Vol. 52, No.

4, 1973 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

CARBON-13 NUCLEAR YiGNETIC RESONANCE SPECTRA

OF LIGNINS

H.-D.Liidemnnn
Lehrstuhl fiir Physik, Fachbereich Biologic, Universitgt
Regensburg, D-84 Regensburg, UniversitatsstraBe 31
and H.Nimz
Polymer-Institut der Universitat Karlsruhe,
D-75 Karlsruhe, Hertzstr.16
Germany
Received April 5, 1973
SUMMARY: From the 'SC-nmr spectra of a large number of
dimeric and monomeric lignin model compounds the chemical
shifts of the carbon atoms of the Ce-units in lignin with
different substitution patterns were determined. The
absorption peaks of the carbon-13 spectra of two lignins
(beech and spruce) could be assigned by comparison (Table 3).

After cellulose, lignin is the most abundant polymeric

natural product, consisting of p-hydroxy-phenyl propane
structural units. ' Its complicated structure results from
the various means through which the "Ca-units" are connected
with one another by carbon-carbon or by carbon-oxygen bonds,
giving a cross-linked three dimensional high polymera From
the isolation of about thirty dimeric and oligomeric lignin
degradation products it could be shown that at least ten
different kinds of bonds exist between the Ca-units in
lignin.3 Based on the yields of degradation products a
structural scheme of beech lignin was recently proposed.4
In this paper 13C-nmr spectroscopy was chosen as a physical
method of completing our chemical results on the consti-
tution of beech and spruce lignins. In comparison with
Vol. 52, No, 4,1973 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

'H-nmr spectroscopy of lignins, 5 'XC!-nmr offers convincing

advantages regarding a twenty fold wider chemical shift

range and line sharpening by proton decoupling.

MATERIALS AND METHODS

Milled wood lignins (MWL) of beech (Fagus silvatica) and

spruce (Picea excelsa) were prepared according to Bjijrkman.'
In order to get the MWLs free from carbohydrates, the lignin-
-carbohydrate-complexes were precipitated on alumina from
the solutions of the crude MWLs in dioxane/water (9:l) by
gradually adding discrete amounts of benzene to the solutions,
with agitation by means of a vibro mixer. -,Dimeric lignin
model compounds were prepared according to standard proce-
dures or obtained by lignin degradation.
The 'X-spectra were taken of 2oo/o W/W solutions of the
lignins in deuteroacetone/heavy water mixtures (9:l) (resp.
deuterodioxane/heavy water, 9:l) by means of a Varian XL-
-100-15 spectrometer operating at 25.2 MHz by the Fourier-
-transform-technique. The protons are noise decoupled. 100

blocks of 1000 transients were accumulated in the long-

-term-averaging-mode. The resolution of the spectra is
+3 Hz (approximately 0.2 ppm). The spectrometer was locked
to the -CD, groups of the acetone (resp. -CDs- of dioxane).
TMS was added to the solutions as an internal reference.
Shift values quoted are given in the TMS scale.

RESULTL AND DISCUSSION

Figs. 1 and 2 show the 13C-nmr spectra of spruce and

beech lignin, respectively. In order to assign the absorption
lines of the lignins, the '%-spectra of eighteen dimeric and

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12 3 L S-10 11-16l7-222232L 25-30 31-3435 36 @ 37 @

IL ,,,I,,, I I,,, I I, , I
200 150 100 50 0
-6K

Fig. 1: 'SC-nmr spectrum of beech lignin. (S) = aceton,

(R) = TMS.

seven monomeric lignin model compounds were taken for com-

parison. From these spectra the values for the chemical
shifts of carbon atoms with a ligninlike substitution pat-
tern, as tney exist in the Cg-units (I-III), were determined.
There are two main regions in the spectra, one covering the
aromatic (C-l to C-6) andolefinic (C-a and C-S) sp2 carbon
atoms ranging from 160 to 105 ppm, and the other covering
the saturated aliphatic sp3 carbon atoms of the propane side
chains (C-a to C-y) and the methoxy groups with chemical shifts
between 88 and 20 ppm downfield from TMS.

Y-F-
B-y-
a-C-
I: R' = R2 = H

II: R' = OCH,, R2 = H

OH III: R' = R2 = OCH3

The chemical shifts of the aromatic carbon atoms (C-l to

C-6) depend on the substituents in the aromatic ring, on the

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12 3 I s-10 n-i6i7---22 25-30 31-3C3536 w I

Fig. 2: 'SC-nmr spectrum of spruce lignin. (S) = aceton,

(R) = TMS;

C-a substituents and on the fact, whether the phenolic hydroxy

group is alkylated or not (Table 1).

From Table 1 it can be seen that the methoxy group in II

has no influence on the chemical shifts of both meta-positions
(C-l and C-5)) relative to I, but causes an upfield shift to
the ortho carbon atoms C-2 (-16 ppm) and C-4 (-11 ppm) and
to the para position (C-6, -8 ppm), while the absorption of
C-3 is shifted downfield (+32 ppm). Similarly the second
methoxy group in III causes upfield shifts at the carbon
atoms 6 (-15 ppm), 4 (-11 ppm), and 2 (-7 ppm), and a down-
field shift of +33 ppm at C-5, relative to II.7

Smaller effects at the ring carbon atoms C-l to C-6 are

observed when altering the substituents at C-a. When C-a
is the carbon atom of a carbonyl group instead of a HCOH-
or a HCOR-group in II, the chemical shifts of the carbon
atoms 2, 4, and 6 move downfield (+2 to +5 ppm) and that
of C-l upfield (-6 ppm). The same effects are observed, but
even smaller in value, when C-a is an olefinic or a tertiary

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Table 1: Chemical shifts and differences in chemical shifts

of the aromatic carbon atoms (C-l to C-6) of the Cg-units
(I-III) in ppm. For further details see text.

C-u in: a-HCOH(R) a-C=0 a-H&C a-HC-C C-4-OR

C9-unit I II III II III II III II III
C-l 134 134 134 -6 -4 -3 +3 +4
c-2 128 112 105 +2 +2 -1 +3 +I 0
c-3 116 148 149 +3 +I +I 0 0 +5
c-4 158 147 136 +5 +2 0 +3 +3
c-5 116 116 149 -2 +I 0 0 -1 +5
C-6 128 120 105 +5 +2 +2 +3 0 0

Table 2: Chemical shifts of the propylic side chain carbon

atoms (C-a to C-y) of the &-units (I-III) in ppm.

Structural ty-pe c-a C-6 c-y

Ar-YH-YH-CH20H (IV) 73 87 62
OH O-C-4'
Ar-CH-CH-CH20H 0) 65 64 75
&H k-1'
Ar-YH- YH-CH2011 01) 88 54 65
O-C-4' C-5'

Ar-CH- CH-YHp (VII) 87 55 72

&c-,, &+ O-C-a '
Ar-CII2-$-YHp (VIII) 40 47 73
C-6 ' o-c-y '
Ar-YH-YH-C02R (IX) 59 43 173
C-6' C-a'

Ar-yH-CH2-CO2R co 52 38 173
c-p '
Ar-CH=CH-C02H (XI > 146 116 170
Ar-CH-CH-CHO (XII > 154 132 195
Ar-CH=CH-CH20H (XIII) 131 127 63
Ar-CO-CH-CH20H (XIV) 195 83 64
b-C-4'

Ar-CHO on 192

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Table 3: Assignment of absorption lines in the lignin spectra of

Figs. 1 and 2.

Intensity +>
Assignment
beech spruce
(1) 195.2 w W a-C=0 in XIV, y-CHO in XII
(2) 192.7 ‘W W a-CHO in XV
(3) 171.8 m VW O-C=0 in aliphatic esters
(4) 162.0 VW VW C-4 in I (alkylated at C-4)
(5) 154.5 s W C-3 and C-5 in III (alkylated at C-4) C-a in XII
(6) 152.9 VW m C-4 in II with a-C=0
(7) 150.6 w S C-4 in II (alkylated), C-i; in II (a-C=O)
(8) 148.5 m S C-3 in II, C-3, C-5 in III, C-l in biphenyls
(9) 146.6 w In C-4 in II, C-a in XI
(10) 144.8 v-w m C-4' in VI
(11) 138.7 m VW C-4 and C-l in III (alkylated)
(12) 136.0 m m C-l in II (aikylated), C-4 in III
(13) 133.6 w m C-l in I-III
(14) 132.7 w VW C-B in XII
(15) 130.2 w W C-l with C-a in HC=C or HC-C (= C-l' in V)
(16) 129.0 w m C-2 and C-6 in I
(17) 126.7 7JW VW C-8 in XIII
(18) 120.1 m S C-6 in II
(79) 117.5 w m C-6' in VI
(20) 115.6 m S C-5 in II, C-3 and C-5 in I, C-8 in XI
(21) 113.8 VW W C-2 in II (with a-C=O, a-HC-C)
(22) 112.0 m S C-2 in II
(23) 107.1 m C-Z and C-6 in III (with a-C=O, a-HC-C)
(24) 105.1 vs C-2 and C-6 in III
(25) 88.1 VW W C-a in VI
(26) 86.8 s W C-p in IV, C-a in VII (syringaresinol-type)
(27) 85.8 w m C-a in VII (pinoresinol-type)
(28) 81-83 w W C-8 in XIV
(29) 75.0 w VW C-y in V
(30) 73.3 vs s C-a in IV, C-y in VII and VIII
(31) 63.8 m m C-a and C-8 in V, C-y in XIII and XIV
(32) 61.3 vs S C-y in IV
(33) 56.3 vs vs OCH,
(34) 54.5 w W C-8 in VI and VII
(35) 52.3 VW VW C-a in X
(36) 46.8 VW VW C-P in VIII
(37) 20.8 m acetoxy-CH3
+)Intensity: w=weak, m=medium,s=strong, vw=very weak, vs=very strong
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carbon atom (cf. Table 1). - Alkyiation of the phenclic

hydroxy group in II mainly causes a weak downfield shi.X of
about +3 ppm both for C-i+ and C-l, relative to II (cf. last
column in Table 1).

The chemical shifts of the side chain carbon atoms C-d,

C-R, and C-y, which depend on typical lignin substitution
patterns, are listed in Table 2.
From the data of Tables 1 and 2 a preliminary assignment
for 37 absorption lines in the two lignin spectra of Figs.
1 and 2 is proposed in Table 3.
The most characteristic difference between the two lignin
spectra is the very strong absorption peak at 105 ppm (24)
with a shoulder at 107 ppm (23) in the spectrum of beech
lignin (Fig. I), which is completely missing in the spectrum
of spruce lignin (Fig. 2). As this absorption peak is assigned
to C-2 and C-S 0-f syringyl units (III) in lignin (cf. Table 3),
the absence of such units in spruce lignin is clearly indica-
ted. Therefore j3C-nmr spectroscopy offers an outstanding
method for distinguishing between hardwood and softwood
iignins.
The rather weak but distinct peaks at 195.2 (1) and 192.7

ppm (2) indicate at least two different types of carbonyl

groups to be present in both lignins (cf. Table 3). The
absorption peak at 171.8 ppm (3) in the spectrum of beech
lignin, which is assigned to aliphatic ester carbonyl carbon
atoms, should be regarded with some caution, because it could
possibly result from polyuronic ester groups contaminating
the beech lignin. Similarly the absorption line at 20.8 ppm
(37) in beech lignin may be caused by substances other than
lignin. Further details of lignin '3C-nmr spectra are under
research and will be published later.
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REFERENCES

1. K.V.Sarkanen and C.H.Ludwig, "Lignins, Occurrence,

Formation, Structure,and Reactions", WiLey-Interscience,
New York - London - Sydney - Toronto 1977.
2. K.Preudenberg and A.C.Neish, "Constitution and Biosynthesis
of Lignin, Springer-Verlag, Berlin - Heidelberg - New York
1968.
3. H.Nim:z and K.Das, Chem. Ber. 104, 2359 (1971).
4. H.Nim,z, TapDi, in press.
5. C.H.Ludwig, B.J.Nist, and J.L.McCarthy, J.Amer. Chem. Sot.
86, 1186, 1196 (1964).
6. A.BjGrkman, Svensk Papperstidn. 59, 477 (1956).
7. For comparison see: P.C.Lauterbur, J.Amer. Chem. Sot.
3, 1846 (1961).

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