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Lidocaine Base and Hydrochloride: Groningsson, Lindgren, Lundberg, Sandberg
Lidocaine Base and Hydrochloride: Groningsson, Lindgren, Lundberg, Sandberg
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
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
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
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)
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 ) .
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.
Sol vent
water 95% ethanol chloroform n-hexane
-
base 0.004 0.76 0.79 0.12
hydrochloride 0.68
n-octanol - phosphate
buffer a t pH 7.4 46 4
methylene chloride - water (base) 5200 3
-
toluene water (base) 160 3
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
1 , , ~ , 1 , , 1 , , , , 1 , , , , ~
75 50 2.5 0 Ppm
t l r , , l l l r , , ' " , , l , , ;
75 50 25 0 ppm
I , I I I II
80 60 40 20 100 80 60 40 20 0 ppm
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
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
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
Peak-find
L 262 = 0.421 4
L271 =0.3417
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.
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.
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.
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
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
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,
1
(GW
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.
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
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
Packed Nm Na
n 3 NO 75
-1 * Internal
standard
Techniqur (hrivatlvr
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).
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
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
-
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 .
REFERENCES
1. N. L o f ren, A r k i v f o r kemi, mineralogi och geologi,
-
22A, (981, l U t r 9 4 b 1
2. N. Lofgren, Studies on l o c a l anaesthetic. Xylocaine. A
new s y n t e t i c drug. Monography (1948)
3. P-A. Johansson, Acta. Pharm. Suec., -
19, 137 (1982)
4. Astra Sweden, unpublished data.
5. A. Brodin, A. Nyqvist-Mayer, T. Wadsten, B. Forslund and
F. Broberg, J. Pharm. Sci. -73, 481 (1984)
6. N. I. Nakano, J. Pharm. Sci., -
68, 667 (1979)
7. U.S. Pharmacopeia, USP XX, (1980)
8. European Pharmacopoeia, I I (1971
9. B r i t i s h Pharmacopoeia, Vol. I (1980)
10. K. Bullock and J. Grundy, J. Pharm. Pharmacol., Vol.
-
V I I , 755 (1955)
11. P. A. Johansson U. Stefansson and G. Hoffman, Anal.
-
Chim. Acta, 151, 49 (1983)
12. N. Lofgren Studies on Local Anaesthetics, Dissertation,
Stockholm 11948)
13. G. T. Tucker and L. E. Mather, Br. J. Anaesth., -
47,
213 (1975)
14. G. T. Tucker and L. E. Mather, Clin. Pharmacokinet., -
4,
241 (1979)
15. N. L. Benowitz and W. Meister, Clin. Pharmacokinet., -
3,
177 (1978)
16. D. B. Scott, D. G. Littlewood, B. G. Covino and G.B.
Drummond, B r . J. Anaesth., -
48, 899 (1976)
17. D. P. Cruikshank, D. W. Lambe and L. J. De Bacher,
Obstet. Gynecol., -
42, 127 (1973)
18. D. B. Scott, P. J. R. Jebson, B. Orten r e n and P.
Frisch, B r . J. Anaesth., -
44, 1040 (1979)
LIDOCAINE BASE AND HYDROCHLORIDE 239