Thermo Scientific Acclaim Surfactant Product Manual

37 Pages·2012·1.1 MB·English

by  Thermo Scientific

Thermo Scientific
Acclaim Surfactant
TM
Product Manual
P/N: 065087-03 February 2012
Part of Thermo Fisher Scientific
PRODUCT MANUAL
for the

ACCLAIM SURFACTANT GUARD COLUMN
4.3 x 10mm, P/N 063215
2.1 x 10mm, P/N 069693
3.0 x 10mm, P/N 071991
4.6 x 10mm, P/N 069701

ACCLAIM SURFACTANT ANALYTICAL COLUMN
5µm, 4.6 x 150mm, P/N 063201
5µm, 4.6 x 250mm, P/N 063203
5µm, 2.1 x 150mm, P/N 068123
3µm, 3.0 x 150mm, P/N 070084
3µm, 2.1 x 150mm, P/N 070085

©2012 Thermo Fisher Scientific
Document No. 065087
Revision 03
February 2012
Acclaim Surfactant Product Manual Page 2 of 36

Table of Contents
SECTION 1 – INTRODUCTION TO THE ACCLAIM SURFACTANT COLUMN ......................................... 4 
1.1  FEATURES OF THE ACCLAIM SURFACTANT COLUMN ................................................................................... 4 
1.2  ACCLAIM SURFACTANT OPERATING LIMITS AND SPECIFICATIONS ............................................................. 6 
1.3  FORMATS OF THE ACCLAIM SURFACTANT COLUMN ................................................................................... 6 
1.4  ACCLAIM SURFACTANT OPERATING CONDITIONS ....................................................................................... 7 
1.5  KEY APPLICATIONS OF THE ACCLAIM SURFACTANT COLUMN .................................................................... 7 
SECTION 2 – CHROMATOGRAPHY SYSTEM .................................................................................................. 8 
2.1  DETECTORS ................................................................................................................................................. 8 
2.1.1   Evaporative Light Scattering Detection (ELSD) ........................................................................... 8 
2.1.2   Suppressed Conductivity Detection (SCD) .................................................................................... 8 
2.1.3   UV Detection ................................................................................................................................. 8 
2.1.4  Mass Spectrometric Detection (MS) .............................................................................................. 8 
2.1.5  Refractive Index Detection (RI) ..................................................................................................... 9 
2.2   PUMPS ......................................................................................................................................................... 9 
2.3   INJECTORS ................................................................................................................................................... 9 
SECTION 3 - INSTALLATION ............................................................................................................................. 10 
3.1  SYSTEM REQUIRMENTS ............................................................................................................................. 10 
3.2   SYSTEM VOID VOLUME ............................................................................................................................ 10 
3.3   THE INJECTION LOOP ................................................................................................................................ 10 
3.4   COLUMN INSTALLATION ........................................................................................................................... 10 
3.5   COLUMN GUARD ....................................................................................................................................... 10 
SECTION 4 – OPERATION ................................................................................................................................... 11 
4.1  MOBILE PHASE SELECTION ....................................................................................................................... 11 
4.1.1   Evaporative Light Scattering Detection (ELSD) ......................................................................... 11 
4.1.2   Suppressed Conductivity Detection (SCD) .................................................................................. 11 
4.1.3   UV Detection ............................................................................................................................... 11 
4.1.4  Mass Spectrometric Detection (MS) ............................................................................................ 11 
4.1.5  Refractive Index Detection (RI) ................................................................................................... 11 
4.2  VALIDATING COLUMN PERFORMANCE ...................................................................................................... 11 
4.2.1  Procedure for Validating Column Performance: ........................................................................ 12 
4.3  EQUILIBRATING THE COLUMN ................................................................................................................... 12 
4.4  QUICKSTART FOR ACCLAIM SURFACTANT COLUMN ................................................................................ 12 
4.4.1  Using the chromatographic conditions for Column Quality Assurance Report: ......................... 12 
4.4.2  Column Quickstart ....................................................................................................................... 12 
4.5  CARING FOR THE COLUMN ......................................................................................................................... 13 
4.6  CHEMICAL PURITY REQUIREMENTS ......................................................................................................... 13 
4.6.1  Inorganic Chemicals .................................................................................................................... 13 
4.6.2  De-ionized Water ......................................................................................................................... 13 
4.6.3  Solvents ........................................................................................................................................ 14 
4.7  MOBILE PHASE PREPARATION ................................................................................................................... 14 
4.7.1  0.1 M Ammonium Acetate Buffer (1 L) ........................................................................................ 14 
4.7.2  20 mM Sodium Borate Buffer (1 L) ............................................................................................. 14 
4.7.3  20 mM Acetic Acid Buffer (1 L) ................................................................................................... 14 
4.7.4  Mobile Phases Containing Solvents ............................................................................................ 14 
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SECTION 5 – METHODS DEVELOPMENT ....................................................................................................... 15 
5.1  IONIC STRENGTH ....................................................................................................................................... 15 
5.2  MOBILE PHASE PH .................................................................................................................................... 16 
5.3  ORGANIC MODIFIER ................................................................................................................................... 17 
5.4  ION-PAIRING AGENT ................................................................................................................................. 17 
5.5  TEMPERATURE .......................................................................................................................................... 17 
SECTION 6 – APPLICATIONS ............................................................................................................................. 18 
6.1  ANIONIC SURFACTANTS ............................................................................................................................. 18 
6.1.1  Linear Alkylbenzene Sulfonates ................................................................................................... 18 
6.1.2  Sodium Lauryl Sulfate .................................................................................................................. 19 
6.2  CATIONIC SURFACTANTS ........................................................................................................................... 21 
6.2.1  Cationic Surfactants with ELS Detection .................................................................................... 21 
6.2.2  Cationic Surfactants with Suppressed Conductivity Detection .................................................... 22 
6.2.3  Benzalkonium Salts ...................................................................................................................... 23 
6.2.4  Quaternary Imidazolium Compounds .......................................................................................... 24 
6.3  NON-IONIC SURFACTANTS ........................................................................................................................ 25 
6.3.1  Ethoxylated Quaternary Compounds ........................................................................................... 25 
6.3.2  PEG Monoethyl Ether (MW-550) ................................................................................................ 26 
6.3.3  Triton X-100 ................................................................................................................................. 27 
6.3.4  NEODOL 25-7 ............................................................................................................................. 28 
6.3.5  ZONYL FSO Perfluorosurfactant ................................................................................................ 29 
6.3.6  Polyethylene Glycols (PEG) ........................................................................................................ 30 
6.4  SURFACTANTS IN COMMERCIAL PRODUCTS .............................................................................................. 31 
6.4.1  Laundry Detergent ....................................................................................................................... 31 
6.4.2  Shampoo ...................................................................................................................................... 32 
6.4.3  Fabric Softener ............................................................................................................................ 33 
SECTION 7 – TROUBLESHOOTING GUIDE .................................................................................................... 34 
7.1  HIGH BACKPRESSURE ............................................................................................................................... 34 
7.1.1  Finding the Source of High System Pressure .............................................................................. 34 
7.2 HIGH BACKGROUND OR NOISE .................................................................................................................. 35 
7.2.1 Preparation of Mobile Phases ........................................................................................................... 35 
7.2.2 A Contaminated Guard or Analytical Column .................................................................................. 35 
7.2.3 Contaminated Hardware ................................................................................................................... 35 
7.3 POOR PEAK RESOLUTION ........................................................................................................................... 35 
7.3.1 Loss of Column Efficiency ................................................................................................................. 35 
7.3.2   Poor Resolution Due to Shortened Retention Times .................................................................... 36 
7.3.3 Loss of Front End Resolution ............................................................................................................ 36 
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SECTION 1 – INTRODUCTION TO THE ACCLAIM SURFACTANT COLUMN
1.1 FEATURES OF THE ACCLAIM SURFACTANT COLUMN
The Acclaim Surfactant column is a specialty silica-based column that incorporates a proprietary bonding
chemistry developed specifically for the separation of surfactants. The separation mechanism of the Acclaim
Surfactant is a combination of reversed phase, anion-exchange, and dipole-dipole interactions. The Acclaim
Surfactant column offers ideal selectivity and unprecedented capability for separating cationic, nonionic, and
anionic surfactants in a single chromatographic run, using simple and volatile (MS compatible) mobile phases.

Acclaim Surfactant columns consist of 5-μm,or 3-µm, high purity spherical silica particles with 120 Ǻ diameter
pores bonded with proprietary functional groups, which provide unsurpassed resolution and peak symmetry for
a variety of surfactants, as shown in Figure 1. The Acclaim Surfactant column provides several features:

• Ideal selectivity for simultaneous separation of anionic, nonionic and cationic surfactants
• Excellent peak shapes for cationic surfactants
• Improved resolution for oligomers of ethoxylated surfactants
• Compatibility of highly aqueous mobile phase conditions
• Proven chromatographic conditions suitable for various detection methods
(ELSD, UV, conductivity detection, etc.)
• Broad range of applications

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Column: Acclaim Surfactant, 5 µm
Dimensions: 4.6 x 150 mm
Mobile phase: A –ACN, B- 0.1 M NHOAc, pH 5.4
4
Gradient: 25% to 85% A in 30 min., then hold for 10 min
Temperature: 30 °C
Flow rate: 1 mL/min
Injection vol.: 25 µL
Detection: ELS Detector

Analytes:
1. Xylene sulfonate
3
2. Lauryldimethylbenzyl
ammonium chloride
3. Octylphenoxyethoxyethyl
mV dimethylbenzylammonium
2 chloride
4. Triton X-100
5. Decylsulfate
6
6. Dodecylsulfate
7-10. Dodecylbenzene sulfonate
8
1 5
9
4 7
10
0 5 10 15 20 25 30 35 40
Minutes

Figure 1
Separation of Cationic, Nonionic, and Anionic Surfactants

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1.2 ACCLAIM SURFACTANT OPERATING LIMITS AND SPECIFICATIONS
Column Max Recommended
Stationary Phase Particle size P/N Typical Flow Rate
Dimensions Pressure
2.1x150 mm 070085 5500 psi 0.2 - 0.5 mL/min
3 µm
3.0x150 mm 070084 5500 psi 0.4 - 1.0 mL/min
Surfactant 2.1x150 mm 066866 5000 psi 0.2 - 0.5 mL/min
5 µm 4.6x150 mm 063201 5500 psi 0.8 - 2.0 mL/min
4.6x250 mm 063203 5500 psi 0.8 - 2.0 mL/min

Shipping Solution: Acetonitrile / 0.1% Acetic Acid v/v 80/20
Storage Solution: Acetonitrile / 0.1% Acetic Acid v/v 80/20
Buffer pH Range: pH 2.5–7.5
Solvents: Acetonitrile, Methanol
Temperature Range: <60 °C

Physical
Bonding: Proprietary
Endcapping: Yes
% C: 12
Pore size: 120 Å
Surface area: 300 m2/g
Particle size: 5 µm

1.3 FORMATS OF THE ACCLAIM SURFACTANT COLUMN
Column
Acclaim Surfactant Particle size P/N
Dimensions
2.1x150 mm 070085
3 µm
3.0x150 mm 070084
Analytical 2.1x150 mm 068123
5 µm 4.6x150 mm 063201
4.6x250 mm 063203
Requires Holder P/N
4.3 x 10mm 063215
059456
Requires Holder V-2
2.1 x 10mm 069693
P/N 069580
Guard 5µm
Requires Holder V-2
3.0 x 10mm 071991
P/N 069580
Requires Holder V-2
4.6 x 10mm 069701
P/N 069580

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1.4 ACCLAIM SURFACTANT OPERATING CONDITIONS
The recommended pH range for Acclaim Surfactant columns is between pH 2.5 – 7.5. They are compatible
with mobile phases containing 0–100% HPLC solvents, such as methanol or acetonitrile. The Acclaim
Surfactant can be operated at any flow rate, as long as the backpressure remains below its recommended
maximum pressure (see section 1.2). When setting up the analytical system, please be sure to check the special
precautions listed in Section 3, “Installation”.

1.5 KEY APPLICATIONS OF THE ACCLAIM SURFACTANT COLUMN
The Acclaim Surfactant column is a valuable tool for the analysis of all types of sample matrix, including food
and beverages, pharmaceuticals, chemicals, and environmental samples.

Applications by Class of Compounds:
• Anionic surfactants
• Cationic surfactants
• Non-ionic surfactants
• Amphoteric surfactants

Application by Industries:
• Food and beverages
• Pharmaceuticals
• Chemicals
• Semiconductors
• Environmental

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SECTION 2 – CHROMATOGRAPHY SYSTEM

Typical Flow Rate: Please refer to Section 1.2
Injection Volume: 5–50 µL for the 4.6mm ID, 2-20 µL for the 3.0mm ID,
and 1-10 µL for the 2.1mm ID
System Void Volume: Minimize the lengths of all connecting tubing and remove all unnecessary switching
valves and couplers.
Pumps: Any HPLC pump capable of delivering gradients
Detectors: UV, Conductivity, ELSD, MS, RI

2.1 DETECTORS
The Acclaim surfactant column is compatible with a variety of detectors and UV, ELSD and RI detectors. Example
applications using each of these detectors are shown in Section 6 “Applications”.

2.1.1 Evaporative Light Scattering Detection (ELSD)
When different surfactants are present in the sample with concentrations of no less than 50 ppm for each individual
surfactant, ELSD is a good option. ELSD is a universal detection method that is compatible with gradient methods
and is far more sensitive than RI. In addition, methods developed with ELSD can be easily transferred to LC-ESI-
MS applications with little or no modification, because both detectors share the same mobile phase requirements.
A UV detector can serve as a complementary tool and, if chosen, should be placed between the analytical column
and ELSD.

2.1.2 Suppressed Conductivity Detection (SCD)
Suppressed conductivity detection can also be used for surfactant analysis and provides certain advantages for
analyzing trace levels ionic surfactants in complex matrices. In fact, when analyzing ionic surfactants, especially at
low concentrations (below 10 ppm), conductivity detection using a Dionex PEEK system, equipped with a gradient
pump, electrochemical detector, MMS suppressor, and conductivity cell, is the best approach. Eluents for the
separation of surfactants using conductivity detection include borate buffer and acetic acid. If required, a UV
detector can serve as a complementary tool that should be placed between the analytical column and the MMS
suppressor.

2.1.3 UV Detection
UV absorbance is the most popular detection method in HPLC, due to its ease of use and sensitivity. The drawback
with this approach is that the analyte must have a chromophore to be detected and many surfactants do not. The
UV detector can be used alone or with other detectors.

2.1.4 Mass Spectrometric Detection (MS)
Mass spectrometry (MS) is an inherently sensitive and universal method and has become the widely accepted tool
for characterization of organic compounds. The soft ionization techniques, such as electrospray ionization (ESI),
have greatly increased the applicability of MS detection to surfactant analysis. MS detectors are the detector of
choice when less than 1 ppm individual surfactant is present and/or identification is required. Mass spectrometry
compatible methods using ammonium acetate and acetonitrile have been developed. They are shown in Section 6,
“Applications” in this manual.
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MS detection can be used in series with other detectors, such as UV. Place the UV detector between the column
and the MS or ELSD.

2.1.5 Refractive Index Detection (RI)
Although refractive index (RI) detection is a universal detection method capable of detecting all analytes, it is
incompatible with gradient methods, exhibits low sensitivity, and thus is typically only used when other detection
methods are not applicable or available.

2.2 PUMPS
Any HPLC pump can be used for the separation of surfactants with the Acclaim surfactant column. However, if
suppressed conductivity detection is chosen, a PEEK pump from Dionex is recommended.

2.3 INJECTORS
Either an autosampler or a manual injector can be used for sample injection.
Document 065087 Rev. 03 © Thermo Fisher Scientific February 2012