HPLC
Analysis of Garlic Clove Homogenates
(continued)
Results
The method employed provided optimum
separation of thiosulphinates with a short analysis
time and avoided peak-broadening of late peaks.
The HPLC contours indicate ten peaks (in agreement
with Naganathan
although the separation of methyl 2-propenethiosulphinate
and (Z)-1-propenyl methanethiosulphinate is inadequate
principally due to the trace amounts of (Z)-1-propenyl
methanethiosulphinate present in the sample -
see Figure 1.
Each
sample clone was the subject of three separate
extractions. Individual peak analysis is given
in Appendix 1 and HPLC
contours are shown in Appendix 2.
In his study of Allium extracts, Naganathan
prepared synthetic thiosulphinate standards and
his published data has been used in this study
for tentative identification (Fig 2).
Further identification was afforded by comparison
of UV spectra with published
data (Figure 2) and examples are shown in
Appendix 3.
Data
Analysis
The coefficient
of variation for total thiosulphinate averaged
3.1% for all eighteen analyses and this level
of accuracy is reflected in the values for the
major components - 2-propenyl methanethiosulphinate
(4.2%) and diallyl thiosulphinate (3.4%) (Appendix
1). Where thiosulphinates were present in very
small amounts the coefficient of variation was
often higher (>10.0%) and this was most likely
due to the unstable nature of the extracted compounds
in organic solvents.
The results obtained from the HPLC analyses (Appendix
1) are summarised below.
Clone
Group
1-propenyl
allyl + allyl 1-propenyl
allicin (diallyl)
methyl
1-propenyl +1-propenyl methane
2-propenyl
methane + methyl 2-propenyl
dimethyl
Total
Area (equiv.)
Yugoslavian
1
10.32
53.98
2.72
31.35
1.63
815595
Israeli
1
12.22
53.14
2.78
30.43
1.43
648415
French Red
1
9.62
53.51
2.4
32.6
1.86
742744
Yerina
1
11.17
49.8
3.06
33.96
2
740736
German Red
1
12.3
47.08
3.68
32.21
4.74
728148
Dominics
1
12.14
64.55
0.98
21.57
0.76
719903
Roja
1
10.35
63.58
1.33
24.25
0.76
789610
Carpathian
1
8.28
61.95
0.91
27.49
1.38
604524
Music
3
1.24
47.85
0.6
42.5
7.8
974971
Georgia Fire
3
3.73
60.49
0.38
33.95
1.45
883232
Leningrad
3
2.91
57.67
0.97
33.79
4.66
1115679
Red Rezan
3
5.32
56.43
0.77
35.62
1.87
758803
Freudenberger
4
4.29
62.36
0.42
31.49
1.44
1073418
Morado
4
3.94
72.39
0.26
22.65
0.75
1030958
Red Janice
4
5.23
64.2
0.48
28.56
1.52
527125
California
Late
4
3.13
57.79
0.4
35.49
3.2
857919
Cristo
4
3.84
59.78
0.31
34.21
3.51
960630
Inchelium
4
2.2
53.8
1.12
36.42
5.28
514311
Summary of thiosulphinate
content of garlic clones (Concentration of thiosulphinates
in Mole % of total)
Single factor analysis of variance (ANOVA) of the
data (Table 3) indicates
that there are significant differences in the 1-propenyl
allyl/allyl 1-propenyl and 1-propenyl methyl/methyl
1-propenyl contents between isozyme groups.