Eicom ENO-30 Manual de usuario

ENO-30 ENG, #04v2,2014

NOx Analyzer (ENO-30)

User’s Guide

Distributed by Eicom USA: 7098 Miratech Dr, Ste 100, San Diego, CA 92121, USA

Phone: (888) 680-7775 FAX: (858) 560-8040 Email: info@eicom-usa.com

Distributed by Eicom Europe: Hilton House Ardee Rd, Ground Floor, Rathimines Dublin 6, Ireland

Phone: +353 1 902 2700 FAX: +353 1 443 0784

Manufacturered by Eicom Corporation: 113 Kita Enmenden-cho, Shimotoba, Fushimi-ku Kyoto, Japan, 612-8497

Phone: +81-75-622-2112 FAX : +81-75-622-2114

Contents

1. INTRODUCTION ---------------------------------------------------------------------------------------------------------------------------1

1-1. About this user’s guide -----------------------------------------------------------------1

1-2. Important Safety Information ---------------------------------------------------------2

2. OVERVIEW ----------------------------------------------------------------------------------------------------------------------------------3

2-1. Principle of Measurement -------------------------------------------------------------3

2-2. Schematic of Parts and Functions --------------------------------------------------4

2-3. Tubing Flow Diagram-------------------------------------------------------------------6

2-4. Pump Seal Wash ------------------------------------------------------------------------7

2-5. Fittings and Tubing----------------------------------------------------------------------8

3. INSTALLATION--------------------------------------------------------------------------------------------------------------------------- 10

3-1. Location ----------------------------------------------------------------------------------10

3-2. Power Plug------------------------------------------------------------------------------ 10

3-3. Electrical connections (Detector Unit side panel)----------------------------- 11

3-4 Electrical Connections (Pump Unit Signal Terminal)--------------------------12

3-5. Manual injector Tubing Connections --------------------------------------------- 13

3-6. Connect Tubing to Autosampler (optional)--------------------------------------13

4. PUMP UNIT OPERATION-------------------------------------------------------------------------------------------------------------- 14

4-1. Pump overview-------------------------------------------------------------------------14

4-2. Power On --------------------------------------------------------------------------------15

4-3. Key Pad ----------------------------------------------------------------------------------16

4-4. Display Screen -------------------------------------------------------------------------18

4-5. Log Function---------------------------------------------------------------------------- 18

4-6. Adjusting the Settings ---------------------------------------------------------------- 19

4-7. Priming the Pump/ Purge Valve Operation -------------------------------------22

4-8. Pump error messages---------------------------------------------------------------- 23

5. DETECTOR/OVEN OPERATION ---------------------------------------------------------------------------------------------------- 24

5-1. Power Switch ---------------------------------------------------------------------------24

5-2. Setting Temperature ------------------------------------------------------------------24

5-3. Detector Readings--------------------------------------------------------------------- 24

5-4. Autozero Function --------------------------------------------------------------------- 24

6. ANALYSIS---------------------------------------------------------------------------------------------------------------------------------- 25

6-1. Accurate Analysis---------------------------------------------------------------------- 25

6-2. Water Quality ---------------------------------------------------------------------------25

6-3. Reagent Quality------------------------------------------------------------------------ 25

6-4. Carrier and Reactor Solutions Preparation ------------------------------------- 26

6-5. Standard Solution----------------------------------------------------------------------27

6-6. Waste-------------------------------------------------------------------------------------27

6-7. Prepare for Analysis-------------------------------------------------------------------27

6-8. Start up ----------------------------------------------------------------------------------- 28

7. SAMPLE INJECTION ------------------------------------------------------------------------------------------------------------------- 29

7-1. Manual Injector Operation-----------------------------------------------------------29

7-2. Sample Amount ------------------------------------------------------------------------30

7-3. Autosampler Operation (optional) -------------------------------------------------30

8. SHUTDOWN AFTER ANALYSIS----------------------------------------------------------------------------------------------------- 31

8-1. Short Shutdown, less than 2 weeks ----------------------------------------------31

8-2. Complete Shutdown, over 2 weeks----------------------------------------------- 31

9. RESTART ---------------------------------------------------------------------------------------------------------------------------------- 32

9-1 After complete shutdown procedure----------------------------------------------- 32

9-2. After short shutdown procedure----------------------------------------------------32

10. PRECOLUMN --------------------------------------------------------------------------------------------------------------------------- 34

10-1. Determining the Precolumn Condition------------------------------------------34

10-2. How to Repack the Precolumn---------------------------------------------------35

10-3. Filter Exchange-----------------------------------------------------------------------37

11. SEPARATION COLUMN (NO-PAK) ----------------------------------------------------------------------------------------------- 38

11-1. NO-PAK--------------------------------------------------------------------------------- 38

11-2. Identification of Separation Quality----------------------------------------------38

11-3. Washing the Separation Column-------------------------------------------------38

12. REDUCTION COLUMN (NO-RED) ------------------------------------------------------------------------------------------------ 39

12-1. Structure and Maintenance--------------------------------------------------------39

12-2. Timing to Exchange -----------------------------------------------------------------39

12-3. Damage --------------------------------------------------------------------------------39

13. TROUBLESHOOTING ---------------------------------------------------------------------------------------------------------------- 40

13-1. Pump Problems ----------------------------------------------------------------------40

13-2. Checking the Flow Rate------------------------------------------------------------ 40

13-4. Removing Air Bubbles --------------------------------------------------------------40

13-4. High Carrier Pump pressure ------------------------------------------------------41

13-5. Unstable Baseline-------------------------------------------------------------------- 41

13-6. Peak Shape --------------------------------------------------------------------------- 41

14. MAINTENANCE------------------------------------------------------------------------------------------------------------------------- 42

14-1. Pump Head Parts--------------------------------------------------------------------42

14-2. Changing the Piston Seal and Piston------------------------------------------- 43

15. APPENDICES --------------------------------------------------------------------------------------------------------------------------- 47

15-1. Autosampler and Data Processor (EPC-700) Connection ----------------47

15-2. Specifications ------------------------------------------------------------------------- 48

15-3. Sample Preparation -----------------------------------------------------------------49

1 | P a g e

1. Introduction

Thank you for purchasing the Eicom NOx Analyzer (ENO-30). Please read this user’s guide

before use.

The ENO-30 is an HPLC-based system designed to perform high sensitivity analysis of Nitrite

and Nitrate in biological samples. The lower half of the system is the pump unit. It has two

independent pumps and a two channel online degasser. The pump has s self-learning pulse

damping flow control which ensures stable liquid delivery, and is made of inert, non-metallic

parts for better salt and acid resistance.

The upper unit uses forced air and a Peltier device to maintain the columns, reaction loop, and

detector cell at the correct temperature for the analysis. The detector signal is taken from the

side terminals into a data process (EPC-700) and digitized for analysis in a PC with the Envision

software.

1-1. About this user’s guide

Latest-breaking information may be supplied separately.

We reserve the right to change the content of this document with or without notice.

No part of this guide may be reproduced by any means without the prior written permission

The ENO-30 requires a thorough understanding of this document for proper operation.

Please contact Eicom directly if you have any unanswered questions after reading this

document.

2 | P a g e

1-2. Important Safety Information

We insist that users observe the following procedures in order to prevent accidents:

1) Be sure to read and understand this manual completely prior to operating the ENO-30.

2) Follow all warnings listed herein very closely.

3) Do not alter the ENO-30.

4) Never attempt to repair or dismantle the ENO-30 on your own.

This device has been produced for experts who have knowledge of chemical

analysis and handling research reagents. Failure to follow instructions may

not only lead to poor quality data, but could result in a safety hazard such as

fire, electric shock, injury or other damage. DO NOT OPERATE the ENO-30

until reading and understanding this instruction manual completely.

WARNING

Before you operate the ENO-30 using harmful chemical reagents,

be sure to understand its handling methods, physical and chemical

characteristics and MSDS (material safety data sheets). Mishandling

harmful chemical reagents might result in death or serious injury to the user.

To avoid health hazards, wear proper protective gloves, goggles and mask,

and be sure there is adequate ventilation. Never allow leakage from any

connection points of the tubing.

WARNING

Flammable chemical reagents must be kept away from sources of

ignition and may give off flammable vapors if left uncovered.

WARNING

3 | P a g e

2. Overview

2-1. Principle of Measurement

The ENO-30 is a high sensitivity instrument for measuring nitrite and nitrate ion level in

biological fluids. This is achieved by combining a colorimetric diazo coupling method (Griess)

with the advantages of HPLC.

After the sample is injected it is filtered by the guard column. Then the separation column

interacts with the ions in such a way that they flow though the column at different rates. Nitrite

exits the column first, and then some minutes later, the Nitrate leaves the column. The NO2-and

NO3-next flow into the reduction column made is cadmium and copper.

Inside the reduction column, the NO3-is reduced to NO2-through a reaction with the cadmium

and reduced copper. The NO2- that exited first will not react. So now there are two Nitrite peaks,

the first is from the Nitrite in the sample and the second is from the Nitrate in the sample. Both

are mixed with a Reactor solution supplied by a second pump at a 3-way joint, and then flow

into a reaction loop. This gives time for the reaction come to the right temperature and complete

formation of a light absorbing diazo compound that can measured at the detector.

In the detector cell, 540 nm (green) light passes through the sample, and the absorption is

proportional to the amount of diazo compound, and thus to the amount of Nitrite or Nitrate.

The response of the detector is transformed to a voltage which it generates at the CPU terminal.

The time/voltage change is traced by a data processor such as the optional Eicom EPC-700. By

comparing peak area or height shown on a chromatogram with a standard one, the exact

concentration Nitrite and Nitrate can be calculated.

4 | P a g e

2-2. Schematic of Parts and Functions

1. Pump control panel

Set the flow rate of each pump and the upper pressure limit. Then start and stop the pump.

2. Degasser

This degasser removes dissolved air from the Carrier and Reactor solutions before they enter

the pumps. It does not work for large bubbles in the inlet tubing. The degasser prevents bubbles

from forming in the pumps, which will cause them to stop. The internal volume of each degasser

channel is 300 µL.

3. Carrier Pump

It supplies carrier solution to the separation column and reduction column.

4. Reactor Pump

5 | P a g e

It supplies the reactor solution to the reaction coil.

5. Reactor Backpressure Coil

It maintains high enough pressure that the reactor pump can work efficiently.

6. Manual Injector (and/or Autosampler)

Manual injection of up to 50 µL of sample using a blunt ended Hamilton syringe can be made

here.

7. Mixer

Mixes the column effluent with the Reactor solution before entering the reaction coil.

8. Reaction Coil

Gives time for the reaction to be heated and go to completion. It also helps to reduce mixing

noise.

9. Detector Cell

Detects absorbance at 540 nm.

10. Separation Column

Separates the Nitrite and the Nitrate from other biological compounds.

11. Reduction Column

Reduces the Nitrate to Nitrite so that it can react with the Greiss reagent.

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