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Diseases caused by Mycobacterium avium (M. avium) and Mycobacterium tuberculosis (M.
tuberculosis) have reached pandemic proportions with some strains being resistant to existing
chemotherapies. Complex therapies requiring four to six drugs are sometimes required to prevent
the emergence of resistant strains. There is a need for the discovery of new drugs or compounds
that are potential drug templates that can be used to treat diseases caused by these bacteria. The
research reported in this paper describes the isolation of 6-, 8-, and 10-gingerol from fresh ginger
rhizome and the identification of 10-gingerol as the most active inhibitor of M. avium and M.
tuberculosis in vitro. The gingerols were isolated by fractionation of a crude methylene chloride
extract of fresh ginger rhizome by normal phase HPLC. Identification was based on mass spectral
data. The identification of 10-gingerol was confirmed by synthesis.
Keywords: Zingiber officinale; ginger; 6-gingerol; 8-gingerol; 10-gingerol; AIDS; Mycobacterium
avium; Mycobacterium tuberculosis
INTRODUCTION
Isolation of Gingerols
mobile phase A
mobile phase B
0
10
45
80%
80%
20%
20%
20%
80%
Hiserodt et al.
gingerol
retention time
(min)
area %
at 282 nm
(M + H)+
(amu)
base peak
(amu)
10-gingerol
8-gingerol
6-gingerol
9.8
10.5
11.6
15
11
72
351
323
295
333
305
277
87
85
95
65
0
58
0
95
0
0
13
0
73
48a
0
0
0
42
0
0
Outlier.
0
27
0
0
0
0
0
0
0
0
0
0
0
0
0
The crude residue isolated from the methylene chloride extract of fresh ginger rhizome was a clear golden
oil that was 0.55% of the fresh ginger weight. It had a
characteristic ginger aroma and consisted of a complex
mixture of components that were fractionated by normal
phase HPLC. Samples of the crude residue from the
initial methylene chloride extract and residues from
semicrude fractions 1-4 were analyzed to determine the
percent inhibition of M. avium and M. tuberculosis. At
100 g/mL the crude residue was inhibitory for both M.
avium (Table 1) and M. tuberculosis (Table 2) in vitro.
This activity was increased in semicrude fraction 2 and
provided the justification for further fractionating semicrude fraction 2 to purified 6-, 8-, and 10-gingerol. Since
the residues from semicrude fractions 1, 3, and 4 did
not show an increase in activity, nothing further was
done with these residues.
The residue from semicrude fraction 2, which was
0.20% of the fresh ginger weight, was analyzed by APcILC-MS and was found to consist of three major compounds. The area percent and LC-MS data are listed
in Table 3. APcI is a soft ionization technique which
generates protonated molecular ions in the positive ion
mode. The mass spectra for the three main components
in semicrude fraction 2 showed weak protonated molecular ions, at 5% relative intensity, and indicated the
molecular weights for the three components to be 350,
322, and 294 amu in the order of the retention time.
10-gingerol
8-gingerol
6-gingerol
10-gingerol
8-gingerol
6-gingerol
84
2
nd
6
86
nd
10
12
99
nd ) none detected.
Isolation of Gingerols
gingerol
yielda (ppm)
10-gingerol
8-gingerol
6-gingerol
120
93
880
To confirm the identity of 10-gingerol, it was synthesized using an Aldol reaction. The direct probe EI-mass
spectra for synthesized 10-gingerol (Figure 4) correlated
well with the direct probe EI-mass spectra for 10gingerol extracted from fresh ginger rhizome which
confirmed the identification.
CONCLUSION
Table 6. Minimum Inhibitory Concentration (MIC) for
M. avium and M. tuberculosis
MIC (g/mL)
gingerol
M. avium
M. tuberculosis
10-gingerol
8-gingerol
6-gingerol
25
50
>100
50
50
>100
10-gingerol to be the most active compound for inhibition of M. avium with a MIC of 25 g/mL. This
represents a significant increase in activity over the
crude residue which by definition was greater than 100
g/mL. 10-Gingerol was less active for inhibition of M.
tuberculosis with a MIC of 50-100 g/mL.
Direct probe EI-mass spectra were obtained for 6-, 8-,
and 10-gingerol which confirmed the APcI-LC-MS data.
The EI-mass spectrum for 10-gingerol is shown in
Figure 3. The ions at m/z ) 322 and 294 are due to the
presence of 8- and 6-gingerol as impurities in this
sample.
We acknowledge the Center for Advanced Food Technology (CAFT) mass spectrometry facility for providing
instrumentation support. CAFT is an initiative of the
Hiserodt et al.
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Received for review November 7, 1997. Revised manuscript
received May 6, 1998. Accepted May 7, 1998. We acknowledge
support from the National Institutes of Allergy and Infectious
Diseases, Intra-agency Agreement Y02-AI-50101-00.
JF970948L