954
the concentrations of 2,3-dimercapto-l-propancland 1.3-dimer-
rapto-2-propanol were kept constant and the manganous ion
concentration was varied (Table 111). Here it was observed that
a t large nianganous ion concentrations the fading of the complex
formed from the dithjol mixture was such that the effect of the
1.3-dimercapto-2-propanol was apparently minimized.
On the basis of these observations, the dithiol test, as applied
to 2,3-dimercapto-l-propanolcontaining possible admixtures of
1,3-dimercapto-2-propanol,was revised to use a high concentra-
tion of manganous acetate in spite of the increased rate of fading
caused thereby. I n this way, 2,3-dimercapto-l-propanolcon-
taining as much as 40% of the isomer gave absorbances attribu-
table onlv to its 2,3-dimercspto-l-propanol content (Table 11).
Furthermore, the curve for absorbance versus 2,3-dimercaptc1-1-
propanol concentration was strictly linear for solutions of the pure
substance over the concentration range studied. I n Figure 3 the
absorbances obtained, 1 minute after mixing, n ith solutions of
constant dithiol concentration but varying ratios of 2,3-dimer-
capto-1-propanol to 1,3-dimercapto-2-propanol, are plotted along
with absorbances for varying concentrations of 2.3-dimercapto-
1-propanol alone (data from Table 11),with 0.122.1f manganous
acetate as the reagent. With this high concentration of manga-
nous acetate the observed molar absorptitivity is about 11 to
12% greater than that shown under Linearity of the Test.
DISCUSSION
Vicinal dithiols can be estimated via their slightly dissociated
colored manganous complexes in pyridine-containing solvents
while monothiols, being unable to chelate, do not form such com-
plexes. I n the case of 1.3-dimercapto-2-propanol it appears that
in addition to the probable possession of a lower stability con-
stant, the manganous complex is destroyed much more rapidly
Quantitative Elution of Morphine from Ion Exchange Resins
CECIL H. VAN ETTEN
Northern Utilization Research Branch, Agricultural Research Service,
In studying the use of ion exchange resins in determina-
tion of morphine, the compound was not always com-
pletely eluted from the resin. Consequently, a study
was made of the effects of degree of cross linkage of the
resin and of the pII and ionic strength of the elutriant
on the elution of morphine from strong cation and
anion resins. Conditions were found w-hich gave
quantitative elution from resins of both types. Be-
havior of other ampholytes and bases has been ex-
amined under the same conditions of elution. In-
formation presented is of value in determining condi-
tions under which these resins may be used to give
quantitative separation of morphine prior to anal?sis.
B EC.4C:SE of the ampholytic properties of morphine, its sepa-
ration from acids, baqes, and neutral molecules should he
possible by successive exchange from strong cation and anion
exchange resins. Separation of codeine from morphine based on
this principle has been reported ( I , 6). LIorphine n as exchanged
on an anion resin column. whereas codeine passed through because
it contained no functional group that acted as an anion a t high
pH. ?inion exchange repins have also been applied to the libera-
tion of morphine and similar bases from their salts, permitting the
subsequent titration of the free base in the effluent ( 3 . 9, l O , l / t ) .
I n this paper results are reported n-hich show the effect of degree
of cross linkage of the exchange resin on the elution of morphine,
ANALYTICAL CHEMISTRY
by dissolved oxygen than are the vicinal dithiol compleues, but we
cannot infer that other nonvicinal dithiols 15 o d d behave simi-
larly. One aspect of the interference of 1.3-dimercapto-2-pro-
panol in the test for 2,3-dimercapto-l-propanolthat must be con-
sidered in future applications is the possibility that with other
dithiol combinations the formation of mixed complexes might
limit the range of usefulness of the test.
I n the present analyses it n a s possible to minimize the inter-
ference of the nonvicinal dithiol component of a mixture by tak-
ing advantage of its oiidizability. K i t h other mixtures, dif-
ferent approaches might prove more suitable. For euample,
solvent conditions could be varied, differences in absorption
spectra could be euploited. or other heavy metals could be ap-
plied. Conductometric titration with heavy metal acetates in
pyridine also offers hitherto unexplored possibilities.
LITERATURE CITED
Aldridge, W. S . ,Biochem. J . , 42, 52 (1945).
Barron, E. S. G., Rliller, 2 . B., and Kalnitsky, G., Ibid., 41,
62 (1947).
Hughes, H. K., and coworkers, ASAL.CHEY.,24, 1349 (1952).
Jaffe, E., Ann. chzm. (Rome),41, 397 (1951).
Mellon, AI. G., "dnalytical Absorption Spectroscopy," p. 45,
Wiley, Kew York, 1950.
hliles, L. W. C., and Owen, L. N., J . Chem. Soc., 1950, 2943.
Rosenblatt. D. H.. and Jean. G. S . .J . Phus. C h m . . in Dress.
Sjoberg, B., Ber., 7 5 , 13 (1942).
Spray, G. H., Biochem. J . , 41, 360 (1947).
Spray, G. H., Stocken, L. A., and Thompson, R. H. S., Ibid., 41,
362 (1947).
Stocken, L. A,, and Thompson, R. H. S., Physiol. Revs., 29, 168
(1949).
Welcher, F. J., "Organic Analytical Reagents," VoI. IV, p. 161,
Van Nostrand, Kew York, 1948.
lbid., pp. 117, 192.
RECEIVED
for review September 2, 1954. -4ccepted January 19, 1955.
U. S. Department of Agriculture, Peoria, Ill.
and the effect of p H and ionic concentration of the elutriant. Bn
objective of the experiments was to find conditions under which
morphine could be quantitatively removed from strong anion and
cation exchange resins using micro ion eschange columns and
samples of about 10 mg. The behavior of several other am-
pholytes and bases was examined under similar conditions of
exchange.
EXPERIMENTAL
Resins. Dowex 50 (a strong, sulfonic acid, cation exchange
resin) and Dowex 1 (a strong, quaternary ammonium anion
exchange resin) of different degrees of cross linkage were used in
this study. I n particular, Dowex 50 X 1, X 2. X 4, X 8, and
X ,l6 and Dowex 1 X 1, X 2, x 4, x 8, and X 10 were ex-
amined. Particle size of each resin ranged from 50 to 100 mesh.
Xumbers following the X indicate the percentage of divinyl-
benzene used in their preparation. With larger amounts of
divinylbenzene, the resins become more highly cross-linked, swell
less in water, and show less volume changes with variation of
solvent concentration.
Exchange resins were conditioned by heating 5 bo 30 grams on
a steam bath with excess 1 S sodium hydroxide. decanting, wash-
ing with distilled water, and then heating with excess 1.Y hy-
drochloric acid. This cycle ivas repeated until no color was ob-
served in the supernatant. Decanting removed a port'ion of the
smaller resin particles which did not settle out. The final stock
resin was stored in dii;t,illed water in eit,her the hydrogen or
chloride form.
Procedure. Columns 8 min. in diameter, similar in design to
those previously described ( l e ) ,xvere filled to a depth of 4 cm.
with 0.2 to 0.6 gram of exchange resin, the xveight depending on
V O L U M E 2 7 , N O . 6, J U N E 1 9 5 5
the density of the resin. Resins thus were compared on a volume
basis. Because of greater density of the more highly cross-linked
resins, the volume of those used had a greater total exchange
capacity. This does not affect the conclusions reached from the
study, because a large excess of elutriant and resin n a s used in
every case. Except as noted below, cation resins were used in
the hydrogen form. anion resins were converted to the hydrox-
ide form by passing 10 ml. of lh' sodium hydroxide through the
column, followed by a wash n ith distilled water t o remove excess
alkali.
AA
A The difference between the first titration of the loosely held hy-
20
1
A .
I
1 . 1 .AJ
1% 2%
4% 8% 16%
EXCHANGE RESIN CROSS LINKAGE
Figure 1. Elution of morphine from strong cation
exchange resins of different degrees of cross linkage
0 NHaOH elutriant
A NaOH elutriant
I incomplete elution was obtained even a t high normalities. Ap-
1% 2%
' 4% 8% IO%
EXCHANGE RESIN CROSS LINKAGE
Figure 2. Elution of morphine from strong anion
exchange resins of different degrees of cross linkage
0 Acetic acid elutriant
A HCI elutriant
Ten milliliters of solution containing morphine sulfate or the
other compounds examined, equivalent in amount t o 10 mg. of
free base, were pipetted on these columns. Morphine sulfate
solutions Tyere prepared from U.S.P. XIV morphine sulfate,
which gave moisture, carbon, and hydrogen values that agreed
with those calculated from the formula for the pentahydrate.
Complete exchange of morphine from the original solution was
confirmed by testing the effluent. I n case of anion resins, sulfate
ions were exchanged from the original solution in addition to the
morphinate iom. After morphine was exchanged on the column,
it was washed n-ith 5 to 10 ml. of water. The column then was
eluted with 50 ml. of elutriant. Under gravity flow, the average
elution time was about 30 minutes. Elutriant passed through the
columns of low-cross-linked resins more slo\vly than through the
columns of higher-cross-linked resins.
Eluate Tyas collected in a volumetric flash and morphine was de-
termined in suitable aliquots by a modification of the colorimetric
method reported by Adamson and Handisyde ( 3 ) . A test of
the accuracy and precision of this method on 10 consecutive
samples of 10.0 mg. of morphine showed an average recovery of
10.05 mg. with a standard deviation of 0.22.
Sarcotine, codeine, and thiamine concentrations were deter-
mined from their ultraviolet absorption in acid solution measured
with a C a r r spectrophotometer. Xarcotine was measured a t
3120 A., codeine a t 2840 A , , and thiamine at 2460 A. Narcotine
was exchanged on resins from 50% acetone solution; i t was neces-
95s
sary, however, to remove the acetone before making ultraviolet
absorption measurements.
Lysine \vas measured by direct titration. The ammonium hy-
droxide eluate from the cation column on which lysine had been
eachanged n a s evaporated to dryness on a steam bath. The
residue of lysine, as the free base, was dissolved in distilled water
and titrated to the monohydrochloride with 0.01S hydrochloric
acid using methyl red as a n indicator. The pH of an aqueous
solution of lysine monohydrochloride was found to be between 4
and 5 , which is the pH a t which methyl red changes color. The
end point n a s sharp. I n the case of the anion column, the hydro-
chloric acid eluate was evaporated to dryness, and the residue,
which contained less than two equivalents of hydrochloric acid
per molecule of lysine, was titrated to the methyl red end point
n i t h 0.01L\7sodium hydroxide. This titration measured the hy-
drochloric acid held by the weaker basic group of lysine, which,
because of its n e & nature, held only about 0.8 equivalent of
hydrochloric acid under the conditions of evaporation. The
solution then was passed through a Dowex 50-H column and the
effluent n-as titrated for the total hydrochloric acid present.
drochloric acid and the titration of the cation effluent gave a
quantitative measure of the lysine present.
RESULTS
Extent of elution of morphine from resins of varying degree of
cross linkage by different elutriant,s is shown for the cation resins
in Figure 1, and for the anion resins in Figure 2.
Complete elut,ion was obtained from the 1, 2, and 4y0 cross-
linked cation resins with either ammonium or sodium hydroxide,
but incomplete elution was obtained from the 8 and l6Y0 cross-
linked resins.
Conditions for complete elution from t,he anion resin were more
restricted. Only for the 1% cross linked rcsin with acetic acid
as the elutriant, n-:-:ts quantitative elution always obtained.
Figure 3 s h o w how readily sodium hydroxide and sodiuni
citrbonat,e removed the morphine. even a t loiv normalities, pre-
sumably because t,heir pH was above the isoelectric point of
morphine [pH 8.94 (II)]. \ W h mdiuni chloride as elutriant,
parently morphine has such a ptrong affinity for the cation ex-
change resin, as long as it has a positive charge, that it elutes
with difficulty.
60b
2
i
0
I I Ail
0 0.05 0.I 0.15'b0.5
I
NORMALITY OF ELUTAIANT
Figure 3. Elution of morphine from Dowex 50 X 1
Na by charge reversal and mass action of sodium
ion
Likewise, in the case of the anion resin (Figure A), acetic acid
&as a very good elutriant because it lowered the p H of the me-
dium below the isoelectric point of morphine. At this pH,
morphine was no longer in anionic form and was completely
eluted from the column. Results on the anion resin showed that
morphine was completely removed by sodium hydroxide, but n-as
not displaced by ammonium hydroxide even in concentrated
solutions. This map be attributed to the relatively higher con-
centration of hydroxyl ions in sodium hydroxide solution, which
was sufficient to displace morphine from the resin t5ithout its
charge reversal.
Table I shows results obtained with compounds of different
956 ANALYTICAL CHEMISTRY

molecular weight. Molecular weight had little effect on the ex-

tent of elution from resins of low degree of cross linkage, as is
shonn by the almost complete recovery of lysine, morphine,
codeine, and narcotine from Dowe\: 50 X 1 cation exchange resin.
I n contrast, recovery of these compounds from the higher cross-
linked resin, Dowex 50 X 16, decreased regularly as the molecular
weight of the compounds increased. Similar results also were
obtained for lysine and morphine with the anion exchange resins
Dowex 1 X 1 and X 8. For both the anion and cation exchange 93‘ I I I
resins, lysine showed the same recovery regardless of degree of 0.1 0.2 0.30.4 0.5
cross linkage, which indicates it was sufficiently small in size to NORMALITY OF ELUTRIANT
reach all exchange sites. Figure 5. Recovery of morphine
from Dowex 1 X 1 OH with strong
and weak acid
100
ap
d
W
80
the influelit solution, and contracted the resin to a greater extent.
5
w
60 Kressman (12) and Kunin and l I ~ . e r s( 1 3 ) have investigated the
exchange of ions of different molecular weights by eschange resins
w 40
z
r
of different degrees of cross linkage. In their studies on exchange
n 20 they found that molecular size affected the equilibrium eschange
K and the rate of attainment of equilibrium. They, as well as
0
I Deuel. Solms, and .lnyas-Weisz ( 5 ) , shoived it was possible to
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 separate molecules of the same charge, but different size, by the
NORMALITY OF ELUTRIANT “screening” action of the appropriate ion eschange resin. How-
Figure 4. Elution of morphine from Dowex 1 X 1 ever. they did not determine conditions under which the exchange
OH by charge reversal and mass action of hydroxyl ion could be completely recovered.
ion Don-es 1 X 1 OH contracted more vihen elut,ed with hydro-
chloric acid than when eluted n.ith acetic acid. Gregor, Gutoff,
and Bregman ( ? ) have shown that the volume of an exchange
Table I. Elution of Organic Ions of Different Molecular resin contracts as the ionic concentrat,ion of t,he solution in which
Sizes it is placed increases, and such volume changes of the resin are
Molec- more marked as the cross linkage decreases. Such behavior may
ular Dowex % be t,he explanation for the incomplete elution of morphine from
Compound Weight Resin Elutriant Eluted
Dowes 1 X 1. using hydrochloric arid (Figure 5 ) . If the mor-
Lysine 146 Dowex 50 X 1 0 5 3 NHaOH 89
50 X 16 0 5N NHiOH 90 phine molecules are only slightmi!-smaller than the openings in the
1 X 1 0 05.V HC1 100
1X 8 0 05,V HCl 95 exchangr rpsin, any contraction of t,he resin will make their com-
31orphine 285 50 X I 0 5J-NHIOH 100 plete elution difficult. Further evidence supporting this ex-
50 X 16 0 5 sNHIOH 5Y planation is shown in Table I. Oiily 10% of the codeine was re-
1 X 1 0 3.”HCI 94
1 X 8 0 5.VHC1 83 covered from Dowex 50 x 16. using highly ionized sodium .
Codeine 299 50 X 1 0 5.” NHhOH 98 i hydroxide, but 44% \vas displaced with ammonium hydroxide as
$50 X
50 X
1
16
0 05.V XaOH
0 5,Y KHnOH
100
44 elutriant .
50 X 16 0 05,V NaOH 10 Lrsine n-as incompletely eluted from t,he cation exchange
Narcotine 413 50 X I 0 5 s XHIOH 98 resin3. Since about 90% recovery vias obtained from both the
.iO X 16 0 ~YXHIOH 30
1 and 16% cross-linked resin, it appears that incomplete eh1-
Thiamine 263 30 X 2 0 j.V KHaOH 110
50 X 2 3.” HCl <I tion was not caused by excessive cross linkage of the resin,
but by some other factor. Incomplete elution of basic amino
acids from Dowes 50 a t high pH has been observed by Moore
and Stein (15).
DISCUSSIOY
Poor recovery of thiamine was surprising. It’s incomplet’e
Results presented in Figures 1 and 2 shoiy that, as the degree of elution by hydrochloric acid from a phenol-formaldehyde-sul-
cross linkage of the exchange resin increased above a certain fonic acid-type eschange resin n-as reported by Herr (81, using
minimum (4% with Dowex 50 and 1% with Dowex l), morphine acid concent,rations up to 37%. Molecular adsorption, rather
was not completely eluted. illthough fewer resins were in- than ionic, on the exchange resin may be the explanation.
vestigated, with compounds other than morphine, data of Table Davies and Thomas (4),using cation eschange resins and am-
I indicate that elution of codeine and narcotine also \vas in- matic and aliphatic acids, showed that molecular adsorption
complete from resins having more than a certain degree of cross occurs. They found that adsorption increased with molecular
linkage. As extent of swelling and, hence, molecular-pore size tyeight arid was greater with compounds containing an aromatic
in a resin decrease with increasing cross linkage, incomplete elu- group.
tion mag result from trapping of large ions on exchange sites. This study has shon-n that n-it,h appropriat,e selection of ion
For a given resin and elutriant. larger ions would be expected to exchange resins and conditions of elution, morphine ca’n be
be less completely eluted. This is borne out by the regular de- quantitatively exchanged from cat,ion and anion exchange resins.
crease in recovery with increase in molecular size in the series- These results, as well as those obtained with several other organic
lysine, morphine, codeine, and narcotine-on elution from Don ex ions, indicate that the following factors are critical in det,ermining
50 X 16. Attributing irreversible adsorption to trapping in pores conditions for complete elution of organic ions from ion exchange
of the resin implies that certain pores, or regions of the resin avail- resins: degree of cross linkage of the resin and the size of the ion,
able for exchange, are contracted by the elutriant so that ex- p H of t.he elutriant, especially in case of ampholytes, and effect
changed ions can no longer diffuse out. I n the present experi- of the ionic concentration of the elutriant on volume changes of
ments the ionic strength of the elutriant was higher than that of the eschange resin. On the basis of the incomplete recover?’ of
V O L U M E 2 7 , N O . 6, J U N E 1955 957
lysine and the very poor recovery of thiamine under all condi- (6) Grant, E. W., and Hilty, W.W., J . A m . Pharm. Assoc., 42, 150
tions examined, it may be concluded that other factors influence (1953).
(7) Gregor, H. P., Gutoff, F., and Bregman, J. I., J . Colloid Sei., 6 ,
the completeness of elution of some compounds. 245 (1951).
(8) Herr, D. S . , I n d . Eng. CRen?., 37, 631 (1945).
ACKNOWLEDGMENT (9) .Jindra, A., J. Pharm. Phar?nacoZ., 1, 87 (1949).
(10) Jindra, A , , and Pohorsky, J.,I h i d . , 2,361 (1950).
The author thanks Clara E. McGrew for technical assistance (11) Kolthoff, J. hl., Biochcm. Z., 162, 338 (1925).
and Bettye L. Wilson for spectrophotometric assays. (12) Kressman, T. R. E., J . Phys. Cliem., 56, 118 (1952).
(13) Kunin, It., and Myers, R. J., DZ:scussio?~sFaraday SOC., 7, 114
(1949).
LITERATURE CITED (14) Levi, L., and Farmilo, C. G., Can. J . Chem., 30, 793 (1952).
(15) Aloore, S.. and Stein, W. H., J . Riol. Chem., 192, 663 (1951).
(1) .ichor, L. B.,and Geiling. E. 11. K., ANIL. CHEM.,26, 1061 (16) VanEtten, C. H., and Wiele, 11. B., ANAL.CHEM.,25, 1109
(1954). (1953).
( 2 ) Adamson, D . C. AI., and Handisyde, F. P., Andust, 70, 305
(1945). RECEIVEDf o r review November 86, l Y X . Accepted February 26, 195.5.
(3) Baggesgaard. 11. H.. Fuchs. D., and Lundberg, L., J . Pharnz. Presented a t the 16th Midwest Regional Meeting of the AIIERICANCHEMI-
Pharmacol., 4, 566 (1952). CIL SOCIETY, Omaha, Xeb., November 4 and .5, 1954. Mention of firm
(4) Davies, C . W., and Thomas. G. G., J . C'henz. SOC., 1951, 2624. names or commercial products under a proprietary name or names of their
( 5 ) Denel, H.. Solnis. J . . and Anyas-Weiss, L., Hell'. Chim. Acta, 33, manufacturer does not constitute an endorsement of siich firins or products
2171 (1950). by the U. S. Department of Agriculture.

Test for Establishing Residual Safe Life of Stabilized Solid Propellants

CARL BOYARS and W. G. GOUGH
Research and Development Department, U. S. Naval Powder Factory, Indian Head, Md.

Accelerated decomposition of propellants subsequent to
depletion of stabilizer results i n extensive deterioration
and possible spontaneous ignition. Chemical analysis
is n o t a practical method of measuring t h e loss in
stabilizing power because of t h e stepwise formation
during storage of niany nitrated stabilizer derivatives.
.i test has been deiisetl which does measure t h e stabi-
lizer loss i n terms of t h e proportion of original safe life
remaining. I t has been evaluated with propellants sub-
jected to varying amounts of high-temperature storage.
This stability test involves heating t h e volatile-free
propellant a t constant temperature i n a constant-
volume system containing an oxygen atmosphere. The
time required to reach a predetermined positit e pres-
sure correlates well with residual safe life as measured
by t h e length of preliminary storage of the propellant
a t high temperature. Data are presented comparing
the variation of pressure with time i n tests carried o u t
under atmospheres of nitrogen, air, and oxygen. IIy-
potheses about the niechanism of degradation of pro-
pellants, based on this work, are offered.
complicated by the separations :inti interpretations required.
A good stability test which gives a precise measure of degree of
deterioration is desirahle, and this requirement, together with
the need for more inforniation about the phenomena of degrada-
tion of propellants, inspired this investigation. The stability
test devised by Taliani (6) and modified by Kiggam and .Good-
year ( 8 ) differentiates bet\\-ecn ,stable and unstable propellants
by observing the increase of pressure \\-ith time in a constant-
temperature, constant-volume system containing 'the propellant
and air. However, this test provides no means of differentiating
h e t w e n the various degrees of residual stability.
VACUUM PUMP AND MANOMETER
NITROGEN SUPPLY
II F
HELIX II E

N EARLY all solid propellants based on nitrate esters contain

a uniformly distributed stabilizer. The function of the F
stabilizer is the removal of nitrogen dioxide, which is formed as
a primary decomposition product during storage of thc, propel- E
lant ( 2 ) and which would othern-ise autocatalytically accelerate
the decomposition. Accelerated decomposition subsequent to
depletion of the stabilizer results in spontaneous ignition if heat
-
H E A T I N G BLOCK
BALL JOINT
1
k
is generated by the exot,hermic reaction more rapidly than it can
be dissipated. Even where ignition does not occur, extensive
deterioration of the propellant follow stabilizer depletion.
I t is important for an organization that uses formulations con-
taining nitrate esters and stabilizer to know how much of the
stabilizing power incorporated in manufacture remains after MERCURY
RESERVOIR
storage for any length of time. The problem is complicated by
the fact that the common stabilizers, which are secondary aro-
matic amines or urea derivatives containing AV-aromaticlinkages, MERCURY
go stepwise to their higher nitrated forms, each of the less com- LEYELING
DEVICE
pletely substituted nitro compounds retaining some stabilizing
power (1, 4, 5'). An analytical technique would thewfort. he Figure 1. Diagram of Taliani test equipment