description
Pack
Size
Part
No.
1
205000
Pack
Size
Part No.
Guard Cartridge Holder
Guard Cartridges
Description
Column ID
(mm)
Exsil™ ODS - 3 micron
4.0 and 4.6
3*
2050010
Exsil™ ODS - 5 micron
4.0 and 4.6
3*
205001
Exsil Silica - 5 micron
4.0 and 4.6
3*
205002
Exsil™ C8 - 5 micron
4.0 and 4.6
3*
205003
Exsil Amino - 5 micron
4.0 and 4.6
3*
205004
Exsil™ SCX - 5 micron
4.0 and 4.6
3*
205005
™
™
™
Exsil SAX - 5 micron
4.0 and 4.6
3*
205006
Exsil™ Cyano - 5 micron
4.0 and 4.6
3*
205007
Exsil Phenyl - 5 micron
4.0 and 4.6
3*
205008
Nucleosil® ODS - 5 micron
4.0 and 4.6
3*
2050014
™
* Holder not included
HPLC Supplies | ProteCol™ Accessory Kits and Guard Cartridge Systems
HPLC Supplies | Solvent Filter
• PTFE and porous glass membrane filter.
• Efficiently filters particles down to 1.2 µm
with minimal pressure drop.
• Filters solvent as it is drawn from the
reservoir.
Description
Solvent Filter (1 µm)
Pack Size
Part No.
1
204000
HPLC Supplies and
Accessories
HPLC Accessories | SilFlow™ - New Splitter Technology
Using SGE’s SilFlow™ technology of
microchannels in wafers, the next generation
of splitters is now available.
There are two configurations of wafers
developed to enable connecting flow lines
to external components in a GC or HPLC
system.
SilFlow™ offers low dead volume connections,
that are deactivated and can be easily
installed. For further information on SilFlow™
technology see page 182.
Configuration Types
SilFlow™ Splitters are available in 3 port
configuration for HPLC applications, and as
3 and 4 ports for GC. See page 185 for GC
applications.
Kit Contents
Each kit comes complete with: wafer,
fingertite
tool,
mounting
bracket,
appropriately sized ferrules and nuts, and
blanking ferrules to assist with set up.
Tubing Dimensions
Description
Port A
Port B
Port C
For
Tubing
Pack
Size
Part No.
SilFlow™ HPLC 3 Port Splitter
LC Kit (1/32)
1/32"
1/32"
1/32"
1/32
Kit
123740
LC Wafer (1/32)
1/32"
1/32"
1/32"
1/32
1
123741
Part No.
Replacement Parts
Description
For Tubing
Pack Size
SilFlow™ LC Nut 1/32"
1/32" OD
10
123708
SilTite™ Ferrule 1/32"
1/32" OD
10
073473
HPLC Supplies | Solvent Filter and SilFlow™
HPLC Accessories | ProteCol™ MicroFlow Meter
Accurate, stable flow rates are the key to
reproducible retention times, and using
the optimum flow rate for a column is
very important to achieve the best possible
performance.
Whether conducting research, or doing
routine analysis, good lab practice requires
documentation of flow rates for your
records or publication.
HPLC Supplies and
Accessories
The ProteCol™ MicroFlow Meter is a fast,
simple and reliable way to accurately
measure flow rates in Capillary HPLC and
for other low flow applications. The direct
measurement of flow provides definitive
confirmation of the actual flow rate through
the column and is the perfect HPLC system
companion whether pumps require flow
splitters or deliver low flow directly.
• Universal principle does not require
calibration for different solvents.
• Two versions cover the flow rate range for
HPLC columns ranging from microbore
to nanobore:
– Choose 0.2-10 µL/min meter for
columns 300 µm ID and below.
– Choose 2.5-100 µL/min meter for
columns 300 µm ID and above.
• Valuable tool for troubleshooting when
retention times fluctuate.
Description
Pack Size
Part No.
ProteCol™ MicroFlow Meter - 2 µL
flow path volume (0.2 - 10 µL/min)
1
222013
ProteCol™ MicroFlow Meter - 25 µL
flow path volume (2.5 - 100 µL/min)
1
222014
HPLC Accessories | MicroFlow™ Meter
• Designed to accurately measure flow
rates from the HPLC pump and column.
• Flow cells supplied are traceable to
international standards allowing the
calibration of HPLC pumps for GLP, GMP
and other protocols.
Description
Pack Size
Part No.
1
206370
MicroFlow Meter 50 µL, 250 µL
and 1 mL cells
1
206372
Replacement Flow Cell 250 µL
1
206377
Replacement Flow Cell 1 mL
1
206379
MicroFlow™ Meter 1 mL cell
™
HPLC Supplies | ProteCol™ MicroFlow Meter and MicroFlow™ Meter