Semi-micro organic SEC (GPC) Columns: KF-400HQ

Shodex organic SEC (GPC) columns have a history of more than twenty years and have been used numerous customers all over the world. The current series is of an improved type based on the long term experience and the latest technology.
Five types of GPC columns packed with different solvents are available to avoid solvent replacement. Moreover, custom columns packed with other solvents such as quinoline, o-dichlorobenzene, ethyl acetate, tetrachloroethane and dimethylacetamide are available uponyour request.

The following columns are availble as the standard column.

1) In-column solvents

THF KF-800, KF-600, KF-400HQ, LF series
Chloroform K-800 series
DMF KD-800 series
HFIP HFIP-800, HFIP-600 series
Toluene UT-800, HT-800, AT-806MS series

2) Types

Mixed gel columns Mixed gel type columns are packed with mixture of gels which are different in pore size are to have wide linear range in molecular weight calibration curves from oligomers to macromolecules.
The mixed gel columns have “M” at the end of the column name.
Linear type
columns (1)		 Linear type columns are specially designed to have wider linear range in molecular weight calibration curves than that of mixed gel columns. Linear type columns are highly recommended for correcting non-linear sections of molecular weight calibration curves of ordinary columns.
It is highly recommended that linear type columns than ordinary type columns or mixed gel columns be used when both of them are available.
The linear type columns have “L” at the end of the column name.
Linear type
columns (2)		 Linear type columns with special designed packing material.
Semi-micro GPC Columns Small volume type columns are of 4.6mm ID column. The internal volume of 4.6mm ID column is about one third of 8.0mm ID column.
Smaller particle size enables higher-resolution fractionation. Detection sensitivity is about four times higher than that of standard GPC columns. A single column can substitute for conventional analysis using two or more coupled columns, which shortens analysis time.
Downsized GPC Columns Small volume type columns are of 6.0mm ID column. The internal volume of 6.0mm ID column is about one third of 8.0mm ID column.
With small volume type column, by reducing the flow rate to one third, the same separation as with 8.0m ID columns can be obtained. And, the consumption of eluent will be also one third.
Small volume type columns are good for reducing the consumption of potentially-environmentally-damaging organic solvents.
High Temperature Columns The high temperature columns can be used for the GPC analysis at elevated temperature up to 230 °C.
Solvent-Peak Separation Column Solvent-peak separation columns delay the elution of low-molecular components and can be used to separate a solvent peak or other troublesome peak to realize accurate measurement of molecular weight distribution. The use of the columns with 805L, 806L, 806M and 807L columns is recommended.

KF-400HQ · About 1.5-times better separation performance than standard columns, with higher resolution
· About 4-times better sensitivity than that of standard columns, supporting high sensitivity analysis
· the amount od solvent used is reduced to about a third
· Improved applicability of solvent repracement

*Please refer to Replacement of In-Column Solvent
*The columns with ‘L’ at the end of the column name are mixed-gel columns capable of a wide molecular weight distribution range of samples.
*Use of the KF-400HQ series with semi-micro type devices is recommended.

(High performance semi-micro columns)
KF-400HQ series shipping solvent : Tetrahydrofulan (THF)

Product code Product name Plate number
(TPN/column)		 Exclusion limit
(Polystyrene)		 Particle size
(micron)		 Size(mm)
ID x Length
F6028111
F6028112
F6028114
F6028116
F6028118
F6028119
F6028122
F6700300		 GPC KF-401HQ
GPC KF-402HQ
GPC KF-402.5HQ
GPC KF-403HQ
GPC KF-404HQ
GPC KF-405LHQ
GPC KF-406LHQ
GPC KF-G
> 25,000
> 25,000
> 25,000
> 25,000
> 25,000
> 10,000
> 10,000
Guard column
 1,500
5,000
20,000
70,000
400,000
4,000,000
*(20,000,000)
–		 3
3
3
3
3
10
10
8		 4.6 x 250
4.6 x 250
4.6 x 250
4.6 x 250
4.6 x 250
4.6 x 250
4.6 x 250
4.6 x 10

*( ): Estimated value
For all products
Max. usable pressure: (KF-401HQ to KF-404HQ) 7MPa, (KF-405LHQ, KF-406L) 0.5MPa
Recommended flow rate: 0.2 to 0.3mL/min
Max. usable flow rate: 0.5mL/min
Max. usable temperature: 45deg-C

Please refer to the following tables for the classification of solvents in terms of capability of being replaced with original in-column solvent. However, frequent replacement of in-column solvent may shorten column life, and is, therefore, not recommended. We can replace the in-column solvent with another solvent such as o-dichlorobenzene, ethyl acetate, dimethylacetamide or methylpyrrolidon upon your request.

(Standard Organic SEC (GPC) columns)

(Downsized GPC columns and Semi-micro GPC columns)

(Linear Type : LF series)

These are calibration curves of GPC KF-401HQ to KF-404HQ.

Sample : PS, Polystyrene

Columns      : Shodex GPC KF-401HQ , KF-402HQ, KF-403HQ, KF-404HQ (4.6mmID*250mm each)
Eluent       : THF
Flow rate    : 0.3mL/min

These are calibration curves of GPC KF-405LHQ andKF-406LHQ.

Sample : PS, Polystyrene

Columns      : Shodex GPC KF-405LHQ x 2, KF-406LHQ x 2 (4.6mmID*250mm each)
Eluent       : THF
Flow rate    : 0.3mL/min
Column temp. : 40deg-C

Polystyrene oligomer was analyzed using organic solvent SEC (GPC) semi-micro columns, 2x GPC KF-402HQ. Polystyrene standards separated had average MW of 580, contained dimer (MW:266) to octamer (MW1098).

Sample : Styrene oligomer

Columns      : Shodex GPC KF-402HQ (4.6mmID*250mm) x 2               
Eluent       : THF               
Flow rate    : 0.3mL/min
Detector     : Shodex RI(small cell volume)
Column temp. : 25deg-C

Shodex GPC KF-400HQ series are used for “Semi-micro organic Size Exclusion Chromatography” and these are best suited to determine molecular weight distribution and analysis of low molecular weight materials in polymer. Semi-micro column, GPC KF-402HQ and conventional column KF-802 are comapred using styrene oligomer. In this data, we adjusted the linear velocity of two columns. From these chromatograms, each peak heights of KF-402HQ has increased by 3 to 4 times. It means the detection limits for them has decreased to one third or one fourth. Generally KF-400HQ series have 2 strong points.
1) The plate number has increased by 1.5 times or more than that of conventional column.
2) The consumption of eluent has decreased to less than one fourth.
It is one of important things to reduce consumption of eluent, this Shodex GPC KF-400HQ series is useful very much.

Sample : Styrene oligomer, 5micro-L

Columns      : Shodex GPC KF-402HQ (4.6mmID*250mm), KF-802 (8.0mmID*300mm)            
Eluent       : THF    
Flow rate    : 0.3mL/min (KF-402HQ), 1.0mL/min (KF-802)
Detector     : UV(254nm)(KF-402HQ; small cell volume, KF-802; conventional cell)
Column temp. : 25deg-C

Figure 1 shows the relationships between the flow rate and the theoretical plate number (TPN) when using an organic solvent SEC (GPC) semi-micro column, GPC KF-402HQ. With flow rate of over 0.2 mL/min, TPN of over 30,000 was obtained. While with flow rate less than 0.2 mL/min, TPN decreases due to the analyte being spread-out. Figure 2 shows the comparison between KF-402HQ and conventional GPC KF-802 on the height equivalent theoretical plate (HETP) vs. linear flow rate. The packed gels filled in KF-402HQ and KF-802 are 3 um and 6 um respectively. As shown in Figure 2, with smaller gel KF-402HQ has less influence from the linear velocity. This is especially beneficial for the separation of bigger molecules such as EPON1001.
Please refer to Effect of flow rate (2) (KF-402.5HQ + KF-403HQ) for an additional information.

HETP (Column length)/(Theoretical plate number)

Sample : 0.1% Dicyclohexyl phthalate, 5micro-L

Columns      : Shodex GPC KF-402HQ (4.6mmID*250mm), KF-802 (8.0mmID*300mm)             
Eluent       : THF              
Detector     : UV(254nm) (KF-402HQ; small cell volume, KF-802; conventional)
Column temp. : 25deg-C

The effect of flow rate was examined for organic solvent SEC (GPC) semi-micro columns, GPC KF-402.5HQ and KF-403HQ. Improved separation is observed at lower flow rate, especially for the analysis of large MW compound such as EPON1001.
Please refer to Effect of flow rate (1) (KF-402HQ) for an additional information.

Sample : 0.2% EPON1004, 10micro-L

Columns      : Shodex GPC KF-403HQ + KF-402.5HQ (4.6mmID*250mm each)
Eluent       : THF              
Detector     : Shodex RI (small cell volume)
Column temp. : 40deg-C

The effect of injection volume on theoretical plate numbers (TPN) is investigated for organic SEC semi-micro columns GPCKF-402HQ and KF-406LHQ. Injection volume of 5µL or less is appropriate for KF-402HQ and injection volume of 20µL or below is adequate for using 2 KF-406LHQ columns.

Sample : Dicyclohexyl phthalate

Columns      : Shodex GPC KF-402HQ, KF-406LHQ x 2 (4.6mmID*250mm each)
Eluent       : THF              
Flow rate    : 0.3mL/min
Column temp. : 25deg-C

Figure presents the relationship between the concentration of sample and the height equivalent theoretical plate (HETP) for organic solvent SEC (GPC) semi-micro columns, GPC KF-402HQ. Lower HETP is obtained with sample concentration < 2.5% for the analysis of a lower MW compounds such as dichlorohexylphthalate.
Please refer to Effect of Sample Concentration (2) (KF-403HQ + KF-402HQ) for the analysis of compound having a wide MW distribution, such as EPON1001.

Sample : Dicyclohexyl phthalate, 10micro-L

HETP (Column length)/(Theoretical plate number)

Column       : Shodex GPC KF-402HQ (4.6mmID*250mm)         
Eluent       : THF       
Flow rate    : 0.3mL/min
Column temp. : 40deg-C

The effects of sample concentration on separation of EPON1001 polymer were examined using organic solvent SEC (GPC) semi-micro columns, GPCKF-403HQ and KF-402HQ. No considerable decline in resolution was observed up to analysis of 5% (m/v) EPON1001.
Please refer to Effect of Sample Concentration (1) (KF-402HQ) for the analysis of sample with single component.

Columns      : Shodex GPC KF-403HQ + KF-402HQ (4.6mmID*250mm each)           
Eluent       : THF       
Flow rate    : 0.3mL/min
Column temp. : 40deg-C

The relationships between the sample concentration and their elution volumes were examined using organic solvent SEC (GPC) semi-micro column, GPC KF-406LHQ. The figure shows increased elution volumes when sample concentrations were increased. Also it shows that this influence is bigger for the larger compounds analysis even at the lower concentration ranges. The table shows recommended maximum sample concentrations and injection volumes based on analytes MW.

Sample : PS , 10micro-L
Maximum sample concentration and maximum sample load for KF-406

MW range Max.Sample conc.
(%)		 Max. Sample load
(micro-g)
～ 1,000
 2.0 200
10,000 ～ 50,000
 0.6 60
50,000 ～ 200,000
 0.2 20
200,000 ～ 500,000
 0.1 10
500,000 ～ 2,000,000
 0.05 5
2,000,000 ～ 5,000,000
 0.03 3
5,000,000 ～ 0.02 2 
Columns      : Shodex GPC KF-406LHQ (4.6mmID*250mm) x 2             
Eluent       : THF       
Flow rate    : 0.3mL/min
Column temp. : 40deg-C

Refer to Effect of Sample Concentration (KF-402HQ) and Effect of Sample Concentration (2) (KF-403HQ + KF-402HQ).

The effect of column temperature on HETP was examined for organic solvent SEC (GPC) semi-micro column, GPC KF-402HQ. There is not a considerable temperature influence for the analysis of relatively small MW compound such as dicyclohexylphthalate. However, improved separation can be expected for the analysis of larger MW compound analysis at higher column temperature conditions.

Sample : 0.05% Dicyclohexyl phthalate, 5micro-L

HETP　=　 (Column length)/(Theoretical plate number)

Columns   : Shodex GPC KF-402HQ (4.6mmID*250mm) x 2             
Eluent    : THF       
Flow rate : 0.3mL/min
Detector  : UV(254nm)

The chromatograms show the cell volume of UV and RI detectors affected the performance of the separetion. We set the cell volume of the detectors smaller for down sized chromatography. When the UV detector whose cell volume is 2.4 micro-L is used for the analysis, the plate number is ca. 10,000 higher than when the UV detector whose cell volume is 17.7 micro-L is used. (Fig.1)
We can say the same thing regarding RI detector. The cell volume of RI detector affects the plate number of the peaks. The plate number when RI-101 detector is used is a half of that when RI-104 designed for down sized chromatography is used. (Fig.2)
* When the detector for down sized chromatography is used, it is needed to prepeare the instruments including the columns and the injector designed for the down sized chromatography to get good performance.

Sample :
1. PS, Polystyrene (MW: 66,000)
2. PS, Polystyrene (MW: 3,250)
3. PS, Polystyrene (MW: 580)
4. Propylbenzene
5. Benzene

               (Fig.1)               (Fig.2)                 
Columns      : Shodex GPC KF-402HQ   Shodex GPC KF-402HQ (4.6mmID*250mm) x 2
Eluent       : THF                   THF               
Flow rate    : 0.3mL/min             0.3mL/min
Column temp. : 40deg-C               25deg-C

MW distribution of SRM 706 was measured using organic solvent SEC (GPC) semi-micro column, GPC KF-406LHQ and conventional organic solvent SEC (GPC) column, GPC KF-806M. Both columns provided good agreements even using different detector systems (Light scattering and sedimentation equilibrium) that provide absolute MW information as well as column fractionation method using Mw/Mn measurements.

MW distribution analysis of SRM706

Analysis method Mw x 104 Mn x 104 Mw/Mn
KF-406LHQ x 2
(4.6mmID*250mm)
 26.22 12.26 2.13
26.46 12.43 2.13
KF-806M
(8.0mmID*300mm)
 26.00 12.40 2.10
28.00 12.90 2.17
Light scattering 25.78 – –
Sedimentation equilibrum 28.81 – –
Column fractionation 2.10

A mixture of polystyrene standards was analyzed with GPC KF-401HQ to KF-404HQ each other and their chromatograms were compared.

Sample : 5micro-L
PS, n-Propylbenzene, Benzene

Columns      : Shodex GPC KF-401HQ, KF-402HQ, KF-403HQ, KF-404HQ (4.6mmID*250mm each) 
Eluent       : THF
Flow rate    : 0.3mL/min
Detector     : Shodex RI
Column temp. : 40deg-C

Antioxidant (Irganox1010) in Polystyrene was analyzed with GPC/MS using GPC KF-402HQ (a semi-micro organic SEC (GPC) column). Only a low molecule weight domain can be poured into a mass spectrometer (MS) by using GPC mode. Therefore, it is not necessary to pretreat.
Please refer to Simultaneous Analysis of Various Additives of Polymer in Polystyrene (GPC/MS)

Sample : 1ppm Irganox 1010 in 1000ppm Polystyrene, 5micro-L
1. Polystyren
2. Irganox 1010

(HPLC)
Instrument   : Agilent 1100
Column       : Shodex GPC KF-402HQ (4.6mmID*250mm) x 2
Eluent       : THF
Flow rate    : 0.3mL/min
Detector     : UV(210nm)
Column temp. : 40deg-C
(MS)
Instrument      : Thermo Quest LCQ Advantage
Ionization      : APCI
Capillary temp. : 180deg-C
Vaporizer temp. : 450deg-C
Source voltage  : 6000V
Sheath gas flow : 80L/min

Irganox 1010 in styrene foam cup for instant noodles was analyzed with GPC/MS using KF-402HQ (a semi-micro organic SEC (GPC) column). Only a low molecule weight domain can be poured into a mass spectrometer (MS) by using GPC mode. KF-402HQ can provide the high sensitivity analysis without pretreatment of samples.
Refer to Simultaneous Analysis of Various Additives of Polymer in Polystyrene (GPC/MS).

Sample : 0.1%, 5micro-L
1. Polystyren
2. Irganox 1010

(HPLC)
Instrument   : Agilent 1100
Column       : Shodex GPC KF-402HQ (4.6mmID*250mm) x 2
Eluent       : THF
Flow rate    : 0.3mL/min
Detector     : UV(210nm)
Column temp. : 40deg-C
 
(MS)
Instrument      : Thermo Quest LCQ Advantage
Ionization      : APCI
Capillary temp. : 180deg-C
Vaporizer temp. : 450deg-C
Source voltage  : 6000V
Sheath gas flow : 80L/min

Various additives of polymer in polystyrene were analyzed simultaneously with GPC/MS using GPC KF-402HQ (a semi-micro organic SEC (GPC) column).

Sample : 5micro-L
Irganox 1010
Irganox 1076
Irgafos 168
Tinuvin 234
BHT dimer
BHT

(HPLC)
Instrument   : Agilent 1100
Column       : Shodex GPC KF-402HQ (4.6mmID*250mm) x 2
Eluent       : THF
Flow rate    : 0.3mL/min
Column temp. : 40deg-C
 
(MS)
Instrument      : Thermo Quest LCQ Advantage
Ionization      : APCI
Capillary temp. : 180deg-C
Vaporizer temp. : 450deg-C
Source voltage  : 6000V
Sheath gas flow : 80L/min

Fatty acids were analyzed using GPC KF-402HQ ((a semi-micro organic SEC (GPC) column)).

Sample : CnH2n+1COOH, 10micro-L
1. Melissic acid (n=29) 0.1%
2. Cerotic aid (n=25) 0.1%
3. Behenic acid (n=21) 0.1%
4. Arachidic acid (n=19) 0.1%
5. Stearic acid (n=17) 0.1%
6. Palmitic acid (n=15) 0.1%
7. Myristic acid (n=13) 0.1%
8. Lauric acid (n=11)
9. n-Decanoic acid (n=9) 0.1%
10. n-Octanoic acid (n=7) 0.1%
11. n-Hexanoic acid (n=5) 0.3%
12. n-Valeric acid (n=4) 0.3%

Columns      : Shodex GPC KF-402HQ (4.6mmID*250mm) x 2
Eluent       : THF
Flow rate    : 0.3mL/min
Detetor      : Shodex RI(small cell volume)
Column temp. : 40deg-C

Phthalates and alkylbenzenes were analyzed using GPC KF-401HQ (a semi-micro organic SEC (GPC) column).

Sample :
1. Dioctyl phthalate
2. Dibutyl phthalate
3. Dipropyl phthalate
4. Diethyl phthalate
5. Dimethyl phthalate
6. n-Propylbenzene
7. Ethylbenzene
8. Toluene
9. Benzene

Columns      : Shodex GPC KF-401HQ (4.6mmID*250mm) x 2
Eluent       : THF
Flow rate    : 0.3mL/min
Detector     : UV(254nm)(small cell volume)
Column temp. : 40deg-C

The calibration curves for downsized organic SEC (GPC) columns using PMMA are shown. In the case of the analysis for polymer whose MW is 1,000,000 or less, GPC LF-604 or LF-404 is suitable. When it is needed to separate low MW polymer and high MW polymer and low MW polymer is the purpose for the analysis, GPC HFIP-603 is suitable.
Please refer to Comparison of Analysis of PMMA with Down-sized SEC Columns

Sample : PMMA, Poly(methyl methacrylate) standards, 50micro-L

Columns      : Shodex GPC LF-604 (6.0mmID*150mm) x 2
               Shodex GPC LF-404 (4.6mmID*250mm) x 2
               Shodex GPC KF-405LHQ (4.6mmID*250mm) x 2
               Shodex GPC KF-406LHQ (4.6mmID*250mm) x 2
               Shodex GPC HFIP-603 (6.0mmID*150mm) x 2
               Shodex GPC HFIP-606M (6.0mmID*150mm) x 2    
Eluent       : 5mM CF3COONa in HFIP
Flow rate    : 0.3mL/min
Detector     : Shodex RI(small cell volume)
Column temp. : 40deg-C

The calibration curves for downsized organic SEC (GPC) columns using PMMA are shown. In the case of the analysis for polymer whose MW is 1,000,000 or less, GPC LF-604 or LF-404 is suitable. When it is needed to separate low MW polymer and high MW polymer and low MW polymer is the purpose for the analysis, GPC HFIP-603 is suitable.
Please refer to Comparison of Calibration Curves for Down-sized SEC Columns (HFIP Eluent: PMMA)

Sample : 50micro-L
1. PMMA, Poly(methyl methacrylate) (MW: 1,944,000)
2. PMMA, Poly(methyl methacrylate) (MW: 281,700)
3. PMMA, Poly(methyl methacrylate) (MW: 79,250)
4. PMMA, Poly(methyl methacrylate) (MW: 13,300)
5. PMMA, Poly(methyl methacrylate) (MW: 1,960)

Columns      : Shodex GPC LF-604 (6.0mmID*150mm) x 2
               Shodex GPC LF-404 (4.6mmID*250mm) x 2
               Shodex GPC KF-405LHQ (4.6mmID*250mm) x 2
               Shodex GPC KF-406LHQ (4.6mmID*250mm) x 2
               Shodex GPC HFIP-603 (6.0mmID*150mm) x 2
               Shodex GPC HFIP-606M (6.0mmID*150mm) x 2    
Eluent       : 5mM CF3COONa in HFIP
Flow rate    : 0.3mL/min
Detector     : Shodex RI(small cell volume)
Column temp. : 40deg-C

Liquid paraffin was analyzed using GPC KF-401HQ (a semi-micro organic SEC (GPC) column).

Sample : 1% Liquid paraffin, 5micro-L

 Column       : Shodex GPC KF-401HQ (4.6mmID*250mm)
 Eluent       : Chloroform
 Flow rate    : 0.3mL/min
 Detector     : Shodex RI(small cell volume)
 Column temp. : 40deg-C