LIDOCAINE BASE AND

HYDROCHLORIDE
K. Groningsson, J-E. Lindgren, E. Lundberg,
R. Sandberg, and A. W a h l h
Astra Lakemedel AB
Siidertalje, Sweden

1. Foreword 209
2. Description 210
2.1 Structure 210
2.2 Name 210
2.3 Formula, Molecular Weight 210
2.4 Colour, Odour, Crystal Forms 210
3. Physical Properties 210
3.1 Melting Ranges 210
3.2 Solubility 213
3.3 Acid Dissociation Constant 213
3.4 Distribution Ratios 213
3.5 Infrared Spectra 213
3.6 Nuclear Magnetic Resonance Spectra 213
3.7 Ultraviolet Spectra 219
3.8 Mass Spectrum 222
4. Synthesis 222
5 . Methods of Analysis 224
5 . 1 Elemental Analysis 224
5.2 Identification Tests 224
5.3 Titrimetry 225
5.4 Gas Chromatography 225
6. Stability-Degradation 226
7. Pharmacokinetics 227
7.1 Plasma Concentrations after Different Routes of Administration 227
7.2 Protein Binding 228
7.3 Placental Transfer 228
7.4 Biotransformation and Elimination 228
8. Methods of Analysis-Biochemical Applications 230
8. I High Performance Liquid Chromatography (HPLC) 230
8.2 Gas Chromatography (CC) 230
8.3 Gas Chromatography -Mass Spectrometry, Selected Ion Monitoring 230
8.4 Homogeneous Enzyme Immunoassay, EMIT@ 234
9. Identification and Determination in Pharmaceuticals 234
9.1 Identification Tests 234
9.2 Extraction Analysis 235
9.3 Gas Chromatography 235

ANALYTICAL PROFILES OF DRUG SUBSTANCES Copyright 0 1985

VOLUME 14 207 by the American Pharmaceutical Association
ISBN 0-12-260814-3
208 K . CRONINGSSON ET A L .

4,4 Capilldq hotachophorem 236

9 Li Liquid Chromatography 236
Reference$ 238
LIDOCAINE BASE AND HYDROCHLORIDE 209

1. FOREWORD
I n 1935 Swedish chemists engaged i n t h e search f o r l o c a l
anaesthetics a t the i n s t i t u t e f o r Organic Chemistry,
U n i v e r s i t y o f Stockholm were i n v e s t i g a t i n g i n d o l e
derivates. Several organic compounds w i t h l o c a l
anaesthetic e f f e c t were synthesized. Further s y n t h e t i c
work b Lofgren, r e s u l t e d i n 1943 i n the l o c a l
$I
anaest e t i c compound 1 idocaine ( 1 , 2 ) .

R
The c l i n i c a l t r i a l s performed i n co-o e r a t i o n w i t h
Astra, Sweden, showed t h a t l i d o c a i n e ad c l e a r
advantages over t h e a v a i l a b l e drug o f choice, procaine.
The documentation was presented t o the Swedish
r e ulator u t h o r i t i e s i n 1947, i n c l u d i n g t h e trademark
Xyfocai n e w
Lidocaine was launched i n Sweden i n 1948 followed by
France and Canada i n the e a r l y f i f t i e s .
210 K . GRONINGSSON ETAL.

2. DESCRIPTION

@
2.1 STRUCTURE

/ACH3 NH-co-cH2-N / C 2 H 5

'CZH5
CH3
2.2 NAME
1-Diethy1 amir
1-Diethyl ami no-2' ,6 '-acetoxyl i d i d e

2.3 FORMULA, MOLECULAR WEIGHT

Lidocaine base: 1qH 2 2N 23'
molecular weight 234.34
Lidocaine hydrochloride hydrate: C1,H2,N20 HC1 H,O,
molecular weight 288.82

2.4 COLOUR, ODOUR, CRYSTAL FORMS

Both t h e base and t h e hydrochloride a r e w h i t e odourless
substances. The base c r y s t a l l i z e s from n-hexane as f i n e
needles w h i l e t h e hydrochloride i s obtained as a micro-
c r y s t a l 1i n e powder from aqueous acetone

3. PHYSICAL PROPERTIES
3.1 MELTING RAgGES
The measurements were performed i n a M e t t l e r FP-5 m e l t i n g
p o i n t apparatus ( h e a t i n g r a t e l"C/min) ( 4 ) .
Lidocai ne base 68 - 69.0"C
Lidocaine hydrochloride hydrate 76 - 79.0"C
I n Figures 1 and 2 a r e shown t h e d i f f e r e n t i a l scanning
c a l o r i m e t r y (DSC) thermal curves f o r t h e base and t h e
h drochloride respectively.
T i e anhydrous hydrochloride me1t s a t 125°C.
LIDOCAINE RASE AND HYDROCHLORIDE 21 1

2- 57.9 Jlg
67.4”C
h
0-

-2 -
z
-
U
5
-4:
-
c

I” 6-

-0-
-
- 10-
68.3%

-12 1 1 ~ ~ ~ ~ ~ ~ 1 1 1 1 1 1 , , 1 , , 1
20 30 40 50 So 70 80 90 100 110 120 130

Fig. 1. DSC curve of lidocaine base

 K . GRONINGSSONE T A L .

10 '
c?n 40 ' 20 ;O $0 $0
1
do
I
$0 I
I
100
1
1jO
I
IhO
I l
130
l i
140
Temperature ( C)

F,g 2 DSC [ urve of lidocame hydrochloride hydrate

LIDOCAINE BASE AND HYDROCHLORIDE 213

3.2 SOLUBILITY
The s o l u b i l i t data given i n Table 1 have been determined
photometrical y ( 4 ) .
The temperature de endence f o r t h e s o l u b i l i t y o f t h e base
i n water i s descriged by a AH o f -7.6 k 0.69 (12) kJ/mol,
k SE ( d f ) (5). This de endence i s i n agreement w i t h
7
previously reported so u b i l i t i e s ( 6 ) .

3.3 A C I D DISSOCIATION CONSTANT

The a c i d d i s s o c i a t i o n constant pK' defined by t h e
equation K I H A = a"+ x [ A ] x [HA+]- YA has been determined by
p o t e n t i o m e t r i c t i t r a t i o n ( 3 ) . A t 25'C t h e value 7.84 was
obtained

3.4 DISTRIBUTION RATIOS

The constants obtained a r e shown i n Table 2.

3.5 INFRARED SPECTRA

The I R spectra a r e shown i n F i ures 3 a n t 4.
They were taken as K B r discs wyth a Perkin-Elmer 298
spectrophotometer ( 4 ) . The assignments a r e given i n
Table 3.

3.6 NUCLEAR MAGNETIC RESONANCE SPECTRA

nF P
l H NMR spectra are shown i n F i ures 5 and 6 and t h e
NMR spectrum ( o f t h e base) i n i g u r e 7.
A l l t h e spectra were obtained on a Jeol FX 200 a t
200 MHz ( 4 ) . As solvents were used chloroform f o r t h e base
w i t h tetrameth l s i l a n e as i n t e r n a l standard and deuterium
o x i d e f o r t h e t y d r o c h l o r i d e w i t h 1 % dioxan as i n t e r n a l
standard (Tables 4 and 5).
214 I<. CRONINGSSON ET AL.

Table 1. S o l u b i l i t y ( / m l , -t 25’C) of lidocaine base and

1 idocaine hylrochloride hydrate

Sol vent
water 95% ethanol chloroform n-hexane
-
base 0.004 0.76 0.79 0.12
hydrochloride 0.68

Table 2. Distribution r a t i o s a t + 25’C.

Sy s tern p h a s d C water phase Ref.

n-octanol - phosphate
buffer a t pH 7.4 46 4
methylene chloride - water (base) 5200 3
-
toluene water (base) 160 3

Table 3. Infrared S e c t r a l Assignments f o r Lidocaine base and

Li docaine Rydrochl oride hydrate.

L i docai ne base Lidocai ne hydrochloride

3240 amide N-H 3430, 3360, 3160 amide N-H

2960, 2790 C-H 2960 C -H
1660 amide I 2450 NH+
1590 c=c 1645 amide I
1490 amide I1 i530 amide I1
760 aromatic C-ti 775 aromatic C-H
LIDOCAINE BASE AND HYDROCHLORIDE 215

1
40Qn
I
3500
1
3000
I
2500
I
2000
I
1800
I
1600
I
1400
I
1200
I
1000
I
800
I
600
t
400
wave rlumbol (ea ‘1

Fig 3 IR spectrum of lidocaine base

216 K . GRONINGSSON E T A L

WaYe number (cm )

-
600 400

Liq .1 IR iprcirurn ot Itdocamehydrochloridehydrate

LIDOCAINE BASE AND HYDROCHI.OKII>E 217

1 , , ~ , 1 , , 1 , , , , 1 , , , , ~
75 50 2.5 0 Ppm

Fig 5 'H NMR spectrum of lidocatne base CCoClJ

t l r , , l l l r , , ' " , , l , , ;

75 50 25 0 ppm

Fig 6 'H NMR spectrum of lidocaine hydrochloride hydrate (D,O)

 K . GRONINGSSON ET AI,.

I , I I I II
80 60 40 20 100 80 60 40 20 0 ppm

Fig. 7 IJCNMR spectrum of lidocaine base (CDCIJ

LIDOCAINE BASE AND HYDROCHLORIDE

Table 4. 'H NWf Assignncntr for Lidocalne base and

Lidocaine hydrochloride hydrate

Group CH2CI$ CHJli3 9-CH3

iCH.,NH Amtic H

Chemical s h i f t ( p p )
base 1.11 2.69 2.23 3.21 7.07
hydrochloride 0.30 2.30 1.13 3.24 6.14

~ H - ~spin
H coupling
(J W t(7) q(7) s S S
base and hydrochloride

Numkr of protons 6 4 6 2 3

Tabla 5. 13C Nm A s s i q ~ n t sfor Lidocalne base

Chemical s h i f t
(PP)
base 12.6 49.0 18.5 57.7 127.0;128.21U.1;135.1 170.1

3.7 ULTRAVIOLET SPECTRA

The U V s ectra were taken with a Hewlett-Packard 8450 A
R
spectrop otometer and are shown i n Figures 8 11 ( 4 ) . The
data are sunmarized in Table 6.
-
Table 6. Data from UV-spectra lidocaine base and
1 idocaine hydrochloride hydrate
~ ~~~~ ~

Sol vent 1 E Conc.

nm mol/l it e r

Base H2O 262 420 1.089.10-3

Hydrochloride 262 455 0.926.10-3
Base EtDH, 99.5% 262 348 1.052.10-3
Hydrochloride I1 263 377 0.921*10'3
 K . GRONINGSSON ET AL

10

09

oa

07
Peak-find
5 06 L262 04570
L 270 = 0 3367
p;
P 05

zr 03

03

O?

01

0
I I I 1 I I I I I i
220 240 260 280 300 320 340 360 380 400
Wavelength (nrn)
Fig 8 UV spectrum 01 lidocaine base in water

Peak find

- L262 03661

F 05-
2
0’-

oi-

’2-

$1’ -

I I I I I I I I I 1
710 x o 260 280 300 370 340 360 380 400
iVavelength In ni

F .? 9 UL s o e r t r w of 1idocail.e base in ethanol

LIDOCAINE BASE AND HYDROCHLORIDE 22 1

Peak-find
L 262 = 0.421 4
L271 =0.3417

Fig. 10. UVspectrum of lidocaine hydrochloride hydrate in water

Peak-find
L 263 = 0.3468
L 271 = 0.2596

Wavelength (nrn)
Fig 11 UV spectrum of lidocaine hydrochloride hydrate in ethanol
722 K . GRONINGSSON ETAL.

3.8 MASS SPECTRUM

The mass s ectrum o f 1 idocaine was o b t a i n e d w i t h an LKB
R
2091 gas c romato r a h - mass spectrometer ( 4 ) . I t i s
shown i n Figure 18. ?he fragmentation p a t t e r n l e a d i n g t o
ions of d i a g n o s t i c value is o u t l i n e d below.

'I 'I i----

I
m/z- 72
--

FMy is
orms the
a s s i g n e d by the ion a t m/z 234. The ion a t m/z 86
base peak.

4. SYNTHESIS
Lidocaine i s prepared according t o t h e f o l l o w i n g scheme:

.
CH3 CH3

2,6-Xyl i d i n e i s a c y l a t e d w i t h c h l o r o a c e t y l c h l o r i d e i n the
presence of a s u i t a b l e base, and t h e r e s u l t i n g x y l i d i d e
r e a c t e d w i t h diethylamine. The product, 1 i d o c a i n e b a s e , i s
e x t e n s i v e l y p u r i f i e d and, where a p p r o p r i a t e , converted t o
hydrochloride which i s further p u r i f i e d .
LIDOCAINE BASE AND HYDROCHLORIDE 223

100

I
80

20

0
100 200
* 40
Fig. 12. Mass Spectrum of lidocaine
‘24 K. GRONINGSSON E T A L .

5. METHODS OF ANALYSIS
5.1 ELEMENTAL ANALYSIS
The r e s u l t s from elemental analysis are sunmarized i n
Table 7.

Table 7. Elemental analys s o f l i d o c a i n e and l i d o c a i n e

hydrochloride.

Theoretical Found (1) I

base C 71.75 % 71.5 %

(C14H22N20) H 9.46 % 9.39%
N 11.95 % 11.93 %

hydroc h l o r ide c1 13.09 % 13.15 % (Mohr)

(C 14H 23N ,OC1 1

5.2 IDENTIFICATION TESTS (7,8,9)

The i n f r a r e d absorption spectrum o f a potassium bromide
dispersion o f the sample (1 idocaine base o r hydrochloride)
e x h i b i t s maxima only a t t h e same wavelengths as t h a t o f a
s i m i l a r preparation o f a chemical reference standard.
To about 0.1 g o f l i d o c a i n e base dissolved i n 1 m l o f
alcohol i s added 10 drops o f a 2 %, w/v, aqueous s o l u t i o n
o f cobaltous c h l o r i d e containing 1 %, v/v, o f hydrochloric
acid. The mixture i s shaken f o r about 2 minutes when a
b r i g h t green c o l o u r develops, and a f i n e p r e c i p i t a t e i s
formed.
To a l i d o c a i n e hydrochloride s o l u t i o n (0.2 g i n 10 m l o f
water) i s added 10 m l o f a 1 %r w/v, p i c r i c a c i d solution.
The p r e c i p i t a t e , when washed w i t h water and dried, has a
me1t i n g p o i n t o f about 230°C.
From 1 idocaine hydrochloride t h e base is i s o l a t e d e i t h e r
9
by a l k a l i z a t i o n o f an a ueous s o l u t i o n o f t h e s a l t
followed by i s o l a t i o n o t h e base or by making an a l k a l i n e
e x t r a c t i o n i n t o e.g. chloroform where t h e base i s i s o l a t e d
by evaporation and drying. The i d e n t i f i c a t i o n procedure
f o r t h e base can then be used.
LIIIOCAINE HASE A N D HYDROCHLORIDE 225

An aqueous s o l u t i o n o f l i d o c a i n e hydrochloride y i e l d s a
w h i t e p r e c i p i t a t e , i n s o l u b l e i n n i t r i c acid, by a d d i t i o n
o f s i l v e r n i t r a t e . This p r e c i p i t a t e i s however s o l u b l e i n
a sl i g h t excess o f ammonium hydroxide ( i d e n t i f i c a t i o n o f
c h l o r i d e ) . Another c h l o r i d e t e s t i s performed by m i x i n g
equal weights o f t h e sample and manganese dioxide, moiste-
n i n g w i t h s u l p h u r i c a c i d and then heating. Chlorine i s
then evolved and i s recognizable by i t s odour and by
p r o d u c t i o n o f a b l u e c o l o u r w i t h moistened s t a r c h i o d i d e
paper.

5.3 TITRIMETRY
Lidocaine base can be determined by d i s s o l v i n t h e Sam l e
i n an excess o f 0.1 M hydrochloric a c i d o r 0. 5 M sulp u-
r i c a c i d followed by back t i t r a t i o n w i t h 0.1 M sodium
8 i
hydroxide. A m i x t u r e o f bromocresol reen and meth 1 r e d
i s a p p r o p r i a t e as i n d i c a t o r (7,lO). ion-aqueous ti r a t i o n
u s i n g a s o l u t i o n o f t h e base i n anhydrous g l a c i a l a c e t i c
acid, c r y s t a l v i o l e t as i n d i c a t o r , and 0.1 M p e r c h l o r i c
a c i d as t i t r a t o r i s a l s o possible.
For t h e assay o f l i d o c a i n e hydrochloride 0.3
7
sa l e i s dissolved i n anhydrous g l a c i a l a c e t c acid, 6 t o
7 m o f mercuric acetate s o l u t i o n ( 7 8,9) i s added and t h e
9 o f the

s o l u t i o n i s t i t r a t e d w i t h 0.1 M p e r c h l o r i c a c i d u s i n g
c r y s t a l v i o l e t as i n d i c a t o r . An emerald-green c o l o u r
i n d i c a t e s t h e end-point.
Amines and amine s a l t s can be determined by automatic
p o t e n t i o m e t r i c two-phase t i t r a t i o n (11). Lidocaine hydro-
c h l o r i d e (0.2400 g) i s weighed i n t o a t i t r a t i o n vessel and
20.0 m l o f dichloromethane and 20.0 m l o f 0.05 M
hexadecylpyridini um c h l o r i d e a r e added. T i t r a t i o n i s
performed w i t h 0.1 M sodium hydroxide w i t h vigorous
stirring.

5.4 GAS CHROMATOGRAPHY

The p u r i t y o f l i d o c a i n e can be determined by as chroma-
tography u s i n a ca i l l a r y column of c r o s s l i n 8 e d SE 54 on
fused s i l i c a ?lengtR 25 m, i.d. 0.3 mm, f i l m thickness
B
0.5 m) and a flame i o n i z a t i o n detector. (For instrument
s e t t ngs see Table 8).
The sample (0.20 -
0.25 g o f l i d o c a i n e ) i s d i s s o l v e d i n 1 0
m l o f dichloromethane and 2 p l i s i n j e c t e d i n t o t h e gas
chromatograph. The chromatogram i s evaluated using
i n t e r n a l normalization. For determination o f 1 idocaine
hydrochloride a base e x t r a c t i o n i n t o dichloromethane i s
performed before i n j e c t i o n .
226 K . GRONINGSSON ET AL.

Table 8.
Instrument settings: I n i t i a l oven temperature 140°C
I n i t i a l oven time 4 min.
program r a t e 3"C/mi n.
Final oven temperature 180°C
Final oven time 23 m i n .
Injector temperature 220:c
Detector temperature 270 C
Carrier gas He
In1 e t pressure 0.7 kg/cm
Spl i t vent 20 ml/min

6. STABILITY - DEGRADATION
In solution lidocaine would be expected t o decompose by
hydrolysis as follows:

Lidocaine

2,6-Xyl i d i n e Diethyl aminoacetic acid

I t a m e a r s t h a t 1 idocaine i n aoueous solution i s extremelv
r e s i s t a n t t o heat, acid and a l k a l i , b u t when decomposi tio'ir
does occur i t i ? by the hydrolysis a s shown above. The
high s t a b i l i t y i s due t o the s t e r i c a l hindrance towards
attack on the amido grou e x h i b i t e d by the two ortho
E Y
methyl grou s. However, idocaine i s more r e a d i l y
hydrolysed y acid than by a l k a l i . This can b e a t t r i b u t e d
t o the i n h i b i t e d mesomerism - due t o l o s s o f planarity
between the benzene nucleus and the amide group - w h i c h
3
gives rise t o h i her electron densities a t the amide
nitrogen and acy carbon atoms, t h u s i n h i b i t i n g
nucleophilic attack. T h i s steric inhibition o f mesomerism
a
i n lidocaine has been demonstrated by L o f ren (10). Acidic
hydrolysis, i n i t i a t e d by protonation o f t e carbonyl
oxygen, should occur somewhat more readily (12).
LIDOCAINE BASE AND HYDROCHLORIDE 227

S t a b i l i t y studies have shown t h a t a 2 per c e n t s o l u t i o n o f

l i d o c a i n e h d r o c h l o r i d e made a l k a l i n e t o pH 7.3 and heated
f
i n an autoc ave a t 115°C f o r 3 hours showed only 0.05 p e r
c e n t decomposition. By h e a t i n g i n an autoclave w i t h 50 p e r
c e n t s u l p h u r i c a c i d f o r 5 hours a t 116°C only 3 p e r c e n t
decomposed w h i l e a s i m i l a r treatment w i t h 20 p e r c e n t
ethanol i c potassium hydroxide s o l u t i o n caused
approximately 0.5 p e r c e n t decomposition (10). Under more
vigorous c o n d i t i o n s such as heating f o r 24 hours i n
constant1 b o i l i n g h y d r o c h l o r i c a c i d s o l u t i o n (108"C,
6.5 M H C l j 50 p e r c e n t o f t h e l i d o c a i n e remained
unhydrolysed.

i
Solutions f o r i n ' e c t i o n c o n t a i n i n g 1 and 2 p e r c e n t
l i d o c a i n e hydroc l o r i d e (pH 6.7) s t o r e d f o r 5 ears a t
9
room temperature contained o n l y 0.5 p / m l and 6.7 pg/ml
r e s p e c t i v e l y o f t h e h y d r o l y s i s produc 2,6-xyl i d i n e .

7. PHARMACOKINETICS
The 1i t e r a t u r e concernin pharmacokinetics o f
1idocaine is extensive. o f t h e drug have
been sunmarized i n

7.1 PLASMA CONCENTRATIONS AFTER DIFFERENT ROUTES OF

ADMINISTRATION
The major determinants o f t h e systemic absor t i o n o f
l i d o c a i n e a r e t h e dose (concentration and vo urnel, t h e Y
!
s i t e o f i n j e c t i o n , t h e e r f u s i o n and t i s s u e b i n d i n g a t t h e
s i t e o f i n j e c t i o n and w e t h e r o r n o t adrenaline has been
added. The absorption r a t e decreases i n t h e order: i n t e r -
c o s t a l block > caudal block > epidural block > b r a c h i a l
plexus block > s c i a t i c and femoral block (14).
The plasma concentrations a f t e r endotracheal administra-
t i o n a r e comparable t o those a f t e r c e n t r a l and p e r i p h e r a l
nerve blocks (16).
I n t r a p e r i toneal i n f u 2 i o n o f 500 t? 1000 mg o f 1idocaine is
accompanied by re1a t i v e l y 1ow maximum plasma drug
concentrations (17).
Adrenaline reduces t h e peak plasma concentration o f 1 i d o -
c a i n e a t a l l i n j e c t i o n s i t e s b u t t o a v a r y i n g degree a t
t h e d i f f e r e n t s i t e s (18).
Lidocaine i s n o t used o r a l l y due t o extensive f i r s t - p a s s
metabolism. Observations i n p a t i e n t s r e c e i v i n g 1idocaine
by pro1 onged i n f u s i o n f o r suppression o f v e n t r i c u l a r
dysrhythmi as support t h e g e n e r a l i z a t i o n t h a t s u b j e c t i v e
e f f e c t s a r e associated w i t h plasma concentrations of
3-5 pg/ml and t h a t o b j e c t i v e signs appear a t 6-10 pg/ml
(15).
228 K. GRONINGSSONE T A L .

7.2 PROTEIN BINDING

Lidocaine i s bound t o plasma p r o t e i n s t o about 60 % a t
therapeutic dru l e v e l s . The f r a c t i o n bound decreases t o
8
about 40 % a t 1 pg/ml plasma, which may c o n t r i b u t e t o t h e
Z
t o x i c i t a s t h e f r a c t i o n o f f r e e drug increases (13,191.
Fetal p asma b i n d i n g i s approximately 50 % l e s s than
b i n d i n g i n maternal lasma (20). Lidocaine i s
predominantly bound t o al-acid g l y c o p r o t e i n (21).

7.3 PLACENTAL TRANSFER

Lidocaine passes t h e placenta and t h e r a t e o f d i f f u s i o n i s
mainly a f u n c t i o n o f t h e concentration g r a d i e n t between
maternal and f e t a l blood, t h e pH-gradient between mother
and f e t u s and the maternal p r o t e i n b i n d i n g (20,22,23)
The r a t i o between t h e l i d o c a i n e concentrations i n t h e
u m b i l i c a l and maternal v e i n i s 0.52 -
0.69 (24).

7.4 BIOTRANSFORMATION AND ELIMINATION

metabolised i n man

and arnide
The major m e t a b o l i t e i n u r i n e i s 4-hydroxy-2,6-x l i d i n e .
The b i o l o g i a l h a l f - l i f e o f l i d o c a i n e i s 1.5 h (14,281
The estimates o f plasma clearance range from 0.54 t o 1.44
1/min (15) and have been shown t o be dependent on 1i v e r
blood f l o w (29).
Both MEGX and GX have pharmacologic e f f e c t s b o t h as
a n t i a r r h y t h m i c s and i n terms o f t o x i c i t y (36,311. The
h a l f - l i f e o f MEGX appears t o be about t h e same as o r
s l i g h t l y g r e a t e r than t h a t o f 1idocaine, b u t t h e p o t e n t i a l
o f MEGX f o r t o x i c i t y would be more-apparent i n p a t i e n t s
w i t h h e a r t f a i l u r e , i n whom i t s e l i m i n a t i o n r a t e i s
reduced (32,331. GX i s accumulates i n p a t i e n t s with r e n a l
f a i l u r e , which should be considered d u r i n g continuous
a d m i n i s t r a t i o n (34).
The clearance o f 1 idocaine does n o t d i f f e r between e l d e r l y
(mean age 65 years) and young (mean age 24 years) subjects
(35).
Studies i n neonates have shown t h a t although t h e plasma
h a l f - l i f e i s prolonged, clearance does n o t d i f f e r from
t h a t i n a d u l t s ( 3 6 ) . Metabolism o f l i d o c a i n e by t h e
f e t u s h e o n a t e has a l s o been confirmed (37).
LIDOCAINE BASE AND HYDROCHLORIDE 229

&J /H
/C2H5
NHCOCHZN
\
Q$'LC-H~N,
CH3 C2H5 CH3 CZH,

lidocaine (2.8) monoethylglycylxylidide (3.7)

2,6-xylidine (1.0) glycylxylidide (2.3)

1
(GW

conjugates -HO &LH2 [@iH2 COOH

4-hydroxy-2,6-xylidine 2-amino-3-methylbenzoic acid

(72.6)

Fig. 13. Pathways for the biotransformation of lidocaine in man. Values in

parentheses indicate percentages of dose found in urine.
330 K . GRONINGSSON ET AL.

8. METHODS OF ANALYSIS - BIOCHEMICAL APPLICATIONS

8.1 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
HPLC i s probably t h e most v e r s a t i l e method f o r t h e r a p i d
and s p e c i f i c determination o f l e v e l s o f l i d o c a i n e and i t s
active, p o l a r metabolites. Recently, several HPLC methods
u s i n u l t r a v i o l e t (UV) detection have been employed (Table

1 i
9) (38-51). Unfortunately, as these com ounds c o n t a i n no
chromo hore which absorbs strongly i n t e v i s i b l e o r
near-U regions, t h e wavelength employed f o r d e t e c t i o n
must be approximately 200 nm. I n recent years,
amperometric electrochemical d e t e c t i o n f o l l ow1 n
chromatography has become increasing1
T! s
opul a r oliquid
r the
q u a n t i t a t i o n o f e a s i l y o x i d i z a b l e ana y es. I t s p r i n c i p a l
advantages include uniformly high s e n s i t i v i t y and a unique
s e l e c t i v i t y towards compounds t h a t can be e l e c t r o l y z e d a t
t h e a p p l i e d detector p o t e n t i a l . The anodic e l e c t r o -
chemistry of 1 idocaine, MEGX and GX a t graphite electrodes
and the develo ment o f an LC-ED procedure f o r t h e i r
determination Rave r e c e n t l y been described (52). The
absolute d e t e c t i o n l i m i t s f o r these compounds were 2 ng,
5 ng and 4 ng injected, respectively.

8.2 GAS CHROMATOGRAPHY (GC)

Numerous gas chromatographic approaches have been d e s c r i -
bed f o r monitoring o f l i d o c a i n e alone o r together w i t h i t s
metabolites MEGX and GX (Table 10),(53-76). These methods,
employing flame i o n i z a t i o n o r n i trogen-phosphorous detec-
t i o n have been shown t o possess e x c e l l e n t s e n s i t i v i t y f o r
lidocaine, generally i n the 1-10 ng/ml range. But, i n many
procedures, t h e metabolites MEGX and GX e i t h e r a r e n o t
determined a t a l l o r are n o t separately distinguished from
the parent compound. More importantly, t h e a p p l i c a t i o n o f
a l l these a proaches t o the r o u t i n e analysis o f r e a l serum
samples i s f i m i t e d by t h e sample preparation time taken up
by t h e lengthy d e r i v a t i z a t i o n , extraction, o r evaporation/
preconcentrati on procedures required.

8.3 GAS CHROMATOGRAPHY - MASS SPECTROMETRY ( GC/MS) , SELECTED

I O N MONITORING
As proposed i n r e f 77 the term “selected i o n monitoring”
i s used. I n t h i s a p p l i c a t i o n the mass spectrometer i s used
t o monitor ions o f c e r t a i n mass normally i n t h e gas
chromatographic e f f l u e n t . The technique i s a l s o commonly
r e f e r r e d t o as “mass fragmentography’ i n a d d i t i o n t o
various other names.
LIDOCAINE BASE AND HYDROCHLORIDE 23 1

Table 9. HPLC methods for measuring lidocaine and ncrabolites i n biological m t e r i a l

Saaple Internal standard Extracting oatstor Analytical

solvent PMlsion
(CV X)
%I'W thaKOa1 ad- uv 6-10 No 38
sorption
Plasm Ethyl acetat. uv wo 39
Senm Ethylmthyl - Chlomfon-
- MGX
Urine q&$iylidide h e x a m iso-
propanil uv 0.9 - 5
ot
40

Plasm MOM hthylrm

chloride uv < 10 NO
PI.- NOW uv mot
GX
PlasM Ethyl acetat. uv 2-1 n€Gx
GX
SWU
<S NO

Plasm hthy1e M uv <7 MEGX

chlorida GX
Sew IlrPivacain Chlomfom
CSF hexane-iso- MGX
propanol uv 3-5 Gx 46-47
SIM 2-hino-&
Urine -(2.6-xylyl)- UEGX
butyrmide Ethyl acetate UY 2-1 GX 48

Plasm
nyocardial MEGX
samles GX 09
(rabbit) EHGX Ethyl acetate uv
UEGX
Senm EHGX Chlomfon uv (2 GX 50

MGX
Plasm EHGX Dichlom- GX 51
methlM uv <5

Senr Bupivacaim Sep-Pak Electm- MEGX

chemical GX 52
detection 5-10
232 K . GRONINGSSON ET A L .

Table 10 0 s chrortographlc methods for m s u r l n g l { d o c a f m

a d .rtabolit.s in b i o l a g l u l r b r i a l

knDlS Colul, Dctcctor k r i v a t i ve hlytical Wstabolites Ref

Prcciflon measured
ICY 5)
Blood Packed FIO now 5 NO 53
Plasm Packed wPD wow no 54
Pins Acstyl- HEW.
Urtn Packed )(PD CrivatIve 3-8 GX
4-hydmxy-2.6-
xylidim 55
Plasm Packed wm won 2-18 HEGX 56
Plasm Packed wm worn no 51
Plasm Packed FIO NOW 1-7 Kcx 50
Tissue Packed FIO wow NO 59
Plasm Packed FIO uon No 60
PlOU Packed FIO NOm 6 No 61
Plasm cap4 1l a r y wm Trifluoro- nECX
a c e t y l d w i v a t i v e 4-7 GX 62
81wd
P18W Packed FIO llorn 2 no 63
BlWd
Plasm capi 11ary nm now NO 64-65

Blmd
PlOsm
5.- Packed FIO NOW 2-6 NO 66

various
biolcqlcal
fluids Packed wm non. NO 67

Plasm
Packed FID MOW NO 68
kru
Plasm
Urim
Packed FIO Ron NO 69
(h0-l)
Pla- Packed FIO NOW (5 no 70

Plasm patted urn WW 1-7 acx

bx 71

Packed FID Now 2 no 12

(cat)
Packed FIO IIOW (5 ((0 73
Plau
mst crtl
fluids a d
tissun sv 52% Packed wm NOW 3 nEcx ?4

Packed Nm Na
n 3 NO 75

Packed wm n w 1-11 nEGX

GX 76
LIDOCAINE BASE AND HYDROCHLORIDE 233

I n t h r e e studies (78, 79, 81) t h e gas chromatographic

peaks o f MEGX and GX.were asymmetrical and-the lower l i m i t
o f a n a l y t i c a l s e n s i t i v i t y f o r both metaboli t e s was about
0.5 pg/ml. Another .mass spectra! method bas9d on . e a r l i e r
work u s i n g s t a b l e isotope l a b e l i n g and chemical i o n i z a t i o n
procedures i n v o l v e s t h e use o f deuterated 1idocaine MEGX
and GX as i n t e r n a l standards (80). Samples c o n t a i n i n g
these standards a r e e x t r a c t e d and then analyzed by d i r e c t
probe i n t r o d u c t i o n and isobutane chemical i o n i z a t i o n mass
spectrometry. The range o f d e t e c t i o n was from 5 n t o 4 g
f o r l i d o c a i n e and 0.1 ug t o 1 pg f o r MEGX. GX cougd n o t be
a c c u r a t e l y q u a n t i f i e d due t o i n t e r f e r i n g substances.
A method u s i n t h e p r o y l d e r i v a t i v e s o f MEGX and G X
3
formed d i r e c t y i n e i tp1e r plasma o r u r i n e by treatment
w i t h propionaldeh de and sodium cyanoborohydride has been
pub1 ished (82). T i e d e r i v a t i v e s and unchanged 1 idocaine
Y
a r e extracted, se a r a t e d by gas chromatography and
q u a n t i t a t e d by se ected i o n monitoring, u s i n g mepivacaine
as t h e i n t e r n a l standard. Q u a n t i t a t i o n o f these com ounds
t o l e v e l s as low as 50 ng/ml bod f l u i d has been a c h e v e d
f
w i t h a c o e f f i c i e n t o f v a r i a t i o n ess than 10 % (Table
11).

Table 11 %lscted {on mnltorlng mthodr for masurlng

l i d w i n e and mtaboliter in blologicil mtarlal

-1 * Internal
standard
Techniqur (hrivatlvr

Plasm Trfmrin Electron

f*Ct NOM 3-7 MEGX 10

Plasm Electron HEI

Urim Trlmaln inpact Nom 3-9 GX 79

Plasm 2H4-L1 douf IN ISObubM

chrnicrl
2H3-M6X lonlzation
Direct In-
tertion pmb. Now RGX 80
Plasm Trimaim MEW(
Urim 2.4 .S-lrfmtyl- Gx
anlllm Electron 4-hydk~y-2.6-
t-naptho1 lnplCt NOIN 3-9 xyl idine 81
Plasm Electron PTYl mGx
Urlm I*plVaufM l*ct derlvativr 4-10 GX 82
BlMld
Urlne SKF-525A Electron
TlSSW imct WOW 3-6 MfGX 74
13.1 K GRONINGSSON ET A L .

8.4 HOMOGENEOUS ENZYME IMMUNOASSAY, EMIT @

EMIT
8 ( S va Corp. Palo Alto, CA 94304) i s a noniso-
t o p i c tecgnique t h a t i s used t o measure l i d o c a i n e
B r i e f l y , t h e procedure i s based on c o m p e t i t i v e p r k e i n
b i n d i n g w i t h g l ucose-6-phosphase dehydrogenase t o 1abel
the drug and antibody t o l i d o c a i n e as t h e s p e c i f i c
b i n d i n g p r o t e i n . The enz e a c t i v i t y , monitored as a
change i n absorbance a t %O nm due t o conversion o f
NAD' t o NADH, c o r r e l a t e s d i r e c t l y w i t h t h e c o n c e n t r a t i o n
of l i d o c a i n e i n the sample. This technique i s simple and
very r a i d and r e q u i r e s o n l y 50 1 o f serum. However,
Y
s i n c e cRromatograph i s n o t i n v o ved i n t e r a c t i o n s can
occur w i t h t h e a n t i i o d y and cause f a f s e l y elevated drug
1 eve1 s.
Several s t u d i e s have been performed
accuracy and s e n s i t i v i t y o f t h e EMIT 8
evaluate the
l i d o c a i n e assay
b comparing i t w i t h the r e s u l t s obtained by gas
cgromatographic methods (83 86). -
The technique has a l s o been used f o r i n v e s t i g a t i o n o f
t h e importance o f bl ood-coll e c t i o n tubes i n plasma
1 idocaine determinations (871, a n t i b o d s e l e c t i v i t y
(88) curve s t a b i l i t y (89) and pharmacoiinetics i n
thermally i n j u r e d r a t s (90). Recently a procedure
f o r adapting the EMIT procedure t o t h e Cobas B i o
c e n t r i f u g a l analyzer has been described which avoids
t h e decrease i n p r e c i s i o n a t t h e upper end o f t h e
therapeutic range reported i n o t h e r adaptations (91).
t:
The l a t e s t rogress i n t h e area o f enzyme imnunoassa
techniques ave appeared i n several a b s t r a c t s from t e
National meetin s f o r the American Association f o r
K
C l i n i c a l Chemisfry (92 -
97).

9. IDENTIFICATION AND DETERMINATION I N PHARMACEUTICALS

9.1 IDENTIFICATION TESTS
A sample ( a l k a l i z e d i f c o n t a i n i n g l i d o c a i n e hydrochlor-
i d e ) i s e x t r a c t e d w i t h n-hexane o r chloroform followed
by evaporation t o dryness. An i n f r a r e d absorption
spectrum i n a potassium bromide d i s p e r s i o n o f t h e
residue e x h i b i t s maxima only a t t h e same wavelengths as
t h a t o f a s i m i l a r preparation o f a chemical reference
standard.
Apart from i n f r a r e d spectroscopy t h e a d d i t i o n a l
i d e n t i f i c a t i o n t e s t presented i n Section 5.2 may be
performed on t h e e x t r a c t i o n residue.
LIDOCAINE BASE AND HYDROCHLORIDE 235

Comparison of retention data ( e g retention times i n gas

chromato raphy and l i q u i d chromatography and R -values
9
i n t h i n - ayer chromatography (98) w i t h those i f a
reference standard may serve a s identity. In a d d i t i o n ,
when u s i n g l i q u i d chromatography and a variable wave-
1ength detector absorption r a t i o s between two wave-
l e n ths (e g 254 nm an 265 nm) a r e valuable
9
con ributions t o identity.

9.2 EXTRACTION ANALYSIS

To an a l i q u o t ( n o t more than 30 ml) of injection solu-
t i o n corresponding t o about 0.2
3 he
of lidocaine h dro-
chloride i s added 2 ml of 2 M so ium hydroxide.
s o l u t i o n is extracted w i t h two 20-ml portions of
chloroform. To the combined chloroform extracts are
added 10 m l of dioxane and f i v e dro s of a mixed
indicator (2 p a r t s of 0.2 %, w/v, BftL-blue and 3 p a r t s
o f 0.1 % w/v, alcoholic solution of methyl red).
T i t r a t i o n i s performed w i t h 0.1 M perchloric a c i d t o a
reddi sh-violet endpoint.
Lidocaine hydrochloride i n Lidocaine Injection and J e l l y
and Lidocaine i n Lidocaine Ointment are determined by
base extraction of a d i l u t e d sample w i t h chloroform.
After evaporation of the combined chloroform e x t r a c t s ,
sulphuric acid i n excess i s added. The excess 1 s titra-
ted w i t h sodium hydroxide usin a potentiometric
determination of the endpoint ?7).
The acid-dye technique has been used for Injections and
Creams containing Lidocaine (99). The sample, buffered
t o pH 3.6, i s extracted w i t h chloroform usin the ion-
9
p a i r i n g dye reagent bromophenol blue. The ch oroform
phase i s rendered a l k a l i n e and the b l u e colour i s
measured photometrically a t 600 nm.

9.3 GAS CHROMATOGRAPHY

Determination of 1idocaine, a f t e r removal from the
sample matrix by base extraction e.g. into dichlorome-
thane, can be performed by a s chromatography prefera-
9
bly u s i n g mepivacaine as i n ernal standard.
The chromatographic s stem may be identical t o t h a t
P
presented i n Section 5.4, i.e. u s i n g a ca i l l a r y column
and fl ame ionization detection. Add1 t i o n a columns t h a t
have been used contain 3 % OV-17 on 80 - 100 mesh
s
Chrgmosorb W HP (detector andoinjector t e e r a t u r e s
250 C, column temperature 190 C ) (72) and % OV-101 on
80 - 100 mesh Chromosorb W HP (detector temperature
275°C i n j e c t o r temperature 310°C, column temperature
190°Cf (71).
236 K. GRONINGSSON E T A L

9.4 CAPILLARY ISOTACHOPHORESIS

C a p i l l a r y isotachophoresis has been used for t h e q u a l i -
t a t i v e and q u a n t i t a t i v e sirnul taneous determi n a t i o n o f
1 idocaine- procaine- and t e t r a c a i n e hydrochlorides i n
drugs (loof. The recovery o f t h e method i s about 99.7 %
and acids, vitamins and nonionised organic compounds
present i n the drugs do n o t i n t e r f e r e w i t h t h e
1 so tac hophore t i c separation.

9.5 LIQUID CHROMATOGRAPY

L i q u i d chromatography (HPLC) i s t h e most widely used
technique f o r determination o f 1idocaine i n pharmaceu-
9-
t i c a l s . The chromato raphy has been performed i n
straight-phase a we1 as i n reversed-phase modes using
UV-detection a t 230 254 nrn.
Suitable chromatographic systems a r e given i n Table 12.

ACKNOWLEDGEMENT
We wish t o express our sincere thanks t o Mrs C h r i s t i n
.
Andersson f o r her assistance i n the preparation o f the
manus c r i p t
LIDOCAINE BASE AND HYDROCHLORlDE 237

Table 12 Llquld U l m t o p r r p h i c S P t r for Lldocain

Elumt ~ l ~ raw
lr Rrtmtla t i n RIf
(rllmln) (.In)

MUclWSll 5 110, dP 5 I0
(mclunY*91?)
1.o 1.1 1

.
200 a 1I l.d.
R u c l w a i l 7 OH. dp 1.1 p
5w
mCkreY-ua911 1
I x 1 Il.d. n-H.xan - ethanol (97:3) 1.o 6.9 4

--
n-Heptm d i c k l o m r t h a n -
r c m t o n i t r l l e pmp ldn
(50 : 75 : M : O.l{ 1.o 3.6 101

n-Weptam - d l c h l o m t l u w -
acetonitrlh - pmpylmlm
(54 : 15 : 5 : 0.1) 1.o 1.1 101

- -
A c e t o n i t r i l e phorplut. buffer
pn 3 . 0 . p 0.1, contalntng
0.1 I. u/v. l-octannulpkonlc
0.8 7.9 1
acid, $ o d f U Salt (10 : w))

pbondapk QI, dp
(watrn)
- 10 )I 0.01 M Idctanesulphonic acld.
sodim salt, 2 I r/r. aceClc acid.
.
2 I rlr. r c r t o n l t r l l e . and 1 I.
2.0 6.0 101

300 a 4 m l.d.
rlv.~rthamlt n wter
Trtrahydrofuran 50 ml/l-
@msphat. buffer a d j u s t d to
2.0 5.6 46
x 4 Il.d. pn 3.5 ( 3 : 97 )
LlChrosorb RP-8 dp = 10 p k r t o n l t r i l e - phmphata b u f f e r
E:ki I 1.d.
pn 8.0 (p 0 . 5 ) . ( M)o : b a l ) 1.o 3.1 1

A c e t a t f t r f l t - phosphatr buffrr
pn 8.0. )I 0.05 (600 : 400) 1.o 3.0 4

S-rl 10 R? 8 k r t o n l t r l l e - phosphate buffer

I 00
E r m m l n Labr)
x 4.6 rn t.d. rlth
Spheri 10 R? 1,
pn 8.0
p = 0.05, ( 500 : 5W) 0.8 6.5 1
#) 1 b.6 I l.d..
guard c o l m
octadecyl s f l a m , c l v i u l l y
b0nd.d to s111cI -
Pkosphat. buffer pn 7.0
wUr r c c t o n l t r i l r
-
(50 : rm : 5%) 2.0 7
I*thaml- ptmphata buffer 1.o 3.5 1
pn 8.0
p * 0.05 ( 700 : 300)
ketonitrlle
--
phosphta buffrr
pn 8.0. p 0.05 (600 : 400) 1.4 1.2 4

-
A c r t o n l t r l l e phosphata buffrr
Pn 6.2. p.- 0.05. contrln1ng
2 . 9 a 10~’ R t r t r m p n t y l u s n i u
chlor1d. (200 : Sm) 1.1 1.1 1
-
k e t o n l t r 1 l e phosphtm buffrr
pn 6.2, p-- 0.05, contalnlng
2.9 x R Utrabutylraanlu
h ~ m g m u 1 p h a t . a(200 : 800) 1.6 5.5 1
238 K . GRONINCSSON E T A L .

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LIDOCAINE BASE AND HYDROCHLORIDE 239

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Anaesthesiology, 48, 4b9 (1978)
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