o 1615-

14 ~ ~3:~

12-

..a ua

10-

/~

p5-

,

,

1

2

, 3

,

,

,

,

,

4

5

6

7

8

LAYER

DEPTH

[rnm]

Fig. 2. Variation of the wavelength of the structure versus the shallow layer depth

I

12

I

Fig. 3. Solvent evaporation influence on pattern formation (mercury dithizonate in toluol). I Petri dish with solution, 2 large Petri dish, 3 light source (visible), 4 tall beaker, 5 air stream, a) Pattern formation, b, c) no pattern formation, d) pattern formation origin seems to be quite independent of the mechanisms involved in the

chemical reactions. According to Nicolis and Prigogine [7] the occurrence of spatial dissipative structures corresponds to the presence of non-linearities in the differential equations describing the phenomenon. The kinetics of the photochemical reactions we studied are quite simple, hence the nonlinearities are to be sought among physical causes. Solvent evaporation and diffusion would seem to play an important role here.

"V-"

Received November 2, 1982 1. M6ckel, P. : Naturwissenschaflen 64, 224 (1977) 2. Kagan, M., Levi, A., Avnir, D.: IX Symp. Photochem. (IUPAC), Pau 1982; Naturwissenschaften 69, 548 (1982) 3. Meriwether, L.S., Breitner, E.C., Sloan, C.L.: J. Am. Chem. Soc. 87, 4441 (1965) 4. White, D.M., Sonnenberg, J.: ibid. 88, 3825 (1966) 5. Guglielmetti, R., Meyer, R., Dupuy C. : J. Chem. Educ. 50, 413 (1973) 6. Brown, G.H. : Techniques of Chemistry, Vol. III. New York: Wiley-Interscience 1971 7. Nicolis G, Prigogine I. : Self Organisation in Non-Equilibrium Systems. New York: Wiley-Interscience 1977

T

o:5

lih

"

Fig. 1. Analytical enantiomer resolution of exo- and endo-brevicomin (Ii). Conditions: 42 m x 0.3 mm glass capillary column coated with IIIb (0.06 m in OV 101), oven temp. 52 ~ injection temp. 200 ~ carrier 0.8 bar He, split ratio 1:100, injection 0.1 gl Ila + lib diluted in CC14 and exo- and endo-brevicomin (IIa and IIb) on manganese(II)-bis(1R-3-heptafluorobutyrylcamphorate) (IIIb, of. Fig. 1). R, 0

I." R1 = R2 = Me, R3 = R4 = H lla: R 2 = Me, R 3 = Et, R 1 = R 4 = H lib: R 2 = M e , R 4 = E t , R I = R 3 = H

V. Schurig, R. Weber and G.J. Nicholson Institut ffir Organische Chemie der Universit/it, D-7400 Tiibingen A.C. Oehlschlager, H. Pierce, Jr. and A.M. Pierce Department of Chemistry, Simon Fraser University, Burnaby, British Columbia, Canada, V5A 1S6 J.H. Borden Department of Biological Science, Simon Fraser University, Burnaby, British Columbia, Canada, V5A 1S6 L.C. Ryker Department of Entomology, Oregon State University, Corvallis, Oregon, U.S.A.

92

exo

M.G. gratefully acknowledges a scientific and technical grant from the European Communities Committee.

Enantiomer Composition of Natural e x o - and endoBrevicomin by Complexation Gas Chromatography] Selected Ion Mass Spectrometry

Complexation gas chromatography [1] as an efficient tool for analytical enantiomer resolution is preferentially applied to racemates devoid of suitable chemical functionalities for diastereo-

endo

meric derivatization, e.g. pheromone acetals [2]. Thus, for the first time, frontalin (/) has now been quantitatively resolved on manganese(II)-bis(1R-3pentafluorobenzoylcamphorate) (IIIa)

As the methodology neither requires substrate derivatization nor purification and, when compared to chiral shift N M R spectroscopy [3], exhibits high sensitivity (1-ng sample) and high precision (_+0.5% e.e. over the entire range of 0-99% e.e.) we attempted the direct screening of the enantiomeric composition (e.e.) of natural exo- and endo-brevicomin (II). Exo- and endo-brevicomin (II) are produced by males of Dryocetes confusus and by males or females of three species of Dendroctonus beetles, among them the mountain pine beetle D. ponderosae, the major insect pest in Western

Naturwissenschaften 70 (1983)

9 Springer-Verlag 1983

Table 1. Enantiomeric composition of exo- and endo-brevicomin (IIa +IIb) determined by complexation GC on IIIb (experimental conditions see legend of Fig. 2) Volatiles from emerged males of

( + )-exo-brevicomin (lla) b

(+)-endo-brevicomin (IIb) b

Mode a % e.e.

Mode a % e.e.

Dryocetes confusus A, B (extract prepurified by prep. GC) D. ponderosae A, B (Brit. Columbia) (extract prepurified by prep. GC) D. ponderosae (Oregon) B (steam-distilled abdomens)

ratio exo-/endobrevicomin ~

98.3 _+0.6

A, B

63.1 __+1.0

19-17/1

98.3 -+ 0.6

A, B

64.8 + 1.0

14-10/1

97.2_+0.6

B

74.7_+ 1.0

18-16/1

a A: digital integration of gas chromatographic peak eluates, B: weighing of the peak area of the selected ion (m/e = 98 and 85) mass spectrum b Assignment of sign of optical rotation by coinjection &synthetic (+)-brevicomins [6] c As determined by analysis on a 28 m x 0.66 mm glass capillary column coated with SP-1000 North America. Response of D. ponderosae to racemic exo-brevicomin (IIa) varies from rejection to attraction depending on pheromone release rate and population [4]. This observation underlines the r61e of enantioselectivity in pheromone perception. The e.e. of natural IIa and lib (cf. Table 1) was determined on a 42 m x 0.3 m m glass capillary column coated with IIIb dissolved in OV 101. The identity of the eluted peaks as brevicorain configuration isomers [5] was confirmed by GC-MS. Enhancement of sensitivity and simplification of analysis of complex mixtures by using selected ion MS as specific detector [7] permitted the determination of e.e. of H in an unpurified extract of biological material (entry 3, Table 1, Fig. 2). Agreement ofe.e, obtained by G C with F I D detector [graphical peak evalua-

I Ion current

[i00

tion and digital peak integration (Spectra Physics Minigrator)] and by selected ion MS ( m / e = 8 5 and m / e = 98) as detector (Varian M A T 112S, entry 1-2, Table 1) was better than 1%. The results of Table 1 reveal differences of e.e. for Ila and lib between different locations of D. ponderosae. The incomplete enantioselectivity of pheromone production which is specially pronounced for Ilb raises questions as to the biosynthetic origin and its evolutionary implication and to the mechanisms of pheromone perception. The concise knowledge of configurational composition of natural brevicorain (I/) available by complexation G C - M S m a y aid biological pest management. Supported by the Fonds der chemischen Industrie and the Deutsche Forschungsgemeinschaft, and by the Natural Sciences and Engineering Research Council of Canada, grants A0851 & A 3881, the Science Council of British Columbia, and the National Science Foundation, U.S.A. Received November 18, 1982 1. Schurig, V.: Chromatographia 13, 263 (1980); Schurig, V., Bfirkle, W.: J. Am. Chem. Soc. (in press) 2. Weber, R., Hintzer, K., Schurig, V. : Naturwissenschaften 67, 453 (1980); Weber, R., Schurig, V. : ibid. 68, 330 (1981) 3. Stewart, T.E., et al. : J. Chem. Ecol. 3, 27 (1977) 4. Ryker, L.C., Rudinsky, J.A. : ibid. 8, 701 (1982); Conn, J.E., Borden, J.H.: unpublished 5. Silverstein, R.M. : J. Chem. Educ. 45, 794 (1968) 6. Oehlschlager, A.C., Johnston, B.D.: J. Org. Chem. (in press) 7. Frank, H., Nicholson, G.J., Bayer, E. : J. Chromatogr. I46, 197 (1978)

{103 (min~

Fig. 2. Mass fragmentogram of natural endo-brevicomin enantiomers (lib). Conditions: 42 m x 0.3 mm glass capillary column coated with IIIb (0.08 m in OV 101), oven temp. 48 ~ injector temp. 100 ~ carrier 0.6 bar He, interfaced by open-split to a Varian MAT 112S MS used as selective detector for m/e = 98 (ionizing voltage 70 eV, interface temp. 170 ~ ion source temp. 200 ~ resolution 1000). Splitless injection of steam-distilled abdominal volatiles of male D. ponderosae from Oregon (U.S.A.) diluted in n-pentane, injection volume 2 gl Naturwissenschaften 70 (1983)

Poly-ADP-Ribosylation is not Required for Excision of a DNA Damage H. Klocker, H.J. Burtscher, B. Auer, M. Hirsch-Kauffmann, and M. Schweiger Institut fiir Biochemie (nat. Fak.) der Universit~t, A-6020 Innsbruck Poly-ADP-ribosylation apparently is involved in D N A excision repair [1-3]. However, the step of excision repair which depends on conversion of N A D + into poly-ADP-ribose is not

9 Springer-Verlag 1983

known yet. Incision of D N A [2] as well as repair D N A synthesis [3] do not depend on the function of poly-(ADPribose)polymerase. We show in this communication that the exonuclease 93