1874 CANADIAN JOURNAL O F CHEMISTRY. VOL.

43, 1965

1. J. F. KINGand T. DURST. J . Am. Chem. Soc. 86, 287 (1964).

2. J. F. KING,P. DE MAYO,E. MORKVED, A. B. M. A. SATTAR, and A. STOESSL. Can. J. Chem. 41,100
\---.-,.
11 462)
3. N. S. HAMand A. N. HAMBLY. Australian J . Chem. 6, 135 (1953).
4. N. S. HAMand A. N. HAMBLY. Australian J . Chem. 6, 33 (1953).
5. A. SIMON,H. KRIEGSMANN, and H. DUTZ. Chem. Ber. 89, 1990 (1956).
6. G. GEISELERand I<. 0. BINDERNAGEL.Z. Elecktrochem. 63, 1140 (1959).
7. N. S. HAM,A. N. HAMBLY, and R. H. LABY. Australian J. Chem. 13, 443 (1960).
8. G. MALEWSKI and H.-J. WEIGMANN.Spectrochim. Acta, 18, 725 (1962).
9. R. W. TAFT,E. PRICE,I. R. FOX,I. C. LEWIS,I<. K. ANDERSEN, and G. T. RITCHIE. J. Am. Chem.
SOC.85, 709 (1963).
10. R. J . GILLESPIEand E. A. ROBINSON. Can. J. Chem. 39, 2171 (1961).
11. A. LocHERand H. E. FIERZ. Helv. Chim. Acta, 10, 642 (1927). A. LOCHER.Doctoral Thesis, Swiss
Can. J. Chem. Downloaded from www.nrcresearchpress.com by 186.155.238.196 on 08/03/17

Federal Institute of Technology, Zurich, Switzerland. 1925.

12. K. A. FREEMAN and C. D. RITCHIE. J. ASSOC.Offic. Agr. Chemists, 40, 1108 (1957).
13. F. ULLMANN and P. KERTESZ. Ber. 52, 545 (1919).
14. L. R. BLAINE,E. I<. PLYLER,and W. S. BENEDICT. J. Res. Natl. Bur. Std. A, 66, 223 (1962).
15. F. G. BORDWELL, C. M. SUTER,J . M. HOLBERT, and C. S. RONDESTVEDT.J. Am. Chem. Soc. 68, 139
(1946).
16. S. SMILESand T. P. HILDITCH. J. Chem. Soc. 91, 519 (1907).
17. W. DAVIESand J. H. DICK. J. Chem. Soc. 2104 (1931).
18. W. DAVIESand J. H. DICK. J. Chem. Soc. 483 (1932).
19. R. L. HINMAN . ,
and L. LOCATELL.TR. T. Am. Chem. Soc. 81. 5655 (19591.
20. H. MEYERand K. SCHLEGL. ~ & a t s G .34, 561 (1913).
21. C. ZIEGLERand J . M. SPRAGUE. J. Org. Chem. 16, 621 (1951).
22. E. B. EVANS,E. E. MABBOTT, and E. E. TURNER. I. Chem. Soc. 1159 (1927).
23. B. HELFERICH and H. GRUNERT. Ber. 74, 1531 (1941).
24. D. A. MACHOUL. Ber. 13. 692 (18801.
. ,
26. R. OTTOand H. OSTROP. Ann. 141, 365 (1867).
27. R. OTTOand 0. VON GRUBER. Ann. 142, 92 (1867).
For personal use only.

28. F. C. WHITMORE and N. THURMAN.J. Am. Chem. Soc. 45, 1068 (1923).

GLUCOPUTRANJIVIN

Long known as a synthetic product (I), isopropyl isothiocyanate was first isolated
from natural sources by Puntambekar (2) who obtained it1 from seeds of the Indian tree
Putranjiva roxbz~rghii Wall. (Euphorbiaceae). Puntambekar named the (assumed)
glucosidic precursor of this mustard oil glucoputranjivan (2).
Although- P . roxbz~rghii
- still remains the only well-established example of the occur-
rence of mustard oils in the family E u p h o r b i a ~ e a e isopropyl
,~ isothiocyanate was sub-
sequently found to be quite widely distributed (5), particularly within the Cruciferae,
and in 1959 Kjaer succeeded in isolating glucoputranjivin from seeds of Lunaria annua
L.3and characterized it as the crystalline tetraacetate (6).
'Together with (+)-2-butyl isothiocyanate and a third mustard oil originally identified ( 2 ) as phenyl isothio-
cyanate, but later shown by Kjaer and Friis ( 3 ) to be (S)-2-methylbtrtyl isothiocyanate.
2The family Euphorbiaceae i s normally regarded as taxonomically remote from the order Rkoeadales, which
contains the families Cruciferae and Capparidaceae. (Mustard oil glzrcosides are widely distributed amongst the
species of these two families.) However, Hutchinson (4) regards the Euphorbiaceae as a heterogeneous family,
probably derivedfrom several stocks, including the Bixales, which he also regards as the ancestral stock from which
the Capparidaceae were derived; i.e. the occurreizce in the Euphorbiaceae of species containing mustard oil
glucosides i s not to be entirely unexpected.
3Synonym, L. biennis Moench.
Canadian Journal of Chemistry. Volume 43 (1965)
 NOTES 1875

T h e now classic studies of Ettlinger and Lundeen (7,8) established the general structure
of the mustard oil glucosides and the mechanism by which they are enzymatically de-
composed t o mustard oils; thus glucoputranjivin is assigned the structure (V, R = H).
We have confirmed this structure by synthesis, using the same approach as that which
we developed for the synthesis of glucotropaeolin (9) and subsequently successfully
applied t o the synthesis of a number of other mustard oil glucosides (10).
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Isobutyraldoxiille (I) was chlorinated; the isobutyrohydroxarnoy1 chloride (11) was

converted t o the nitrile oxide (111) and condensed, i n situ, with tetra-0-acetyl-/3-D-
glucopyranosyl mercaptan t o give tetraacetylproglucoputranjivin (IV). Sulfonation of
For personal use only.

this with pyridine -sulfur trioxide, followed by isolation of the product as the potassium
salt, gave tetraacetylglucoputranjivin (V, R = COCH3), identical with a sample of
natural origin. We deacetylated this material, but although the product, glucoputranjivin
(V, R = H ) was chro~natographicallyhomogeneous it refused to crystallize a s either the
potassium or tetramethylammonium salt.4
Our original and main interest in the mustard oil glucosides was (and remains) in their
biogenesis. Inspection of the structures of the glucosides, general structure (VIII),
reveals that in many cases the aglycone skeleton is that of con~monlyencountered
a-amino acids,5 from which the carboxyl groups have been lost. Other aglycones cor-
respond t o unknown a-amino acids,'j implying either that these amino acids remain to
be discovered or that alternative biosynthetic pathways exist, which do not necessarily
involve a-amino acids.
NH2 NOSOI-
I //

\
S-8-D-glucopyranosyl
(VI) (VII) (VIII)
T o date, experiments have revealed that some of the nlustard oil glucosides are indeed
derived from a-amino acids7 Glucotropaeolin (VI I I, R = PhCH2) is derived fro111
"Tlte
- . tetramethylanzntonium
. salts of the mustard oil glucosides often crystallize better than the potassiunt salts
(7, 8,IUc).
5For e.va?nple glucocapparin ( V I I I , R = C H I ) and alanine ( V I , R = C H I ) , glzrcocochlearin ( V I I I ,
R = C H % C H 2 C H ( C H 3 ) ) ,and isoleucine ( V I , R = C H 3 C H 2 C H ( C H 3 ) )with the same conjiguration at the
asyntnzetric center i n the side chain ( 1 1 ) , and several other examples zn addztion to those cited in the text (6).
GForexantple glucohirs2rtin ( V I I I , R = C H ~ S O ( C H Z )and ~ ) a series of sulfides, szrlfoxides, and sulfones of
closely related st~uctzire( 5 ) .
? I n additiorz, glziconastzrrtiin ( V I I I , R = C G H ~ C H ~ Ci sHpartly
~ ) derived from phenylalanine ( I S ) . However,
the best Precursor yet discovered for sinigrin ( V I I I , R = CH2=CHCH2-) - there was sonte
i s acetate although
conflict ;?L the reszi?ts reported b j the twogroups ( I S , 17).
 18'76 CANADIAN JOURNAL O F CHEMISTRY. VOL. 43. 1965

phenylalanine (VI, R = PhCH2) (12, 13, 14), glucobrassicin (VIII, R = P-indolylmethyl)

from tryptophan (VI, R = P-indolylmethyl) (15) and sinalbin (VIII, R = p-HOCsH4CH2)
(16). Interestingly, there appear t o be two pathways from a-amino acids, one in which
the nitrogen of the a-amino acid is lost (16), biosynthesis proceeding via the keto acid
(VIII), and the other in which the nitrogen of the a-amino acid is incorporated directly
into the aglycone (14).8
In terms of the "a-amino acid hypothesis", glucoputranjivin can be visualized as
derived from valine (VI, R = (CH3)zCH) (5, 19).
T. peregrinumgplants fed ~ ~ - v a l i n e - 4 -were
l ~ C worked up and the mustard oil aglycones
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isolated as the thiourea derivatives. Paper chromatography revealed that a spot identical
with isopropylthiourea contained the bulk of the radioactivity, corresponding to 0.127,
incorporation of activity into the mustard oil. This initial result strongly suggests that
glucoputranjivin is in fact biosynthesized in T. peregrinum from valine.1°

EXPERIMENTAL
Tetraacetylproglucoputranjivin ( I V )
Isobutyrohydroxamoyl chloride, freshly prepared from isobutyraldoxime (2.0 g) (20) and dissolved in
ice-cold ether (50 ml), was added t o a solution of 2,3,4,6-tetra-0-acetyl-8-D-glucopyranosyl inercaptan
(1.0 g) in cold, dry ether (100 ml) and was followed by triethylamine (2.8 ml) dissolved in dry ether
(10 ml). Triethylamine hydrochloride precipitated and the blue color of the solution was discharged. After
30 min the reaction mixture was washed with ca. 0.6 N dilute aqueous sulfuric acid (50 ml); the ether
extract was separated, dried, and evaporated t o yield a colorless oil which crystallized froin concentrated
ethereal solution upon addition of petroleum ether. Recrystallization from ether - petroleum ether gave
For personal use only.

colorless needles of tetraacetylproglucoputranjivin, m.p. 93-95" (0.75 g, 60%). Further recrystallization

raised the m.p. t o 98-100°. [ a ] ~ ~ ~ - 2(c,
9 " 3 in CHC13). Anal. Calcd. for C I ~ H ~ ~ N O I O
C,S48.10;
; H , 6.06;
N, 3.12. Found: C, 48.31, H , 6.09; N, 3.29.
Tetraacetylglucoputranjivin ( V , R = CHaCO)
Tetracetylproglucoputranjivin (0.37 g) was dissolved in dry pyridine (5 ml), reshly prepared sulfur
trioxide - pyridine complex (0.42 g) added and the reaction mixture stirred a t room temperature overnight,
with exclusion of moisture. T h e reaction mixture was poured into water (ca. 20 ml) containing potassium
bicarbonate (0.75 g). After effervescence ceased the aqueous solution was extracted with ether (2 X 20 ml),
the combined ether extracts washed with water (2 X 20 ml), and the combined aqueous extracts freeze
dried. T h e residual solids were extracted with boiling 95% aqueous ethanol (v/v) and filtered hot. On
cooling the filtrate deposited fine colorless needles, m.p. 180.5-182" (0.220 g, 4 7 7 3 , [C~]D"- 18.3' (c, 1.6, H20) ;
lit. (6), [Ct]DZ3-15.4' (c, 1.2, H20). T h e infrared spectrum in a I<Br disc was superimposable on t h a t of a n
authentic specimen of natural origin.
Glucoputranjivin ( V , R = H ) .
Tetraacetylglucoputranjivin (0.93 g) was dissolved in anhydrous methanol, previously saturated with
ammonia, and kept a t room temperature overnight. T h e solution was evaporated t o dryness under reduced
pressure t o give a gum which refused t o crystallize. T h e gum was talcen up in water and passed through a
Dowex-50 ion exchange column (in the tetramethylammonium form) and the aqueous eluates freeze dried.
Again, the gummy residue of tetramethylamn~oilium glucoputranjivin which was chromatographically
homogeneous (21) refused t o crystallize.
Biosynthesis of Glucopzitranjivin from DL- Vali~1e-4-'~C
T h e freshly cut stems of the above-ground portion of young T . peregrinz~mplants (27 g) were dipped into
a n aqueous solution of D~-valine-4-~~C(50 pcuries). After 50 h the uptake of radioactivity amounted t o
3 3 Fcuries T h e plants were removed, washed, homogenized in a Waring Blendor, and, after standing a t
awe have also performed experiments with doubly labelled phenylalanil~e-2-"C,W similar to those described by
Underhill and Chisholm ( I d ) , as well as with phenylpyruvic a ~ i d~ x i m e - k ' " C , ' ~ Nand
, also concluded that the
nitrogen of the phenylalanine is retained during incorporation into glucotropaeolin.
9Synonym T . canariense Hort., the canary bird vine. The seeds of this plant are reported to produce gluco-
coclrlearin in addition to glzicoputranjivin, bzit both compounds are only present in small amounts (18).
loWe had insuficient material to carry out the degradation of the thiourea necessary to establish this absolzitely.
Further, as yet, we have no evidence which permits discrimination between the two possibilities of the nitrogen atom
of the valine being lost or retained during incorporation into glucoputranjivin. Since the plant i s in the same genzis
as T . nzajus i n which glucotropaeolin i s biosynthesized from phenylalanine with retention of the nitrogen atom,
we favor the same process here. However, it i s quite possible that in the Cruciferae, e.g. L. biennis, the biosynthesis
might proceed by the alternative pathway via the keto acid ( V I I , R = (CH3)zCH).
 NOTES 1877
room temperature for 1 h, steam distilled into ethanolic ammonia. The distillate was kept a t room tempera-
ture overnight and then freeze dried. The residue (a very small amount of dark-brown solid) was taken up
in ethanol and examined by paper chromatography (22). No spot appeared on spraying with Grote's
reagent (23) but a radioactive spot could be detected with a Nuclear-Chicago Actigraph I1 chromatogram
scanner in the approximate position expected for isopropylthiourea. The chromatography was repeated with
the addition of authentic isopropylthiourea. T h e chromatogram was examined with the scanner prior t o
spraying and 95% of the activity was located in the position exactly corresponding to the spot which appeared
on spraying. The activity of this spot was further measured by cutting out the area on the paper chromato-
gram and, after a complete wet combustion (24), counting as carbon dioxide, using a Dynacon 6000 ionization
chamber system. The total activity of the isopropylthiourea was calculated t o be 0.04 pcuries, a n
incorporation of 0.12%.
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ACKNOWLEDGMENTS
We wish to thank the National Research Council for financial support of this research
and Professor Anders Kjaer for a n authentic sample of tetraacetylglucoputranjivan. We
are also grateful t o the Golden West Seed Co. of Calgary for a generous gift of T.
peregrinz~mseeds.
1. H. JAHN. Monatsh. 3, 165 (1882).
2. S. V. PUNTAMBEKAR. Proc. Indian Acad. Sci. Sect. A, 32, 114 (1950).
3. A. KJAERand P. FRIIS. Acta Chem. Scand. 16, 936 (1962).
4. J . HUTCHINSON.The families of flowering - plants.
. 2nd ed. Vol. I. Dicotyledons. Clarendon Press,
Oxford. 1959.
5, A. KJAER. Fortschr. Chem. Org. Naturstoffe, 18, 122 (1960).
6. A. KJAER. Acta Chem. Scand. 13, 851 (1959).
7. M. G. ETTLINGER and A. J. LUNDEEN. J. Am. Chem. Soc. 78, 4172 (1956).
8. M. G. ETTLINGER and A. J . LUNDEEN. J . Am. Chem. Soc. 79, 1764 (1957).
For personal use only.

9. M . H. BENN. Can. J . Chem. 41, 2837 (1963).

10. M. H. BENN. (a) Can. J. Chem. 42, 163 (1964); (b) J. Chem. Soc. 4072 (1964); (c) Can. J. Chem. 43,
----,.
- \(19fifil~
1
11. '4. KJAERand S. E. HANSEN. Acta Chem. Scand. 11, 898 (1957).
12. M. H. BENN. Chem. Ind. London, 1907 (1962).
13. E . W. UNDERHILL, M. D. CHISHOLM, and L. R. WETTER. Can. J. Biochem. Physiol. 40, 1505 (1962).
14. E. W. UNDERHILL and M. D. CHISHOLM. Biochem. Biophys. Res. Commun. 14, 425 (1964).
15. M. ICUTA~EK, 2. PROCHAZKA, and K. VERES. Nature, 194, 393 (1962).
16. H. KINDL. Monatsh. 95, 439 (1964).
17. M. MATSUOand M. YAMAZAKI.Chem. Pharm. Bull. Tokyo, 11, 545 (1963).
18. P. G. DELAVEAU and A. KIAER. Acta Chem. Scand. 17, 2562 (1963).
19. A. KJAER,J. CONTI,and 1.-LARSEN. Acta Chem. Scand. 7, 1276 (1953).
20. M. H. BENN. Can. 1. Chem. 42. 2393 (1964).
21. 0.-E. SCHULTZ and R. GMELIN. 'z. ~ a & r f o r s c h .8b. 151 (1953).
22. A. KJAERand K. RUBINSTEIN. Acta Chem. ~ c a n d . ' 7 ,528 (1953).
23. I. W. GROTE.J. Biol. Chem. 93, 25 (1931).
24. D. D. VANSLYKE,J. PLAZIN,and J. WEISIGER. J. Biol. Chem. 191, 299 (1951).

THE HEAT OF FORMATION OF BORON TRIBROMIDE- ACETONITRILE

Laubengayer and Sears (1) were the first to suggest that BF3 is not the strongest
electron-pair acceptor in the boron trihalide series. They found that heats of forination
of the solid adducts from their gaseous components were 33.8 and 26.5 lical mole-I for
CHBCN.BC13 and CHsCN.BF3, respectively, in good agreement with their heats of
Canadian Journal of Chemistry. Volume 43 (1965)
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