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Flow Analysis with Spectrophotometric and Luminometric Detection

Flow Analysis with Spectrophotometric and Luminometric Detection, 1st Edition

Flow Analysis with Spectrophotometric and Luminometric Detection, 1st Edition,Elias Ayres Guidetti Zagatto,Claudio Oliveira,Alan Townshend,Paul Worsfold,ISBN9780123859242

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229 X 152

This book describes the amazing developments in the last decade of flow analysis, the approach to automated wet chemical assays.

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Key Features

  • Detailed historical and theoretical background
  • Various modes of operation
  • Spectrophotometric and luminometric detection
  • Strategies for solution handling
  • Large number of applications


With the ever increasing number of samples to be assayed in agronomical laboratories and servicing stations, fertilizer and food industries, sugar factories, water treatment plants, biomedical laboratories, drug quality control, and environmental research, the interest for automated chemical analysis has been increasing.

In this context, flow analysis is very attractive, as they the flow-based procedures are characterized by enhanced analytical figures of merit. Moreover, the flow analysers do not usually require sophisticated and expensive instrumentation, are amenable to full automation and to miniaturization, and are well suited for in situ analyses.

The tendency to carry out traditional methods of analysis in the flow analyser has becoming more pronounced, especially in relation to large-scale routine analyses. The technology of solution handling has become more and more improved, leading to enhanced strategies for chemical assays. Consequently, different modalities of flow analysis (e.g. SFA, FIA, SIA) have been conceived, developed and applied to solve real problems. Most of the flow-based analytical procedures presently in use, however, do not exploit the full potential of flow analysis.

The main object of the book is then to provide a scientific basis and to familiarise a wide community of researchers, students, technicians, etc with the uses of flow analysis. Emphasis is given to spectrophotometric and luminometric detection, in relation to agronomical, geological, industrial, pharmaceutical and environmental applications.

The book includes historical and theoretical aspects, recent achievements in instrumentation, guidelines for methodology implementation, and applications. It serves also as an applications-oriented text book.


Researchers, students, consultants and practitioners in flow analysis, spectrophotometry, luminescence, flow injection analysis, sequential injection analysis and segmented flow analysis.

Elias Ayres Guidetti Zagatto

Affiliations and Expertise

CENA, University of São Paulo, Brazil Universidade de Sao Paulo, Centro de Energia Nuclear na Agricultura, Brazil

Claudio Oliveira

Affiliations and Expertise

University of Maringa, Brazil

Alan Townshend

Alan Townshend holds B.Sc., Ph.D. and D.Sc., degrees from the University of Birmingham, where he lectured in analytical chemistry from 1964-80. He moved to the University of Hull in 1980, and became Professor of Analytical Chemistry in 1984. He has served twice as Dean of the School of Chemistry, and currently holds the G.F. Grant Chair of Chemistry. He is also Deputy Dean of the Faculty of Science, and Director of the Institute for Chemistry in Industry. He has published ca. 300 scientific papers, his main research interests currently being in analytical applications of chemiluminscence and of immobilised reagents (especially enzymes) and in flow injection analysis.

Professor Townshend is a senior editor of Analytica Chimica Acta, and was Editor-in-Chief of the Encyclopedia of Analytical Science (Academic Press, 1985) and the Dictionary of Analytical Reagents (Chapman and Hall, 1993). He was President of the Analytical Division of the Royal Society of Chemistry (RSC) (1996-98) and is currently Chairman of the RSC's Committee for Accreditation and Validation. He serves on the Steering Group of the UK Analytical Partnership and has recently completed a 4-year stint on the Council of the Analytical Chemistry Division of the Federation of European Chemical Societies.

Affiliations and Expertise

University of Hull, UK University of Hull, Department of Chemistry, United Kingdom

Paul Worsfold

Affiliations and Expertise

University of Plymouth, UK

Flow Analysis with Spectrophotometric and Luminometric Detection, 1st Edition

1. Introduction

1.1. General

1.2. The advent of Flow Analysis

1.3. The development of Flow Analysis

1.4. Main features of Flow Analysis

1.4.1. Sample insertion

1.4.2. Sample dispersion

1.4.3. Reproducible timing

1.4.4. Other attractive features

Transient analytical signal

Low susceptibility to biased results

Improved system design

    1. Aims and scope of the monograph

Appendix 1.1. Important monographs related to Flow Analysis

Appendix 1.2. Journal Special Issues dedicated to Conferences on Flow Analysis

2. Historical view

2.1. Early developments

2.2. Segmented flow analysis

2.3. Flow injection analysis

2.4. Sequential injection analysis

2.4.1. Bead injection analysis

2.4.2. Lab-on-valve

2.5. Multi-commuted flow analysis

2.5.1. Multi-syringe flow injection analysis

2.5.2. Multi-pumping flow analysis

2.6. Other flow systems

2.7. Commutation in flow analysis

2.8. Flow pattern

2.9. Instrument characteristics

2.10. Trends

3. Fundamentals

3.1. The flowing sample

3.1.1. Flow pattern Flow regime Composition of the flowing stream

Tandem stream

Mono-segmented stream Temporal variations in flow rates

Constant flow

Pulsating flow

Sinusoidal flow

Reversed flow

Linearly-variable flow

Intermittent flow

Pulsed flow Alterations to the flow pattern

3.1.2. Sample dispersion Dispersion inside a tubular reactor Dispersion inside a mixing chamber

Improved mixing conditions

High sample dispersion

Exponential dilution

Establishment of fluidized beads

Manifold components behaving as mixing chambers Dispersion by confluent streams Practical indices for expressing sample dispersion

The volumetric fraction.

Experimental determination

Practical situations

Worked examples

3.1.3. Visualisation of the dispersing sample zone

3.2. System configurations

3.2.1. Single-line flow systems

3.2.2. Confluence flow systems

3.3. The detector response

3.3.1. Flat peaks

3.3.2. Bell shaped peaks Peak height Peak area Peak width

3.3.3. Gathering the calibration model

4. Interaction of radiation with the flowing sample

4.1. Fundamentals

4.1.1. UV-Visible spectrophotometry Losses of radiation

Radiation losses at interfaces

Radiation losses at the cuvette walls

Radiation losses inside the sample

Compensation of radiation losses The Lambert-Beer law Practical aspects of the Lambert-Beer law

Concentration of the radiation absorbing species

Monochromaticity of the radiation

Sample homogeneity

Chemical deviations


Stray radiation Special strategies

Dual-wavelength spectrophotometry

Solid phase spectrophotometry

4.1.2. Turbidimetry Losses of radiation Relationship between turbidance and analyte concentration Practical aspects

Characteristics of the particles

Monochromaticity of radiation

Sample uniformity

Chemical deviations (including co-precipitation)

Rate of turbidity formation

Stray radiation Special strategies Final remarks

4.1.3. Nephelometry

4.1.4. Fluorimetry and phosphorimetry

4.1.5. Chemiluminescence and bioluminescence

4.2. The Schlieren effect

4.2.1. Physical principles

4.2.2. Occurrence

4.2.3. The Schlieren effect in flow analysis Historical testimonies The two components of the Schlieren effect

Poor mixing conditions

Good mixing conditions Applications Emergence

Differences between sample and carrier solutions

Pulsed sample inlet into the flow cell

Pulsed addition of merging streams

Exploitation of intermittent streams

Addition / removal of manifold components Minimizing the Schlieren effect

Improvement of system design

Subtraction of monitored signals

4.3. Presence of immiscible phases

5. Flow Analysers

5.1. The segmented flow analyser

5.1.1. Characteristics

5.1.2. Sample dispersion

5.1.3. Controlling sample dispersion

5.2. The flow injection analyser

5.2.1. Characteristics

5.2.2. Sample dispersion

5.2.3. Controlling sample dispersion Dispersion parameters


Sample viscosity

Diffusion coefficient

Composition of the sample and reagent solution

Others Dispersion parameters - system geometry

Sample volume

Injection mode

Dimensions of the analytical path

Tubing inner diameter

Flow rate

Confluent stream additions

Site of confluence stream addition

Artefacts in the analytical path

5.3. The sequential injection analyser

5.3.1. Characteristics

5.3.2. Sample dispersion

5.3.3. Controlling sample dispersion Large sample volume Limited sample dispersion

5.4. The multi-commuted flow analyser

5.4.1. Characteristics

5.4.2. sample dispersion

5.4.3. Discretely actuated devices Automated devices Devices with feedback mechanisms

5.5. Other flow analysers

5.5.1. The mono-segmented flow analyser

5.5.2. The discontinuous flow analyser

5.3.3. The Lab-on-Valve and Lab-on-Chip flow analysers

5.6. Describing the flow analyser

5.6.1. Establishment of the flowing stream

5.6.2. Sample introduction

5.6.3. Manifold characteristics

5.6.4. Sample processing

5.6.5. Detection

5.6.6. Performance of the flow system General figures of merit





Detection limit

Dynamic range Figure of merit specific to flow based procedures


Sampling rate



6. Instrumentation

6.1. Fluid propulsion

6.1.1. Peristaltic pumps

6.1.2. Syringe (piston) pumps Use of large volume pistons (syringes) Use of small volume pistons

6.1.3. Diaphragm pumps Solenoid pumps Piezoelectric pumps

6.1.4. Gas pressurized reservoirs

6.1.5. Osmotic pumps

6.1.6. Gravity

6.2. sample handling

6.2.1. Tubes

6.2.2. Sample introduction Sampler Time-based introduction Loop-based introduction Hydrodynamic injection Nested injection

6.2.3. Reactors Coiled reactors / mixing coils Packed bed reactors Single bead string reactor Knitted (or knotted) reactors Reactor-like artefacts

6.2.4. Accessories Connectors

Connectors for linking tubes together

Connectors for linking manifold tubes with other components Solution containers

6.3. Detection and data processing

6.3.1. Flow-cells Classical and Z-shaped flow-cells Spiral flow-cells Long optical path length flow-cells

6.3.2. Detectors

6.3.3. System control, data acquisition and data treatment

6.4. Miniaturisation of the flow system

7. Special strategies for flow manipulation

7.1. Merging zones

7.1.1. Implementation Merging zones relying on different convergent carrier streams Merging zones relying on a single carrier stream Merging zones relying on an intermittent stream

7.1.2. Applications Reduction of reagent consumption Matching the analytical signal with the dynamic concentration range Single point determinations incorporating blank determinations Speciation Reducing undesirable adsorption processes Sequential determinations Standard addition method

7.2. Zone sampling

7.2.1. Implementation

7.2.2. Applications High sample dispersion Simultaneous determinations Variable dispersion Detailed study of dispersion

7.3. Stream splitting

7.3.1. Segmented flow analysis

7.3.2.Unsegmented flow analysis Stream splitting / stream merging

Expansion of the dynamic concentration range

Differential kinetic analysis

Simultaneous determinations Stream splitting without stream merging

7.3.3. Sample removal from the analytical path

7.4. Zone splitting

7.4.1. Implementation

7.4.2. Applications

7.5. Sample incubation: zone trapping (general) and sample stopping

7.6.Prior assay

7.7. Multi-detection (with a single detector)

8. Sample handling

Quotes and reviews

"The book is clearly written and comprehensive. This monograph could be used as a textbook both for advanced courses and for experienced researchers in different fields who would like to obtain detailed information about flow techniques."--Anal Bioanal Chem, 2012, Volume 403
"The aim [of this book] is to provide the reader with the most important basic aspects of flow techniques, according to the experience of the authors. In this sense, a detailed overview of the use of spectrophotometric and luminometric detection is presented with some details which may be scarcely found in other similar books. Some injection devices developed by the Brazilian Center for Nuclear Energy in Agriculture (CENA) are presented in detail. All the chapters end with an interesting and useful list of references. Summary The book is clearly written and comprehensive. This monograph could be used as a textbook both for advanced courses and for experienced researchers in different fields who would like to obtain detailed information about flow techniques."--Analytical and Bioanalytical Chemistry, (2012) 403:1465-1466

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