Tohoku

J. exp.

Med.,

Infrared

1975,

116, 259-265

Absorption

Spectroscopy

of Pure

Pigment

Gallstones NORIYOSHI SUZUKI, YASUSHI NAKAMURA and

TOSHIO SATO

Departmentof Surgery,Tohoku UniversitySchool of Medicine, Sendai

SUZUKI, copy

of

N.,

Pure

„Ÿ Pure

pigment

appearance, these

stones

constituents

but

of

the

It

associated and

such

as

that

order

was

calculated

cm-1

at

was

found

to

they

which

the to

gallstone;

of stone

calcium

in

From

this

commonly and

is due with

pigment and

at to

stones

synthetic

bilirubinate

the

three

pyrrole

"blackness"

calcium

calcium

rings

over

bile

the positions,

0.30

bilirubinate.

for

Q all

were of

being

65

components stones. quotient

Q

including The

stone,

bilirubin

stones

bilirubinate

of bilirubin. the

main

incidence

organic

spectrum,

determined

of

and

with

of

an

of

the

the

at

of

respective spectra

polymers

calcium

than

of

of

findings

as

black

spectra

than

other

that

of

acids

position aspect

and

associated

fatty

readings

well

general

259-265 by

The

in

identified

phosphate

(3),

characterized

spectroscopically

"smoothness"

absorption

pure

been

Spectros

116

spectroscopy.

smoother

acids

1975,

bilirubinate

have

Absorption

Med.,

gallstones

were

less

intensity

correlate

percent

bilirubinate

bile

of

calcium

revealed

were

Infrared

exp.

absorption

and/or

quantitate from

infrared

stones

carbonate

T.

J.

stones.

also

cholesterol,

In

91

black

SATO,

variety

of

they

was

with

minor

those

bilirubinate

derivatives.

and Tohoku

by

resembled

calcium

percent,

a

analyzed

bands,

usual

Y.

Gallstones. stones,

were

absorption

for

NAKAMURA,

Pigment

1624

"smoothness" less

specimens

than of

pure

0.30

calcium pigment

gallstone

The pure pigment stone which is characterized by black appearance is a minor subtype of gallstones of bilirubin line (Maki 1966). This type of stones comprised about 9 percent of the 803 cases undergoing operation in the last 10 years at our Department (Table 1). They are formed almost exclusively in the gallbladder, and about 45 percent of them reveal positive shadows on a plain abdominal film. Patients with hemolytic jaundice are complicated by such stones at an incidence of about 40 percent. Suzuki (1965a, b, 1966) found that black pigments of the pure pigment stone consisted mostly of polymers of some bilirubin derivatives, with possible contribu tion of metal complexes. It was also found spectroscopically that pure pigment stones contained such metal elements as Ca, Cu, Mg, Mn and Fe at higher concentrations than calcium bilirubinate stones and cholesterol stones. In succession to those investigations, this paper deals with results of infrared absorp tion spectroscopy accomplished on a large number of pure pigment stone specimens. Received

for publication,

April

18, 1975. 259

260

N. Suzuki

et al.

TABLE1. Classification of gallstonesand incidence in 803 surgical cases

* The

numbers

in brackets

represent

MATERIALS Forty mens, of

pure

were

gross

pigment

Group ‡T:

very

cases.

which

Such

on

number small

the

AND METHOD

them

derived

classified

from

into

the

40

sets

of

following

recent

three

surgical

groups

speci

on

the

basis

10

was

in

15

surface

stones

(about

surface

seen

of

milimeters

the

or

but

less

number

in

patient in

varied

of

stones

from

in

the a

globules

Such two

to

as

smooth-surfaced

portion of

thorny-surfaced

intersections.

diameter)

a central

The

diameter)

also

a single

mm

black

cases.

in

in

and

inside

seen

many

as

spherical

had

patient

were

dark

ranged

in 300.

stones

brownish

between

hue. two

and

200. Group ‡V:

shaped

relatively

black

brownish stones

in

whenever gallstones;

a patient

stone

portions

56

bilirubinate,

photometer

method spectra

purely of

10

one

several

in

a mortar

(Maki

et were

al.

was

gross of

black

portions

1964),

measured

was

at

82

into

in

somewhat

stratified

17

powder

whereas were

and

the

KBr-disk

for

irregularand

cases,

the

included number

were 26

thus

and

control

a stone and

of

black

semisynthetic

supply

Group

by

the II

or

separately

from

brownish 17

In of

pulverized

prepared

from

commercial

method

spectroscopy.

with

isolated

stones of as

or or

seen

fine

specimens

bilirubin served

were

pulverized,

bilirubinate

by

spherical

amorphous

most.

appearance

A total

from

diameter)

Such

or

stone

calcium

made

in

surface

extent.

different

feasible. from

specimens

Infrared

various

whole

mm

cut

pulverized the

with

technically

(10-30 the

only

were I

of

described

of

being

Pulverized calcium

which

portions

a Group III

large

of

gallstones

of

Group

stones,

black

The case

of

were

(several

black

The

were

about

each

stones

small

uniformly

Group ‡U: in

The

.

appearance.

appearing eight

stones,

utilized.

percentages

the

40

black

portions. specimens

the

previously

materials. with

Hitachi

Infrared

Spectro

EPI-S2.

RESULTS

Typical examples of infrared absorption spectra of the purely black specimens are illustrated in Figs. 1-3. They resembled spectra of calcium bilirubinate stones and synthetic calcium bilirubinate (Suzuki and Toyoda 1966, 1967; Toyoda 1966) in the position of principal absorption bands, but they differed from the latter in that the respective bands were broader and the whole spectral curves were less sharply shaped. These basic spectra were occasionally associated with absorption bands of calcium carbonate (Fig. 2) and/or calcium phosphate (Fig. 3). Such were seen in

Infrared

Fig.

1. Infrared absorption spectra examples of infrared absorption pigment

Fig.

from

26

than

in

the alkanes

absorption

at

about

to

these in

of those

the of

specimens

were

purely

black

the

absorptions As

261

of

the

1702

in

vicinity

cm-1,

40

salts

Unlike in

the

calcium

pigment indicate

the of

stones associated with the specific absorption

black was

stones. a

spectra 3000

probably

little of

was

carboxyl

shown

in

frequent

calcium

cm-1

of

As more

absorption bands of

Table

in

bilirubinate

2,

Group ‡U stones,

usually

very

weak

groups,

was

usually

the

and

the

feeble

absent.

Spectra similar

of pure Arrows

percent)

inorganic others.

by

even

(65

of

absorption

or

Gallstones

stones.

association

stones

of Pigment

of pure pigment stones. A, B and C are typical spectra of the purely black specimens in the pure

2. Infrared absorption spectra bands of calcium carbonate. calcium carbonate.

specimens the

Absorption

of mentioned

such

specimens the

from

purely

black

associated

with

ones. organic above,

Some components the

basic

brownish

black

specimens. inorganic of

the as

spectral

portions

However, calcium brownish

cholesterol pattern

as

salts

more

black and of

were shown

essentially in

commonly

specimens fatty the

Table

3, than

exhibited

acids. pure

pigment

stone

262

N. Suzuki

et al.

Fig. 3. Infrared absorption spectra of pure pigment stones (A, B and C) associated with absorption bands of calcium phosphate. An arrow indicates the specific absorption band of calcium phosphate. TABLE 2.

*

B

(bile

t •õC

Infrared

pigments

(calcium

analyses of 40 pure pigment stones

and/or

it's

carbonate);

$

derivatives); P

(calcium

phosphate).

was characterized by smoother aspect as compared with the spectrum of calcium bilirubinate. In order to evaluate such a difference in objective terms, an index Q was defined as a measure of the "smoothness' of spectrum as follows:

where

In

stands

centimeter

(Fig.

(91

percent)

0.30

for

of all

bilirubinate. order

of

for

the

4). the

the

As 56

intensity shown

black

specimens

Among

of Fig.

specimens of

pure

Groups ‡V, ‡U

in

calcium

pigment and ‡T

absorption 5,

the

of

pure

at Q

stones

wave less

pigment

bilirubinate

(Fig.

the was

value

the

Q

stones, stones

and

value

tended

number than

n

0.30

while

it

synthetic

per

for was

51 over

calcium

to

decrease

in

6).

DISCUSSION

pure

Group ‡T

stones

pigment

stones.

of

the Infrared

above-mentioned absorption

gross spectra

classification of

these

stones,

represent and

also

typical those

of

Infrared

Absorption

of Pigment

Fig. 4. Illustration showing position of transmission text for explanation.

black

specimens

differed

from

the

bilirubinate. bands; the spectra

They but

former of

they being black

isolated

from

spectra

of

resembled differed more stones

Group ‡U calcium each

from

each

smoothly also

resembled

reading

263

for calculation

of Q.

See

Fig. 6. Distribution of Q for each gross classification of pure pigment stones.

Fig. 5. Distribution of Q in various gallstone materials and synthetic calcium bilirubi nate.

the

Gallstones

or ‡V

stones,

bilirubinate other

in

other

in

shaped those

resembled

stones

the

position

general than of

aspect the black

or of

but

apparently

synthetic principal

of

the

calcium absorption

spectral

latter.

Moreover,

substances

which

curve, the Suzuki

264

N. Suzuki

et al.

TABLE3. Infrared analyses of purely black specimens and brownish black specimens of the pure pigment stone

(1965a, is

b)

82 specimens

were prepared

purely

portions

prepared

confirmed

of

a

mixture

the

of

ratio

quotient

spectra

of

Q