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IonPac NS1 Manual

Document No. 034024-06

Page 22 of 35

4.2.1 Water Soluble Vitamins by Ion Pairing / Ion Suppression Chromatography

Peaks

1.

Ascorbic acid

2.

Nicotinic acid

3.

Nicotinamide

4.

Riboflavin

5.

Pyridoxine

6.

PABA

7.

Folic acid

8.

Pyridoxamine

9.

Thiamine

10. Cyanocobalamin

Injection: 5 nmoL each

Figure 8

Water Soluble Vitamins by

Ion Suppression / Ion Pair Chromatography

4.2.2 Water Soluble Vitamins by Ion Suppression / Ion Pair Chromatography

Peaks

1.

Ascorbic acid

2.

Nicotinic acid

3.

Nicotinamide

4.

Riboflavin

5.

Pyridoxine

6.

PABA

7.

Folic acid

8.

Pyridoxamine

9.

Thiamine

10. Cyanocobalamin

Injection: 2 nmoL each

Column:

IonPac NS1-5µm

Eluent Conditions:

12 to 48% Acetonitile in 35 minutes
5 mM Octane Sulfonic Acid

Flow Rate:

1.0 mL/min

Detection:

UV 254 nm

Column:

IonPac NS1-5µm

Eluent Conditions:

12 to 56% Acetonitile
0 to 35 minutes
5 mM TBAOH

Flow Rate:

1.0 mL/min

Detection:

Suppressed Conductivity

Figure 7

Water Soluble Vitamins by

Ion Pairing / Ion Suppression Chromatography

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Summary of Contents for IONPAC NG1

Page 1: ...eluent for the IonPac NS1 is 3 mM Tetrabutylammonium hydroxide 28 Acetonitrile See Section 3 4 Preparing Eluents with Solvents Make the required stock and working solutions for eluents See Section 3 O...

Page 2: ...irements 9 3 3 1 Inorganic Chemicals 9 3 3 2 Deionized Water 9 3 3 3 Solvents 9 3 3 4 Acid Modifiers 10 3 3 5 Base Modifiers 10 3 4 Preparing Eluents that Contain Solvents 10 3 4 1 Eluents for Reverse...

Page 3: ...2 3 Separation of Benzoic Acid and o nitrobenzoic Acid 23 4 2 4 Gradient Separation of Aliphatic Carboxylic Acids 24 SECTION 5 TROUBLESHOOTING 25 5 1 High Back Pressure 26 5 1 1 Finding the Source of...

Page 4: ...eluent suppressor designed for use with ion exclusion and ion suppression separation modes of ion chromatography Many analytes of interest are neither ionizable nor UV detectable Organic and inorgani...

Page 5: ...Back Pressure Flow Rate Flow Rate psi MPa mL min mL min NS1 Analytical 900 6 21 1 400 9 66 1 0 3 0 NG1 Guard 200 1 38 300 2 07 1 0 3 0 NS1 NG1 columns 1 100 7 59 1 700 11 73 1 0 3 0 NS1 5 m analytica...

Page 6: ...192 The IonPac Trap Column should be installed in place of the high pressure gradient mixer between the gradient pump and the injection valve The IonPac Trap Column is filled with high capacity ion ex...

Page 7: ...usingsuppressedconductivitydetection The ion pair reagents such as tetrabutylammonium hydroxide TBAOH are used in concentrations ranging typically from 1 0 to 5 0 mM For detailed information on the op...

Page 8: ...onPac NS1 Manual Document No 034024 06 Page 8 of 35 2 7 Detector Requirements See Section 2 Ion Chromatography System Operation Summary for 4 mm system detector cell and thermal stabilizer requirement...

Page 9: ...er possible inorganic chemicals that meet or surpass the latest American Chemical Society standard for purity should be used These inorganic chemicals will detail the purity by having an actual lot an...

Page 10: ...rength of the eluent and the flow rate used The column back pressure will vary as the composition of water methanol and water acetonitrile mixture varies The practical back pressure limit for the IonP...

Page 11: ...nts Eluent A 10 solvent 90 water Eluent B 90 solvent 10 water Then by programming the gradient pump properly you can go from 100 Eluent A to 100 Eluent B in a prescribed time This will avoid outgassin...

Page 12: ...ity to increase which can result in a noisy background Most solvents have no affect on the cartridge lifetime In all cases the ionic strength of the eluent determines the lifetime of the AutoRegen Reg...

Page 13: ...ranteed high purity use Dionex Cation Regenerant Concentrate P N 039602 to make regenerant for the CSRS ULTRA II CAUTION Acetonitrile is not compatible with the CSRS ULTRA II in the Chemical Suppressi...

Page 14: ...ese ion pair reagents can be anionic or cationic Here is a partial list of typical ion pair reagents A Cationic Tetramethyl ethyl propyl and butyl ammonium salts of the general form R R N R R Counter...

Page 15: ...ng of the IonPac NS1 To guarantee that all IonPac NS1 analytical columns meet high quality and reproducible performance specification standards all columns undergo the following production control tes...

Page 16: ...Detection and Solvent Ion pairing can separate monovalent anions such as chlorate and nitrate which differ in hydration energy Because ion pairing has low selectivity for higher valency ions selected...

Page 17: ...furic acid regenerant Aliphatic sulfonic acids are UV transparent but well detected by suppressed conductivity Column IonPac NS1 10 m Eluent 2 mM Tetrabutylammoniumhydroxide 24 to 48 Acetonitrile in 1...

Page 18: ...atic sulfonic acids Column IonPac NS1 10 m Eluent 2 mM Tetrabutylammonium hydroxide 24 to 48 Acetonitrile in 10 min Flow Rate 1 0 mL min Injection Loop 50 L Detection Suppressed conductivity ASRS ULTR...

Page 19: ...nitrile increases a decrease in the background conductivity is observed Column IonPac NS1 10 m Eluent 2 mM Nonafluoropentanoic acid 20 to 80 Acetonitrile in 10 min Flow Rate 1 0 mL min Injection Loop...

Page 20: ...rate counter ion displaces the hydroxide counter ion from the fully ionized alkali and alkaline earth metals which reduces the observed conductance for these ions Column IonPac NS1 10 m Eluent 2 mM No...

Page 21: ...4 Basic molecules can be deprotonated at high pH s 9 O OH H Anionic R C O R C O pH 4 OH Cationic R NH3 R NH2 H2 0 pH 9 Ionic Neutral Additionally selectivity can be effected by working within one pH u...

Page 22: ...uble Vitamins by Ion Suppression Ion Pair Chromatography Peaks 1 Ascorbic acid 2 Nicotinic acid 3 Nicotinamide 4 Riboflavin 5 Pyridoxine 6 PABA 7 Folic acid 8 Pyridoxamine 9 Thiamine 10 Cyanocobalamin...

Page 23: ...acid 4 2 3 Separation of Benzoic Acid and o nitrobenzoic Acid Figure 9 Separation of Benzoic Acid and o Nitrobenzoic Acid Using Ion Suppression Chromatography BEFORE AFTER Eluent 3 mM HCl 32 Acetonit...

Page 24: ...ydroxide Column IonPac NS1 Flow Rate 1 0 mL min Eluent 1 24 ACN 6 MeOH 0 03 mM HCl Eluent 2 60 ACN 24 MeOH 0 05 mM HCl Gradient 0 100 E2 in 20 min Temperature 42 C Detection Suppressed Conductivity AM...

Page 25: ...dules Disconnect Replace System Module Manual High Background Conductivity Bad Eluents Remake Eluents 5 2 5 2 1 Contaminated Columns Clean Column 5 2 2 5 2 3 Contaminated ASRS or AMMS Clean Suppressor...

Page 26: ...nstall two adaptor assemblies on the filter On the end of the filter with the 1 4 28 male fitting place a 1 4 28 to 10 32 union P N 042806 On the end of the filter with the 1 4 28 female port place a...

Page 27: ...into the end fitting IonPac NS1 Column 4 mm Part P N Analytical Column 10 m 035321 Analytical Column 5 m 039568 Guard Column 039567 Bed Support Assembly 042955 End Fitting 052809 CAUTION If the column...

Page 28: ...minatethehardwareasthesourceofthehighbackgroundconductivity bypasstheSRSsuppressorandpumpdeionizedwater with a specific resistance of 18 2 megohm cm through the system The background conductivity shou...

Page 29: ...lumn must be replaced 5 3 2 Poor Resolution Due to Shortened Retention Times Even with adequate system and column efficiency resolution of peaks will be compromised if analytes elute too fast A Checkt...

Page 30: ...n the system prior to the analytical column and test for front end resolution after every system change 5 4 Spurious Peaks A The column may be contaminated If the samples contain an appreciable level...

Page 31: ...Regenerating Suppressor SRS to the detector conductivity cell at the suppressor end of the line C Install a 10 32 Luer adaptor fitting with a plastic syringe in the ELUENT OUT port of the Self Regene...

Page 32: ...n Suppressed Conductivity ASRS ULTRA AutoSuppression External Water Mode with 10mN Sulfuric Acid Range 10 SFS Background Conductivity 3 15 S Injection Volume 10 L Peak Information Found Components Pea...

Page 33: ...n Suppressed Conductivity ASRS ULTRA AutoSuppression External Water Mode with 10 mN Sulfuric Acid Range 10 SFS Background Conductivity 3 15 S Injection Volume 10 L Peak Information Found Components Pe...

Page 34: ...lush the column for a minimum of 10 minutes with the storage solution Cap both ends securely using the plugs supplied with the column B 4 Column Cleanup The following column cleanup protocols have bee...

Page 35: ...umn Cleanup Procedure A Prepare a 500 mL solution of the appropriate cleanup solution using the guidelines in Section B 4 1 Choosing the Appropriate Cleanup Solution B Disconnect the Suppressor from t...

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