Agricultural and Biological Chemistry

ISSN: 0002-1369 (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/tbbb19

Anti-inflammatory Active Constituents of Aloe

arborescens Miller

Masatoshi Yamamoto, Toshio Masui, Kiyoshi Sugiyama, Masami Yokota,

Kazuya Nakagomi & Hiroyuki Nakazawa

To cite this article: Masatoshi Yamamoto, Toshio Masui, Kiyoshi Sugiyama, Masami Yokota,
Kazuya Nakagomi & Hiroyuki Nakazawa (1991) Anti-inflammatory Active Constituents
of Aloe�arborescens Miller, Agricultural and Biological Chemistry, 55:6, 1627-1629, DOI:
10.1080/00021369.1991.10870794

To link to this article: https://doi.org/10.1080/00021369.1991.10870794

Published online: 08 Sep 2014.

Submit your article to this journal

Article views: 462

View related articles

Citing articles: 2 View citing articles

Full Terms & Conditions of access and use can be found at

https://www.tandfonline.com/action/journalInformation?journalCode=tbbb20
Agric. Bioi. Chem., 55 (6), 1627-1629, 1991 1627

Note constituents have been isolated from Aloe,3.4) but no one

has correlated the constituents to an anti-inflammatory
activity. We have previously reported 5 ) that the major
Anti-inflammatory Active Constituents constituents of Aloe, barbaloin and aloenin, showed
of Aloe arborescens Miller inhibitory effects on rat mast cell degranulation. This report
describes the inhibitory effects of the constituents of Aloe,
including barbaloin and aloenin, on carrageenan-induced
Masatoshi YAMAMOTO, Toshio MASUI, rat-paw inflammatory edema.
Kiyoshi SUGIYAMA,*l. Masami YOKOTA,*l The procedure for isolating the anti-inflammatory
Kazuya NAKAGOMI*2 constituents of Aloe is shown in Chart 1. The structures of
and Hiroyuki NAKAZAWA*3 the isolated active constituents, aloenin, barbaloin,
aloe-emodin, J'l-sitosterol and a mixture of straight-chain
Shizuoka Prefectural Institute of Public Health higher alcohols (C 26 , 3.5%; C 28 , 57.7%; C 30 , 25.9%; C 32 ,
and Environmental Science, 4-27-2 Kita-ando, 8.8%), are shown in Fig.!. The anti-inflammatory activity
Shizuoka 420, Japan of fractions from Aloe and of the 5 isolated active
*lSchool of Pharmaceutical Science, constituents toward carrageenan-induced edema in rats is
University of Shizuoka, 395 Yada, Shizuoka 422, Japan summarized in Table I. The original MeOH extract (MeOH
*2 Fermentation Research Institute, 1-1-3 Higashi, ext.) showed significant inhibitory activity toward the
Tsukuba, Ibaraki 305, Japan edema. From partially purified fractions of the MeOH ext.,
*3 The National Institute of Public Health, the anti-inflammatory activity was detected mainly in the
4--0-1 Shirokanedai, Minato-ku, Tokyo 108, Japan n-BuOH extract (n-BuOH ext.). Frs. I and 2 from the
n-BuOH ext. exhibited similar activity, 45.9% and 58.2%,
Received July 16, 1990
respectively. Aloenin, barbaloin and aloe-emodin, which
were isolated from Fr. I, also significantly inhibited the
edema. Among the fractions separated from Fr. 2, only
Aloe arborescens Miller (Aloe; "Kidachi-aroe" in Fr. 2- I showed significant inhibitory activity toward the
Japanese) has been widely used as a folk remedy for edema. The constituents isolated from Fr. 2- I, higher
constipation, bites, burns, etc. in Japan. 1.2) Many alcohols and J'l-sitosterol, also inhibited the edema by

Aloe leaf (800g)

Iextracted with ~eOH

MeOH ext. (l18g)

0 dissolved in H,O
1 2) extracted ,,·ith a-BuOH

I
a-BuOH ext. (56.2g) H,O ext. (50.7g)

Ipartitioned with CHCl./~eOIl/H,O (19:19:12)

I
Fr. 1 (22.7g) Fr. 2 (28.1g)
Upper layer LO\ller layer

ILH-20 C.C. (70%MeOH) ISiOe C.C. (benzene/CHCL/MeOH) .

Fr. I-I (9.6g) Fr. 1-2 (6.8g) Fr. 1-3(3.2g) Fr. 2-1 (l1.2g) Fr, 2-2 (4.8g) Fr. 2-3 (lOg)
Kd'O.75-0.84 Kd'l.lZ-1.21 Kd-1.79-2.79 (2:3:0) (O:100:5) (0:0: 100)

I recryst. from MeOIl I recryst. from MeOH 1) SiO, C.C. (EtOAc)

2) recryst.
ISi0 2 C.C. (benzene/CHCj,=1: 1)
f - .- - - - - - - - - - - - - ,
1
aloenin (4.8g) barbaloin (l.Bg) from toluene
Fr. 2-1-1 (5.6g) Fr. 2-1-2 (4.8g)
aloe-emodin (0.8g)
1) SiO, C.C. (benzene/CHC]-=2:1) II active charcoal (EtOAc)
1 2) recryst. from benzene I 2) recryst. from EtOAc
higher alcohols (0.8g) f3 -sitosterol (2.4g)

Chart 1. Isolation Procedure for the Constituents with Anti-inflammatory Activity from Aloe arborescens
Miller by Monitoring Carrageenan-induced Edema in Rats.
The yield of each fraction obtained from 800 g of Aloe arborescens Miller is designated in parentheses.

Kd (distribution constant) = (elution volume~void volume)/internal volume

1628 M. YAMAMOTO et al.

HO We have disclosed for the first time that aloenin,

o barbaloin, aloe-emodin, p-sitosterol and a mixture of
straight-chain higher alcohols have an inhibitory action
on rat paw edema. Furthermore, we found that these
OCH, compounds play an important role in the anti-inflammatory
action of Aloe.
p·litosterol

~
~CH20H CH3 -(CH2 Jn- OH
Experimental

Aloe leaves. Aloe arborescens Miller was purchased from

GIe
(n~25. 27.29.31 ) Aloe Seiyaku Co., Ltd., Shizuoka, Japan.
Barbaloin
ltraight·chain higher alcohols

~
Measurements. 'H-NMR spectra were recorded at
90 MHz on a JEOL FX-90 NMR spectrometer with TMS

~~CH20H as an internal standard. Mass spectra were taken with a

Hitachi M-80A GC-MS mass spectrometer, and IR spectra
o
on a JASCO A-220 IR spectorophotometer. TLC was
!loe·mdin
performed on precoated Kieselgel 60 F 254 , and the spots
Fig. 1. Structures of 5 Isolated Constituents with were visualized by spraying with a mixture of ammonium
Anti-inflammatory Activity from Aloe arborescens Miller. molybdate, ceric sulfate and 10% H 2 S0 4 reagent. Column
chromatography was carried out with Sephadex LH-20
Table I. ANTI-INFLAMMATORY ACTIVITY OF THE (25-100 !lm, Pharmacia LKB Ltd.) and Wakogel C-200
FRACTIONS AND ISOLA TED CONSTITUENTS FROM (74-149 !lm, Wako Pure Chemical Industries Ltd., Osaka,
Aloe arborescens MILLER Japan).

Dose % Swelling" Inhibition" Assay for anti-iriflammatory activity. The carrageenan-

Test sample
(mg/kg/i.p.) (mean ± S.E.) (%) induced rat inflammatory edema described by Winter et
al. 6 ) was used in this experiment. Ten male Wistar strain
Control 53.6±3.0 rats weighing 130-150 g were used at each dose level, the
MeOH ext. 250 25.6±3.2* 52.2 samples tested being administered 30 min before the
n-BuOH ext. 150 21.5 ± 0.9*** 59.9 carrageenan treatment. A subplantar injection of 0.1 ml of
H 2 0 ext. 150 38.0±2.0** 29.1 1% carrangeenan (lambda carrangeenan type IV, Sigma
Fr. I 100 29.0± 5.4** 45.9 Chemical Co. Ltd.) in saline solution was given, the volume
Fr. 2 100 22.4 ± 3.4*** 58.2 of the foot was measured with a Ugo Basile plethysmom-
Fr. 2-1 50 30.1 ± 0.9*** 43.8 eter every I hr for 5 hr, and the percentage swelling (foot
Fr. 2-2 50 41.0±4.6 23.5 edema) was calculated. The swelling of the paw reached a
Fr. 2-3 50 51.1±6.1 4.7 peak 3 hr after injecting the carrageenan. The results are
-------------------------------------
expressed as the inhibition of swelling at 3 hr, relative to
Control 60.6±4.8
the control group given a saline solution. The significance
Aloenin 50 35.5 ± 5.3* 41.7
of difference between the means of the treated and control
Barbaloin 50 43.1 ±3.3* 28.8
Aloe-emodin groups in swelling was assessed by Student's t test.
100 36.4±4.4* 39.9
Higher alcohols 50 23.1 ± 3.7*** 61.9
P-Sitosterol 50 18.6±0.6** 69.3 Structural elucidation. The structures of aloenin,
barbaloin, aloe-emodin and p-sitosterol were each
Indomethacin 10 31.4± 1.0* 48.2 confirmed by comparing their MS, MNR and IR spectra
Aspirin 100 23.2±0.9** 61.7 with those of authentic samples. The higher alcohols were
acetylated with acetic anhydride and pyridine at 140°C for
a Significantly different from the control at *p < 0.05; 2hr, subjected to GC-MS in a column (3mm i.d. x 1m)
** p<0.02, and *** p<O.OOI. packed with 2% OV-I, and identified as a mixture of
b Inhibition 3 hr after injecting carrageenan. straight-chain higher alcohols by comparing their mass
spectra with those of authentic samples (obtained from
61.9% and 69.3%, respectively. The isolated active Gasukuro Kogyo Co., Ltd., Tokyo, Japan).
compounds showed almost the same inhibitory activity as
that of aspirin (Table I). On the basis of their yield and Acknowledgment. We are very grateful to Professor T.
activity, each of these 5 constituents is considered to ac- Hirata of Hiroshima University for kindly supplying the
count for the anti-inflammatory action of the MeOH ext. data for aloenin and for evaluating our spectral data for
 Anti-inflammatory Active Constituents of Aloe arborescens M.
aloenin. We thank Aloe Seiyaku Co., Ltd. for providing
authentic barbaloin and aloe-emodin.
References
1) T. Namba, "Coloured Illustrations of Wakan-
Yaku," Vol. II, Hoikusha Publishing Co., Ltd.,
Osaka, 1980, pp. 218-221.
2) S. Kameyama and K. Sugimoto, Fragrance J., 60,
101 (1983).
1629
3) K. Makino, A. Yagi and I. Nishioka, Chern. Pharrn.
Bull., 21, 149 (1973).
4) T. Hirata and T. Suga, Bull. Chern. Soc. Jpn., 51, 872
(1978).
5) K. Nakagomi, M. Yamamoto, H. Tanaka, N.
Tomizuka, T. Masui and H. Nakazawa, Agric. Bioi.
Chern., 51, 1723 (1987).
6) C. A. Winter, E. A. Risley and G. W. Nuss, Proc.
Soc. Exp. Med., 111, 544 (1962).