Вы находитесь на странице: 1из 7

22

trends in analytical

chemistry,

vol

Ij

no.

1, 1981

Enthalpimetry

- a change

of emphasis

vol Ij no. 1, 1981 Enthalpimetry - a change of emphasis J. Keith Grime Department of

J. Keith Grime

1, 1981 Enthalpimetry - a change of emphasis J. Keith Grime Department of Chemistry, University ofDenver,

Department

of Chemistry,

University ofDenver,

University Park,

Denver,

CO 80208,

U.S.A.

ofDenver, University Park, Denver, CO 80208, U.S.A. A change of emphasis These in enthalpimetric

A change

of

emphasis

These

in enthalpimetric

instrumental

design has widened

the scope

of

this universal

ana-

lytical tool.

recent developments

in enthalpimetric

technology

have

improved

the capabilities

of

the

technique and made possible

a wide variety of clinical,

biochemical

and environmental

applications.

 
biochemical and environmental applications.   Historical development the initial reports, the technique

Historical development

the

initial

reports,

the

technique

went

almost

un-

Enthalpimetric

analysis,

in one form or another,

has

noticed

until

the landmark

paper

of Linde,

Rogers and

been in existence

since the early

1900s. Although

many

Hume3

in 1953, which

introduced

the thermistor

as the

variants

now

exist,

all are

based

on

the

two

classical

temperature

transducer.

The

titration

approach

procedures

of titration

and

injection

(batch

mixing)

dominated

the enthalpimetric

methodology

until

1964

calorimetry.

These

techniques

have

been

termed

when

the

injection

technique,

DIE,

was

introduced4.

thermometric

enthalpy

titrations

(TET)

and direct

in-

DIE

utilizes

the total

temperature

change, engendered

jection

enthalpimetry

(DIE)

respectively.

The

former

by

a

chemical

or

biochemical

reaction

reaching

its

is characterized

by

the

continuous

addition

of titrant

equilibrium

position,

as a measure

of the

number

of

to

the

sample

solution

under

effectively

adiabatic

moles

of sample.

Typically,

the reaction

is initiated

by

conditions.

The

cessation

of the

temperature

change

the injection

of a small

volume

of solution,

containing

(whether

positive

or negative)

associated

with

the titr-

a

stoichiometric

of

reagent,

into

the

sample

ation

reaction

is used

to monitor

the

end-point.

The

solution.

The

excess resultant

enthalpogram

is depicted

in

elegant

simplicity

of the

TET

format

is evident;

since

Fig.

1(b).

Unless

a

series

of analytical

standards

is

most

chemical

reactions

are

exo- or endothermic,

the

used,

implementation

of DIE

requires

prior

knowledge

only

prerequisite

imposed

on

the

choice

of reagent

is

of

AH,

the molar

enthalpy

of reaction

and

electrical

or

that

it

reacts

stoichiometrically

 

and

instantaneously

chemical

calibration

of the

heat

change.

The

original

with the analyte.

A typical

TET

temperature-time

 

plot

motivation

for

the

development

of DIE

was

that

the

or

enthalpogram

is

shown

in

Fig.

1(a).

Cart-’

has

large

excess

of reagent

employed

( 1) assisted

in driving

written

a

comprehensive

 

review

covering

 

the

the

reaction

that

equilibrium

the

lifetime

to

of the

 

and

(2)

quantitative

and

qualitative

features

of TET.

 

ensured

completion, analytical

signal,

AT,

Since

its

introduction

in

19132

progress

in

en-

was short

compared

of heat

loss

from the re-

thalpimetric

analysis

has

been

erratic.

Indeed,

after

action

cell.

The

DIE

to the rate concept

was later

to be a critical

(a)

(b)

 

EOUIVALENCE

POINT

1

-

 

VOLUME

OR

MOL

OF

TITRANT

 

TIME

Fig.

I.

Typical

enthalpograms:

(a)

Thermometric

enthalpy

titration.

(6) Direct

injection

enthal@etry.

0 165.9936/RI/WOO-0000/$02.50

 

0 1981 Elwier

Scientific Publishing Company

trends in an$ytical

chemistry,

vol.

1,

no.

1,

1981

23

factor in the development of enthalpimetric methods

enthalpimetric

determinations,

based

on instruments

for biological

analysis.

in which

the

‘adiabatic

cell’

consisted

of a polythene

There

are

several

reviews,

monographs,

and

books

bottle

enveloped

by polystyrene

insulation,

have

been

which

cover

the

developments

until

19755-s.

reported.

The

contribution

of

this

approach

to

 

thermochemical

determinations,

although

lacking

Fundamental

principles

The

theoretical

basis

of enthalpimetric

in

concept

change

ceeding

classical

q(J)

thermodynamics.

illustrated

with

Thus,

The

is best

associated

to equilibrium.

by considering

a chemical

analysis

lies

fundamental

the total

reaction

heat

pro-

q

=

n,AH

(1)

where

the

An

that

np is the

number

of moles

of product

the

and

AH is

is

molar

enthalpy

of reaction

(J mol-l).

of enthalpimetric

in

important

there

is

no

feature

inherent

methods

selectivity

detection

system.

The

analytical

temperature

change

produced

solution.

Consequently,

 

the introduction

becomes

 

qi =

The corresponding

therefore

be given

by

signal

is composed

by

reactions

reagent,

change

of the

total

the

from all the chemical reactions

engendered

heat

changes

in

with

(1)

(2)

will

occurring

occur

equation

AT

if i simultaneous of the analytical

iZni AHi

temperature

 

AT

= k&r;

AHi

(3)

 

P

where

C, is the

heat

capacity

of the cell and its contents

(J “C-t).

Equation

(3)

illustrates

the

essential

characteristics

of TET

and

DIE

determinations.

The

sensitivity,

as

represented

by

the

magnitude

of

the

temperature

change

for

a given

number

of moles

of

sample,

is given

by the

factor

AH/C,,.

Current

trends in enthalpimetric

Instrumentation

-

the

changing

image

analysis

of

enthalpimetry.

Traditionally,

enthalpimetry

has

been

defined

as the

measurement

of temperature

changes

associated

with

chemical

or

biochemical

reactions

under

conditions

where

the

transfer

of heat

into

and

out

of the reaction

cell is minimized.

Historically,

the

boundary

between

enthalpimetry

and

calorimetry

has

been

defined

by

instrumental

specifications.

 

In

fact,

enthalpimetry

evolved

and

calorimetric

from

simplify

the

need

to increase

instrumentation

techniques

to routine

sample

order

analytical

in

throughput

to

apply

problems.

refinement

instrumentally,

is

significant,

 

since

it

popularized

the concept

of analytical

calorimetry.

The

simplicity

of the

apparatus,

its ability

to tolerate

the

presence

of non-reacting,

insoluble

or colored

matrix

ingredients

and

the

universal

nature

of

a

ther-

mochemical

determination

were

the

primary

motivations

for

its

adoption

in

the

early

stages

of

development.

As

a

result,

this

type

of

apparatus

dominated

the

literature

on

enthalpimetric

 

analysis

in

the

20

years

following

the

introduction

of

the

thermistor.

There

is

no

doubt

that

the

preoccupation

 

with

in-

strumental

simplicity

 

during

this

period

hindered

the

widespread

acceptance

of enthalpimetry

 

as

a viable

alternative

method

of analysis

and

precluded

its

in-

volvement

in

many

important

areas

of

analytical

chemistry.

.

In

the

last

live

years,

however,

there

has

been

a

noticeable

change

in

emphasis

and

direction

in

enthalpimetry.

With

the

trend

towards

trace

analysis

and

particularly

the

increased

prominence

of

biological

analysis,

the

arbitrary

boundary

between

enthalpimetry

and

calorimetry

has

become

vague.

A

new generation

of enthalpimetric

instrumentation

has

evolved

which

represems

a compromise

between

the

rudimentary

‘enthalpimeter’

and

the

sophisticated

adiabatic

calorimeter.

The

increased

acceptability

of

this

form

of instrument

is perhaps

best

illustrated

by

the

fact

that

a

wide

variety

of

commercial

in-

strumentation,

non-existent

a

few

years

ago,

is now

available,

The

most

popular

type

of instrument

in use

is the

‘isoperibol’

calorimeter

shown

in Fig.

2. Isoperibol

is

the

term

used

to describe

a quasi-adiabatic

system

in

which

attention

has

to be given

to the

determination

of the contribution

to the temperature

change

from

the

small

but

finite

transfer

of heat

from

the

reaction

cell

to the

surroundings.

 

The

specifications

of the temperature

measurement,

calibration

and

reagent

addition

systems,

have

not

changed

radically

in

recent

years,

and

the

reader

is

directed

elsewhere

for

informationi+s~lO

on

these

components.

 

Two

major

instrumental

trends,

which

are

in fact

related,

can

be identified.

These

are the diminution

in

This

philosophy

is embodied

in the features

of DIE.

If

the size of the reaction

cell and

the appearance

of com-

one

measures

temperature

changes

associated

with

puterized

data

treatment

facilities.

Cell

volumes

of

effectively

instantaneous

reactions,

heat

transfer

10 cm3

or

greater

are

typical

for

conventional

en-

between

the

cell and

its environment

can

be virtually

thalpimetric

measurements.

It

is

apparent

from

ignored.

Moreover,

if the

analytical

signal

is based

on

Equation

(3) however,

that

all ‘else being

equal,

a re-

a

relatively

large

temperature

change

(>O. l”C),

a

duction

in cell volume

increases

the

sensitivity

of the

modicum

of environmental

temperature

control

is re-

calorimeter

by

decreasing

C,.

 

The

observed

tem-

quired

to

produce

data

with

acceptable

precision

perature

change

is inversely

proportional

to

the

re-

(-

1%).

The

apparatus

required

for such

experiments

 

action

solution

volume

for a given

enthalpy

of reaction

is

simple

and

inexpensive;

many

successful

and amount

of reactant.

Unfortunately,

decreasing

the

inexpensive; many s u c c e s s f u l and amount of reactant.

24

trends in analytical

chemistry,

vol.

1,

no.

1, 1981

I

24 trends in analytical chemistry, vol. 1, no. 1, 1981 I TEMPERATURE MEASSSR~MENT WHEATSTONE \ .

TEMPERATURE

MEASSSR~MENT

WHEATSTONE

\

. v%!‘%!-E

SUPPLY

BRIDGE

. CIRCUITRY

-

iAMPLIFI

ER i-7

[P~ZE~~,F-

I

OUASI-

ADhBATlC

REACTION

CELL

I

L----*_---_C ----

------+lNTERFACE+-

,---a*_

J

L.)--LeTd

-+--------1

CALIBRATION

[

I

 

I

 

THERMOSTAT

-

I

3

d

--------

i

_----

r~iDE0

LDISPLAY I_

~KEYBOARD

1 /HARD@PY:-----

I PRINTER

‘r----!

_,------‘----+-+.+

-lb-----_---_

_,‘,

_

4

-

4

t

---3

:CO;$J$R+:D~SK

1

-----_

(4BK)

~-1Lp_RIv!:

---

I

REPRESENTS

OPTIONAL

FACILITIES

(4BK) ~-1Lp_RIv!: --- I REPRESENTS OPTIONAL FACILITIES   DATA COLLECTION     CORRECTION
 

DATA

COLLECTION

 
 

CORRECTION

SYSTEM

 
 

.

ei

 

Fig.

2.

Comprehensive

enthalpimetric

instrumentation.

 

cell volume

is not without

practical

problems.

The

rate

more

than

ca. 10% of the original

volume

of sample

still

of heat

loss

increases

exponentially

as

the

vessel

size

holds,

if gross

changes

in heat

capacity

and

resultant

decreases

because

there

is a limit

to the extent

that

the

non-linear

 

titration

data

are

to

be

avoided.

measurement

 

and

stirring

facilities

in the

cell can

be

Accordingly,

a more

precise

titrant

delivery

system is

miniaturized.

More

exact

calculations

are therefore

 

re-

required

to

deliver

titrant

volumes

CO.5

cm3 at

a con-

quired

to

correct

for

heat

exchange

 

during

the

stant

rate

(20.

I %)

over

periods

of

up

to

20

min.

experiment

l l.

In

the

titration

format,

the

usual

re-

Finally,

background

stirring

heats

become

more

quirement

that

the

final

titrant

volume

added

is

no

significant.

 

Consequently,

if

a

potentiometric

chart

trends in an&tical

1

chemistv,

recorder

zero

volume

is the method

capability

cm3)

offset

(<5

vol.

I,

no.

1, 1981

ofdata

is necessary.

collection,

cells

The

has

a considerable

use

of small

en-

taken

reaction

thalpimetry

into

the

sphere

of nanomole

determina-

tions.

Several

applications

of

microcell

instruments

have

been

reported

 

recently;

however,

the

pioneering

cell

designs

and

calculations

of

heat

loss

were

published

in 1974lz.

 

The

events

increased

significance

as

enthalpimetric

of background

sensitivity

thermal

are

limits

approached

has

also

resulted

in more

frequent

use of

computerized

data

aquisition

and

treatment

facilities

interfaced

ponents

the author’s

Plus microcomputer.

on

before

with

shown

contemporary

in Fig.

calorimeters.

the

is based

The

com-

used

in

II

effects

of data

2 represent

which

Enthalpograms

corrected

copy

data.

system

laboratory,

screen

and

as hard

on an Apple

can be displayed

a video

for background

type

printout

This

treatment

system

will undoubtedly

become

more

pre-

valent

in the

near

future.

Sensitivity

-

electronic

and chemical

Sensitivity,

or rather

the lack

a major

concern

governing

answers

of it, has always

feasibility

the,

of

been

the

 

25

 

VOLUME

OR MOL

OF TITRANT

ADDED

Fig.

3.

Typical

enthalpogram:

(a)

Thermometric

enthalpy

titration

with

catalysed

end-point

indication.

 

enthalpimetric

approach

to

a

particular

analytical

 

problem.

With

a reaction

enthalpy

of 40 kJmol-1

and

The

enthalpy

of the titration

reaction

can,

and indeed

an

arbitrarily

assigned

precision

limit

of

2%,

the

should,

be

much

smaller

than

that

of the

indicator

sensitivity

limit

of TET

is similar

to a potentiometric

reaction.

Consequently,

much

more

dilute

titrants

and

titration,

i.e. about

1 X

10-3 mol dm-3

or

1 X

10-S mol

samples

can be employed

in comparison

to TET.

It

is,

in

a typical

DIE,

a less precise

procedure,

of course,

highly

desirable

that

the

onset

of the

indi-

has a limit

10 cm3 cell’. of sensitivity

of about

2 X

1V

mol dm-s

at

cator

reaction

occurs

as quickly

as possible

after

the

the

10%

precision

level,

assuming

a

temperature

consumption

of the

sample

and

that

the

temperature

resolution

capability

of

+0.2”

C

(ref.

1).

Enthalpi-

change

associated

with

the

reaction

be

as

large

as

metric

sensitivity

has been improved

both electronically

possible.

This

is usually

achieved

by arranging

the ex-

and

chemically.

perimental

conditions

so that

the titrant

is the catalyst

 

The

most

successful

electronic

adaptation,

 

used

in

for

the

indicator

reaction.

A small

amount

of excess

many

subsequent

a

plications,

 

is

based

on

an

a.c.

titrant

will

therefore

initiate

the

reaction

between

a

Wheatstone

bridge

IP

.

The

incorporation

of a phase-

large

amount

of

indicator

reactants

already

in

sensitive

amplifier

to enhance

the signal-to-noise

ratio

solution.

 

and

a linear

ramp

generator

to offset stirring

heats

and

Limits

of detection

vary

according

to the

indicator’

heat

loss

has

allowed

a

temperature

resolution

of

reaction

used,

but

determinations

of

1 X

1O-7 mol

of

3-4

X

lO-@C.

Ultimately,

the

precision

of this

type

sample

are

not

unusual.

Greenhow

has

written

a de-

of

instrument,

-0.2%,

 

is governed

by the

titrant

de-

tailed

review

on

the

analytical

and

measurement

livery

system.

 

aspects

of catalytic

TET15.

 

A second

approach

to the sensitivity

problem,

which

 

The

simplest

 

and

most

convenient

method

of

has found

considerable

 

success,

is catalytic

TET.

This

improving

sensitivity

is

so-called

‘buffer

technique,

first

introduced

in

1965’*,

is based

on

the

amplification’.

 

Concurrent

buffer

reactions

serve

to

premise

that

the

precision

of a TET

determination

is

enhance

the

heat

effect

associated

with

the

analytical

related

to the difference

between

the titration

slope and

reaction.

This

has

proved

particularly

useful

in

the

post-reaction

slope

in

an

experimental

 

en-

monitoring biochemical

reactions

which

often

involve

thalpogram

(BC

and

CD

respectively

in

Fig.

l(a)).

the

release

of protonsi6.

 

The relative magnitudes of these parameters can be re- gulated by the addition ofa ‘thermochemical indicator’

Selectivity

-

the development

of enzymatic

enthalpimetry

substance

which

reacts

with

the

titrant

when

the

One

of

the

most

often

cited

advantages

of

sample

is consumed.

The

enthalpogram

then

takes

on

enthalpimetry,

its universal

detection

system,

also rep-

the

appearance

of Fig.

3. The

fundamental

 

difference

resents a major limitation, a complete lack of inherent

between

this

technique

and

TET

is that

the

precision

selectivity. The practicability

of titrating mixtures

of

the equivalence

point

is related

to the magnitude

of

depends on the magnitude

of AC

(and

hence

K,,)

and

the

temperature

change

associated

with

the

indicator

AH

for the

pertinent

reactions.

 

reaction

and

hence

to the amount

of indicator

present.

 

The

incorporation

of

the

intrinsic

selectivity

of

26

trends in anabtical

chemistry,

voli i,

no.

I,

1981

   

voli i, no. I, 1981     ‘ Reaction Curves F C a l i b
voli i, no. I, 1981     ‘ Reaction Curves F C a l i b
voli i, no. I, 1981     ‘ Reaction Curves F C a l i b

Reaction

Curves

no. I, 1981     ‘ Reaction Curves F C a l i b r a

F

Calibration

I

I

i

QC

I I

t

-

<

30

min

,-I

I-+tc

4

Substrate,

buffer,

co-substrate

 

I

 

,time

 

Fig.

4. Direction

injection

enthalfio~cram .fir

the determination

of

(a)

substrate

concentration,

S,

by

‘end-point’

or equilibrium

method

or

(b)

the Michaelis

constant,

K,,,.

Curv&

I,

2,3;

SI

>

ST >-

S,

(constant

E),

enzymatic

reactions

into

the

enthalpimetric

method-

catalysed

reactions

have

a

finite

reaction

rate.

 

The

ology

during

the

past

five

years

has

produced

an

sequence

of

addition

of

reagents

and

their

relative

analytical

method

with

considerable

potential,

 

one

amounts

is manipulated

to

an

extent

commensurate

which

embodies

selectivity

and wide applicability.

The

with

the

ultimate

goal

of the

investigation.

 

For

sub-

theoretical

principles,

experimental

procedures

and

strate

determinations,

the

pertinent

signal

is

qi,

the

successful

applications

 

of

this

combination

of

total

heat

change

associated

with

the

reaction

going

technologies,

termed

‘enzymatic

 

enthalpimetry’,

have

to

completion,

calculations

are

therefore

based

on

been

the

subject

of a recent

review16.

 

Equation

(2).

Amounts

 

of

kinetic

determinants,

Experimentally,

the technique

 

is based

on DIE.

The

enzymes

and

inhibitors,

can

be calculated

directly

or

fundamental

difference

 

between

enzymatic

 

en-

indirectly

from

the

time-dependent

 

versions

of

thalpimetry

and

conventional

 

DIE

is

that

enzyme-

Equation

(2) namely,

 
is that enzyme- Equation (2) namely,   Reaction Curve - - -- .- - _ -_-.-
is that enzyme- Equation (2) namely,   Reaction Curve - - -- .- - _ -_-.-
is that enzyme- Equation (2) namely,   Reaction Curve - - -- .- - _ -_-.-

Reaction

Curve

enzyme- Equation (2) namely,   Reaction Curve - - -- .- - _ -_-.- -- time
enzyme- Equation (2) namely,   Reaction Curve - - -- .- - _ -_-.- -- time
enzyme- Equation (2) namely,   Reaction Curve - - -- .- - _ -_-.- -- time

- -

--

.-

-

_

(2) namely,   Reaction Curve - - -- .- - _ -_-.- -- time g Fig.

-_-.-

--

(2) namely,   Reaction Curve - - -- .- - _ -_-.- -- time g Fig.

time

g

  Reaction Curve - - -- .- - _ -_-.- -- time g Fig. 5. Kinetic

Fig.

5. Kinetic

enthalpogram

 

for

the determination

of

(a)

enpe

activity,

EA,

(b)

inhibitor

concentrate,

I,

or

(c)

substrate

concentration,

S, by the ‘initial

slope’

method.

Curves

I,

2,

3

(a)

EA,

>

EA2

>

EA3

(constant

S).

(6)

11

<

12 C

I 3 (constant

S,

E).

(c)

S,

>

S,

>

S,

(constant

E).

trends in nn&tical

chemistry,

vol.

I,

no.

I,

1981

4

= ix

9;.

AHi

At

(4)

By use of appropriate dimensions and with the correct kinetic conditions prevailing, AnilAt can be presented in terms of enzyme activity. The experimental pro- cedures and typical enthalpograms associated with en- zymatic enthalpimetry are summarized schematically in Figs 4 and 5.

Clinical

enthalpimetry

Enzymatic

enthalpimetry

can

be

successfully

applied to almost every facet of clinical analysis,

the

inhibitor

determination

of

enzyme

and

activity,

the

substrate

magnitude

concentration

e.g.

and

of

Environmental

and energy related

Enthalpimetry

has recently

applications

found

application

27

in the

analysis

of airborne

particulates

and the determination

of

sulfur

ecies

associated

.with

the

combustion

of

fossil

s g

fuels2

. In one project,

a combination

ofTET

and

DIE

was

used

to

determine

nanomole

amounts

of

S(IV)

and

S(V1)

extracted

from

filters

containing

airborne

particulates.

S(IV)

was

first

oxidized

to

S(V1)

solution

in a TET was

then

microtitration

injected

with

K2Cr207.

into

the

titrate

BaS04

and