This document provides an overview of flow injection analysis (FIA) presented by Spurti P Patil and G S Narmada. It defines FIA as an automated method where a sample is injected into a continuous flow of a carrier solution that mixes with other continuously flowing solutions before reaching a detector. The key components of FIA instrumentation are described as the propelling unit (typically a peristaltic pump), injector, and detector (often a UV-visible spectrophotometer). The document outlines the principle of FIA and discusses the four main types: flow injection, sequential injection, bead injection, and sequential injection chromatography. It provides examples of FIA applications and notes advantages such as short analysis times and port
Department of Biotechnology Engineering Flow Injection Analysis
1. DEPARTMENT OF BIOTECHNOLOGY ENGINEERING
ACHARYA INSTITUTE OF TECHNOLOGY
(Affiliated to Visvesvaraya Technological University, Belagavi, Recognized by AICTE and Accredited
by NBA, New Delhi)
Subject – PROCESS CONTROLAND
AUTOMATION[18BT61]
Topic - FLOW INJECTION ANALYSIS
PRESENTED BY
Spurti P Patil
(1AY20BT029)
G S Narmada
Submitted to
Dr. Suneetha T B
HOD
3. INTRODUCTION
• FIA IS AN AUTOMATED METHOD IN WHICH A
SAMPLE IS INJECTED INTO A CONTINUOUS
FLOW OF A CARRIER SOLUTION THAT MIXES
WITH OTHER CONTINUOUSLY FLOWING
SOLUTION BEFORE REACHING A DETECTOR.
• FLOW INJECTION METHODS WERE FIRST
DEVELOPED BY RUZICKAAND HANSEN IN
DENMARK IN THE MID 1970.
• A SIMPLE AND VERSATILE ANALYTICAL
TECHNOLOGY FOR AUTOMATING WET
CHEMICAL MANIPULATION OF A DISPERSED
SAMPLE ZONE FORMED FROM THE
INJECTION OF THE SAMPLE INTO A FLOWING
CARRIER STREAM AND DETECTION
DOWNSTREAM.
4. PRINCIPLE
• A SAMPLE (ANALYTE) IS INJECTED INTO A FLOWING CARRIER SOLUTION STREAM THAT
IS FORCED BY A PERISTALTIC PUMP.
• THE CARRIER SOLUTION AND SAMPLE THEN MEET AT MIXING POINTS WITH REAGENTS
AND REACT.
• THE REACTION TIME IS CONTROLLED BY A PUMP AND REACTION COIL.
• THE REACTION PRODUCT THEN FLOWS THROUGH A DETECTOR.
• THE DETECTOR IS A SPECTROPHOTOMETER AS THE REACTIONS USUALLY PRODUCE A
COLORED PRODUCT.
• WE CAN DETERMINE THE AMOUNT OF AN UNKNOWN MATERIAL IN THE SAMPLE AS IT IS
PROPORTIONAL TO THE ABSORPTION SPECTRUM GIVEN BY THE
SPECTROPHOTOMETER.
• AFTER MOVING THROUGH THE DETECTOR, THE SAMPLE THEN FLOWS TO WASTE.
7. PROPELLING UNIT
• THE PROPELLING UNIT MOVES THE CARRIER
STREAM THROUGH THE FLOW INJECTION
ANALYZER.
• THE MOST COMMON PROPELLING UNIT USED IS
A PERISTALTIC PUMP.
• A PERISTALTIC PUMP CONSISTS OF A SET OF
ROLLERS ATTACHED TO THE OUTSIDE OF A
ROTATING DRUM.
• PERISTALTIC PUMPS PROVIDE A CONSTANT FLOW
RATE, WHICH IS CONTROLLED BY THE DRUM’S
SPEED OF ROTATION AND THE INNER DIAMETER OF
THE TUBING.
• FLOW RATES FROM 0.0005-40 ML/MIN ARE POSSIBLE,
WHICH IS MORE ADEQUATE TO MEET THE NEEDS OF
FIA WHERE FLOW RATES OF 0.5-2.5 ML/MIN ARE
COMMON.
8. INJECTOR
• THE SAMPLE, TYPICALLY 5–200 ΜL, IS INJECTED
INTO THE CARRIER STREAM.
• ALTHOUGH SYRINGE INJECTIONS THROUGH A
RUBBER SEPTUM ARE POSSIBLE, THE MORE
COMMON METHOD IS TO USE A ROTARY, OR LOOP
INJECTOR SIMILAR TO THAT USED IN AN HPLC.
• THIS TYPE OF INJECTOR PROVIDES FOR A
REPRODUCIBLE SAMPLE VOLUME AND IS EASILY
ADAPTABLE TO AUTOMATION, AN IMPORTANT
FEATURE WHEN HIGH SAMPLING RATES ARE
NEEDED.
9. DETECTOR
• THE MOST COMMONLY USED DETECTORS FOR FLOW INJECTION
ANALYSIS ARE THE ELECTROCHEMICAL AND OPTICAL DETECTORS
USED IN HPLC.
• A UV-VISIBLE SPECTROSCOPY SYSTEM OR HPLC DETECTOR IS
USUALLY USED FOR THIS PURPOSE.
• FIA DETECTORS ALSO HAVE BEEN DESIGNED AROUND THE USE OF
ION SELECTIVE ELECTRODES AND ATOMIC ABSORPTION
SPECTROSCOPY.
10. TYPES OF FIA
• THERE ARE 4 TYPES OF FIA TECHNIQUES
1. FLOW INJECTION (FI):- IS BASED IN SAMPLE INJECTION INTO A UNIDIRECTIONAL FLOWING CARRIER
STREAM TO WHICH REAGENTS ARE ADDED. SO, THE CONCENTRATION GRADIENT IS FORMED BY
DISPERSION OF THE SAMPLE ZONE ALONE, SIGNAL REFLECTS THE GRADIENT OF THE SAMPLE ZONE
AS IT PASSES THROUGH THE DETECTOR.
2. SEQUENTIAL INJECTION (SI):-SI AND BI ARE BASED ON SEQUENTIAL INJECTION OF
3. BEAD INJECTION (BI) :-SAMPLE AND REAGENTS, INTO CARRIER STREAM, FOLLOWED BY FLOW
REVERSAL THAT PROMOTES MUTUAL DISPERSION OF INJECTED ZONES. THE FLOW OF THE CARRIER
STREAM IS PROGRAMMED, TO OPTIMIZE THE ASSAY PROTOCOL.
4. SEQUENTIAL INJECTION CHROMATOGRAPHY (SIC):-SIC IS EMERGING CHROMATOGRAPHIC
TECHNIQUE THAT USES PROGRAMMABLE FLOW TO SEPARATE COMPONENTS, BASED ON
DIFFERENCE IN MIGRATION VELOCITIES, ON SHORT COLUMNS
11. APPLICATION
• FLOW INJECTION TECHNIQUES HAVE PROVEN VERY USEFUL IN
MARINE SCIENCE FOR BOTH ORGANIC AND INORGANIC ANALYTES
IN MARINE ANIMAL SAMPLES/SEAFOOD.
• FLOW INJECTION METHODS APPLIED TO THE DETERMINATION
OF AMINO ACIDS, DNA/RNA, FORMALDEHYDE, HISTAMINE,
HYPOXANTHINE, POLYCYCLIC AROMATIC HYDROCARBONS.
• FLOW INJECTION ANALYSIS HAS BEEN USED TO ANALYZE A WIDE
VARIETY OF SAMPLES, INCLUDING ENVIRONMENTAL, CLINICAL,
AGRICULTURAL, INDUSTRIAL, AND PHARMACEUTICAL SAMPLES.
12. ADVANTAGES
• SHORTER ANALYSIS TIME IN COMPARISON
WITH CHROMATOGRAPHIC METHOD AND
THE POSSIBILITY OF REAL TIME
CONTINUOUS MONITORING.
• DUE TO MINIATURIZATION THERE ARE
OTHER ADVANTAGES LIKE ROBUSTNESS,
REDUCTION OF REAGENT AND SAMPLE
CONSUMPTION.
• PORTABILITY OF THE SYSTEM ALLOWS
REAL TIME AND IN SITU TO
MEASUREMENT.
• PRODUCING LESS CHEMICAL WASTE
NEEDING DISPOSAL.
DISADVANTAGES
• SHORT REACTION TIME; REACTION MAY
NOT REACH EQUILIBRIUM.
• LOW FLEXIBILITY.
• LESS SENSITIVE THAN THE MANUAL
ANALYTICAL METHODS.