Sedex75 User Manual Revision 7b Complete

SEDEX MODEL 75

EVAPORATIVE LIGHT SCATTERING DETECTOR

OPERATOR’S MANUAL

S.E.D.E.R.E. PARC VOLTA-BP 27

9, Rue Parmentier 94141 Alfortville Cedex

FRANCE

Ph. + 33 1 45 18 05 18 Fax + 33 1 45 18 05 25 E-mail : [email protected] Part Number 76000 Rev. 7 January 2008

© Copyright, 1999 S.E.D.E.R.E., All rights reserved. No part of this manual may be reproduced or transmitted in any form or by any means without the written permission of S.E.D.E.R.E. Printed in France Tygon is a trademark of the Norton Corporation Teflon is a trademark of E.I. Dupont de Nemours, Inc.

Model 75 S.E.D.E.R.E. i PARC VOLTA BP 27 9, Rue Parmentier 94141 Alfortville Cedex FRANCE

Warnings and Safety Precautions The following precautions should be followed to minimize the possibility of personal injury and/or damage to property when using the S.E.D.E.R.E. Evaporative Light Scattering Detector.

1) Maintain a well ventilated laboratory. If the mobile phase typically contains a volatile organic solvent, ensure that the laboratory is well ventilated, so that a buildup of vaporized solvent cannot occur.

2) Avoid open flames and sparks. Do not use an open flame and do not use

any equipment that can cause sparks in the same room as the instrument. 3) The detector must be plugged into a grounded power line. Make certain

that all parts of the system are properly connected to a common ground. 4) If the mobile phase includes an organic solvent, use an inert gas to

nebulize the mobile phase. 5) The exhaust from the detector should be vented into a fume hood or

similar system. Make certain that the output does not escape into the laboratory.

6) The gas pressure should not exceed 4.5 bar (67 psi). Make certain that the

gas flow is maintained while the mobile phase flows through the system. If the gas flow is interrupted for extended periods of time, organic solvents could possibly damage the pressure sensor and/or the photomultiplier.

7) Do not use corrosive materials which could damage the inner metal

cladding of the detector. 8) Access inside the instrument is restricted to a suitably skilled and

competent technician. 9) Do not remove the optical head or the photomultiplier tube while the

instrument is powered up. This can destroy the photomultiplier. 10) The siphon overflow tube must contain liquid at all times. 11) Do not disassemble the nebulizer or touch any components inside the

nebulization chamber. This can lead to the deposition of contaminants that which could affect the signal.

12) Do not adjust any components inside the detector unless specifically

authorized to do so by your dealer.

Model 75 S.E.D.E.R.E. ii PARC VOLTA BP 27 9, Rue Parmentier 94141 Alfortville Cedex FRANCE

13) The instrument includes a Lithium battery CR2032 (3V). When the

battery is to be replaced, replace it with a battery of the same type or an exact equivalent and make certain that it is installed in the same manner (polarity) as the battery to be replaced. The battery must not to be recharged, short circuited, scratched or heated above 100oC. The old battery should be discarded in conformity with local environmental regulations and should not be burned or placed in an incinerator.

14) If the instrument is not used in a manner not specified by the

manufacturer, the protection ensured by the instrument can be ineffective. 15) The user is responsible for decontamination if hazardous material is spilt

on or in the instrument. 16) Warning symbols used on the equipment are : Risk of burn Electric shock risk Warning (Please, refer to user manual for informations)

Model 75 S.E.D.E.R.E. iii PARC VOLTA - BP 27 9, rue Parmentier 94141 Alfortville Cedex FRANCE

Table of Contents Warnings and Safety Precautions ................................................................ i 1 Introduction .......................................................................................... 1-1

1.1 The Evaporative Light Scattering Detector ................................................ 1-1 1.2 Principle of Operation................................................................................. 1-2

1.2.1 Nebulization .................................................................................... 1-3 1.2.2 Evaporation of the Solvent .............................................................. 1-3 1.2.3 Detection ......................................................................................... 1-4

1.3 Contents of this Manual.............................................................................. 1-5 1.4 For Additional Information......................................................................... 1-6 1.5 S.E.D.E.R.E information ............................................................................ 1-7

2 Installation ............................................................................................ 2-1

2.1 Overview..................................................................................................... 2-1 2.2 System Configurations................................................................................ 2-1 2.3 Lifting and carrying the System.................................................................. 2-3 2.4 Unpacking the System ................................................................................ 2-3 2.5 Laboratory Requirements ........................................................................... 2-4

2.5.1 Power Requirements........................................................................ 2-4 2.5.2 Gas Requirements............................................................................ 2-5 2.5.3 Exhaust Venting Requirements ....................................................... 2-5 2.5.4 Location of the Detector in the Laboratory ..................................... 2-5 2.5.5 Environmental conditions................................................................ 2-5

2.6 Installation of the Unit ................................................................................ 2-6 2.6.1 Gas Supply ...................................................................................... 2-6 2.6.2 Vent the Exhaust Line to a Fume Hood .......................................... 2-8 2.6.3 Connecting the Recorder/Integrator ................................................ 2-8 2.6.4 Connecting the Power Cord ........................................................... 2-9 2.6.5 RS-232/Printer Connection ............................................................. 2-9 2.6.6 Installing the Nebulizer/Glass Tube Assembly ............................... 2-9 2.6.7 Connecting the Siphon Overflow .................................................. 2-11 2.6.8 Connecting the Nebulization Gas to the Nebulizer ....................... 2-11 2.6.9 Connecting the Column................................................................. 2-11 2.6.10 Installing the Fluid Circulation Bath (SFC Version Only)............ 2-11 2.6.11 Powering Up the Unit.................................................................... 2-12

Table of Contents

Model 75 S.E.D.E.R.E. iv PARC VOLTA - BP 27 9, rue Parmentier 94141 Alfortville Cedex FRANCE

3 Start-up Procedure............................................................................... 3-1 3.1 Overview..................................................................................................... 3-1 3.2 The Control Panel ....................................................................................... 3-1

3.2.1 Temperature Regulation .................................................................. 3-2 3.2.2 The Signal Display .......................................................................... 3-2 3.2.3 The Pressure Display....................................................................... 3-2 3.2.4 The Gain Display............................................................................. 3-3 3.2.5 The Print Button .............................................................................. 3-3

3.3 Initial Test Procedures ................................................................................ 3-3 3.3.1 Preliminary Activities...................................................................... 3-3 3.3.2 Electronic Noise Test ...................................................................... 3-4 3.3.3 Background Noise (Stray Light) Test ............................................. 3-5 3.3.4 Solvent Noise Test........................................................................... 3-5 3.3.5 Column Noise Test .......................................................................... 3-6

4 Operating the System .......................................................................... 4-1

4.1 Overview..................................................................................................... 4-1 4.2 Preparing the System for Operation............................................................ 4-1 4.3 Zeroing and Auto-zeroing the Unit............................................................. 4-2

4.3.1 Zeroing the Unit .............................................................................. 4-2 4.3.2 Auto Zeroing the Unit ..................................................................... 4-2 4.3.3 External Auto Zero of the Unit........................................................ 4-2

4.4 Routine Operation of the System................................................................ 4-3 4.5 Optimizing Performance............................................................................. 4-3

4.5.1 Selecting the Optimum Temperature .............................................. 4-3 4.5.2 Optimizing the Mobile Phase .......................................................... 4-4 4.5.3 Sample Pretreatment........................................................................ 4-4 4.5.4 Column Treatment........................................................................... 4-4 4.5.5 Selecting the Working Pressure (SFC Mode Only) ........................ 4-5

4.6 Shutting Down the System ........................................................................ 4-5 5 Maintenance and Troubleshooting..................................................... 5-1

5.1 Overview..................................................................................................... 5-1 5.2 Cleaning and decontamination procedure................................................... 5-1 5.3 Maintenance................................................................................................ 5-1 5.4 Troubleshooting.......................................................................................... 5-2

5.4.1 General Troubleshooting Information............................................. 5-2 5.4.2 Initial Troubleshooting Activities ................................................... 5-4 5.4.3 Perform the Noise Tests .................................................................. 5-4 5.4.4 Specific Detector Troubleshooting ................................................. 5-4

5.5 Changing the Lamp..................................................................................... 5-5 5.6 Changing the lithium battery ...................................................................... 5-8

Table of Contents

Model 75 S.E.D.E.R.E. v PARC VOLTA - BP 27 9, rue Parmentier 94141 Alfortville Cedex FRANCE

Appendix 1 Specifications .....................................................................A1-1 Appendix 2 Spare Parts List...................................................................A2-1 Appendix 3 The SEDEX Model 75 Micro-HPLC Detector .....................A3-1 Appendix 4 Application Notes ...............................................................A4-1 Appendix 5 Setting the Date/Time and Selecting the Output Device .A5-1 Appendix 6 HPLC/SFC Configuration Conversion...............................A6-1 Appendix 7 S.O.P. (Standard Operating Procedure) : .........................A7-1

- Installation Qualification (I Q) - Operational Qualification (O Q) - Performance Qualification (P Q) - Original Test Report

Appendix 8 SOFTWARE : …………………………………………………..A8-1

Model 75 S.E.D.E.R.E. vi PARC VOLTA - BP 27 9, rue Parmentier 94141 Alfortville Cedex FRANCE

List of Figures

1-1 The SEDEX Evaporative Light Scattering Detector .................................. 1-1 1-2 Schematic Diagram of an Evaporative Light Scattering Detector ............. 1-2 1-3 Cross-sectional View of the Detector......................................................... 1-3 1-4 The Detection Chamber.............................................................................. 1-4 2-1 Hands placement for carrying instrument ................................................. 2-3 2-2 Rear Panel of the Detector.......................................................................... 2-6 2-3 Inserting the Gas Inlet Tube ....................................................................... 2-7 2-4 Removal of the Gas Inlet Tube................................................................... 2-7 2-5 The Nebulizer/Glass Tube Assembly (HPLC Systems) .......................... 2-10 2-6 The Nebulizer/Glass Tube Assembly (SFC Systems).............................. 2-10 3-1 The Control Panel....................................................................................... 3-1 4-1 Detector Noise as a Function of the Heating Tube Temperature

(1 ml/min flow).......................................................................................... 4-4 5-1 Rear Panel of the Detector ......................................................................... 5-3 5-2 Lamp Removal ........................................................................................... 5-7 5-3 Lamp Alignment......................................................................................... 5-7 A2-1 Identification of Spare Parts- Model 75/Model 75C Configuration ....... A2-2 A2-2 Identification of Spare Parts- Model 75, SFC Configuration ................. A2-4

Model 75 S.E.D.E.R.E. PARC VOLTA - BP 27 9, rue Parmentier 94141 Alfortville Cedex FRANCE

1-1

1.1 The Evaporative Light Scattering Detector The SEDEX Model 75 Evaporative Light Scattering (ELS) Detector (Figure 1-1) is designed to detect materials in the eluent from high performance liquid chromatography (HPLC), micro-HPLC, supercritical fluid chromatography (SFC), gel permeation chromatography (GPC) or counter current chromatography (CCC). ELS is a universal technique which can detect any non-volatile analyte. Detection is not dependent on the absorbtion of radiation and is not affected by the absorbtion characteristics of the solvent; thus solvents which absorb UV radiation can be used.

Figure 1-1: The SEDEX Evaporative Light Scattering Detector

1 Introduction

Chapter 1 Introduction

Model 75 S.E.D.E.R.E. 1-2 PARC VOLTA - BP 27 9, rue Parmentier 94141 Alfortville Cedex FRANCE

There are two versions of the SEDEX Model 75 ELS Detector: Model 75 ELSD, which includes a nebulizer (patent pending) and evaporation tube

that produces an exceptionally uniform droplet size. This leads to a more rapid evaporation of the eluent and maximizes sensitivity.

Model 75C ELSD, which is specifically designed for combinatorial chemistry. This

model includes a unique cell design (patent pending) which minimizes peak broadening and optimizes resolution. It produces peaks with the same peak width as a UV/Vis detector.

The operation of these systems is essentially identical. 1.2 Principle of Operation There are three discrete steps in the operation of the detector; nebulization of the eluent, evaporation of the solvent and detection of the compound(s) of interest (Figure 1-2). NEBULIZATION → EVAPORATION → DETECTION

Figure 1-2: Schematic Diagram of an Evaporative Light Scattering Detector Nebulization involves the conversion of the eluent into a fine mist. The mist is passed through an evaporator to vaporize solvent and a detector to measure the degree of light scattering; which is related to the concentration of the compound of interest in the sample. A cross sectional view of the system is presented in Figure 1-3.



Figure 1-3: Cross-sectional View of the Detector 1.2.1 Nebulization The eluent from the chromatograph is nebulized by the inlet gas (which is typically air or nitrogen). At the outlet of the nebulizer, the aerosol travels through a chamber. Large droplets in the spray from the nebulizer are drawn to a siphon while the fine mist travels to the evaporation tube. 1.2.2 Evaporation of the Solvent A heated tube is used to evaporate the solvent. The exit of the heated tube leads directly into the detector cell. In liquid chromatography, water and organic solvents with low boiling points are typically employed (e.g. CH3OH, CHCl3, CH3CN). A typical mobile phase for a reverse phase separation using evaporative light scattering detection might be CH3OH/H2O (60/40) while a typical mobile phase for normal phase separation might be C6H14/CHCl3 (60/40). In supercritical fluid chromatography, CO2 based mobile phases are commonly employed. These may contain volatile organic modifiers; typical mobile phases include CO2/CH3OH (95/5) and CO2/CH3OH/(C2H5)3N (79.8/20/0.2).



If acids, bases and salts are used to modify mobile phase to provide the desired separation, they should able to be readily evaporated, sublimed or decomposed into gases in the evaporation tube. Mobile phase modifiers that are commonly used when an evaporative light scattering detector is employed include NH4OH, (C2H5)3N, NH4OAC, HCOOH, CH3COOH, CF3COOH and HNO3. 1.2.3 Detection The carrier gas transports the microparticles from the heating tube into the detection chamber (Figure 1-4).

Figure 1-4: The Detection Chamber The detector chamber contains a tungsten/halogen lamp and a photomultiplier that is positioned at an angle of 120o with respect to the light beam (Figure 1-4). When the carrier gas contains microparticles, the light is scattered and is detected by the off axis photomultiplier. The intensity of the scattered light is a function of the mass of the scattering particles and generally follows an exponential relationship, which is shown in equation 1-1.

I = k mb Eq. (1-1)

where: I is the intensity of light m is the mass of the scattering particles k and b are constants



A plot of log I versus log m provides a linear response. The value of the constants (k and b) are dependent on a variety of experimental conditions (e.g. the temperature and the nature of the mobile phase). An inlet to provide additional gas is located immediately before the detector chamber to provide a concentric shield for the carrier gas. This serves to eliminate diffusion of the carrier gas and eliminate contamination of the detector cell. 1.3 Contents of this Manual This manual is designed to describe the installation, operation, maintenance and troubleshooting of the SEDEX Model 75 Evaporative Light Scattering Detector. It includes:

• Chapter 2 - Installation of the Unit, describes suitable laboratory conditions, for the detector and includes information about interfacing the unit to other devices.

• Chapter 3 - Start Up Procedure, describes the role of the various

controls and displays on the detector. In addition, this chapter discusses a number of activities to prepare the unit for routine data collection.

• Chapter 4 - Operating the System, describes how to operate the

Evaporative Light Scattering Detector. It includes information about starting the unit on a routine basis, collecting data and shutting the unit down.

• Chapter 5 - Maintenance and Troubleshooting, describes a series of

activities which should be performed on a periodic basis to ensure maximum performance. In addition, this chapter includes a protocol which can be used to determine the cause of problems that are observed with the instrument.

• A series of appendices are provided which include product specifications,

a list of spare parts, a list of applications and information about converting a system from the HPLC format to the SFC format (or vice versa) or from the HPLC format to the micro-HPLC format (or vice versa), a S.O.P., a software.



1.4 For Additional Information The detector is used as a part of a high performance liquid chromatography system or a supercritical fluid chromatography system. Manuals supplied with other components in the system (e.g. pumps, data system) should be used to obtain information about these components. For additional information about Evaporative Light Scattering Detection, the following may be of interest:

Papers:

• Use of Light Scattering as a Detector Principle in Liquid

Chromatography, A. Stolyhwo, H. Colin, G. Guiochon; J. Chromatogr., 265, 1-18 (1983).

• Study of the Qualitative and Quantitative Properties of the Light

Scattering Detector, A. Stolyhwo, M. Martin, H. Colin, G. Guiochon; J. Chromatogr. 288, 253-275 (1984).

• Detect Anything your LC Separates, P.A. Asmus, Research and

Dev., February 1986, pp. 96-98. • Supercritical Fluid Chromatography/Light Scattering Detectors: A

Promising Coupling for Polar Compound Analysis with Packed Columns, P. Carraud, et. al.; J. Chrom. Sci. 25, 395-398 (1987).

• Optimization of High Performance Liquid Chromatographic

Analysis with UV Detection: Light Scattering Detection to Establish the Co-elution of UV and non UV Absorbing Compounds, C. Elfakir, M. Lafosse, M. Dreux, J. Chromatogr. 513, 354-359 (1990).

Books:

Journal of Chromatography Library (Elsevier) • Carbohydrate Analysis HPLC and CE. Edited by Z. El Rassi.

Chapter 13, Light Scattering Detection of Carbohydrates, M. Dreux. M. Lafosse, Volume 58: 515-539 (Year).

Chromatographic and Electrophoretic Analyses of Carbohydrates,

Edited by S. Honda, Chapter xx, Supercritical and Electrophoretic Analyses of Carbohydrates, M. Lafosse, B. Herbreteau, L. Morin-Allory.



1.5 S.E.D.E.R.E Information S.E.D.E.R.E company has two locations in France :

• Administration is located in Paris. • Production is located in Orléans.

S.E.D.E.R.E, S.A. 9 Rue Parmentier Parc Volta - BP 27 94141 Alfortville, France Phone: 33 (0)1 45 18 05 18 Fax: 33 (0)1 45 18 05 25 e-mail: [email protected] S.E.D.E.R.E 6 rue du carbone Technoparc 45072 Orléans La Source France Please, visit our web site for any information : www.sedere.com



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2-1

2.1 Overview This chapter describes how the laboratory should be prepared to optimize the performance of the Evaporative Light Scattering Detector and indicates how the unit is interfaced to other devices such as the column and the data recording device. When you have successfully installed the unit, refer to Chapter 3 for start-up procedures. 2.2 System Configurations The SEDEX Model 75 Evaporative Light Scattering Detector is provided in a variety of configurations as described below. HPLC Version 230 V (part number 75000); 115 V (part number 75001)

• Main Unit • Nebulization Cell (part number 75002), consists of nebulization chamber

(part number 45002-13) and nebulizer (part number 75003) • Instruction Manual (part number 76000) • Test Report • Nebulizer Leakage Control Sheet • Start-up Kit (power cable, autozero cable, signal cable, siphon drain tube,

2 pieces of 6 mm (o.d.) gas tubing Supercritical Fluid Chromatography (SFC) Version 230 V (part number 75100); 115 V (part number 75101)

• Main Unit • SFC Nebulization Cell (part number 55104), consists of Nebulization

Chamber with heating jacket (part number 55102-13) and nebulizer with restrictor (part number 45019)

• Instruction Manual (part number 76000) • Test Report • Nebulizer Leakage Control Sheet • Start-up Kit (power cable, autozero cable, signal cable, siphon drain tube,

2 pieces of 6 mm (o.d.) gas tubing

2 Installation

Chapter 2 Installation


HPLC and SFC Version 230 V (part number 75200); 115 V (part number 75201)

• Main Unit • SFC Nebulization Cell (part number 55104), consists of Nebulization

Chamber with heating jacket (part number 55102-13) and nebulizer with restrictor (part number 45019)

• HPLC Nebulizer (part number 75003) • Instruction Manual (part number 76000) • Test Report • Nebulizer Leakage Control Sheet • Start-up Kit (power cable, autozero cable, signal cable, siphon drain tube,

2 pieces of 6 mm (o.d.) gas tubing Micro HPLC Version 230 V (part number 75600); 115 V (part number 75601)

• Main Unit • Micro-nebulizer (part number 55123) • Instruction Manual (part number 76000) • Test Report • Nebulizer Leakage Control Sheet • Start-up Kit (power cable, autozero cable, signal cable, siphon drain tube,

pieces of 6 mm (o.d.) gas tubing. Combinatorial Chemistry Version 230 V (part number 75400); 115 V (part number 75401)

• Main Unit • Nebulization Cell (part number 75005), consists of nebulization chamber

(part number 45002-13) and CC nebulizer (part number 75008) • Instruction Manual (part number 76000) • Test Report • Nebulizer Leakage Control Sheet • Start-up Kit (power cable, autozero cable, signal cable, siphon drain tube,

pieces of 6 mm (o.d.) gas tubing

S.E.D.E.R.E. provides a wide range of accessories (e.g. Gas Regulator with Filter and Manometer [part number 45100], RS-232 cable [part number 55038] and replacement parts (e.g. Replacement Halogen Lamp, [part number 45800]). A complete listing of all spare parts and accessories is included as Appendix 2.



2.3 Lifting and carrying the System Before the system is unpacked, it must be lift by two persons to ensure easy transport and avoid body injuries. Once unpacked, to carry the instrument, firstly ensure that no cables or tubing are connected. Then place hands under the instrument. Two persons are needed to ensure easy transport and avoid body injuries.

Hands placement for carrying instrument 2.4 Unpacking the System Carefully inspect all cartons and components against the packing slip to ensure that you have received all items. The nebulizer cell assembly is packed in a foam container which is placed in the lower compartment of the detector for shipping. If there is any damage to a carton or to any components or any components appear to be missing, contact both the shipping agent and your local S.E.D.E.R.E. agent immediately. If there is any evidence that the main unit has been damaged, do not plug the unit into the power line. Contact your local S.E.D.E.R.E. agent immediately. It is recommended that the shipping carton be retained as it can be used if it should become necessary to transport the system.

GOOD GOOD

BAD BAD



2.5 Laboratory Requirements 2.5.1 Power Requirements The equipment is powered by 115 V AC/60 Hz or 230 V AC/50 Hz, depending on the country. Ensure that the voltage value indicated on the power connector on the rear panel corresponds with the line voltage. Ensure that power lines accept enough current to ensure proper operation for the equipment. The current consumption is : - 1,4A for a Sedex55-115V and 1,2A for a Sedex55-230V - 1,5A for a Sedex75-115V and 1,7A for a Sedex75-230V The detector must be connected to a properly grounded three prong plug to ensure proper operation of the system. If a two prong outlet is used, make certain that the ground wire is used to ground the instrument. It is recommended that all components of the HPLC system are connected to a common ground. The detector should not be connected to an electrical line which also serves units with a large power drain or which may be subject to power surges. Such units include refrigerators, ovens, centrifuges and fume hoods.



2.5.2 Gas Requirements A supply of filtered, oil free inert gas (e.g. N2 or air) is required to operate the detector. Do not use gases that support combustion with combustible solvents. The gas supply should include a pressure gauge. A filter (0.025 µm) and manometer (part number 45100) is available as an option. Replacement filter cartridges are available as part number 45007. 2.5.3 Exhaust Venting Requirements The exhaust from the detector should be directed into a fume hood or exhaust vent. It should not be allowed to enter the laboratory atmosphere. 2.5.4 Location of the Detector in the Laboratory All components of the system (e.g. HPLC pump, detector) should be located on a sturdy table. It should be placed in an area that is free from drafts or significant temperature changes. Do not place it near air conditioning vents, windows, ovens, etc. When placing the detector, the access to the disconnect device (the appliance coupler or the mains plug) must be kept accessible at all time. The detector should be placed close to the outlet of the column to minimize extra-column band broadening which will reduce the resolution of the chromatogram. 2.5.5 Environmental conditions This instrument has been designed for following conditions :

• Use inside buildings • Altitude until 2000 meters. • Ambiant temperature from 5°C to 40°C • Maximum humidity of 80% for temperatures under 31°C, with linear

decrease to 50% at 40°C • Maximum variations for main power voltage : ±10% from nominal

voltage. • Transitory overvoltage of class II • Pollution degree 2.



2.6 Installation of the Unit 2.6.1 Gas Supply

The unit is connected to the gas supply via the 6 mm plastic tubing (supplied) using the fitting on the back of the unit (Figure 2-1).

Figure 2-1: Rear Panel of the Detector



To insert the tubing on the gas fitting, refer to Figure 2-2.

Figure 2-2: Inserting the Gas Inlet Tube Two pieces of tubing are provided. If you are using the system with an external filter, connect the gas source to the filter and then connect the filter to the back of the unit. To remove the gas inlet tube (if necessary) refer to Figure 2-3.

Figure 2-3: Removal of the Gas Inlet Tube

Note: For all units except the combined HPLC and SFC Version (230 V, part number 75200; 115 V, part number 75201) a gas regulator is inside the control unit and is set during the manufacture of the instrument.



For the combined HPLC and SFC Version; a gas regulator is mounted on the rear panel. The unit is shipped in the HPLC configuration, to change the configuration, refer to Appendix 6. 2.6.2 Vent the Exhaust Line to a Fume Hood The black exhaust line on the back of the unit should be vented to a fume hood. Make certain that the fume hood withdraws gas from the detector (i.e. there should be a positive pressure between the detector and the hood).

Note: If gas from the hood enters the detector (i.e. a negative pressure exists between the detector and the fume hood), it is possible that foreign material from the hood could contaminate the detector.

Install the vent tube so that it cannot become blocked, bent or otherwise restrict the flow of gas from the detector to the hood. If an extension tube is required (i.e. the supplied tube is not long enough) a suitable length of Tygon tubing can be fitted over the exhaust tubing. 2.6.3 Connecting the Recorder/Integrator If a recorder is employed, connect the recorder input to the 10 mV output terminal on the rear panel of the unit (Figure 2-1). If an integrator is employed, connect the integrator input to the 1 V output on the rear panel of the unit. If the external autozero function is to be employed, plug the cable that is supplied into the appropriate socket.



2.6.4 Connecting the Power Cord Place the ON/OFF switch to the OFF position and plug the power cord into the socket on the rear panel of the detector. Do not turn on the power at this time. The power cord of this system contains three wires which must be connected to a grounded line. All components of the chromatographic system should be connected to a common ground. If a two hole outlet is used, make certain that an adapter is used to connect the third wire to ground. 2.6.5 RS-232/Printer Connection If a printer or personal computer is used with the detector, the device should be connected via the RS-232 port. Appendix 5 describes how the port is configured for the printer or personal computer and how the time/date is entered (if a printer is used). 2.6.6 Installing the Nebulizer/Glass Tube Assembly Remove the plastic screen on the front of the unit. Parafilm is used to cover various openings inside the compartment and on the nebulizer cell to prevent dust particles from entering the system during shipment. These protective coverings should be removed at this time. Fill the siphon overflow on the nebulizer/glass tube assembly with the mobile phase that will be used for the separation. If you are using a very volatile solvent (e.g. hexane or CH2Cl2), use water. For SFC systems, fill the siphon with any solvent that will not react with the sample.

Note: The liquid should fill the bent part of the siphon, but should not pool in the bottom of the condenser tube.

Position the nebulizer/Glass Tube assembly inside the lower compartment of the system so that the large hole at the bottom of the tube fits over the opening for the drift tube on the back panel of the compartment. The nebulizer/glass tube assembly should be at an angle of approximately 30o so that it fits in the compartment as shown in Figure 2-4 (HPLC systems) and Figure 2-5 (SFC systems).



When the assembly is in the appropriate position, finger tighten the large black fitting that is on the drift tube opening.

Figure 2-4: The Nebulizer/Glass Tube Assembly (HPLC Systems)

Figure 2-5: The Nebulization/Glass Tube Assembly (SFC Systems)



2.6.7 Connecting the Siphon Overflow Attach the Tygon drain tube assembly to the end of the siphon tube using the tapered hose connector. Make certain that the drain tube is led to a collection bottle in such a way that condensed solvent can flow freely from the condenser and ensure that the end of the tube is not be immersed in the collected liquid. If the solvent that you are using is not compatible with Tygon (e.g. THF), use a piece of Teflon tubing in its place. 2.6.8 Connecting the Nebulization Gas to the Nebulizer Attach the fitting on the left side of the compartment to the gas inlet fitting on the nebulizer. 2.6.9 Connecting the Column Attach the fitting from the inlet of the nebulizer to the end of the column. Route the tubing through the indentation in the upper left corner of the compartment and replace the plastic screen (unless step 2.6.10 must be performed). 2.6.10 Installing the Fluid Circulation Bath (SFC version only) For supercritical fluid chromatography, the glass cell includes a heating jacket as shown in Figure 2-5. Connect the heating tubes on the right side of the compartment to the heating jacket and connect the other end (at the bottom of the unit) to the circulating bath. The temperature of the circulating fluid should be set at some temperature between 30 and 40o C. The exact value of this setting will be determined during the optimization procedure. Replace the plastic screen.

Note: The plastic screen must be replaced before heating to avoid the possibility of burns.



2.6.11 Powering Up the Unit Place the ON/OFF switch to the OFF position and plug the unit into the wall. Turn on the unit via the ON/OFF switch and ensure that the temperature, signal and pressure are indicated. Refer to Chapter 3 to prepare the unit for routine operation.

Model 75 S.E.D.E.R.E. PARC VOLTA BP 27 9, Rue Parmentier 94141 Alfortville Cedex FRANCE

3-1

3.1 Overview This chapter:

• describes the role of the controls and data display devices on the control panel

• describes the start-up test procedure

• describes how to prepare the system for operation

3.2 The Control Panel All user controls and data display devices are on the control panel (Figure 3-1).

Figure 3-1: The Control Panel

3 Start-up Procedure

Chapter 3 Start-up Procedure

Model 75 S.E.D.E.R.E. 3-2 PARC VOLTA-BP 27 9, rue Parmentier 94141 Alfortville Cedex FRANCE

3.2.1 Temperature Regulation The present temperature of the evaporation tube is indicated on the display panel. To set the temperature: a) Press the SET button until the TEMPERATURE LED is illuminated. The

TEMPERATURE field will indicate the present set point. b) Press the UP or DOWN button until the desired temperature is set. When the

desired temperature is set, wait approximately 10 sec. The set point will be entered into the system and the present temperature of the evaporation tube will be presented again.

c) The system will automatically increase (decrease) the temperature to reach the

new setpoint. 3.2.2 The Signal Display The SIGNAL (mV) display indicates the present intensity of the signal (minus the intensity that has been subtracted via the manual control as described below). To set the signal:

a) Press the SET button until the SIGNAL LED is illuminated.

b) Press the UP or DOWN button until the desired signal is set. When the desired signal is set, wait approximately 10 sec. The set point will be entered into the system and the present signal will be presented again.

When you have adjusted the signal intensity to zero, press the AUTO button to reset the circuit (e.g. autozero the detector). 3.2.3 The Pressure Display The PRESSURE (bar) display indicates the pressure of the nebulizer gas.



3.2.4 The Gain Display The GAIN is displayed in the pressure field when the SET button is used to select the GAIN LED. To set the gain:

a) Press the SET button until the GAIN LED is illuminated. The PRESSURE field will indicate the present set point (1-12).

b) Press the UP or DOWN button until the desired gain is set. After the gain

is set, wait approximately 10 sec. The set point will be entered into the system.

3.2.5 The Print Button The PRINT button is used to send the present Date, Time, Temperature, Gain and Pressure to an external device such as a printer or personal computer (this information can be used for archiving of data). The operator can set the present time and data and the output type as described in Appendix 5. 3.3 Initial Test Procedure 3.3.1 Preliminary Activities This section presents a protocol which can be used to ensure that the system is functioning in the proper manner. When the instrument is first installed and you have completed the preliminary steps; the procedures described in Sections 3.3.2 through 3.3.5 should be performed to determine the specific characteristics of your unit. We suggest that you save the results in a permanent location, as they may be very useful when you are performing troubleshooting activities.

a) Power up the instrument. The TEMPERATURE, SIGNAL and PRESSURE fields will indicate the present status of the unit.

b) Set the GAIN to 1 and set the TEMPERATURE to 40o C. The

TEMPERATURE display will present the ambient room temperature. After a short period of time, it will begin to rise.



Note: When the system is initially powered up (or if you change the temperature), the temperature will first overshoot the temperature set point and will stabilize at the set-point after a short period of time. The initial overshooting of the set point is due to the Proportional Integral and Derive regulation of the system and should not be a concern.

If you are using an SFC unit, start the circulating bath.

c) Make certain that the siphon contains mobile phase (if necessary, turn on the pump for a short period to fill the siphon).

Provide gas to the detector and adjust the gas flow so that the pressure is 3.5 bar.

Note: MAKE CERTAIN that the pressure of gas is less than 4.5 bar (67 psi). If the pressure is allowed to increase above 4.5 bar, the sensor may be damaged.

d) Set the signal to zero and press the AUTO button. The SIGNAL display should read close to zero and should remain near zero.

3.3.2 Electronic Noise Test To determine the Electronic Noise: Check to ensure that the gas is flowing and the temperature is set to 40o C. Set the GAIN to 12 over a period of two minutes (i.e. increase the gain by 2 units every 20 seconds and monitor the baseline. When the gain is set to 12, Monitor the display for a period of five (5) minutes). The variation in the signal should be less than +/- 3 mV. There may be some spiking of the signal Record the level and autozero the detector.



3.3.3 Background Noise (Stray Light) Test To determine the background noise, change the GAIN to 1 and record the level. Increase the gain and monitor the signal (which will increase). The table below provides the approximate stray light to be expected at various gain settings.

Gain Signal (mV) 1 0 6 2 7 2 8 5 9 12 10 29 11 58 12 110 Stray light table example.

3.3.4 Solvent Noise Test To determine the solvent noise:

a) Check to ensure that the gas is flowing and the temperature is set to 40°C, and the pump is off.

b) Set the gain to 12 (do not autozero). The signal may be negative. c) Pump the mobile phase through the detector at a rate of 1 mL/min. The

column should be bypassed during this operation. For the micro-HPLC detector, the flow rate should be set as indicated in Table 3-1.

Column ID (mm)

Flow Rate (µL/min)

4.6 1000 2.1 208 1.0 47 0.8 30 0.32 4.8

Table 3-1: Flow Rates for the Micro-HPLC Detector d) When the flow is initiated, the signal will increase. After a few minutes,

the signal will fall. The intensity of the signal will depend on the solvent that you are using.



e) Monitor the baseline for a few minutes, it should be flat with a 0.5 to 1.0% noise level.

For water, the signal should be 10 mV or less, while it should be less than 200 mV for an organic solvent. The signal level is approximately linear with respect to the concentration of organic phase in the solvent (e.g. a water:organic solvent [50:50] will provide a signal level around 100 mV). The solvent noise is a critical factor in the sensitivity that is obtained from the detector. The sensitivity of the system is inversely proportional to the solvent noise (equation 3-1). Sensitivity = Signal/Noise 3-1 The purity of the solvent is a critical issue in the noise. Filtering of the solvent is not recommended, as the solvent may extract contaminants from the filter. In most cases, distilled water and HPLC grade solvents are satisfactory. When you are comparing solvents, the critical parameter is the Residue after Evaporation; this should be less than 1 ppm to maximize the sensitivity of the detector. The mobile phase should not contain non-volatile solvent modifiers. Volatile solvent modifiers (e.g. CF3COOH, CH3COOH, (C2H5)3N, can be used, but they may increase the noise level at high gain settings. In addition, preservatives such as BHT (which is commonly found in Tetrahydrofuran) should not be used. 3.3.5 Column Noise Test To determine the Column Noise:

a) Turn off the pump and reconnect the column. b) Re-start the pump and allow the mobile phase to flow through the system.

It is suggested that you flush the column with your strong solvent for a few minutes before attaching it to the detector.

c) Set the gain to 12, attach the column to the detector and monitor the

baseline. A “good column” will give a baseline that is 20-50 mV above the solvent baseline.

Note: If the mobile phase contains acidic modifiers (e.g. TFA), disconnect the detector and wash the LC system for 12 hours before starting to analyze unknowns.


4-1

4.1 Overview This chapter describes operations that should be performed on a routine basis when you want to collect chromatographic data. It is assumed that you have determined the system parameters described in Section 3.3 and the general chromatographic conditions for the desired separation. 4.2 Preparing the System for Operation To prepare the system for operation:

a) Power up the instrument by pressing the switch on the rear panel. b) Ensure that the overflow siphon of the nebulization chamber contains

sufficient liquid. If necessary, pump a few mL of solvent through the system.

c) Open the gas distribution valve and adjust the pressure to 3.5 bar. d) Select the desired temperature as described in Section 4.5.1

e) Start the mobile phase flow through the system and allow the system to

operate for at least 15 minutes to ensure that all components are equilibrated and a stable baseline is obtained.

Important : Do not forget to operate the solvent noise test described in section 3.3.4 – Record the value in your file.

Note: If the gas flow stops, turn off the pump. The pressure sensor and/or the photomultiplier may be permanently damaged if organic solvent is allowed to flow through the Nebulization chamber for an extended period of time.

4 Operating the System

Chapter 4 Operating the System


4.3 Zeroing and Auto-zeroing the Unit 4.3.1 Zeroing the Unit To set the detector Zero:

a) Select the desired gain for the analysis using the GAIN control. b) Adjust the zero signal by adjusting the SIGNAL control.

4.3.2 Auto Zeroing the Unit To auto-zero the unit:

a) Set the GAIN to 1. b) Press the Up and Down buttons at the same time to set the signal level to

the center of the signal range.

c) Press the AUTO ZERO button. d) Change the GAIN selector to the gain setting that is to be used for the

analysis. e) Monitor the SIGNAL value. If it is greater than 10 mV, adjust the value to

0 with the SIGNAL control. f) Press the AUTO ZERO button.

Note: If you change the GAIN selection, make certain that steps d and e are repeated before taking measurements.

4.3.3 External Auto Zero of the Unit If desired, the auto zero command can be initiated via an external device such as an HPLC system controller. To employ this feature, a cable from the external device is plugged into the EXT AUTO ZERO socket on the rear panel. To auto zero the system, a contact closure signal is used to short circuit the contacts. The signal should be at least 1 sec long, with a maximum current of 20 mA at 5 V.

Note: Do not use a 5 V TTL logic signal.



4.4 Routine Operation of the System During operation of the system, the following points should be considered:

a) Make certain that the exhaust from the detector is led into a fume hood or other device. The exhaust gas should not be allowed to enter the laboratory.

b) Ensure that the siphon is filled with liquid at all times. The overflow from

the siphon should be collected in a suitable container. c) Never exceed a gas pressure of 4.5 bar. d) Avoid the use of solvent or samples which could corrode the detector.

4.5 Optimizing Performance 4.5.1 Selecting the Optimum Temperature When you are setting up a system, the temperature should be set to 40o C if an aqueous mobile phase is used and 35o C if an organic based mobile phase is used (flow rate of 1 mL/min). These values can be adjusted during the method optimization process. If you suspect that your compound is thermally labile, a lower temperature could be used to improve the sensitivity to minimize thermal decomposition. For a given solvent and flow rate; there is, however, a point at which the noise in the chromatogram increases dramatically because not all of the eluant is vaporized. As an example, if a 1 mL/min flow of water is employed, the noise level increases significantly as the temperature is lowered from 33o C to 31o C (Figure 4-1).



33oC 31oC

Figure 4-1: Detector Noise as a Function of the Heating Tube Temperature (1ml/min flow) At higher flow rates, higher temperatures are required to minimize the noise (e.g. at 2 mL/min, Tmin = 35o C; at 3 mL/min, Tmin = 40o C). 4.5.2 Optimizing the Mobile Phase Particulate matter in the mobile phase will increase the background and the noise. In most cases, distilled water and HPLC grade solvents are satisfactory. When you are comparing solvents, the critical parameter is the Residue After Evaporation; this should be less than 1 ppm. Filtering of the solvent is not recommended, as the solvent may extract contaminants from the filter. The mobile phase should not contain non-volatile solvent modifiers. Volatile solvent modifiers (e.g. CF3COOH, CH3COOH, (C2H5)3N) can be used, but they may increase the noise level at high gain settings. In addition, preservatives such as BHT (which is commonly found in Tetrahydrofuran) should not be used as they will increase the noise level, especially at higher gain settings. The wetted parts of the detector are made from Teflon, Stainless Steel, Glass and Nickel. Make certain that solvents will not react with these materials. 4.5.3 Sample Pretreatment If the sample contains any particulate matter, it should be filtered through a 0.2 µm or 0.45 µm filter before injection. 4.5.4 Column Treatment The chromatographic column contains microparticles to separate the compounds of interest. Under certain circumstances, the column packing will undergo chemical and/or mechanical breakdown; this may lead to the introduction of particulate matter into the detector which may lead to an increase in the noise.



The breakdown of the column is dependent on a variety of factors including the particle size, the type of column that is used, the manufacturer of the column, the nature of the mobile phase (e.g. high pH will degrade silica based columns). When you install a new column, it is suggested that you pump mobile phase through it for a few minutes before connecting it to the detector. This will flush out any microparticles that remained in the column after manufacture of the column. After you install a new column, perform the Column test (Section 3.3.5) to obtain the baseline signal value for this column. 4.5.5 Selecting the Working Pressure (SFC Mode Only) The minimum and maximum pressures for a variety of flow rates using the standard restrictor (part number 45022) are presented in Table 4-1. Flow Rate Minimum Pressure Maximum Pressure ∆ Pressure 1 mL/min 80 bar 130 bar 50 bar 2 mL/min 100 bar 290 bar 190 bar 3 mL/min 150 bar 370 bar 220 bar 4 mL/min 220 bar > 400 bar

Table 4-1: Working Pressure Range for SFC 4.6 Shutting Down the System To shut down the system:

a) Turn off the pump. b) Allow the nebulization gas to flow though the detector for a few minutes

to drain the evaporation tube and detection chamber. c) Turn off the gas flow. (If the SFC version is being used, turn off the power

to the circulating bath). d) Turn off the power to the detector.

Note: If you are using a mobile phase which contains salts, acid or bases, pump a few mL of water or methanol through the system before turning off the system so that salts do not deposit on any components of the detector.




Chapter 5 Maintenance and Troubleshooting


5.1 Overview This chapter describes:

• the cleaning and decontamination procedure that should be performed to maintain instrument performances. Section 5.2)

• the maintenance procedures that should be performed by the operator on a routine basis. (Section 5.3)

• a guide to troubleshooting activities that should be useful in determining the cause of erratic or erroneous results (Section 5.4).

5.2 Cleaning and decontamination procedure

a) Instrument cleaning In case of cleaning need, the instrument must be turned off and all connection cables must be unplugged (power cable, signal cable, autozero cable, RS-232 cable, instrument gas input and nebulizer tubing). The user must allow the detector to cool before cleaning. The instrument should be cleaned with a non-abrasive rag. The user must ensure no liquid can enter in the detector when cleaning. b) Instrument decontamination S.E.D.E.R.E suggests the following actions :

• Set the temperature to 90°C during 3 hours with gas flow • Use the appropriate solvent to decontaminate the instrument. The

choice of the solvent will be determinated by the nature of samples previously analyzed by the user.

5.3 Maintenance The SEDEX Evaporative Light Scattering detector is designed to require a minimum of maintenance activities. If preventive maintenance activities are followed, the system should provide high sensitivity measurements without intervention by the operator.

5 Maintenance and Troubleshooting



The following actions are suggested:

• maintain the detector in a clean laboratory environment • if the system is not going to be used for a period of time, flush out any

mobile phase that contains acids, bases or salts to prevent the deposition of crud on components or corrosion of the system

• only use filtered non-oil pumped gas • Inspect all cables and tubes to prevent injuries due to bad cables. This

includes the main cord set, the signal cable, auto-zero cable, RS-232 cable if applicable, nebulizer inlet tubes and main gas inlet tube.

In routine operation it is not necessary to access the interior of the instrument. The lamp has a long (but finite) lifetime (~ 2000 hours) and should be replaced periodically. In typical use, the lamp should be replaced on an annual basis. The procedure to replace the lamp is provided in Section 5.5. Conversion of the system between the HPLC and SFC format, or between the HPLC and Micro HPLC format is described in Appendix 5 and Appendix 3.3 respectively. 5.4 Troubleshooting 5.4.1 General Troubleshooting Information The SEDEX Evaporative Light Scattering detector is incorporated into a liquid chromatography or SFC system. It is important to note that the detector response reflects the overall performance of the system, and a “problem” that is seen on the detector output may not necessarily be a “detector problem”. In almost all cases, there is one and only one cause for a problem. As an example of this point, if the user observes a noisy baseline, it is possible that the problem is due to:

• the pump (e.g. a defective check valve) • the mobile phase (e.g. improper degassing) • the column (e.g. elution of strongly retained components) • the detector (e.g. an electronic problem)

It is very unlikely that two problems will occur at the same time. The role of the troubleshooting activities is to determine the cause of the problem. In this discussion, we will assume that the operator has determined that other components of the system are operating in an appropriate fashion.



WARNING: DO NOT DISASSEMBLE THE NEBULIZER. THIS WILL VOID THE WARRANTY.

Note: The control panel and system electronics do not contain any user replaceable components. If the suggestions provided in this chapter do not remedy the problem, contact your nearest S.E.D.E.R.E. representative.

If the digital displays and the temperature and signal LED’s do not illuminate when the system is powered up, turn the unit off and inspect the fuses on the power connector at the rear of the detector. Main fuses holder

Figure 5-1: Rear Panel of the Detector

Changing the fuses must be carried out by suitably skilled technician. If necessary, replace the fuse with a fuse of the same rating as the original fuse. Main fuse : T 2AL / 250V (part number 45300-2) If the detector is a high temperature GPC system use a T 3.15AL / 250V fuse, (part number 45300-3.15). If the main fuses are not blown, please contact your S.E.D.E.R.E representative.



5.4.2 Initial Troubleshooting Activities

a) Make certain that the instrument and all components of the system are grounded to a true ground.

b) Check to ensure that the liquid level in the siphon is appropriate, and there

is no liquid pooling in the nebulization chamber. c) Check that the gas pressure is sufficient and is stable. The gas filter should

be clean and in place. d) Ensure that the flow rate of the pump is constant and check that there are

no leaks in the chromatographic system. 5.4.3 Perform the Noise Tests Repeat the tests described in Section 3.3 and compare the observed data to the results that were obtained when the unit was initially installed. These tests can be very valuable to isolate the problem. As an example of this point, if the Electronic Noise test (Section 3.3.2), Background Noise test (Section 3.3.3) and Solvent Noise test (Section 3.3.4) provide results that are similar to that obtained when the unit was initially installed, but the Column Noise test (Section 3.3.5) provides a significantly different value than what was obtained at installation, it is likely that the problem is in the column (e.g. highly retained compounds are being eluted). 5.4.4 Specific Detector Troubleshooting

a) The mist from the nebulizer should be fine and homogeneous. If it is not fine and homogeneous, the nebulizer, the needle or the Teflon tube may be obstructed. To remove the obstruction, pump a solvent that can dissolve the foreign material. As an alternative, the nebulizer can be placed in an ultrasonic bath to dissolve the foreign material..

WARNING: DO NOT DISASSEMBLE THE NEBULIZER. THIS WILL VOID THE WARRANTY.



b) If the sensitivity of the system is low, ensure that there are no leaks in the system. In some cases, a small increase in the gas pressure (e.g. 0.1 or 0.2 bar) may solve the problem. Alternatively, the lamp may need to be replaced or the nebulizer could be obstructed.

c) If the detector signal is saturated or if there is a decrease in the dynamic range of the system, it is possible a residue is passing through the detector cell, which leads to an intense signal due to a significant amount of light scattering. This residue may be derived from the elution of strongly retained materials from the column, or may be derived from the solvent. To determine the cause of the problem, bypass the column and observe the signal intensity:

• if the signal returns to normal, strongly retained materials are

eluting from the column. Flush the column with a strong solvent to elute all material.

• if the signal does not return to normal, the solvent contains a

residue material and is not suitable for use with the detector.

d) If the noise of the system without solvent is high or if ghost peaks occur, it is possible that foreign material is present in the drift tube. In this situation, increase the temperature to 95o C and pump solvent at the rate of 2 mL/min, using a gas pressure of 3 bar.

5.5 Changing the Lamp

Note: The Lamp is very hot when power is applied. Allow the lamp to cool before touching it.

Changing the lamp must be carried out by suitably skilled technician. To change the lamp:

a) Turn off the instrument power, disconnect the main power cable and remove the cover

b) The lamp sits at the end of a black tube which can slide in and out of the

unit and can rotate about the longitudinal axis. Mark the present position of the tube with respect to both axes. These alignment marks will be useful when you replace the tube (step g).



c) Remove the set screw that holds the lamp/tube assembly in place (Figure 5-2).

d) Pull out the lamp assembly/tube from the detector. e) Remove the two screws that attach the lamp assembly to the tube.

Note: Do not touch the new lamp with your bare fingers. The oil from your finger can greatly reduce the lifetime of the lamp. If fingerprints do get on the lamp, remove them with a lint free tissue saturated with methanol.

f) Attach the new lamp assembly onto the tube and replace the two screws.

After you mount the lamp, look down the tube to ensure that the filament is visible in the center of the tube, and is centered.

g) Replace the lamp assembly/tube into the instrument. When replacing the

tube, use the alignment marks that were made in step c. Turn on the detector. Set the gain to 1, autozero the detector and turn the gain to 12.

Note: The lamp produces a significant amount of energy. Shield your eyes and do not look directly at the lamp.

h) Determine the correct position (see the arrow labeled 1 in Figure 5-3). The

correct position is that which minimizes the intensity of the signal.

i) Rotate the lamp/tube assembly around the central axis of the tube to determine the correct position (see the arrow labeled 2 in Figure 5-3). The correct position is that which minimizes the intensity of the signal.

j) Tighten the set screw.

k) Replace the detector cover.



Figure 5-2: Lamp Removal

Figure 5-3: Lamp Alignment



5.6 Changing the Lithium battery

Note: Please refer to Warning and safety precautions chapter to ensure safety with the lithium battery.

Changing the lamp must be carried out by suitably skilled technician. Lithium battery type : CR2032 (3V)

a) Turn off the instrument power, disconnect the power cable and remove the cover. b) The lithium battery is located on the left of the display board. This is a round metallic component. Informations on battery type (lithium, 3V and CR2032) are written on the battery.

Model 75 S.E.D.E.R.E. A1-1 PARC VOLTA - BP 27 9, rue Parmentier 94141 Alfortville Cedex FRANCE

Appendix 1 Specifications Detection High Sensitivity Photomultiplier Light Source Polychromatic Tungsten Halogen Lamp Flow Rate 0.1 to 5 mL/min (standard HPLC) 10 to 150 µL/min (micro-HPLC) Signal Drift Less than 1 mV/hr Signal Output 0-10 mV, 0-1 V Zero Control Manual, Auto Zero and Remote Auto Zero Gas Flow Control Nebulizer and Auxiliary Gas Flow (Patent Pending) Gas Consumption Less Than 3 L/min Temperature Range Ambient to 80o C (standard unit) Microprocessor Controlled via Keypad Interface RS-232 Serial Output Power 115V/60Hz or, 230V/50Hz Dimensions 360 mm (14”) W x 504 mm (20”) H x 500 mm (20”) D Weight 20 kg (44 lb.)




A2-1

Appendix 2 Spare Parts List

For assistance in identifying parts, please refer to Figure A2-1 (HPLC System) or A2-2 (SFC System). In these tables, the numbers in brackets (e.g. [1] refer to the appropriate figure SEDEX Model 75/Model 75C Detectors Part Part Number HPLC Nebulizer Assembly [1] 75003 * HPLC Glass Cell [2] 45002-13 Complete HPLC Nebulization Cell 75002 (consists of 75003 and 45002-13) CC Nebulizer Assembly [1] 75008 * Complete HPLC Nebulization Cell 75005 (consists of 75008 and 45002-13) Black Plastic Nut for Nebulization Chamber (Nebulizer Side) diam 30 mm [3] 55018 Black Plastic Nut for Nebulization Chamber

(Heating Tube side) diam 13 mm [10] 45700-13 Drain Assembly [4] 45200 (includes fitting, part number 45108) Drain Assembly Fitting [5] 45108 PTFE Inlet Tube for Nebulizer [6] 45013 (includes stainless steel nut, part number 45014) Pneumatic tube (diameter 4 mm) for Nebulizer 55070 (includes stainless steel fitting) Tube Fitting for Nebulizer (4 mm diameter) [8] 55015 Wall Mounting Fitting (4 mm diameter) [9] 55014 Stainless Steel Nut 1/16” with Ferrule 45014 Tungsten Lamp 45800 Gas Regulator with Filter and Manometer 45100 Tube Fitting (6 mm diameter) for Gas Regulator 45018 (part number 45100) Cartridge for Gas Regulator 45007 Power Cable 45600 Autozero Cable 55500 Signal Cable 55400 Fuse (2 amp) 45300-2 Fuse (3.15 amp) 45300-3.15 Conversion Kit to SFC Detection 55027 Conversion Kit to Micro HPLC Detection 55023 * If desired, the nebulizer assembly can be exchanged for a new unit (For Model 75 HPLC version, part number 75004, For Model 75 CC version, part number 75007)



1

2

3

4

5

6

7

8

9

10

Figure A2-1: Identification of Spare Parts - Model 75/Model 75C Configuration



SEDEX Model 75 SFC Detector

Part Part Number SFC Nebulizer with Restrictor [1] 45019 SFC Nebulization Cell with Heating Jacket [2] 55102-13 Complete Nebulization Cell 55001 (consists of part number 45019 and part number 55102-13) Stainless Steel Inlet Tube for Nebulizer [3] 55013

(includes stainless steel nut, part number 45014) Black Plastic nut for Nebulization Chamber (Nebulizer Side) Diam 30 mm [4] 55018 Black Plastic Nut for Nebulization Chamber [4]

(Heating Tube Side) diam 13 mm [10] 45700-13 HPLC Nebulizer Assembly 75003 * Drain Assembly [5] 45200 (includes fitting, part number 45108) Drain Assembly Fitting [6] 45108 Pneumatic tube (diameter 4 mm) for Nebulizer 55070 (includes stainless steel fitting) Tube Fitting for Nebulizer (4 mm diameter) [8] 55015 Wall Mounting Fitting [9] 55014 Stainless Steel Nut 1/16” with Ferrule 45014 Restrictor for SFC (Type I) 45022-1 Restrictor for SFC (Type II) 45022-2 Tungsten Lamp 45800 Gas Regulator with Filter and Manometer 45100 Tube Fitting (6 mm diameter) for Gas Regulator 45018 (part number 45100) Cartridge for Gas Regulator 45007 Power Cable 45600 Autozero Cable 55500 Signal Cable 55400 Fuse (2 amp) 45300-2 Fuse (3.15 amp) 45300-3.15 Conversion Kit to Micro HPLC Detection 55023 * Used for modification to HPLC detection



1

2

3 4

5

6

7

8

9

10

Figure A2-2: Identification of Spare Parts - Model 75, SFC Configuration



SEDEX Model 75 HPLC Detector-Micro HPLC Configuration

Part Part Number Nebulizer Assembly (Micro HPLC) 55123 Silica Tube-325 µ 55223 Socket (16 x 325 µ) 55323-325 Socket (16 x 280 µ) 55323-280 (All other components are the same as the SEDEX Model 75 HPLC Detector)





A3-1

Appendix 3 The SEDEX Model 75 Micro HPLC Detector

A3.1 Overview Micro HPLC refers to the use of narrow bore columns (e.g. 1 mm I.D.) and low flow rates (e.g. 10 µL). The decrease in the column diameter reduces the limit of detection of the detector; as an example if the column diameter is changed from 4.6 mm ID to 300 µm ID, the limit of detection will decrease by a factor of approximately 100. A3.2 Installation of the Detector Installation of the SEDEX Model 75 Micro HPLC Detector (Part Number 75600/75601) is identical to that for the standard HPLC detector. The temperature and gas pressure values that should be employed are the same as that for the standard HPLC detector. A3.3 Conversion of the Standard HPLC Detector to the Micro HPLC Detector The Micro-HPLC kit (part number 55023) can be used to convert a standard HPLC detector to the micro HPLC detector. This kit includes the following parts: Nebulizer Assembly (Micro HPLC) 55123 Silica Tube-325 µ 55223 Socket (16 x 325 µ) 55323-325 Socket (16 x 280 µ) 55323-280 To convert the standard detector to the micro-HPLC detector:

a) Remove the Nebulization Cell (which consists of the nebulization chamber and the nebulizer).

b) Fit the Micro-Nebulizer into the heating tube and tighten the screw. c) Connect the nebulization gas inlet tubing, the column inlet tubing and the

drain tube. To check the flow rate of the pump, connect the silica tube (part number 55223) instead of the micro-nebulizer.

Appendix 3




A4-1

Appendix 4 Application Notes Please consult our Website: http://www.sedere.com for more applications (more than 140 are provided). To obtain more details about an application, please contact your local representative, or request information from: [email protected]

Page Application Chemical Family Application number

A4-3

Fructose Glucose Lactose Sucrose Raffinose Stachyose

Carbohydrates 75

A4-3 Maltodextrins DP1 to DP17 Polysaccharides 35

A4-4 Alkyl Glucosinolates Carbohydrates Derivatives 73

A4-4 Vitamins A, D2, E1, K1 β-carotene Fatty acids 18:3, 18:2, 16,0, 18:0

Vitamins 101

A4-5 Ginseng Extract Saponins Saponins 60

A4-5

Cerebroside Hydroxylated Cerebroside Phosphatidyl Ethanolamine Phosphatidyl Inositol Phosphatidyl Serine Phosphatidyl Choline Phosphatidyl Acid Sphingomyeline Lysophosphatidyl Choline

Phospholipids 5

Appendix 4 Application Notes


A4-6

Trioleine α-Tocopherol Oleic Acid β-Tocopherol γ-Tocopherol δ-Tocopherol Diolein Cholesterol

Oleins, Tocopherols Cholesterol 116

A4-6 Polyoxyethylene Alcohols (C12OE9)

Polyoxyethylene Alcohols 18

A4-7 PEG 400 Polyethylene Glycols 23

A4-7 21 Amino Acids sample Amino Acids 114

A4-8 D & L Amino Acids (Trp, Val, Asn) Amino Acids 140

A4-8 Di, Tri & Tetrapeptides Peptides 142

A4-9 Inorganic Anions Inorganic Anions 109

A4-9

Tergitol type B Octyl B glucoside Sodium Dodecyl sulfate N Lauryl sarcosine Zwittergent 3 14 Triton X 100 Lubrol PEG 10 Tridecyl ether

Detergents 61

A4-10 Low Temperature Operation: Ethylene Glycol Quantitation Ethylene Glycol 141

A4-11 Low Temperature Operation: Thermosensitive Urea Urea 94

Table A4-1 : Applications table



Carbohydrates HPLC Gradient F: Fructose G: Glucose L: Lactose S: Sucrose R: Raffinose St: Stachyose

Gradient: Acetonitrile, 25 % to 50 % in 15 min

Analyzed by: LCBA 45000 Orleans FRANCE

Maltodextrines HPLC Gradient

Gradient:

60% to 80% Acetonitrile in 20 minutes

Refractive Index detector can detect concentrations of 50,000 of DP 1 to DP 10

SEDEX Evaporative Light Scattering Detector can detect concentrations of 1000 ppm of DP 1 to DP 17



Alkyl Glucosinolates (R-GI) HPLC Gradient

Peaks 1. C10 GI 2. C11 GI 3. C12 GI 4. C13 GI 5. C14 GI

Gradient:

Methanol, 40 to 75% in 10 min Hold at 75% Methanol for 8 min.

Vitamins and Fatty Acids Standard HPLC Gradient 1 = 18:3, 14 ppm 2 = Vitamin A, 80 ppm (all trans retinol) 3 = 18:2, 42 ppm 4 = 18:1, 56 ppm 5 = 16:0, 98 ppm 6 = 18:0, 70 ppm 7 = Vitamin D2, 92 ppm (Ergocalciferol) 8 = Vitamin E1, 42 ppm (α-Tocopherol) 9 = Vitamin K1, 76 ppm 10 = β-Carotene, 50 ppm Gradient:

85:10:5 to 0:70:30 in 15 min. 0:70:30 to 0:30:70 in 5 min.

Analyzed by: Richard Scientific, Novato CA 94949 USA



Ginseng Saponins HPLC Gradient R = D GLU D GLU

R = D GLU D GLU 1

= L Ara D GLU 2 Furanose = L Ara D GLU 3 Pyranose = D GLU 4 Gradient: 20-40% Acetonitrile in 15 min Detection limit (S/N = 3) = 120 ng Analyzed by: J. Becart - Parfums Christian Dior

Phospholipids HPLC Gradient 1. Apolar lipids 2. CR : cerebroside – fatty acid not hydroxylated 3. CROH: cerebroside – fatty acid hydroxylated 4. PE : phosphatidyl ethanolamine 5. PI : phosphatidyl inositol 6. PS : phosphatidyl serine 7. PC : phosphatidyl choline 8. PA : phosphatidyl acid 9. Sph : sphingomyeline 10. LPC: lysophosphatidyl choline Gradient Profile Analyzed by: Mr. Becart, Parfums C. Dior [J. High Res. Chrom. 13, (1990) 126]

B



Oleins, Tocopherols, Cholesterols HPLC

Analyzed by: Courtesy of Bunge Foods, Bradley, IL

Polyoxyethylene Alcohols HPLC (C12OE9) Gradient Analyzed by: Mr. Becart, Parfums C.Dior

1. Triolein 2. α-Tocopherol 3. Oleic Acid 4. β-Tocopherol 5. γ-Tocopherol 6. δ-Tocopherol 7. Diolein



Polyethylene Glycol PEG 400 HPLC Gradient Analyzed by: LCBA 45100 Orleans FRANCE

Underivatized Amino Acids HPLC Gradient

Analyzed by: Mr Dreux et al. - UNPUBLISHED RESULTS - SP: system peak

Gradient:

Acetonitrile/Water

Gradient: H2O (+volatile ion pairing agent) 0% to 40% organic solvent in 40 min.



D and L Amino Acids HPLC Direct Enantiomeric separation

Analyzed by: CEA SACLAY – Service des molécules marquées – 91191 GIF/YVETTE CEDEX – FRANCE

Underivatized Small Peptides HPLC

Dipeptides:

Tri & Tetrapeptides :



Inorganic Anions HPLC

Analyzed by: LCBA 45100 Orléans FRANCE

Detergent HPLC Gradient Analyzed by: Pasteur Merieux- R & D Laboratory 69280 Marcy L’Etoile FRANCE

1 : Tergitol type 8 2 : Octyl B glucoside 3 : Sodium dodecyl sulfate 4 : N lauryl sarcosine 5 : Zwittergent 3 14 6 : Triton X 100 7 : Lubrol 8 : PEG 10 tridecyl ether

Gradient: 40-100 % Acetonitrile (3% min)



Low Temperature Operation: HPLC Ethylene Glycol

0

0,2

0,4

0,6

0,8

1

1,2

Minutes

Volts

50°C

40°C

35°C

30°C

-0,006

-0,001

0,004

0,009

0,014

0,019

0,024

0,029

0,034

0 1 2 3 4 5 6 7 8 9Minutes

Volts

10 µl injections (2.5 mg/L), mobile phase 100% water

Analyzed by: R. Pennanec, SEDERE Orléans

LOQ = 2.16 ppm LOD = 0.65 ppm

Drift Tube Temperature



Low Temperature Operation: HPLC Urea

Effect of Temperature on Detector Signal

As the temperature of the detector increases, urea decomposes and its contribution to the observed signal decreases.

Analyzed by: Richard Scientific, Novato CA 94949 USA





Appendix 5 Setting the Date/Time and selecting the output Device The Print button on the front panel can be used to send the present time, date, temperature, gain and pressure to a printer or personal computer. To set present time and date :

a) Turn the detector off b) Press the AZ button while powering up the unit. c) The display will indicate the present year (e.g. 99). Use the up or down

Keys to set the desired year and press AZ again d) The display will indicate the present month (e.g. M 10). Use the up or

down keys to set the desired minute and press AZ. e) Repeat the above process to set the day, hour, minute and second.

To select the output device :

c) Turn the detector off.

d) Press the PRINT button while powering up the unit.

e) The left display will present FF and the right display will indicate 1 or 0. 1 indicates Form Feed and 0 indicates Line Feed. To change the setting, press the PRINT key.

If the AZ (or PRINT) button is not pressed within ten seconds of accessing the date/time (or the FF) display, the display will revert to the normal operating screens.

Appendix 5




Appendix 6 HPLC/SFC Configuration Conversion The SEDEX Model 75 HPLC and SFC Version (230V, part number 75200; 115V part number 75201) can be configured as either a HPLC detector or a SFC detector. The unit is shipped in the HPLC mode. To convert to the SFC mode :

a) Install the SFC Nebulizer (part number 45019). b) Open the gas inlet and set the pressure to 0.5 bar c) Set the circulating bath to 35 oC.

To convert the unit back to HPLC Mode:

a) Install the HPLC Nebulizer (part number 75003). b) Open the gas inlet and set the pressure to 3.5 bar.

Note : if the detector has not been ordered as a SFC and delivered from factory with the accessory, the operation to convert to SFC needs an hardware modification to be compliant with the Sedere delivered detector :

The glass cell circulation bath tubings will pass through a hole in the front panel. Depending on the Sedex75 version, you may have to remove a clipped plastic cap.

Appendix 6


Then, after having removed the detector cover, you'll find on the right side a metal optional part on the detector floor, easy to remove.

Remove it, this will be the way the two tubings will get out from the detector, passing under the detector to get out at the rear.

The two tubings will pass through the front panel hole, dive into the new detector floor hole and get out from the rear of the detector. An additional gas valve can be mounted if user desires to adjust easily auxiliary gas without opening detector cover. The standard auxiliary gas valve must be replaced by the supplied one, mounted through a hole (to be drilled) in the detector rear panel.

Model 45-55-65-75-75C S.E.D.E.R.E. Rev. 14, February 26th, 2007 PARC VOLTA - BP 27 9, rue Parmentier 94141 Alfortville Cedex FRANCE

A7-1

Appendix 7 Standard Operating Procedure and I.Q./O.Q./P.Q. A7.1 Overview The Standard Operating Procedure (SOP) is provided to perform the Installation Qualification (I.Q.), Operational Qualification (O.Q.) and Performance Qualification (P.Q.) to validate the system at your site. The original test report is shipped with each instrument to certify that the instrument passed the final test. Installation of the unit is described in Chapter 2. An Installation Qualification checklist is presented on page A7-2. The procedure for the Operational Qualification is described on the worksheet presented on pages A7-3/4. The procedure for the Performance Qualification is described on the worksheet presented on pages A7-5/6. The overall detector Performance report is presented in Section A7.5. Note : Before starting S.O.P for new instruments or after storage, the following operations must be performed : - Set gas pressure to 3.5bars. - Set the temperature to 50°C and wait for stabilization. - Flush detector with 1ml/min of water at least 15 minutes.

Issue 8 : March 1st 2000 Performed by S. Bertoli approved by Ph. Raynaud Issue 9 : March 3rd 2000 Performed by S. Bertoli approved by Ph. Raynaud Issue 10 : October 23rd 2000 Performed by S. Bertoli approved by Ph. Raynaud Issue 11 : March 20th 2001 Performed by S. Bertoli approved by Ph. Raynaud Issue 12 : February 16th 2005 Performed by S. Bertoli approved by Ph. Raynaud Issue 13 : December 20th 2007 Performed by S. Bertoli approved by Ph. Raynaud Issue 14 : February 26th 2008 Performed by S. Bertoli approved by Ph. Raynaud

Appendix 7 Standard Operating Procedure and IQ/OQ/PQ

Model 45-55-65-75-75C S.E.D.E.R.E. A7-2 Rev. 14, February 26th, 2007 PARC VOLTA - BP 27 9, rue Parmentier 94141 Alfortville Cedex FRANCE

A7.2 The Installation Qualification Checklist

PART I - Installation Qualification (I.Q.) for SEDERE ELS Detectors a) Has the instrument been delivered as ordered (e.g. according

to the URS or purchase order) ? YES [ ] NO [ ] b) Has the instrument been checked and verified as undamaged? YES [ ] NO [ ] c) Has the required documentation been supplied? YES [ ] NO [ ] d) Is it of correct issue and appropriately identified by Model Number, Serial Number and Date?

YES [ ] NO [ ] e) Have details of all services and utilities required to operate the instrument been provided?

YES [ ] NO [ ] f) Have methods and instructions for user maintenance been provided along with contact points

for service and spare parts? YES [ ] NO [ ] g) Is the selected environment suitable for the instrument (i.e. is adequate room provided for

installation, operation and servicing)? YES [ ] NO [ ] h) Have appropriate solvent services and utilities (electricity, nitrogen gas, ventilation, solvent

waste recovery, etc.) been provided? YES [ ] NO [ ] i) Has health, safety and environmental information relating to the operation of this instrument

been provided? YES [ ] NO [ ] Name ________________________________________ Title/Affiliation ________________________________ Date ________________ Signature _____________________________________



A7.3 Operational Qualification Protocol

Part II - Operational Qualification (O.Q.) for SEDERE ELS Detectors This procedure checks the proper operation of the detector with respect to stability of the electronic boards, the lamp energy and the sensitivity of the photomultiplier tube (PMT). In all operational qualification tests, the time constant switch (if the option is mounted) must be set to ‘’1’’ Model Number: SEDEX ___ Instrument Serial Number: _________ Location of Detector: _________________________________________________ This procedure is designed to certify the initial conditions of the ELSD and requires the original test report. This report is located in the user’s manual immediately after this section (and before the reference chromatograms).

a) Turn on the detector b) Seal the solvent inlet connection (Teflon tube) with a plug connection or a piece of Parafilm and

fill the glass siphon with water. c) Apply air or nitrogen pressure, monitored by a regulator and checked with a pressure gauge.

Models 45/55/65 use 2.0 bar. Models 75/75C use 3.5 bar. For models 45/55/65 the display should read 2.0 bar + 0.1 bar (29.0 psi). For models 75/75C the display should read 3.5 bar + 0.1 bar (50.8 psi).

RECORD VALUE _______ PASS [ ] FAIL [ ]

d) Set the temperature to 50°C and wait until the temperature stabilizes (60 minutes)

The display should read 50°C + 1°C.

SETTING TEMPERATURE _______ PASS [ ] FAIL [ ] DISPLAY TEMPERATURE _______

e) Set the gain to 12.



f) Measure the noise over ten 1 min segments. For each segment, measure the difference between the lowest noise peak and the highest noise peak.

Segment + Dev. - Dev. Difference. Segment + Dev. - Dev. Difference. 1 6 2 7 3 8 4 9 5 10

Minimum Value: _______mV Maximum Value ________mV 10-minute average _________mV The 1-minute individual noise should be less than 2.5 mV. PASS [ ] FAIL [ ]. The 10-minute average noise should be less than 2.0 mV. PASS [ ] FAIL [ ]. g) Set the Gain to 1 and set the temperature to 50°C (wait until the temperature stabilizes), allow

air and nitrogen to flow through the system at a pressure of 2 bar (for SEDEX 45/55/65) or 3.5 bar (for SEDEX 75/75C) and autozero the detector. Raise the Gain to 12, monitor the signal for 12 seconds and enter the value in the table below. Observed Value ________mV Reference Value ________mV

The observed Stray Light readings must be + 10% of that indicated on the Test Report at Gain 12. PASS [ ] FAIL [ ] h) This test is run at the same conditions as item f. Position the recorder pen near 10 mV and allow

the recorder to collect data for 30 minutes. Initial Signal Level _____mV Final Signal Level ______mV 30 Min Drift ______mV The baseline drift should be less than 2.0 mV. PASS [ ] FAIL [ ]

Note: If any portion of the diagnostics fail, please refer to the service manual. The manufacturer's procedure for the proper Operational Qualification of this instrument was completed by the following certified person.

Name _________________________________________ Title/Affiliation _________________________________ Signature ______________________________________ Date _________



A7.4 The Performance Qualification Checklist

Part IV - Performance Qualification (P.Q.) for SEDERE ELS Detectors Model Number: SEDEX ____ Instrument Serial Number: _________ Location of Detector: _________________________________________________ Before this procedure is performed, it is necessary to complete the Installation Qualification (Part I) and the Operational Qualification (Part II). The test report presented below (page A7-9) or your own appropriate reference test should be used. 1) Turn on the detector and set the conditions as follows:

Temperature : 50oC Gain : 12 Time constant : 1 (if the option is mounted) Gas pressure : 2.0 bar (45/55/65) or 3.5 bar (75/75C) (air or nitrogen) Syphon : filled with water Inlet tube : connected to column (4.6 mm x 30 mm) C18, 5µm (*) or 1/16’’ tubing, 0;005’’ I.D. x 200 cm Solute : Caffeine as specified on the test report. Flow rate : 1 ml/min Mobile phase : 100% of water or 95% of water with 5 % of methanol or acetonitril (*) Remove the plug connector (or Parafilm) from the solvent inlet connector before connecting the inlet tube to the column. Note that when using a loop, it must be placed between the pump and the autosampler.

2) Set the detector signal close to zero and autozero the detector. 3) Make three (3) 20 µl injections of 5 µg/mL of a caffeine standard. The average peak height

should be : SEDEX75 :

At least 50 mV under test report conditions or your internal reference test (column) At least 70 mV under test report conditions or your internal reference test (loop) For CC version deduct 20 mV from the above values.

SEDEX 55 : At least 25 mV under test report conditions or your internal reference test (column) At least 30 mV under test report conditions or your internal reference test (loop)



Column [ ] Loop [ ] Peak height 1 _______ Peak height 2 _______ Peak height 3 _______ Average Peak height ______ PASS [ ] FAIL [ ] 4) Make 6 injections (20 µl) injections of a 250 µg/mL caffeine standard at gain 6 (for SEDEX 55

use gain 8). Deliver 5x the loop volume for manual injections or full loop injection with autosampler. The repeatability should be:

3.0 % RSD maximum.

Peak Area 1: ________ Peak Area 4 _______ Peak Area 2: ________ Peak Area 5 _______ Peak Area 3: ________ Peak Area 6 _______ Avg. Peak Area: _______ Std. Dev. : _______ % RSD : _____

PASS [ ] FAIL [ ]

The manufacturer's procedure for the proper Performance Qualification of this instrument was completed by the following certified person. Name _________________________________________ Title/Affiliation _________________________________ Signature ______________________________________ Date _________



A7.5 Overall Detector Performance After the Installation Qualification, Operational Qualification and Performance Qualification procedures have been completed, the Overall Detector Performance document should be completed to verify the completion of all tests.

Part IV - Overall Detector Performance for SEDERE ELS Detectors Model Number: SEDEX __ Instrument Serial Number: _________ Location of Detector: _________________________________________________ Part I - Installation Qualification (I.Q.) Date ________ PASS [ ] FAIL [ ] Part II - Operational Qualification (O.Q.) Date ________ PASS [ ] FAIL [ ] Part III - Performance Qualification (P.Q.) Date ________ PASS [ ] FAIL [ ] The above instrument was certified by the following certified manufacturer’s representative : Name ________________________________________________________ Title/Affiliation ________________________________________________ Date ________________ Signature _____________________________________________________ The above instrument was certified within presence of the following customer’s representative : Customer name ________________________________________________ Customer title/Affiliation ________________________________________ Customer Signature _____________________________________________





A8-1

Appendix 8 The PC Controller Software Program A8.1 Introduction The SEDEXTM Controller software program has been designed to allow the operator to: • Set operating parameters for manual operation of the SEDEX Model 75, 85 and 754 Low

Temperature Evaporative Light Scattering Detectors from a personal computer. • Generate automated methods that define the operation of the detector. The method may include

temperature and/or gain changes as a function of time. When the run is initiated (either manually or via a signal from the HPLC system), these changes will be executed at the indicated time(s). In this mode, the time and temperature profiles, the present operating conditions and the chromatogram will be displayed on the monitor as shown in Figure A8-1. Figure A8-1 presents the Method Mode screen for the SEDEX Model 754. This screen shows the temperature/gain program as well as the chromatogram (there are slight differences in the program for the three detectors).

Figure A8-1: The SEDEX 754 Method Mode Screen

At the completion of the run, the chromatographic data can be saved as a data file and transmitted to a spreadsheet. The spreadsheet can include the signal intensity/time data and/or peak information that meet a user-selected threshold. If desired, two (or more) methods can be incorporated into a method list. The method lists lets you perform a number of runs using a given method, then switch to another method and perform additional runs. Each method list run can be initiated manually or via a signal from the HPLC system. - Full user manual is available in Software CD-ROM in Acrobat file format. - Demo version is available at Sedere website : www.sedere.com

Appendix 8 The PC software controller program



Documents

Sedex75 User Manual Revision 7b Complete