CHM4400

Advanced Analytical Chemistry

Flow Injection Analysis

Bo Zheng
bozheng@cuhk.edu.hk
Science Center Rm. 165
 Final Exam

• Time: 2015/5/6; 9:30-11:30 am

• Location: Thomas H C Cheung Gymnasium, UC
• You need to bring a calculator with logarithm function
• Only the lecture notes are allowed to be with you
Automation of Serial Assays

The purpose of all

analysis is to perform the
measurements at a
steady state (stable
readout, i.e., “flat top”).
(a) manual handling;
(b) a discrete belt-type
analyzer;
(c) a continuous-flow air-
segmented analyzer, with
a detail of the air and
liquid segments, showing
a mixing pattern that
leads to homogenization
of individual liquid
segments.
 The Simplest Single-Line FIA

(a) Flow injection determination of [Cl-]. The peak absorbance of Fe(SCN)3 is at

around 480 nm. With the sampling valve in the position shown, reagents flow
through the bypass. When a sample has been injected into the loop and the
valve turn 90o, the sample enters the flow as a single, well-defined zone.
Detectors include spectrophotometers, fluorometers, electrochemical, atomic
emission, and atomic absorption detectors.
(b) Record trace of [Cl-] from 5 to 75 ppm. The two curves to the right are high-
speed scans of one of the samples containing 30ppm and another 75ppm Cl-.
 Pumps to Drive the Flow

Peristaltic pump
Syringe pump
Separations in FIA

A dialysis flow module.

Dialysis is often used
in continuous flow
methods to separate
inorganic ions, or
small organic
molecules from high-
molecular weight
species.
 Separations in FIA

A system for the colorimetric determination of an inorganic metal cation by

extracting an aqueous solution of the sample with chloroform (CHCl 3) or CCl4
containing a complexing agent, such as 8-hydroxyquinoline.
(a) Flow diagram of a flow injection system containing an extraction module
(ABC); (b) details of A; (c) details of C

None of the separation procedures in FIA methods are complete.

 Convection and Diffusion

dispersion by dispersion by dispersion

convection convection and by diffusion
radial diffusion

Dispersion coefficient: D = c0/c

low dispersion (D 1-3); medium dispersion (D 3-10); high dispersion (D >10)
 Rules of Dispersion
1. Changing the injected sample volume is a powerful way to change
dispersion.
2. Limited dispersion value D is obtained by injecting a sample volume of
minimum one V1/2 into a manifold consisting of the shortest possible piece
of a narrow tube connecting the injection port and the detector.
3. The dispersion of the sample zone increases with the square root of the
distance traveled through the tubular conduit and decreases with
decreasing flow rate.
4. Any continuous-flow system that includes a mixing chamber generates
larger dispersion and yields a lower measurement sensitivity than a
corresponding channel without a mixing chamber. A system with mixing
chamber will also have a lower sampling frequency, unless the pumping
rates are increased.
5. In order to reduce radial dispersion, the flow channel should be uniform,
and should be coiled, packed, or 3D disorientated, which provides sudden
changes in flow direction.
Effect of Sample Volume and Tubing Length on Dispersion

(a) Response curves as function of injected sample volume;

(b) Dispersion of the injected sample zone as a function of the tubing length traveled.
V1/2: sample volume that is required to reach 50% of the steady-state value.
At V1/2, D=2.
Effect of Sample Volume and Tubing Length on Dispersion

D = 1 / (1 – e-VK)
V: sample volume
K: a constant related to the experimental configuration

In straight tubing, D  L1/2; also D = 1 + KL

L: length of tubing

In straight tubing, D = 1 + KR3/2

R: diameter of tubing

These equations are empirical.

Determining Ca2+ by FIA

Four sets of peaks for 5,

10, 15, and 20 ppm Ca2+
(standards).

Ca2+ + o-cresolphthalein  colored complex

Determining Caffeine in Acetylsalicylic Acid
Preparation by FIA

65%

35%
 Stopped Flow Method Using FIA
LDH
Pyruvate + NADH + H+ Lactate + NAD+

The stopped-flow technique on the assay of the catalytic activity of lactate

dehydrogenase (LDH).
To measure the reaction rate, the reaction mixture within the observation zone is
halted by stopping the flow and monitoring the change of the signal with time. The
dispersed sample zone (left), contains different substrate/enzyme ratios (since
D1<D2<D3), which yield different reaction rate curves.
From a to d (right), the stopped-flow delay time increases and the substrate/enzyme
mixing ratio changes, therefore the slopes for each curve would be different.
 Flow Injection Titration

The dispersed zone (top left) contains two equivalence points (A and B),
the distance between them, Dt, being proportional to the acid
concentration in the injected sample zone.
All FIA titrations are based on peak-width (Dt) measurement.
Flow Injection Titration of HCl with 1 mM NaOH

molarity of HCl

The time interval between the points is a measure of the acid concentration
 Merging Zone FIA

pump
sample

Carrier fluid

Carrier fluid detector

reagent

The main purpose/advantage of merging zone FIA is to economize on

the use of expensive reagents
 Flow Reversal FIA

Initial valve position Valve position during flow reversal

 Flow Recycle FIA

Initial valve position Valve position during flow recycle

 Sequential Injection Analysis

Reagents and sample are

sequentially drawn into the
holding coil and stacked one
after another.

Reagents and sample segments

Syringe pump offers flow through port 5 into the
precise reagent delivery reactor toward the detector (D)
and reversible flow.

CC: communication channel

CL: communication line