Ampicillin

GENERAL

Nases

Ampicillin; D-a-aminobenzy1 penicillin.

Structure

C 16 19N 0 S
H 3 4

mol. wt. 349.41

Poras Available

Free a c l d (anhydrous ampicillin); sodiua salt; ampi-

cillin trihydrate.

Phyaical Properties

Solubility: 1 g d i s s o l v e s i n a b o u t 90 nL o f w a t e r a t
2 5 ° C , a n d 1 g i n 250 mL o f a l c o h o l ( 1 ) . Melting point
202 t o 2 0 3 ° C ( 2 ) .

Stability Summary

A m p i c i l l i n i s s u s c e p t i b l e to 8-lactam r i n g h y d r o l y s i s .
Its pH-rate profile reveáis s p e c i f i c - a c i d - and spe-
c i f i c - b a s e - c a t a 1yzed h y d r o l y s i s . General acid and
g e n e r a l b a s e c a t a l y s i s a r e known t o o c c u r i n c i t r a t e
and p h o s p h a t e b u f f e r systems. T h e pH o f m á x i m u m s t a -
b i l i t y i s 5.8. Owing t o d i m e r i z a t i o n , the r a t e of
degradation increases s u b s t a n t i a l l y with increasing
initial concentration. A d d i t l o n of a l c o h o l to s o l u -
t i o n s o f a m p i c i l l i n has been shown t o e n h a n c e i t s s t a -
bility. I n t e r c o n v e r s i o n o f h y d r a t e s m u s t be p r e v e n t e d

198
 Drug Kinetlcs 199

in order to avoid a l t e r a t i o n of b i o a v a i l a b i 1 i t y char-

acteristics. The r a t e o f d e c o m p o s i t i o n of a m p i c i l l i n
i s e n h a n c e d by i n c r e a s i n g t h e c o n c e n t r a t i o n of dex-
t r o s e and o t h e r c a r b o h y d r a t e s i n s o l u t i o n .

DRUG KINETICS

Reactions and Rate Equation

As i s g e n e r a l l y true f o r the other p e n i c i l l i n s , the

most i m p o r t a n t s o u r c e o f d e g r a d a t i o n o f a m p i c i l l i n i s
the h y d r o l y t i c cleavage of the B-lactam r i n g . In acid
or n e u t r a l s o l u t i o n the f o l l o w i n g products are formed:

a-aminobenzy1penamaldic a-aminobenzy1 penillic

acid acid

A t pH v a l ú e s a b o v e 7, d e g r a d a t i o n o f a m p i c i l l i n i s a
r e s u l t of hydroxide-ion-catalyzed hydrolysis, giving
the i n d i c a t e d p r o d u c t , and a h i g h l y c o n c e n t r a t i o n - d e -
pendent d i m e r i z a t i o n r e a c t i o n ( 3 ) .

a-aminobenzylpeni1loic acid

The p r e s e n c e o f t h e a m i n o s i d e - c h a l n g r o u p o f a m p i c i l -
l i n a p p e a r s t o h a v e a s i g n i f i c a n t e f f e c t on t h e r a t e
of d e g r a d a t i o n . Being amphoteric, a m p i c i l l i n can
e x i s t a s a catión, a z w i t t e r i o n , a n d an anión. The
pK a valúes o f t h e c a r b o x y l a n d a m i n o g r o u p s a r e 2.65
200 Ampicillin

and 7.24, r e s p e c t i v e l y ( 4 ) . T h e s e t h r e e s p e c i e e have

been demonstrated t o undergo the f o l l o w i n g r e a c t i o n s
in dilute solution:
+ kj
H3N-R-COOH + H + H > products (1)

k2
H3N-R-COO- + H + H > products (2)

+ k 3

H3N-R-COOH H > products (3)

k 4

H N-R-C00-
2 Q > products (4)
5 K

H N-R-C00"
2 + "OH H^ > products
Q {5)
This series of reactions i s summarized by the rate
equation
+ +
rate « k [H3N-R-COOH][H ] +
x + k [H N-R-C00"][H ]
2 3 +

+
+ k [H3N-R-COO-]
3 + k [H N-R-C00-]
4 2

+ k [H N-R-C00~][0H~]
5 2 (6)

In t h i s e x p r e s s i o n k j and k are second-order 2 specific

acid catalysis rate constants, k and k are f i r s t -
3 4

order constants f o r h y d r o l y s i s o f z w i t t e r i o n i c and

a n i o n i c a m p i c i l l i n , a n d k$ i s a s e c o n d - o r d e r specific
base c a t a l y s i s constant. At h i g h c o n c e n t r a t i o n s and
pH v a l ú e s a b o v e 7, b a s e - c a t a 1 y z e d a n d u n c a t a l y z e d
d i m e r i z a t i o n r e a c t i o n s h a v e b e e n f o u n d t o be a n a d d i -
t i o n a l important cause of degradation ( 3 ) .

pH-Rate Profile

Figure 1 i s the pH-rate p r o f i l e f o r the h y d r o l y s i s of

a l o w c o n c e n t r a t i o n o f a m p i c i l l i n a t 35°C a n d c o n s t a n t
ionic strength. T h i s p l o t r e p r e s e n t s Eq. (6) w i t h a l l
constants extrapolated to zero buffer concentration.
A t pH l e s s t h a n 1.5 s p e c i f i c a c i d c a t a l y s i s involving
t h e a m p i c i l l i n catión p r e d o m i n a t e s . The s h o u l d e r i s
due t o t h e i o n i z a t i o n o f t h e c a r b o x y l i c a c i d group.
 Drug Kinetics 201

Reactions (1) through (3) a r e r e s p o n s i b l e f o r the

b e h a v i o r o f t h e p r o f i l e o v e r pH 3 t o 5. The l e v e l i n g
o f t h e c u r v e f r o n pH 5.5 t o 6.5 s u g g e s t s predoninance
of r e a c t i o n s (2) and (3) i n t h e d e g r a d a t i o n process.
A t pH g r e a t e r t h a n 7 . 5 r e a c t i o n (5) i s e s s e n t i a l l y
responsible f o r the observed rate, c l e a r l y demonstrat-
i n g the e f f e c t of s p e c i f i c base c a t a l y s i s . The máxi-
mum stability o c c u r s a t pH 5 . 8 , i n d i c a t i n g t h a t t h e
zwitterionic form i s the l e a s t susceptible species
with respect to hydrolysis. T h e tjg a t pH 6.6 i s
a p p r o x i m a t e l y 39 d a y s .

3.8

4.2

4.6

5.0

-* 5.4

5.8

6.2

6.6

1.0 3.0 5.0 7.0 9.0

PH

F I G U R E 1. Ampicillin. pH-rate p r o f i l e f o rh y d r o l y s i s
o f a m p i c i l l i n a t 35°C ( r a t e c o n s t a n t s i n s ) . - 1

Activation Energy

The t e m p e r a t u r e dependence of a m p i c i l l i n h y d r o l y s i s
h a s b e e n r e p o r t e d ( 2 ) a n d i s s h o w n a t pH 4 . 9 3 i n t h e
form o f a n A r r h e n i u s p l o t i n F i g u r e 2. The p s e u d o -
202 Ampicillin

f ir st - o r d e r c o n s t a n t s itere o b t a i n e d under conditions

of c o n s t a n t i o n i c s t r e n g t h . The a c t i v a t i o n e n e r g i e s
obtained f r o i the slopes of such p l o t s are tabulated
i n T a b l e 1.

1/T x 10 3

F I G U R E 2. A m p i c i l l i n . A r r h e n i u s p l o t showing teaper-
ature dependence of r e a c t i o n ratea of h y d r o l y s i s of
a m p i c i l l i n a t pH 4 . 9 3 .

TABLE 1. Activation Energies of

Ampicillin Hydrolysis

pH E a (kcal/aol)

1.35 16.4
4.9 3 18.3
9.78 22.3

Other Data

Ampicillin has a l s o been shown t o undergo general-

acid- and genera 1-base-cata1yzed-hydro 1ysis ( 2 ) .
 Drug Kinetics 203

F i g u r e s 3 a n d 4 show t h e d e p e n d e n c e o f t h e o b s e r v e d
rate c o n s t a n t s on t o t a l b u f f e r c o n c e n t r a t i o n i n c i -
t r a t e and phosphate b u f f e r s , respectively.

i—i—i—i—i—i—i—i—¡—i—r

0 2 4 6 8 10 12 14 16 18 20 22 24
Total citrate x 1 0 / m o l - liter"
? 1

F I G U R E 3. Ampicillin. Plot of observed rate constant

a g a i n s t t o t a l c i t r a t e b u f f e r c o n c e n t r a t i o n át 35°C ( A ,
pH 2 . 2 5 ; B , pH 2 . 7 6 ; C, pH 3 . 1 2 ; D, pH 4 . 7 5 ; E , pH
6.80).

A t 3 5 ° C a n d pH 1.2 a s l i g h t l i n e a r p o s i t i v e s a l t
e f f e c t on a m p i c i l l i n h y d r o l y s i s i s o b s e r v e d ( 2 ) . No
salt effect i s o b s e r v e d a t pH 4 . 9 4 . A t pH 1.2 t h e
addition of alcohol to the solution results in a
decrease i n hydrolysis r a t e ; t h i s has been a t t r i b u t e d
to t h e decrease i n s o l v e n t d i e l e c t r i c constant (4,5).
204 Ampicillin

Ampicillin i n 50* alcohol s o l u t i o n has a half-life

twice that i n p u r e l y aqueous s o l u t i o n .

Total phosphate X 10 /mol • liter

2 -1

F I G U R E 4. Ampicillin. Plot of observed rate constant

against t o t a l phosphate buffer concentration a t 35°C
( A , pH 7 . 1 1 ; B, pH 6 . 6 0 ) .

FORNULATIONS AND COMBINATIONS

Degradation Reactions

A t h i g h t e m p e r a t u r e s and. h u m i d i t i e s , a n d d e s p i t e f i r m
b i n d i n g w i t h i n t h e t r i h y d r a t e c r y s t a l , t h e water mole-
c u l e s a r e s u f f i c i e n t l y f r e e t o particípate i n a s o l i d -
state hydrolytic reaction (6). C a r e must be e x e r c i s e d
t o e x e l u d e w a t e r a s much a s p o s s i b l e f r o m s o l i d d o s a g e
forms and t o maintain t h e storage temperature at a
 Formu1ations and Combinations 205

modérate 1 e v e 1 .
I n t e r c o n v e r s i o n of t h e h y d r a t e d and unhydrated
forms of a m p i c i l l i n can have s i g n i f i c a n t e f f e c t s on
t h e d i s s o l u t i o n r a t e o f f o r m u l a t i o n s and t h u s affect
the b i o a v a i l a b i 1it y ( 7 ) . The a n h y d r o u s f o r m i s more
s o l u b l e t h a n t h e t r i h y d r a t e , but i t s s o l u b i l i t y de-
creases with increasing temperature. Solubility has
b e e n d e t e r m i n e d a t 37°C a s a f u n c t i o n o f pH f o r b o t h
c r y s t a l forms (8). T e m p e r a t u r e and h u m i d i t y control
are e s s e n t i a l t o p r e v e n t t h i s change i n c r y s t a l l l n e
f orm .
Components of commonly used d i l u e n t s f o r i n t r a v e -
n o u s infusión o f s o d i u m a m p i c i l l i n c a n a f f e c t t h e r a t e
of h y d r o l y t i c d e g r a d a t i o n . D e x t r o s e has b e e n shown t o
h a v e a c a t a l y t i c e f f e c t on t h e d e c o m p o s i t i o n o f a m p i -
c i l l i n (9,10). F r e e z i n g of r e c o n s t i t u t e d solutlons
has a v a r i a b l e e f f e c t w i t h r e s p e c t t o l e n g t h of t i m e
required f o r 10* d e g r a d a t i o n i n s e v e r a l diluents
(Table 2).

TABLE 2. Máximum S t o r a g e Time as a Function of

D i l u e n t and T e m p e r a t u r e (9)

Máximum Time of
Solution t <°C) Storage ( t 9 0 )

25* Sodium a m p i c i l l i n -20 48 h

In s t e r i l e water 5 4 h
In r e c o n s t i t u t e d v i a l 27 1 h

1* Sodium ampicillin 5 5 days

In normal sallne 27 24 h

1* Sodium a m p i c i l l i n 4 4 h
In d e x t r o s e 5* 27 2 h

S u c r o s e has been shown t o c a t a l y z e t h e d e g r a d a t i o n o f

a m p i c i l l i n (11,12). A l i n e a r r e l a t i o n s h i p between the
d e g r a d a t i o n r a t e a n d s u c r o s e c o n c e n t r a t i o n up t o 10*
has been o b s e r v e d ( 1 2 ) . O r a l suspensions i n u n i t dose
c o n t a i n e r s m a i n t a i n e d more t h a n 90* a c t i v i t y after
s t o r a g e f o r 60 d a y s a t -10 t o -20°C ( 1 3 ) . Although
206 Ampicillin

differences i n d e g r a d a t i o n r a t e h a v e been observed

among o r a l l i q u i d products manufactured by d i f f e r e n t
companies, a l l tested products maintained acceptable
potency a t room temperature f o r a t l e a s t seven days
(14 ) .

Stabi1ization Methods

To p r e v e n t h y d r o l y s i s o f t h e B - l a c t a m ring, water
s h o u l d be p r e v e n t e d f r o m c o m i n g i n c o n t a c t w i t h s o l i d
ampicillin. T e m p e r a t u r e a l s o p l a y s an i m p o r t a n t role
in rates of degradation i n the s o l i d and s o l u t i o n
states. Because of the l i m i t e d h a l f - l i v e s i n Solu-
t i o n s and s u s p e n s i o n s , s o l i d dosage forms a r e t h e o n l y
f o r m u l a t i o n s s t a b l e over extended periods of time.
Lowering o f t h e d i e l e c t r i c constant of Solutions of
a m p i c i l l i n w i t h a l c o h o l has resulted. i n b e t t e r s t a b i l -
i t y t h a n i n f u l l y a q u e o u s s o l u t i o n s a t l o w pH. Buf-
f e r i n g a t t h e p H - r a t e mínimum a n d s t o r a g e a t r e l a t i v e -
ly low c o n c e n t r a t i o n s a r e v i a b l e a l t e r n a t i v e s f o r
enhancing the s t a b i l i t y of l i q u i d formulations.

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2. J . P. H o u a n d J . W. Poole, J. Pharm. Sci., 58,

447 (1969).

3. H. Bundgaard, Acta Pharm. Suec., 13, 9 (1976).

4. J . P. H o u a n d J . W. Poole,, J. Pharm. Sel., 58,

1510 ( 1969) .

5. G. Scatchard, Chem. Rev., 10, 229 (1932).

6. N. H. G r a n t and H. E. Album, Nature, 207, 645

( 1965 ) .

7. J . W. P o o l e a n d C. K. Bahal, J. Pharm. Sel., 57,

1945 (1968).
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- R i c h a r d A. P y t e r
(Wiscons i n )