Biological activities, chemistry and

identification naturally of occurring

Lignans and neolignans

Introduction
The lignans are a large group of natural polyphenolic
phenylpropanoid chemical compounds found in plants.
Lignans and neolignans are quite widespread in the plant
kingdom, and plants from the families Asteraceae,
Berberidaceae, Piperaceae, Magnoliaceae, Phytolaccacae,
Rutaceae Linaceae and Pinaceae are well known sources,
where they act as antioxidants and defense molecules
against pathogenic fungi and bacteria[1].

Fig. 1: Lignans precursors and basic structure.

Medicinal and General Importance of Lignans

Lignans are known to exhibit a rich structural diversity
and a varied biological activity. This growing interest in
bioactive lignans is motivated mainly by their potential use
as either phytopharmaceuticals or nutraceuticals[2].
In fact, among the several families of secondary
metabolites synthesized by plants, lignans have a wide
range of remarkable biological activities as Antiviral,
 Lignans and neolignans

Anticancer, Cancer prevention, Anti-inflammatory,

Antimicrobial, Antioxidant, Immunosuppressive,
Hepatoprotective, Osteoporosis prevention.
Lignans are one of the major classes of phytoestrogens,
which are estrogen-like chemicals that help in the reduction
in the occurrence of certain types of estrogen-related
tumors, such as breast cancer in postmenopausal women.
In addition, some lignans as: podophyllotoxin, obtained
from plants of the genus Podophyllum (Berberidaceae
family); have been used as chemotherapeutic agents.

Food sources
Plant lignans are co-passengers of dietary fiber, and
therefore fiber-rich food items are often good sources of
lignans. Flax seed and sesame seed contain higher levels of
lignans than most other foods. Other sources of lignans
include cereals (rye, wheat, oat, and barley being the richest
source), soybeans, cruciferous vegetables such as broccoli
and cabbage, and some fruit, particularly apricots and
strawberries[3].

The General Biosynthetic Pathway of Lignans

The pathway starts from hydroxycinnamic acids: p-
coumaric acid, sinapic acid, and ferulic acid which reduced to
corresponding alcohols, called monolignols, that is, p-
coumaryl alcohol, sinapyl alcohol, and coniferyl alcohol

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which are the main precursors of lignans. The lignans are

then obtained by dimerization or oilgomerization to form the
basic chemical structure that consists of two
phenylpropanoid units linked by a C-C bond between the
central atoms of the respective side chains[4].

Fig. 2: Biosynthetic Pathway of Lignans.

Classification of Lignans

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Lignans can be divided into several groups, depending on

other linkages and substitution patterns introduced into the
original hydroxycinnamyl alcohol dimer
Lignans in the strict sense are phenylpropanoid dimers
linked by a C-C bond between carbons 8 and 8' in the
side chain
Neolignans are dimers linked by other carbon atoms
as 8-1', 8-5', 8-O-4', 5-5' and others linkages
Sesquilignans and dilignans are higher oligomers of
lignans
Norlignans are phenolic compounds mainly occurring
with a diphenylpentane skeleton.

The lignans are classified into subgroups based on their

carbon skeleton, cyclization pattern, and the way in which
oxygen is incorporated in the molecule skeleton,
Dibenzocyclooctadiene derivatives, Arylnaphthalene and
aryltetralin derivatives, Dibenzylbutane, dibenzylbutyrolactol
and dibenzylbutyrolactone derivatives, Furofuran derivatives
and Furan derivatives[5].

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 Lignans and neolignans

Fig. 3: Different classes of lignans.

The neolignans also possess many different structures

and are divided into more than fifteen subgroups amongst
them, Benzofurans, Alkyl aryl ethers, Benzodioxanes,
Biphenyl neolignans and other neolignans. The lignans are
mainly present in nature as glycosylated derivatives[6].

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 Lignans and neolignans

Fig. 4: Different classes of neolignans.

Biological activities of some naturally occurring

lignans and neolignans

Classificat Biological
Examples Source
ion activity

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 Lignans and neolignans

Antifungal, anti-
inflammatory,
Sambucus
Furofuran antioxidant, and
williamsii
lignans hypoglycemic
Hance
agent.

(+)-pinoresinol

Furan
Phyllanthus Fungicidal
lignans
niruri L. activities

(+)-lariciresinol

Immunostimulato
ry activity,
Linum
Dibenzylbut antioxidant
usitatissimu
an lignans activity,
m L.
and estrogenic
activity

(-)-secoisolariciresinol

Aryltetralin Podophyllum
Antitumor
lignan peltatum L

(-)-podophyllotoxin

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Anti-platelet
Benzofuran Zanthoxylum
aggregation
s simulans
activity
neolignans Hance

()-simulanol

Magnolia
Biphenyl officinalis Anti-
neolignans Rehder & inflammatory
E.H.Wilson

houpulins G

Miliusa
Benzodioxa Anti-herpetic and
fragrans
nes cytotoxic
Chaowasku
neolignans activities
& Kessler

s (+)-eusiderin A

Spectroscopic Methods for the Structure Elucidation

of Lignans and neolignans
The analytical methods used for the determination of
structures of new natural phenolics includes:

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1. UV-visible spectroscopy that used for the quantitative,

qualitative analysis of phenolic compounds.
2. The Infrared Spectra of Complex Molecules provides a
clear idea about the functions groups present in the
compound.
3. Mass spectrometry was considered as vital for
structural information of molecular fragments and
molecular formulae hence became the essential part of
analysis to be reported in the publications.
4. Optical rotatory dispersion (ORD) and later circular
dichroism (CD) emerged as a tool to determine the
absolute configuration at about the same time as mass
spectrometry.
5. X-Ray crystallography also called as the most direct
method, has advanced through many phases of
development and has been in use for decades as the
only viable method for complex molecules.
6. NMR that have been became the most significant
single tool available to identification of compounds of
high molecular weight, of increasing complexity, low
abundance and low stability[7,8]. The NMR techniques
used for the structure elucidation included 1H and 13
C,
COSY, HSQC, and HMBC NMR experiments[9,10].

Example for identification of furofuran type

neolignans:

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 Lignans and neolignans

UV (MeOH) max 230.4, 280.0 nm, ESIMS m/z 381 [M+Na]

+
[11].
Table 1: 1H NMR (CDCl3, 500 MHz):
H-2, 2' 6.96 (2H, d, J = 1.5 Hz)
H-5, 5' 6.83 (2H, dd, J = 8.0, 1.5 Hz)
H-6, 6' 6.78 (2H, d, J = 8.0 Hz)
H-7, 9' 4.73 (2H, d, J = 4.5 Hz)
Hb-9, 7' 4.25 (2H, dd, J = 9.0, 6.5 Hz)
Ha-9, 9' 3.90 (2H, dd, overlapped)
OCH3-3, 3' 3.87 (6H, s)
H-8, 8' 3.16 (2H, m)

Table 2: C NMR (CDCl3, 125 MHz):

13

C-3, 3' 147.7

C-4, 4' 145.9

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C-1, 1' 132.4

C-6, 6' 118.7
C-5, 5' 114.7
C-2, 2' 109.6
C-7, 9' 86.1
C-9, 7' 71.1
OCH3-3, 3' 55
C-8, 8' 54

Example for identification of benzofuran type

neolignans:
The 1
H NMR spectrum (Fig. 6) of the compound
showed the presence of five aromatic protons due to two
aromatic moieties. One of these moieties showed three
aromatic signals arranged as ABX system at H 6.96 (1H, d, J
= 1.6 Hz), 6.78 (1H, d, J = 8 Hz) and 6.83 (1H, dd, J = 8, 2
Hz) which assigned for H-2, H-5, H-6 with their carbons
resonate at c 110.52, 116.11 and 119.69 respectively,
indicating tri-substituted aromatic moiety. In addition, the
second moiety revealed the presence of two equivalent
protons resonate at H 6.75 (2H, s) with their corresponding
carbons (c 114.18; C-2, 118.06; C-6), showed that the
second aromatic moiety is tetra-substituted.
The C NMR spectrum (Fig. 8) indicated the presence
13

of seven quaternary aromatic carbon atoms, four of which

have an oxygen substituent as judged from their chemical

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shifts at C 149.05, 147.46, 145.14 and 147.42 assigned for

C-3, C-4, C-3 and C-4 respectively.
Moreover, spectral data (Table 1) revealed also the
presence of signals at H 5.49 (1H, d, J = 6 Hz) and 3.47 (1H,
m) assignable to the methine protons H-7 and H-8,
respectively with C 88.96 (C-7) and 55.42 (C-8) of the
dihydrofuran ring fused to a coniferyl alcohol moiety to form
the benzofuran skeleton[12,13]. In addition the signals at H
2.68 (2H, t, J = 7.6 Hz) and 1.91 (2H, m) assignable to the
side chain protons H-7 and H-8, respectively with C 32.95
(C-7) and 32.92 (C-8).
The presence of anomeric proton at H 4.22 (1H, d, J =
7.6 Hz) with the corresponding carbon at C 104.46 together
with the sugar protons at H (3.21-3.86) indicates the
presence of -glucopyranosyl moiety.
The position and mode of the glycosidic linkage were
shown to be 9-O- from the coupling constant (J = 7.6 Hz) of
the anomeric proton, together with the downfield shift of the
signal C-9 (C 69.91), of the coniferyl alcohol side chain
revealed that the glycosidation was at this location. This was
confirmed by HMBC correlations (Fig. 5 &10) between the
anomeric proton and C-9 and C-8.

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Fig. 5: Important HMBC correlations of the compound

The 13
C NMR spectrum also showed the presence of two
carbons at C 56.78 and 56.37 assigned to two methoxy
groups at C-3 and C-3 respectively. This suggestion was
confirmed from the 1H NMR data by presence of two proton
signals each one of three singlet protons at H 3.85 (3H, s)
and 3.82 (3H, s) which further confirmed by HMBC indicate
their correlation with C-3 and C-3 respectively. The absolute
stereochemistry of C-7 and C-8 were shown to be S and R,
respectively, which established from the similarity of the
available NMR data to the previously published one[14].
The complete analysis of the remaining 1H, 13
C NMR and
2D-NMR experiments, together with comparing the above
mentioned spectral data with those reported in the
literature[14], revealed that the compound is
dehyrodiconiferyl alcohol-9-O--D-glucopyranoside
(Glochidioboside) as shown by the structure below.

(7S,8R)-dihydro-dehyrodiconiferyl alcohol-9-O--D-
glucopyranoside (Glochidioboside)
Table 3: 1H (400 MHz) and 13
C NMR data (100 MHz) in CD3OD ( in
ppm, J in Hz).

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No. C H No. C H
134.7 147.4
1 4
9 2
110.5 136.8
2 6.96 (1H, d, J = 1.6 Hz) 5
2 5
149.0 118.0
3 6 6.75 (1H, s)
5 6
147.4
4 7 32.95 2.68 (2H, t, J = 7.6 Hz)
6
116.1
5 6.78 (1H, d, J = 8 Hz) 8 32.92 1.91 (2H, m)
1
119.6 6.83 (1H,dd, J = 8 and
6 9 69.91 3.53 (1H, m)
9 1.6Hz)
3-
7 88.96 5.49 (1H, d, J = 6 Hz) 56.37 3.82 (3H, s)
OCH3
104.4
8 55.42 3.47 (1H, m) 1 4.26 (1H, d, J = 7.6 Hz)
6
3.77 (1H, dd, J = 12,
9 64.97 7.6 Hz) 2 75.15 3.21*
3.87*
3-
56.78 3.85 (3H, s) 3 78.1 3.37*
OCH3
129.8
1 4 71.64 3.29*
1
114.1
2 6.75 (1H, s) 5 77.88 3.31*
8
3.68(1H, dd, J = 12, 2.4
145.1
3 6 62.74 Hz)
4
3.86*
*Signal patterns unclear due to overlapping.

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 Lignans and neolignans

Fig. 6: 1H NMR spectrum of the compound (CD3OD, 400 MHz).

Fig. 7: 1H-1H COSY spectrum of the compound (CD3OD, 400 MHz).

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 Lignans and neolignans

Fig. 8: C NMR spectrum of the compound (CD 3OD, 100

13

MHz).

Fig. 9: HSQC spectrum of the compound (CD3OD, 400 MHz).

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 Lignans and neolignans

Fig. 10: HMBC spectrum of the compound (CD3OD, 400 MHz).

CONCLUSION
Lignans and neolignans are quite widespread in the plant
kingdom
Lignans have a wide range of remarkable biological
activities
Lignans are co-passengers of dietary fiber, and therefore
fiber-rich food items are often good sources of lignans.
The lignans are then obtained biosynthetically by
dimerization or oilgomerization of hydroxycinnamyl
alcohols
Lignans can be divided into several groups lignans,
neolignans, norlignans, Sesquilignans and dilignans

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 Lignans and neolignans

Spectroscopic Methods for the Structure Elucidation of

Lignans and neolignans includes UV-visible spectroscopy,
The Infrared Spectra, Mass spectrometry, Optical rotatory
dispersion (ORD) and later circular dichroism (CD), X-Ray
crystallography and NMR that have been became the
most significant single tool available to identification of
compounds of high molecular weight, of increasing
complexity, low abundance and low stability.

References
1. Rahman, A.-u.; Choudhary, M.I. Applications of nmr spectroscopy.
Volume 2. Applications of NMR Spectroscopy (ISSN 2405-4682 2015,
2.
2. Calvo-Flores, F.G.; Dobado, J.A. Lignin and lignans as renewable
raw materials: Chemistry, technology and applications. John Wiley &
Sons: 2015.
3. Thompson, L.U.; Boucher, B.A.; Liu, Z.; Cotterchio, M.; Kreiger, N.
Phytoestrogen content of foods consumed in canada, including
isoflavones, lignans, and coumestan. Nutrition and cancer 2006, 54,
184-201.
4. Dewick, P.M. Medicinal natural products: A biosynthetic approach.
John Wiley & Sons: 2002.
5. Wink, M. Annual plant reviews, biochemistry of plant secondary
metabolism. John Wiley & Sons: 2011; Vol. 40.

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 Lignans and neolignans

6. Teponno, R.B.; Kusari, S.; Spiteller, M. Recent advances in research

on lignans and neolignans. Natural product reports 2016, 33, 1044-
1092.
7. Whiting, D.A. Natural phenolic compounds 19002000: A bird's eye
view of a century's chemistry. Natural product reports 2001, 18, 583-
606.
8. Garbisch Jr, E.W. Applications of nmr spectroscopy in organic
chemistry. Journal of the American Chemical Society 1965, 87, 3033-
3033.
9. Chang, C.-C.; Lien, Y.-C.; Liu, K.C.C.; Lee, S.-S. Lignans from
phyllanthus urinaria. Phytochemistry 2003, 63, 825-833.
10. Carrier, D.J.; van Beek, T.A.; van der Heijden, R.; Verpoorte, R.
Distribution of ginkgolides and terpenoid biosynthetic activity in
ginkgo biloba. Phytochemistry 1998, 48, 89-92.
11. Park, T.W.; Lee, C.; Lee, J.W.; Jang, H.; Jin, Q.; Lee, M.K.; Hwang,
B.Y. Chemical constituents from buddleja officinalis and their
inhibitory effects on nitric oxide production. Natural Product
Sciences 2016, 22, 129-133.
12. Saracolu, .; varel, m.; ali, .; dnmez, a.a. Neolignan, flavonoid,
phenylethanoid and iridoid glycosides from phlomis integrifolia.
Turkish Journal of Chemistry 2003, 27, 739-748.
13. Pereira, C.; Jnior, B.; Bomfim, C.; Kuster, R.M.; Simas, N.K.;
Sakuragui, C.M.; Porzel, A.; Wessjohann, L. Flavonoids and a
neolignan glucoside from guarea macrophylla (meliaceae). Qumica
Nova 2012, 35, 1123-1126.

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14. Takeda, Y.; Mima, C.; Masuda, T.; Hirata, E.; Takushi, A.; Otsuka, H.
Glochidioboside, a glucoside of (7s, 8r)-dihydrodehydrodiconiferyl
alcohol from leaves of glochidion obovatum. Phytochemistry 1998,
49, 2137-2139.

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