Arch. Oto-Rhino-Laryng.

220, 259--264 (1978)

Archivesof
Oto-Rhino-Laryngology
9 Springer-Vcrlag 1978

Chemical Composition in Various Compartments

of Inner Ear Fluid*
Kazuo Makimoto 1, Teiro Takeda 1, and Herbert Silverstein2
1 Department of Otolaryngology,School of Medicine, Kyoto University,
Sakyo-ku, Kyoto 606, Japan,
2 Sarasota Memorial Hospital,
1901 Arlington Street, Sarasota, Florida 33579, USA

Summary. Sodium, potassium, chloride, glucose and total protein were determined in samples of scala vestibuli perilymph, scala tympani perilymph, CSF,
cochlear endolymph and utricular endolymph from normal cats. Small but significant differences were evident in the concentrations of sodium and potassium
between the scala vestibuli and scala tympani perilymph. It was also apparent
that each compartment of endolymph has different values for sodium and potassium concentrations. Compared with the endolymph, the perilymph was found
to contain a higher concentration of glucose and total protein. These findings are
discussed from the view point of biological significance.
Key words: Inner ear fluid biochemistry - Micro-chemical analysis - Osmotic
pressure in inner ear fluids.
There is a paucity in knowledge concerning chemical composition in inner ear fluids.
In experiment series on inner ear fluid chemistry (Silverstein and Makimoto, 1973;
Makimoto and Silverstein, 1974), we analysed sodium, potassium, chloride, glucose
and total protein in samples of scala vestibuli perilymph, scala tympani perilymph,
cochlear endolymph, utricular endolymph and CSF. 125 ears in 95 cats served as
the materials. The present report is a summarization of the results heretofore obtained.
Sodium and Potassium

Determination of sodium and potassium was done using a microflame photometer

(Makimoto and Silverstein, 1973). The amount used in this analysis was 1.0 ~1 for
perilymph, CSF and serum, and 0.5 p~l for endolymph.
* Read before the 13th Workshop on Inner Ear Biologyin Dfisseldorfon August 23, 1976
This work was supported by Research Grant No. ROI-NS 1026801 from the National Institute of
Neurological Diseases and Strokes, USA
Offprint requests to: Kazuo Makimoto, M.D. (address see alcove)

0302-9530/78/0220/0259/$ 01.20

260

K. Makimoto et al.

These results are shown in Table 1. As demonstrated here, small but significant
differences were evident between scala vestibuli and scala tympani perilymph both in
sodium and potassium concentrations. It was also apparent that each compartment
of endolymph has its own value for sodium and potassium concentrations.
In the perilymph of scala vestibuli, a higher concentration.of potassium was
noted. As to endolymph, a greater value was observed in the sodium concentration
of utricular endolymph. This m a y be regarded as one of the biological features for
utricular endolymph. In Figure 1, data on the concentrations of sodium and potassium in fluids o f extracellular type and serum are graphically illustrated. Regarding
sodium concentration, perilymph of the scala vestibuli was found to be closest to
serum. On the other hand, in potassium concentration, perilymph of the scala tympani showed the closest value to that of serum. When a comparison o f N a / K ratio
between scala vestibuli perilymph and serum was made, it was obvious that a greater
difference existed in this ratio. Accordingly, the perilymph of scala vestibuli m a y not
be a simple blood filtrate.

Chloride
Chloride was determined by the use of a Chloridemeter | With this instrument, the
mEq/1 of chloride in a sample can b e measured by passing a current through the
sample via two generating electrodes. The principle of this measurement is that silver
ions are released from the anode and combine with chloride ions in the sample. The
smallest amount required for this determination is 2.0 ~1.
Table 1. Sodium and potassium concentrations in inner ear fluids, CSF, and serum

Sodium
(mEq/1)

Potassium
Difference

(mEq/1)

Perilymph of
scala vestibuli (Pv)
Perilymph of
~
scala tympani (Pt)
Cochlear endolymph
(Ee)
Utricular endolymph
(Eu)

147.76 (+ 0 . 9 4 ) ~
(N = 54) ~ . > 9.35'
157.11'(+ 1.06) j ~ ( P t - - P v )
(N = 5 8 ) ~
6.07 (+ 0.40) ~
(N=41) " ~ 8 . 7 9 b
14.86 (+ 0.75) ~
(Eu-Ec)
(N = 44)

CSF

152.91 (+ 1.00)
(N = 32)

2.97 (+ 0.08)
(N = 32)

Serum

140.94 (_+ 1.10)

(N = 27)

3.93 (+ 0.16)
(N = 27)

Difference

10.47 (+ 0.31) .
(N = 54) ~
6.68c
3.79 (+ 0.11) / / ~ (Pv--Pt)
(N = 58) :
171.13 (+ 1.64) .
(N = 41) ~ 1 5 . 7 0
a
155.43(+ 1.62) j
(Ec--Eu)
(N = 44) /

a t = 6.55 (significant, P < 0.001)

b t = 10.15 (significant, P < 0.001)
c t = 21.28 (significant, P < 0.001)
a t = 6.81 (significant, P < 0.001)
Each data is expressed as a mean (_+ standard'error); N = represents number of samples

Chemical Composition of Inner Ear Fluid

261
(rnEq/L)
160

150

140

130

I Sodium Concentration
~

Potassium Concentration

Fig. 1. Distribution pattern of sodium and potassium

in scala vestibuli perilymph, scala tympani perilymph,

CSF and serum

Pt : Perilymph of Scala Tympani

Pv : Perilymph of Scala Vestibuli

Table 2. Chloride concentrations in inner ear fluids, CSF, and Serum

Chlorine
(mEq/1)
Perilymph of scala vestibuli (Pv)

138.51 (+_ 3.58)

Perilymph of scala tympani (Pt)

(N = 43) ~
130.00 (_+ 2.31)
(N = 46)

Cochlear endolymph (Ec)

169.82 (+ 5.83)

Utricular endolymph (Eu)

(N = 28) ~
158.88 (+ 6.22)
( N = 24)

CSF

138.97 (_+ 1.97)

(N = 39)

Serum

120.00 (+ 3.59)
(N = 30)

Difference

8.51 a (Pv--Pt)

10.94b (Ec--Eu)

a t = 2.026 (significant, 0.02 < P < 0.05)

b t = 1.283 (not significant, 0.2 < P < 0.3)
Each data is expressed as a mean (_+ standard error); N =.represents number of samples

262

K. Makimoto et al.

As shown in Table 2, the concentrations of scala vestibuli perilymph, scala

tympani perilymph and CSF showed values in a range from 130--140 mEq/1. Compared to these values, relatively high levels of chloride concentration were noted in
both cochlear and utricular endolymph.

Glucose
Glucose determination was carried out by adapting the enzymatic test kit of Boehringer to a micro-chemical technique (Silverstein, 1973; Makimoto and Silverstein,
1974). The principle of this enzymatic method is as follows:
1. Hexokinase catalyses the phosphorylation of glucose by ATP.
Glucose + ATP -~ glucose-6-phosphate + ADP.
2. D-glucose-6-phosphate (G-6-P) is oxidized to 6-phosphogluconate (6-P-G) in
the presence of TPN (NADP) by the enzyme G-6-P dehydrogenase.
G-6-P + TPN -~ 6-PrG + T P N H + H +.
The amount of T P N H arising in Equation (2) is equivalent to the amount of G-6-P
or glucose, respectively.
The amount of the samples used in this measurement was 0.5 ~1 for endolymph
and 1.0 pxl for other fluids.
The results are presented in Table 3. The glucose concentration of cochlear
endolymph was appreciably low, being 11.2 mg/100 ml, whereas that of utricular
endolymph was higher, being 41.9 mg/100 ml. As previously mentioned, utricular
endolymph was found to contain a higher concentration of sodium as compared to
Table 3. Glucose concentrations in inner ear fluids, CSF, and Serum

Glucose
(rag/100 ml)
Cochlear endolymph (Ec)
Utricular endolymph (Eu)
Perilymph of scala Tympani (P0
Perilymph of scala vestibuli (Pv)
CSF
Serum

Difference

11.17 (_+ 1.29)

(N = 28)
(+ 2,23) ~
41.86
(N~ 30)/"
82.77 (+ 3.73)
(N = 66)
79.65 (+ 3.46)
(N = 59)

30.69a (Eu--Ec)

3.12b (Pt--Pv)

81.95 (+ 3.37)
(N = 49)
122.48 (+ 7.12)
(N = 45)

t = 11.71 (significant, P < 0.01)

b t = 0.61 (not significant, 0.5 < P < 0.6)
Each data is expressed as a mean (_+ standard error); N = represents number of samples

263

Chemical Composition of Inner Ear Fluid

cochlear endolymph. Such evidence indicates the difference in chemical composition

between cochlear and utricular endolymph and may suggest that both compartments of endolymph are not freely communicable, and that each possesses its own
independent nature in formation, absorption and circulation of endolymph.
Perilymph of scala vestibuli, perilymph of scala tympani and CSF were almost
identical in glucose concentration, i.e., approximately 80 mg/100 ml. This level is
regarded as being about 40 mg/100 ml lower than that of serum.

Total Protein
Total protein was determined by employing the Folin phenol reaction of Lowry et al.
(1951). As the first step of this method, the sample is treated with copper sulfate in
alkaline solution. Folin reagent is then added to the sample to develop the color
reaction.
According to the results shown in Table 4, the total protein contents of scala
vestibuli perilymph and scala tympani perilymph exceeded those of cochlear and
utricular endolymph. Also in CSF, the content of total protein was noted to be much
lower than those in scala vestibuli perilymph and scala tympani perilymph. This
pattern of concentration gradient in total protein did not coincide with that in glucose. The glucose study revealed that the glucose concentration of CSF was almost
the same level as those of scala vestibuli perilymph and scala tympani perilymph.
In Figure 2, the total amount of electrolytes (sodium plus potassium plus chloride) are illustrated on the left side of this figure. When this value is compared among
various compartments of inner ear fluid, the values for endolymph are greater than
those for perilymph.

Table 4. Amount of total Protein in inner ear fluids and CSF

Total protein
(rag-%)
Perilymph of scala vestibuli (Pv)
Perilymph of scala tympani (Pt)
Cochlear endolymph (Ec)
Utrieular endolymph (Eu)
CSF

233.87 (+ 1 5 . 7 0 ) ~
(N = 23) ~
212.52 (+ 1 0 . 6 6 ) ~
(N = 31)
61.50 (+ 7.49)
(N = 18)
64.33 (+ 7.70)
(N = 9)
37.89 (+ 6.24)
(N = 9)

Difference
a
21 35 (Pv--Pt)
"

2.83b (Eu-Ec)

a t = 1.143 (not significant, 0.2 < P < 0.3)

t = 0.229 (not significant, 0.8 < P < 0.9)
Each data is expressed as a mean (_+ standard error); N = represents number of samples

264

K. Makimoto et al.
I0
[ p

I00

~.,e/ioo~

I mE;q/L

Pt

::::::::::::::::::Na
.........
( m t ::' q /-L ): : . :.:.:.:
.:. . . . .:.:.:-:
.]. . ..

C.S.F.

I:::::::::::::::: J:::;:::::::::::::::::::::::::::::::::?::::::::::]]

Eo

lilllilllllllllll: ;! !: !lIlllIHII IHII

.-

iG'

<mEq,.

!ililll]01111NIllll01111111IIIllllllllllllll
III1
P v : Perilyrnph of Scala Vestibuli

Ec : Cochlear Endolyrnph

P t : Perilymph of Scala Tyrnpani

Eu : Utricular Endolyrnph

P r : Total protein (m E % )

GI : Glucose ( m g / I O O m l )

Fig. 2. Comparison of osmotic concentration among various compartments of inner ear fluid and
CSF

Conversely, the concentration of glucose and that of total protein as shown on

the right side of this figur6, are higher in perilymph than in endolymph. Conceivably,
in terms of total osmotic concentration, the deficit of total electrolytes concentration
in perilymph could be partly overcome by the relatively high concentrations of
glucose and total protein in perilymph. Thus, these two contradictory findings are
likely to be of significance for the osmotic pressure equilibrium between perilymphatic and endolymphatic compartments. For further understanding of the osmotic
pressure equilibrium, other factors such as biocarbonate ion have to be taken into
consideration.
Acknowledgements. Thanks are due to M. Ohara, Kyoto University, for assistance with the
manuscript.

References
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin
phenol reagent. J. biol. Chem. 193, 265-275 (1951)
Makimoto, K., Silverstein, H.: Sodium and potassium concentrations in the endolymph and perilymph
of the cat. Ann. Otol. 83, 174--179 (1974)
Makimoto, K., Silverstein, H.: Effects of insulin on glucose concentrations in inner ear fluids and
cochlear microphonics. Laryngoscope 84, 722--737 (1974)
Silverstein, H.: Biochemical studies of the inner.ear fluids in the eat. Ann. Otol. 75, 48--63
(1966)
Silverstein, H., Schuknecht, H. F.: Biochemical studies of inner ear fluid in man. Arch. Otolaryng. 84,
395--402 (1966)
Silverstein, H.: Inner ear fluids in man. Laryngoscope 83, 79-107 (1973)
Silverstein, H., Makimoto, K.: Superior vestibular and "singular nerve" section. -- Animal and clinical
studies --. Laryngoscope 83, 1414--1432 (1973)
Received October 24, 1977