Biochemical Systematics and Ecology 35 (2007) 454e455

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Alkaloids from the stem bark of Orophea hexandra (Annonaceae)

Hilwan Y. Teruna a,b, Peter G. Waterman a,*
a
Centre for Phytochemistry and Pharmacology, Southern Cross University, PO Box 157, Lismore, NSW 2480, Australia
b
Department of Chemistry, Faculty of Mathematics and Natural Science, University of Riau, Kampus Bina Widya,
Pekanbaru 28293, Indonesia
Received 10 October 2006; accepted 6 December 2006

Keywords: Orophea hexandra; Annonaceae; Alkaloids; Noraporphine; Aristolactam; Morphinandienone; Chemical taxonomy

1. Subject and source

Orophea Blume is a genus of small trees and shrubs distributed from India to Australia and possibly into Africa
(Kessler, 1993). O. hexandra Blume is found in Indonesia (Backer and van den Brink, 1963) and for this study
was collected from Bukit Tigapuluh National Park, Riau Province, Sumatra in September 2001. A voucher specimen
(SA-02) has been deposited at the Herbarium Bogoriense, Bogor, Indonesia.

2. Previous work

In a previous report (Sinz et al., 1999) the leaves of this species collected in Java yielded the following alkaloids:
aporphines (isoboldine, N-methyllaurotetanine), noraporphines (asimilobine, N-nornuciferine), 1-benzyltetrahydroi-
soquinolines (reticuline) and proaporphines (N-methylcrotonosine, pronuciferine).

3. Present study

The dried and powdered stem bark (600 g) was extracted with MeOH to give 13.71 g crude extract. An aliquot of
this (4 g) was sonicated with 1 M HCl for 1 h and filtered. The acid fraction was basified with ammonia and extracted
into dichloromethane to give 91 mg alkaloids. This mixture was separated by semi-preparative HPLC using an RP C-
18 column (5 mm, 10 mm  250 mm, flow rate 2 ml min1) and gradient elution (20% B over 20 min isocratic, then
20e95% B over 15 min, then 5 min isocratic) between A (H2O, 0.05% TFA) and B (ACN, 0.05% TFA). Three pure
alkaloids were isolated: 1 (24.8 mg, tR 8.0 min), 2 (11.2 mg, tR 10.0 min), and 3 (6.0 mg, tR 23.0 min). A minor band
at tR 13.0 min contained 4 mixed with 2. These were separated by further HPLC using a semi-preparative RP C-18
column with the same size and the same elution regime and flow rate to give 4 (4.0 mg).

* Corresponding author. Tel.: þ61 2 6622 3211; fax: þ61 2 6622 3459.
E-mail address: pwaterma@scu.edu.au (P.G. Waterman).

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doi:10.1016/j.bse.2006.12.007
 H.Y. Teruna, P.G. Waterman / Biochemical Systematics and Ecology 35 (2007) 454e455 455

The aqueous residue still contained large amounts of 3 which was separated by preparative HPLC (5 mm,
22 mm  150 mm column, flow rate 25 ml min1) eluting with 20% B isocratic for 2 min followed by a gradient
to 60% B over 15 min to give 3 (28.8 mg, tR 11.5 min).
Compounds 1e4 were characterized as follows.

3.1. Laurolitsine (2,9-dihydroxy-1,10-dimethoxyaporphine) (1)

Mþ 313, C18H19NO4. 1H and 13C NMR in agreement with previously reported data (Guinaudeau et al., 1975, 1983)
and confirmed by HMBC and NOESY studies.

3.2. Norpredicentrine (2-hydroxy-1,9,10-trimethoxyaporphine) (2)

Mþ 327, C19H21NO4. 1H and 13C NMR in agreement with previously reported data (Hocquemiller et al., 1983) and
confirmed by HMBC and NOESY studies.

3.3. Aristolactam AII (2-hydroxy-1-methoxyaristolactam) (3)

Mþ 265, C16H11NO3. 1H and 13C NMR in agreement with previously reported data (Preistap, 1985) and confirmed
by HMBC and NOESY studies.

3.4. Sebiferine (N-methyl-2,3,6-trimethoxymorphinandien-7-one) (4)

Mþ 341, C20H23NO4. 1H and 13C NMR in agreement with previously reported data (Wu et al., 1992) and confirmed
by HMBC and NOESY studies.

4. Chemotaxonomic significance

The occurrence of aporphine alkaloids in this species is typical of the Annonaceae and whilst rarer both morphi-
nanes and aristolactams are not unexpected (Cavé et al., 1987). What is noteworthy is the apparent difference in
alkaloid chemistry compared to the previous study on this species. It is unclear as to whether this reflects the origin
of the material (Java versus Sumatra) or the plant part (leaf versus bark).

Acknowledgements

One of us (H.Y.T.) thanks ADSeAusAID for granting a scholarship and rangers of Bukit Tigapuluh National Park
for guidance in the field.

References

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