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Antec Scientific SenCell User Manual

Electrochemical flow cell
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Antec Scientific
Industrieweg 12
2382 NV Zoeterwoude
The Netherlands
SenCell
TM
Electrochemical Flow cell
User manual
116.0010, Edition 1, 2012
T +31 71 5813333
| info@AntecScientific.com | https://AntecScientific.com

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  • Page 1 Antec Scientific Industrieweg 12 2382 NV Zoeterwoude The Netherlands SenCell Electrochemical Flow cell User manual 116.0010, Edition 1, 2012 T +31 71 5813333 | info@AntecScientific.com | https://AntecScientific.com...
  • Page 2 The information contained in this document is subject to change without notice. ROXY, ALEXYS, DECADE, DECADE II, INTRO, Flexcell, ISAAC, HyREF, SenCell are trademarks of Antec. Whatman™ (word and device) and Whatrnan™ (word only) are trademarks of Whatman lnternational Ltd. SOLVENT IFD™ and AQUEOUS IFD™ are trademarks of Arbor Technologies, Inc.

  • Page 3: Symbols

    INTRODUCTION Table of contents Symbols The following symbols are used in this guide: The warning sign denotes a hazard. It calls attention to a procedure or practice which, if not adhered to, could result in severe injury or damage or destruction of parts or all of the equipment.

  • Page 4: Laboratory Regulations

    SenCell flow cell user manual, ed. 1 (U) HPLC: (Ultra) High Performance Liquid Chromatography (HPLC) is a method for separating substance mixtures, determining substances and measuring their concentration. This device is suitable for high-performance liquid chromatography. It is suitable for laboratory use, for analyzing substance mixtures that can be dissolved in a solvent or solvent mixture.

  • Page 5: Solvents

    INTRODUCTION Table of contents Solvents Organic solvents are highly flammable. Since capillaries can detach from their screw fittings and allow solvent to escape, it is prohibited to have any open flames near the analytical system! Regularly check for leaks and clogged LC tubing and connections. Test back pressure without column.
  • Page 6 SenCell flow cell user manual, ed. 1...

  • Page 7: Table Of Contents

    Reference electrodes 18 ISAAC reference electrode 18 Salt bridge Ag/AgCl reference electrode 20 HyREF reference electrode 20 Installation 22 Introduction 22 Adjusting the SenCell working volume 23 Installation in LC system 24 Maintenance 28 Assembling/Disassembling the Cell 28 HyREF 30 ISAAC 30...
  • Page 8 SenCell flow cell user manual, ed. 1...

  • Page 9: The Electrochemical Flow Cell

    (Patent Pending) like a stepless adjustable working volume (spacerless concept) and toolless assembly. The SenCell is available with a glassy carbon working electrode (WE). The SenCell design eliminates the use of plastic/metal spacers. The working volume of the electrochemical cell can be stepless adjusted without opening the cell by means of a special key, allowing easy optimization of the detection sensitivity for any LC application.

  • Page 10: Three-Electrode Configuration

    Three-electrode configuration In the SenCell flow cell a three-electrode configuration is used (Fig. 2). The working potential is set between the working electrode (WE) and the auxiliary electrode (AUX). The AUX is kept at a precisely defined reference electrode (REF) potential by means of the so-called voltage clamp.

  • Page 11: Working Electrode

    I/E characteristics. This in fact means that a high signal must be obtained at a low working potential. For most applications glassy carbon will be the WE material of choice. The SenCell is currently available with 2 mm diameter Glassy Carbon electrode only. Under certain circumstances other materials are favourable.

  • Page 12: Detection Limit

    SenCell flow cell user manual, ed. 1 Another consideration in choosing a WE is the oxidation or reduction of mobile phase constituents or WE material, that occurs when the potential exceeds the limits as given in Table I. At high positive working potentials the water in the mobile phase electrolyses and results in an strong increase of the background current and noise.

  • Page 13: Cell Working Volume Adjustment

    CHAPTER 1 The electrochemical flow cell times the sigma-noise is 11 pmol/L (see Table II for conditions). Expressing the performance of a detection system by only the peak height makes no sense. A system can easily be changed in a way that a larger peak height is obtained.
  • Page 14 (signal-to-noise ratio) for any LC application. Fig. 4. Example chromatograms of 100 nM standard of catecholamines in 10 mM HAc recorded with the Sencell spacing adjustment set to position 3 and 0.5, corresponding with an approximate spacing setting of 100 µm and 12 µm respectively.
  • Page 15 Fig. 5. Normalized Peak height of Dopamine as a function of spacing setting (red curve) based on chromatograms recorded with a 100 nM standard of catecholamines in 10 mM HAc with a SenCell. The dotted curve is a simulated curve based on the Cotrell equation (Lit ref F.G. Cottrell, Z. Phys.
  • Page 16 Space setting SenCell (µm) Fig. 6. Left side: ASTM noise values as a function of cell spacing. Right side: Noise traces as a function of cell spacing. SenCell spacing position 3 corresponds with approximately 100 ± 10 µm. The spacings used with this particular SenCell under test in this experiment were determined using a stylus profilometer.
  • Page 17 CHAPTER 1 The electrochemical flow cell...

  • Page 18: Reference Electrodes

    SenCell flow cell user manual, ed. 1 C H A P T E R Reference electrodes The SenCell is available with an ISAAC (in situ Ag/AgCl) reference electrode, a salt bridge Ag/AgCl reference electrode and a HyREF reference electrode. ISAAC reference electrode The ISAAC reference electrode is in direct contact with the mobile phase which contains chloride ions.
  • Page 19 CHAPTER 2 Reference electrodes Fig. 7. Dependence of the Ag/AgCl REF potential on the chloride concentration. Table III. Potential of the Ag/AgCl reference electrode, dE is the potential difference with E in saturated KCl. Ag/AgCl (mmol/l) E Ag/AgCl (mV) dE (mV) 3500 2500 1500...

  • Page 20: Salt Bridge Ag/Agcl Reference Electrode

    KCl. This REF for the SenCell is factory filled with KCl. For certain applications another chloride salt is to be preferred. In case of perchlorate containing mobile phases, sodium chloride is mandatory, because potassium perchlorate precipitates and will clog the cotton wool frit.
  • Page 21 CHAPTER 2 Reference electrodes especially in case of high modifier concentrations (i.e. analysis of fat-soluble vitamins) or high pH (analysis of carbohydrates, PAD). Depending on the pH of the mobile phase, the potential setting of the working electrode vs. the HyREF may differ significantly compared to Ag/AgCl. I/E curves show a shift of more than 200 mV at pH 3.1 (e.g.

  • Page 22: Installation

    The SenCell is delivered pre-assembled and ready for installation and use. For instructions about assembling the cell refer to chapter 4. Fig. 9. Left side: photo of assembled SenCell inlet block (green) with In-Situ Ag/AgCl (ISAAC) reference electrode (REF). Right side: bottom side of the SenCell with the Cell working volume adjustment system, Auxiliary (AUX) electrode contact (opening on left side next to inscription ‘3’) and Working...

  • Page 23: Adjusting The Sencell Working Volume

    1. Insert pins of the adjustment key (p/n 116.1015) in the two holes on the bottom side of the SenCell. Note that the diameter of the two pins differ. The larger diameter pin should be inserted in the AUX contact.

  • Page 24: Installation In Lc System

    In our experience, a one-time degassing step of the HPLC buffer is almost never sufficient. Follow the procedure below to install the SenCell, see figure 3 and 4 on the next page for reference: 1. If applicable, install the SenCell clamp (p/n 250.0102A) from the shipkit (p/n 116.0202) in the center position of the DECADE II detec-...
  • Page 25 5. For a SenCell with HyREF (black inlet block) or ISAAC reference (green inlet block): fill the flow cell, by keeping it in an angle of about 45° with the outlet (LC out) on top to force the air through the outlet.
  • Page 26 SenCell flow cell user manual, ed. 1 Fig. 11. Left: SenCell with ISAAC reference mounted under an angle of approximately 45 °C in the DECADE II detector. [1] Cell clamp, [2] Cell Inlet (tubing connection from column–to–cell), [3] Cell outlet (tubing connection...
  • Page 27 In the case of an Intro or DECADE detector the electrical connections should be made as depicted in the figure below: Fig. 13. Installation of SenCell (Intro™ or DECADE™). WORK, AUX and REF are connected using the red, blue and black cell cable. LC out should be...

  • Page 28: Maintenance

    C H A P T E R Maintenance Assembling/Disassembling the Cell Fig. 14. Top: Exploded view of SenCell. The arrows indicate how to assemble the cell. Bottom left: unscrewing the Salt bridge electrode. Bottom right: WE block with O-ring placed in designated O-ring groove.
  • Page 29 2. Check the back side of the WE block and assure that the cell working volume adjustment is set to position 1 or 2. Assure that the SenCell is not set to position 0, because this may cause damage the WE or cell during assembling.

  • Page 30: Hyref

    SenCell flow cell user manual, ed. 1 HyREF The HyREF reference electrode is in principle maintenance free. If not in use it should be stored dry after disassembling the flow cell. ISAAC The ISAAC reference electrode requires maintenance, usually not more than once in 3 months.

  • Page 31: Coating With Isaac Solution

    CHAPTER 4 Maintenance Fig. 15. Polishing: Apply a few drops on the polishing disc(left), and polish the electrode with a ‘figure 8’ motion(right). Coating with ISAAC solution Be careful: take the necessary precautions (gloves, lab coat and glasses) because the ISAAC solution is corrosive. 1.

  • Page 32: Ag/Agcl Salt Bridge

    SenCell flow cell user manual, ed. 1 Ag/AgCl salt bridge Three aspects determine the proper function of an Ag/AgCl reference electrode. The chloride concentration must be kept at a strictly fixed level. This is best guaranteed by using a saturated chloride salt solution at a constant temperature.

  • Page 33: Material

    Material • An over-saturated and thoroughly degassed KCl solution (p/n 250.2004). • SB ref tool (p/n 250.1035). Not included in the SenCell ship kit, should be purchased separately. • Ordinary cotton wool. Procedure Use proper eye and skin protection when working with solvents.

  • Page 34: Maintenance Of The Cotton Wool Frit

    SenCell flow cell user manual, ed. 1 Maintenance of the cotton wool frit Use proper eye and skin protection when working with solvents. Use the SB ref replacement tool (p/n 250.1035) to push out the frit from the outside (Fig. 18).

  • Page 35: Working Electrode

    Set the WE block in position 0 (polishing position) using the adjustment key (p/n 116.1400) and turning it counter-clockwise. Make sure the SenCell is disassembled when setting it in position 0, otherwise it may damage the cell of WE electrode surface.
  • Page 36 SenCell flow cell user manual, ed. 1 Fig. 19. SenCell WE block with working volume set to position 0. 1. Shake diamond slurry (p/n 250.1030) thoroughly before use!! 2. Rinse the polishing disc with demi water before applying the diamond slurry! 3.
  • Page 37 CHAPTER 4 Maintenance...

  • Page 38: Specifications

    SenCell flow cell user manual, ed. 1 C H A P T E R Specifications Cell type Three electrode, wall-jet flow cell 0 – 300 nL (stepless adjustable) Cell working volume (based on 2 mm Ø WE) Total cell volume Approximately 0.5 mL...
  • Page 39 CHAPTER 5...

  • Page 40: Part List

    SenCell flow cell user manual, ed. 1 C H A P T E R Part list In the table below parts and spare parts are listed of the SenCell: Description 116.0010 SenCell manual 116.0014 SenCell Quick-start guide 116.0202 SenCell Shipkit 116.0500...
  • Page 41 In the table below parts and spare parts are listed of the SenCell:...

  • Page 42: Index

    SenCell flow cell user manual, ed. 1 C H A P T E R Index Ag/AgCl reference electrode parts, 29 standard electrode potential, 12 auxiliary electrode, 5 detection limit, 7 flow cell volume, 11 HyREF installation, 16 reference electrode potential, 14...

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