Ethanol metabolism in humans : a thesis presented in partial fulfilment of the requirements for the degree of Doctor in Philosophy at Massey University, New Zealand

Copyright is owned by the Author of the thesis. Permission is given for

a copy to be downloaded by an individual for the purpose of research and

private study only. The thesis may not be reproduced elsewhere without

the permission of the Author.

ETHANOL METABOLIS:t-1

^{IN HlTMANS}

A thesis presented in partial fulfilment of the requirements

for the degree of Doctor of Philosophy

at

Hassey University Nev; Zealand

Kenneth George Couchman 1979

ii ABSTRACT

Thi s the s i s outlines the development of breath alcohol measurement and investigations of the rate of absorPtion,

equilibration and elimination of alcohol from rhc body using breath ana lysi s. After ^a historici¹1 outlillL', tilL rnL·thods of ^a lcohnl

dete

c

^{tion are} rcviL'\.Jed and ^d compariti'.'L' stucty of some· mud�rn breath al

c

^ohol testing in�trumcnts detailed.

Til�

^rvsult!-i shPvl that the gas chromatogr^.::�ph IntoximetL•r ^h'LIS the· r:HJSL rl'producible and nccurate ins t rumen t .

Of Llte

^{fUL·l cell} inslrurnt•nts, tlw Ah·ol:imitcr ^. .;avc reproducible readings but \:ith ^a high..:·r frc·quency of

mechanic.tl

b r eakdown s and the ^Alcometer s failed to hold a calibrated value on r e p e a t e d t e s t i ng . The chemical analysis of the Borke

n

^stein

Br ea tha l yze r o ffe red portability and frel'dlJm from calibration bu t w i th a lowering

of

^accuracy.

No instrument offered the degrL'C of fl,•-.;ibility rc·quired for laborato ry investigation of factors ;Jflvvlin.• breath alcohol

concentrotions. Consequc·ntly a <�<Js chromi!t<J•,raph ^H::lS r.JOdifiL'd for breath sampling at

30 secLmd

intvrv;tl�· ^{. .} 'lhL· !l.trt:iLion coefficients for alcohol bet\h'L'Il ...1ir �1tH.I blood ^V.'L'rv lullt\•l Lobe: relilted Lo the water content of Llw blood �ampll'. Br·,·.llh :tl,·ohol concentrations inc r e a s e d _\vith expiration volume and I·:Lrv re li!ted t o <J risl! in bre_ath temperature. After correcting tll <� st.tnc!.tl ! LL·mpl.' ratun:

or :34°,

^a

linear increasl.' in .1lcolwl conCL'Ittr:Jt i <ll1 ^{r• ,} .. 1inl'd 1.,rhich \·las grl'ater

\vith h i ghe r blood <Jlcohul levels.

Equations for estima ting the dislr ibut i< n volume of alcohol in the bod y \vere derivC'd and the I.Jidm^ark facL<>r 'r' \vas found Lobe:

r ela t ed t o t h e ratio, body water OVl'r bl,lod .·.'ater. ^The b l ood alcohol time curves re s u lting from a f i xed dosl^' of ^.J lcohol g i ven to s em i -fa s t e d s ubj e c t s w e r e analys ed to d etermine the appdrent d i s t r ib u t ion volumes

i n t h e b od y . Vo l um e s exc e e d ing p h

ysi

o

l

og

ica

l l im i t s were fo und

in

some

subj e c t s and

asc

r

ibe

d

to either

a fa s t e r

rate

of me t a bo l i sm d u

ri

ng the

a

b

s o

rpt

i

v

e

p

h a s

e

or

t

o

anomalies in equi1ibrittion. A

ma

rk

e

dly non-linear

a l c oho l e l i

min

a

tio

n

curve

was seen

in one ^alcoh

o l i c

.

F a s t e r r a t e s of

a l c oho l ox i d a t ion \.Jere d isc u s s e d in

rel<�tion to the

H i c h ae l i s -Men t en

k i ne t i c s of enzyma t i c c a t ab o l i sm and

it is sug

ge s t

ed

tha t some s ub j e c t s

h ave a s e co nd e nzyme fo r al coho l m et ab o l i sm wh ich o pe rat e s at a h i gh e r Km th an no rmal .

i i i

The f l u ctuat ions o f b l ood alco ho l le v0l d uri ng t he ab sorpt ive ph ase we re e xami ne d by m eas uri n g the ab und an ce of a t racer do se o f d eut e rat ed al coho l gi ve n o ral l y after a lo ad in g d o s e o f un labelle d al coh o l . The f l uct u at ions we re ascribed to cont ract i on s of the

pyl o ri c s ph inct e r re l 'as i ng alco hol in to tllL' duo d e num in an irre gul ar fas h i o n.

The s t ud ie s we re extend ed

to subj�c·tb

drin ki ng in

a

private b ar.

The rate of al coho l abSLHpt ion

appt'arcd

to keep pace wi th the rate of d ri nkin g wh i ch was s pread ove r at l eas t ^a three h o ur period . The

rat e s o f al coho l e l im i nat i o n from the b lo od we re f as t e r th an in a pre vio us s t ud y wi t h a l owe r d o se of alco hol . Th i s i s e xpl ained b y l o we r b lood al coho l l e ve l s f ro m a sm al l er d os e and i s co ns i st e nt wi th the e nzyme ki ne t i cs o f al coho l cat,1bolism. An equat io n \vas d erived to e nab l e t he e s tim ation o f blo od alcohol levels from amoun t cons um ed wh ich co mpared favourably

\

vi

t

^h traditional m•'thod s for this

cal cul atio n .

Th e accuracy, rapidity and ^{l�.l�.;··} \,•itll

1-1hi.ch

bre at h alcohol analy s e s co uld b e m ad e

to de

termi ^tw a 1 coht) l L"Otwent r:1 t ions i n the bod v 0n ab l ed i ts us e wit h l arge grou ps o f peo pll' con sum in g alcoho l at party

s i t uat ion s or in hotel bars and

t\,'P

examplvs of such stuJics are pre s e n t e d in

the

appen dix.

iv ACKNOWLEDGENENTS

I am iⁿdebt ed to my supe

r

vi^sors^, Professor R. D. Bat t and Dr

R. H.

Gree^m.;ay for their c

o

ntinuing

interest in

^{the study and}

help "'ith the prep^aration of th e manuscript for the thesis. _I also wish to acknowled

g

^{e the} assist�ncc of 1. J. Hraggins and Dr A . R. St m.;e]l ^\vith the social drinking stuc.liL's _and

Miss

C.

Veltman with measurements of hreath volume effc•cts _on breath al co ho l co ncent ratio ns . Hy thanks ^<JrL' UUL' t o the Co mmand i ng Officer of th e Oh akea Air Base for pL'rmission to

cond uct t he so cial d ri nking s t ud ies , to t he Medical Officer, Dr D. St ewart and the Offi cers and men of the Royal New Zealand Air Force at Oh akea and t o st ud ents and staff of Hassey Uni.versity who willingly vo l unt e ered as subjects for study. My appreciation is also acknowl ed ged to Mrs Katherine New]and for a considerable amount of skilful technic^al assistance. The t h es i s was t yped by H.i..ss Hendy Fairc

l

ough an d I am ind('ptecl t o lwr i"or her p:1tience

and

^{care at all s tages in} preparir.g tll< fi:�al m:muscript.

V

TERNINOLOGY

Throughout th e tL!xt, "al cohol" is usc·d as synon ym ous wit h e than ol . All b l ood alcohol concentrations have b e en e xpre s s ed i n m il li grams per lOO _{milJ ilitres}. Ti1is terminology is s t ill common l y

us e d b y worke rs in th e fields of r.Iedicinc, a lcohol ab us e, psych iatry and alcohol and traffic safety to "''l10m tllL' prL'sent study could b e of in tere s t .

Hit h the adoption an d increasing usl' of SI un its j n recent ye ars b y t h e scie n t ific community, b lood alcoho l levels e xpre^ssed in

m il l im ol e s pe r l itre are b e com in g incre as ingly common .

The

equiv al e n t val ue for 10 m il lim ol e s pe r litre

(mmol/1)

is

46

m g

/

100 m l . The

res ults of b re ath al cohol an al ys es haVt', in t he m ain, b e en expre s s e d as b l ood al coh ol con ce n t rat ion s based on the b l ood : b re ath

dis t rib ut ion ratio of 1 ^: 2 , 100 as explained in the t e xt . Recen t l eg is l ation in New Zealand _{has made it} c�n offen ce to drive a mot or vehicle with a b rc>ath .Ilcc1ho1 1<'Vl'1 gn;Itvr than 500 )Jg per lit re, .3.nd ^eventu³lly it is vxpvctcd tlut hrl'.tlll rnlhcr th an b l ood alcohol concentrations wiJl be widelv uscJ.

Chap t er 1

Chap t er 2 2. 1 2.2

2. 2. 1 2.2.2 2.2.3 2.2.4 2. 2. 5

2.3

2. 3. 1 2.3.2 2.3.3 2.3.4 2.3.5

2.4

Chap t e r 3

L I S T O F CONTENTS

Ab s trac t

Acknow l edgements

Term inology List of Contents List of

Fi gures

List

o

^f

Tab l e s

INTRODUCT ION

BREATH ALCOHOL TESTING INSTRlJHENTS

�lEASlJREMENT O F ALCmiOL

^{IN BREATH} TEST ING

PROCEDURES

Breath volume

T e s t s w i th

a br eath alcohol sjt:Htl:Jt,H

Main t a i ned

re l iab i l i ty

Lin ear i ty of r e s po nse

Breath

tes t s on subjects RESULTS

A l c o l imi t er

A lco lme t er s and Alco-Sensor I n t o x ime t e r Mk

IV

Breathalyzer

Resu l ts on s u bjec t s

DISCUS S I O N AND CONCLU S IO NS

B REATH ALCOHOL A NALYSI S

i i

iv

V

v i

X

xii

8 8

10 10 10 14 14

14 15

15 16 20 21 24 27

31

3.0 3. 1 3. 1. 1 3.1. 2 3.

l. 3 3.

^l.

4

3

.l. 5

3. 2 3. 2. 1 3.2.2 3.3

Chapter 4

4 .1.

1 4

^{.l. 2} 4 .1. 3

4 .1.4 4. l. 5 4. 1. 6 4.2 4. 2 .l 4.2.2 4.3 4. 3 .

l

4.3.2

INTRODUCTION M

E

THODS

Alcohol measurement by gas chromatography Alcohol measurement in vitro

Partition c0�fficients Respiration measur�mcnts

Breath alcohol levels in

ciri:1king subjeL·ts

RESULTS

Breath

temperature Breath alcohol levels DISCUSSION

STUDIES ON THE ABSORPTIOt'\, DISTRIBUTION /\NO ELINDlATIO� OF ALCOHOL I:-i Tllf BODY \HTl!

31 31 31 33 35 42 42 46 46 46 62

VOLUNTEERS STUDIED UNDER L\BURATORY CONDITIONS 70 INTRODUCTIO:l

Body Hater compartm\"''ntatiott

Neasurement of the \.Jater C<lmp.trt:•wnt siZL'S using alcohol

Derivation of the \Hdmark f·nttation The kinetics of alcohol metabolism Experimental investigations

HETHODS

Studies on normal volunteers

Studies on alcoholics undergoing detoxication RESULTS

Normal volunteers Alcoholics

70 70 7l

73 76 78 78 78 78 80 80 80

4 . 4

Chapter ⁵

5. 1 - ') .), _ 5.3 5.4

Chapter ⁶

6 .1 6.2

6.3 6.

^{3. l}

6.3.2 6.3.3 6.3.4 6. 3. 5 6.4

Chapter 7

7. 1 7.

2

7.3 7.4

DISCUSSION

STUDIES ON THE ABSORPTION OF' A!.COHOL THROUGH

95

THE GUT \HTII

TRACER

^{DOSES OF} DEUTERATED ALCOHOL 106

I:lTRODUCTIO::

106

�!ETHODS 106

RESULTS 108

DISCUSSIO N ¹⁰⁸

ABSORPTION, DIST RIBUTION ^AND E

L

^{HHNATION OF}

ALCOHOL UNDER NORMAL

S

^OCIAL ^DRINKING CONDITIONS 115

INTRODUCTIOK ¹¹⁵

:lETH^ODS ^{1 15}

RESULTS 1 16

Th� "mouth :ll(ohol" vffL·( l

1 16

Rate llf drjuking 116

1 16

Rat� nf alu,llol

<?1iminati

'I l :zl

\·!idmark ratios Pl

DISCUSSION 127

ESTIMATIO:-JS OF BLOOD ALCOHOL LEVELS AFTER

DRINKING 130

INT RODUCTION ¹³⁰

CALCULATIONS ¹³⁰

RESULTS ¹³¹

DISCUS SION ¹³⁵

Chapter 8

REFERENCES

APrE!\DIX l .

APPENDIX 2.

GENERAL DISCUSSION

Joint

PrnjL·ct,

Department ^,., :)io,·hvm:i!->trv anJ De p ^{a r} l!:: L' n t o t So ^c i o l o g ^{v ,} :! ! :.; �', ". 1.· ^{1 1} i ^{v v}

r

^s i t ^{v .}

"Bloud Alcc.l;,>l

LL'VL·l�

in Y.:.J.lV

:\d1dts Aft,_•r

Free Drinking".

PrPparvd

^{•nr il} Rotarv Yt,tllh Leadership Course.

Breath

^Testing for i\lcohrd >·Jith Hotel Patrons.

A report prep:.�red ior tiw LL<;uor Advisorv Cou!lcll.

137

141

LIST OF FIGU!'�S

2. 1 A range of fuel cell instruments

2.2 The gas chromatograph Intoximet...'r tfk IV \vith mixed hydrogen/nitrogen gas supply and breath s imuLJ tor.

2. 3

2.4

3. 1

3.2

The Bre<:�tlwlyzer 900A an:l !')()() •.·.'itll ^:1 r.!ark IIA breath all·oho1 simulatnt .

Cor:-r'li.ltiuns

bet\·Jl'en

breath t•·st results obtained from a digital Al_colmetcr

and

^a gas chromatograph.

A plan view of ^the modi.fi l'd Ca rle gas chromatograph to show the inter ior.

Alcohol and acetone peaks obtain^ed with the Carle gas chromatograph.

12

l

J

26

32

34

3.3 A plot of the logarithrni� transforrn�tion of partition 37 coefficients fLH alcohol

hl'L\.Jc'Ln

^{air and water at}

3.4

3.5

3.6

3. 7

3.8

various temper<! tu res.

A plot of t11L' �-:atvr cunt,•nr ,l' i>loocl ·1gainst haematocrit.

A

^ploL

ol lllL'

l'artition ,.,,L•tlic· i,·nu; !ur alculllll bet\.;een air

and

^{blood at V+') ag.lillst the \vater}

content of thl' blood.

·;·11<2

mudifiL'J has chrom·ltu1•r.lpl!

\·!i

til ·1 srirometcr, m12rcur\' manorh·tcr and c:hdl't ^rL'< c>r<IL•r.

Frequency diagr.Jm of br•.'alh tLrlp••ratltres .::tt v.Jrious

�xpiratio11 vo lumc•s.

The change of alcohol c^Pn^<·

v

n t r.1 t inn

\.J

^{i th breath}

temperaturL'.

40 43

45

48

51

3.9 a-f Breath alcohol cur

v

es from all sub

j

ects showi n g 52 the effect of temperature corrvc^tion^.

3.10 The reg

r

^{ession of mean} temperature corre cte

d

^{breath 60}

alcohol concentration agains

t

expiration vo lume fo r

each s

u

b

j

ect.

3.11 A plot of the regression coefficien

t

s of breath 61 alcohol con cen t rati o n on e

x

piration volume against

the mean breath alcohol concentration from each subject.

4. 1

4.2

4.3

4.4a

4.4b

4.5

4.6

4.7

4. 8

4.9

The ex

p

ected blood alcohol ^curve resulting from a rapid

i

ntr

a

^venous inj cc t ion of <tlcoho l.

Blood alcohol curves generated �rum the results in t a b l c

4

. 2 i ^ll us t r ^a t i n g t h ^{l' L'} l t l' c t n f

Michaelis-Mentl'n

kinetics.

Blood alcohol curves frun �'U vultt;1teers given an alcohol dose o f

l .0 g/kg.

Raplu fluctu<.Jtions in blood alLnlllll concentration during the absorptive pbCis�· Ln subject ^5.

Ra

p

^id fluctuations in blood J1cohol concentration dur

i

ng the absorptive phase in subject 8 .

A blood alcohol curve in the elimination phase showing the absence of rapid fluctuations.

A blood alcoho

1

^curv�; frl1m .!n .::llculwl ic showing m^ar

k

t.'d non-lin,•arity.

A blood alcohol curve frum ^·m ,tlct>holic· Hhich suggests nc,n-1 im·ar climin.tt iPn.

UmxpL.dnL·d flut·tuations

in

tlw hlc,o,l a lcohol curve from an alcoholic.

Unexplained fluL·tuation:-:; in rh, bloud dl cohol curv^e from an alcoholic.

5 . l^-5 .4 Blood <::llco!1L1l anJ isotop<.: abundancL curves from subjects \.Jho h,Jd consumL·d alcol:ol fo l lowed by a tr<Jccr dost� c,f deutpratcd alcoh(ll.

6. l A representative

sample

^{of the} r.HL'S ^u f alcohol con

s

u

m p

^tion

before

and after a meal.

6. 2 a The rate of increase in bl(nd a l cnlw l level

& b before a meal.

6.3 The rate of increase in blood alcohol level after a meal.

6.4 6.5 7 .1

The rate of decrease i n blood alcohol levels.

A frequency diagram of calculated Widmark ratios.

Observed blood alcohol levels 3 hours after drinking began against alcohol consumption.

72

79

81

90

91

92

96

97

98

99

109- 112

11 7

118-119

120

122 125 134

LIS T

^OF

TABLES

2 . 1 Bre ath al cohol t�sting i n s tr u ments

Page

l 1

2 . 2 Res ults of ten cons ecuti ve

tests

with � s im ul ated 17 b reath al cohol stand ard corres pon ding L•) l OO

mg / 100 ml

o f blo od.

2.3

De vi ation s in

mg/ 100

ml

from

^:1 stand,1rcl simulator

test

¹⁸

of

^lOO

rng/100 ml at

v

ar

i ous intl·rv:tls during

a

on e

mo

nth

per iod.

2. 4

L

i ne a

r i

ty of

i

ⁿ

s

^t

r

u

m

ents^. R0sults u! sr·JrHlard alcohol 18 vapou rs from the simu l ator.

2 . 5 Ab s or b an ce at

440

nm

of dichromate

^solutions

prepa red

2 3 i n

the Brea tha l yzer

9 00A from bre ath al cohol

s ta

n

d

ar

d

s

and read

in

a

Hi tachi s pe ctrophotometer.

2 .6

Ab s orban ce at 440 nm of

d

i^chr

om

^{ate solutions}

p re pa red

^{2 3}

b y p

i

p

e t

t

in

g 50

_{wJ a}l co

ho

^l s oluti on in to 3 . 0 ml

ampoule

s ol u ti on

and read

in a Hitachi spL·(·trophotomete r.

2. 7

Comparative tests

on consecut_iv,•

breath s.!mp1es.

^2.J

2.3 Re su lts of s i �:1ultcnwous b

r

eath saPl!'lL"> t.1ken w i th ^a 28 gas chromatograph and a

digj

ti' l Alt ^,, ! ^{: ·} tt>r.

3 . 1 The alcohol concentrati on of simuLtti't solutions and

36

the

r

^esulting pL•ak l^{t L'}i g

h

ts fr,,m tlll g;t•: chromatogr dphi c an

a

ly

s

i s of thL' ^{v a}p

o

ur

phas� .

3. 2

The range

of al cohol con cen trations

it�

tht=

vap our phase

3 6

f rom simulator

s olutions.

3 . 3 A

l

coh_ol

peak heights

ob tain ed front g.Js chromatograp hic 3 9

a na

l

y

^s

i

s

o f the vapour phases

f

r

^om ·tlcohol conce ntration s

in t

he s i mu l ator

o f 1.2 1 6

g

/

l itr e of ^wat^e

r

^,

p la sm a or b lood a t three d

ifferen

t

haem aLocrits.

3 . 4

Par t i t ion c oeff icien t s f o r

alco

h

^ol between

air and

41

wa ter, p lasma and b l ood a t

three haem Jtocri t value s .

3 . 5 The c a l cula ted

val ue s

o f

wate r con ten t,

alcohol

p

a

r

t

i

t

i

o

n 44

coe f f i

ci

e

n

t

s

and

d i s trib ution

r a tio s for b lood w i th a norma l range o f haema t oc r i t.

3 . 6 Bre a th

te mpe rature s at

the end

of vari ous e xpi rati on

4 7

volume s .

3.7 Alc oho l

concen trati ons i n the b re ath ut

the e nd o f

49

var ious exp ir

ati on

vo lume s .

3.8

Breath alcohol concentrations ;CJfter l·tJrrecting for tem

p

era

t

^ure differences from 34ll.

3.9

Mean breath a

l

c

o

_h

o

l concentrations .Jt L',lch expiration vo ltll1h.'.

50

59

3.10 Distribution ratios at various L'Xpir"lion volumes. 63

3.1 1

The re-equilibration of

alcohol

hett,'t'L'n aqucous and

6L1

vapour phases in ^<l simulated upper rv�piratory tract.

4.1

1-Jidmark ratios

(r)

c.::llculatecJ from knot,'n hodv �r:ater and blood water values.

4 . 2

The velocities of alcohol elimination for two values

of Km an

d

^{various substrate}

(

^{blood alcohol)} concentrations.

4.3 Alcohol distribution concentrations and estimated absor

p

t

i

on times for subjects given al

c

ohol at 1.0 g/kg body weight.

7 5 7 7

89

4.4

Alcohol elimination rates in subjc_•cts )'.l'Jen .1lcohol at

93

1.0 g/kg body weight.

4 . 5 Correlation

^coeff

i

ci

e

^nts for r-cprcs!,i<,ns

of hlood

^alcohol

94

level on time before and after l ogarithmic transformation.

6.1 Alcohol distribution ccmcentr·1t ions itld t•l imination

1 23

rates for beer drinkers.

6.2 Alcohol distribution concL'ntrativns 111d ,]imination rates

124

for spirit drinkers.

6. 3 A co

m

pari

so

n of bera

slopes ..

^Jncl l�idr:.o�r-k ratios obtained

126

by analysing selective portions of

thL·

b load alcohol

curve.

7 . 1

Calculated and observed blood

alcohol l

e

v

el

s

in subjects

133

after the

consumption

of 0.4 or

I .0

g <tlcohol

/

kg body

w

e

i

ght

.