Академический Документы
Профессиональный Документы
Культура Документы
FROM MESQUITE
by
SHOU-JEN R. CHEN, B . S .
A THESIS
IN
CHEMISTRY
Submitted to the Graduate Faculty
of Texas Tech University in
Partial Fulfillment of
the Requirements for
the Degree of
MASTER OF SCIENCE
Approved
)Jd>, l?'?'^
ACKNOWLEDGMENTS
Dr. John N. Marx for the helpful suggestions and advice during the
research period, and to Dr. John A. Anderson for serving as member
of my committee and spending time in analyzing and evaluating the
thesis.
I want to extend my thanks to Mr. Alan Croft for helping me
correct the manuscript.
11
TABLE OF CONTENTS
Page
ACKNOWLEDGMENTS
ii
LIST OF TABLES
vi
LIST OF FIGURES
vii
CHAPTER
I. GENERAL INTRODUCTION
CHAPTER II.
Background
EXPERIMENTAL
General Methods
Infrared Spectroscopy
Ultraviolet Spectroscopy
Column Chromatography
Gas Chromatography
10
n
Page
Column Chromatography
12
Preliminary Separation
12
13
Final Separation
13
15
Decomposition of Compound 1_
15
18
19
Formation of Trimethylsilyl
CHAPTER III.
Ether Derivative
19
20
20
22
22
Heartwood
22
Seasonal Changes
26
27
28
30
35
Decomposition of Compound 1_
35
39
IV
40
Page
CHAPTER IV.
Silylation of Compound 1_
40
Esterification of Compound 1^
42
44
45
52
Summary
52
54
LIST OF REFERENCES
APPENDIX A
B
55
HPLC Chromatograms
56
75
Infrared Spectra.
89
Ultraviolet Spectra,
96
LIST OF TABLES
Page
1. Extraction of Honey Mesquite
2.
3.
14
4.
23
5.
29
6.
7.
46
47
8.
VI
36
LIST OF FIGURES
Page
1.
11
2.
Flow Scheme A
16
3.
Flow Scheme B
17
4.
Flow Scheme C
18
5.
33
6.
49
7.
8.
51
9.
3,3;4;7,8-Pentahydroxyflavan
53
57
10.
11.
12.
50
58
59
60
14.
61
15.
62
16.
63
17.
64
18.
65
19.
66
20.
67
13.
21.
68
Page
HPLC-13, Compound 1^ after Irradiation with the
Light Source for 48 Hours
69
70
24.
71
25.
72
73
22.
23.
26.
27.
74
28.
76
29.
77
30.
78
31.
79
32.
80
33.
81
34.
35.
82
83
84
85
86
87
88
36.
37.
38.
39.
40.
viii
Page
41.
90
42.
91
43.
92
44.
93
45.
94
46.
47.
95
97
48.
98
99
49.
50.
100
51.
101
52.
102
53.
103
IX
CHAPTER
GENERAL INTRODUCTION
Background
If economically valuable
products can be obtained from mesquite, its removal would be considerably more attractive.
An almost totally unexplored area of potential mesquite utilization involves the isolation and use of the organic compounds, other
than carbohydrates, that are present in the mesquite plant.
Although
The extrac-
In this paper,
Table 1.
Fnom Heartwood
Water
6.0%
5.8%
Benzene-ethanol
4.4%
12.2%
Benzene-ethanol-water
10.4%
18.0%
1% NaOH
20.5%
28.9%
Extraction Solvent
animal feed, the Chemical Engineering Department at Texas Tech University investigated a number of thermochemical pretreatments with the
goal of improving the digestibility of harvested mesquite.
These
bility may allow treated mesquite wood to be used as a roughage substitute in animal feed.
Isolated reports of mesquite wood extraction using various
solvents demonstrate that appreciable amounts of organic compounds
2 3
may be removed. *
prepared a report
(edited by Joseph L.
4
concerning organic compounds which can be extracted from mesquite
(Prosopis juliflora).
A few references dealing with the isolation of certain compounds
or classes of compounds such as tannins, waxes, and flavonoids from
specific mesquite plant parts or unspecified mesquite sources were
located.
With a varity of
If unique
The second
CHAPTER
II
EXPERIMENTAL
General Methods
Samples were
Ultraviolet Spectroscopy
Ultraviolet spectra (UV) were recorded with a Gary Model 17
ultraviolet-visible spectrophotometer.
All UV spectral
In all cases,
8
Table 2. Chromatographic Conditions
for HPLC Analysis
High pressure liquid
chromatograph
Detector
Pumps
Programmer
Column
Mobile phase
Recorder
SOLTEC
Model 252A
Chart speed: 0.5 inch/min
or
Linear Instruments Co.
Model 252 INT
Chart speed: 1.0 cm/min
(absorbance versus
retention time)
a clean glass plate (25 cm x 25 cm) and allowed to air dry for
three days.
10
were cut with a hand saw.
The chemically-defoliated mesquite was obtained from the farm
of Mr. D. E. Sosebee which is two miles east of Anson, Texas.
The trees had been sprayed on approximately July 1, 1980 with
Tordon 225 (1 gallon of Tordon 225/100 gallons of water, Tordon
225 is a 1:1 mixture of the triisopropanol amine salt of 4-amino-3,
5,6-trichloropicolinic acid and the propylene glycol butyl ether
esters of 2,4,5-trichlorophenoxyacetic acid).
were sprayed until the spray dripped from the leaves. The chemically-defoliated mesquite was harvested in early January of 1981.
Preparation of Segregated Mesquite
Plant Parts
The freshly-harvested mesquite plants were separated into
component parts (trunk, branches, and leaves) in the laboratory
using a hand saw. The trunk portion was cut further into 25 cm
long sections.
11
Extraction of Different Parts
of the Mesquite Plant
Segregated mesquite plant parts were extracted using a
Soxhlet extraction apparatus (Figure 1).
T-TUBE
>f
COOLING WATER -
--V^COOLING WATER
THIMBLE
SOLVENT
Figure 1.
ROUND BOTTOMED
FLASK
12
The thin shavings of the separated plant part were weighed into
the cellulose extraction thimble (single thickness, 60 mm x 180 mm
or 43 mm X 123 mm or 20 mm x 100 mm, Whatman) of a continuous
Soxhlet extraction apparatus which was protected from moisture
absorption by a calcium chloride filled drying tube. Either one
pure solvent or a mixture of solvents was used for each extraction.
In most cases, the extraction period was 72 hours; but in few cases,
the extraction period was 24 hours. At the completion of the
extraction, the solution was transferred into a tared flask and
the solvent was evaporated in vacuo. A listing of the material
extracted, the solvent employed, the extraction yields is provided
in tabular form in the Results Section.
Column Chromatography
Preliminary Separation
The crude heartwood extracts were initially separated into
non-polar and polar portions using a 60 cm x 5 cm column of silica
gel.
200 ml of methanol.
13
The quantities of non-polar and polar portions of the crude
heartwood extracts which were obtained are presented in the Results
Section.
cap vials. After two hundred and ten fractions had been collected,
a 500 ml round bottomed flask was used to collect the remaining
solution.
at room temperature.
Final Separation
For the final separation, a 60 cm x 2 cm glass column packed
with silica gel was used.
14
Table 3.
Solvent Number
CH CI
CH OH
Percent
Total
(ml)
(ml)
^ ^^3^^
(ml)_
200
200
190
10
200
180
20
10
200
160
40
20
200
140
60
30
200
120
80
40
200
100
100
50
200
80
120
60
200
60
140
70
200
10
40
160
80
200
11
20
180
90
200
12
10
190
95
200
13
200
100
200
15
chromatographed was coated on a small portion of silica gel which
was then applied to the top of the column.
The solvent
Decomposition of Compound 1_
Compound 1_ is the major component in mesquite heartwood polar
extract which was isolated by column chromatography.
A small quantity of Compound 1^ was dissolved in methanol
(anhydrous, MCB, Omnisolv) and kept in a 1/2 dram, screw cap vial.
In order to study the suspected photodecomposition, this sample was
examined by HPLC soon after being prepared and the vial was then
capped and left on the laboratory bench (exposed to overhead
fluorescent light and reflected sunlight) for three weeks before
being analyzed by HPLC again.
laboratory bench and was examined by HPLC every two weeks for a
period of two months.
A more detailed study of the photodecomposition of Compound 1
was performed according to Flow Scheme A in Figure 2.
16
Two samples of Compound 1 were prepared as methanol solutions
by the method given above.
while sample 2 was analyzed by the HPLC. Both samples were examined
immediately after preparation and were then irradiated by a 60 W
light bulb placed 10 cm from the screw cap vial.
Periodically
Sample 1 (UV-1)
Sample 2
(HPLC-11)
Irradiated with
light source
(UV-2)^^
24 hours
-> (HPLC-12)
(UV-3) <-
48 hours
> (HPLC-13)
(UV-4) <-
72 hours
-> (HPLC-14)
17
A small amount of Compound 1_ was dissolved in methanol
(sample 3) and the solution was placed in three 2 dram, shell cap
vials.
Sample 3
Kept in the
dark for
72 hours
(HPLC-16)
(HPLC-15)
Irradiated with
light source
for 72 hours
(HPLC-17)
(HPLC-18)
Figure 3.
Flow Scheme B
18
HPLC chromatograms from the photodecomposition study are presented in Appendix A.
Sample 4 (UV-5)
\f
Add 5 drops of
NaOH solution
Add 3 drops of
HCl solution
Add 3 drops of
NaOH solution
Add 5 drops of
HCl solution
Add 10 drops of
HCl solution
Add an excess
(20 drops) of
NaOH solution
(UV-5)
(UV-6)
-2
The concentration of the NaOH solution was 2.9 x 10
M
b
2
The concentration of the HCl solution was 2.9 x 10
M
Figure 4.
Flow Scheme C
(UV-7)
19
The HPLC chromatograms and UV spectra which resulted from the
experiments conducted with samples 3 and 4 are given in the Appendix
Section.
Structure Determination
of Compound 1_
Formation of Trimethylsilyl
Ether Derivative
A sample (0.328 g) of Compound 1_ was dissolved in 1.0 ml of
pyridine (solvent) in a 10 ml round bottomed flask and a reflux
condenser was attached.
The residue
20
Formation of Ester Derivative
Compound 1^ (0.224 g) was refluxed for 0.5 hr with 1.0 ml of
pyridine (solvent) and 0.5 ml of acetic anhydride. At the end of
the reflux period, the reaction mixture was allowed to cool for
0.5 hr and was then transferred to a 60 ml separatory funnel.
A 10 ml quantity of distilled water, 20 ml of diethyl ether and
10 ml of chloroform were added and the reaction mixture was extracted
for 5 min.
and discarded.
of distilled water and then dried with MgSO, . The resulting organic
layer was transferred to a 50 ml round bottomed flask and the
solvents were evaporated in vacuo.
The effec-
gent was added to the PMR sample tube in small quantities (10-20 mg
each time).
sample tube was weighed and the PMR spectrum was taken.
of each addition was calculated by difference.
shift reagent were added during the experiment.
The weight
Six portions of
21
The PMR spectra are presented in Appendix B.
The influence of
CHAPTER III
Heartwood
Chipped mesquite heartwood was extracted using a Soxhlet
extraction apparatus.
upon the original weight of the heartwood sample) and the extraction
conditions are reported in tabular form in Table 4.
Since the project goal was to obtain useful non-carbohydrate
organic compounds from mesquite, diethyl ether was used initially
as the extracting solvent.
23
<+^
^s
73
0)
o ou
T3 CO
iH
}-i
(U
H
>H
CO
H
fn
}-i
a>
0^
CM
vO
vO
u
CO
3
C
o
o
o
s c
rH
>
>
O
CO
4*
VJ
CO
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
o
CO
H
4-1
o
o
in
c
o
o
o
CN
a
(U
4J
H
cr
c
cu
CO
(U
>
S
na
cu
a
a
CVJ
4J
O
CO
CNI
oCN
oeg
4J
4J
u
W
rg
CN
O
cCNJ
CM
4-1
w
I
CM
a
o
4J
w
CM
W
1
vO f H
ij
,^
W
iH
1
vO CN
ffi
ffi
vO
sd
X\
^-^
SO
CJ
tod
14-1
y-i
4-1 o
-i= o
60 ^
H 4J
(U M
12 CO
o
c
o
H
o
CO
u
CO
00
CTi
CO
CO
C^
CM
00
CN
CN
CO
00
00
00
00
o;
4-1
X
O
H
4J CD
O -U
CO CO
U
Q
CTi
CN
CM
<3^
O
CO
O
00
CO
00
O
00
CO
CM
U
CO
00
00
CM
CN
CO
CO
<r
o
00
00
00
<u o
H
H
4J
0^
(U
c r (u CO
CO . H
o
o
ON
S o
o
sO
O
ON
C3>
O
00
in
CT
o
00
in
00
O
00
CN
CM
CO
CO
00
CJN
00
00
00
00
00
00
CO
4J
c
s
H
M
<U
a.
X
CN
<
CO
vO
CN
^^
24
5>5
-d
W i H
O CO
cd
u u
CM
CM
CO
O
O
-<r
N
4J
CO
0)
J.4
CO
H
KH
-o
JC
JC
<u
H
JC
iJ
5
4-l
O
(U 4J
6 C
CN
0)
'T!
>
cr
CO
o
m
o
o
(U
<
CO
4J
4-1
C
(U
CO
H
rH
O rH
H
V4
> o
4-4
(U
CO
(U
T3
1
cu
X
u
rH
rH
CO
tM
>
33
fH
4J
O.
(U
CJ
4-1
VO
CJ
H
O
CO
0)
X
o
(U
TS
001
00
bO ^
H 4J
(U M
5
CO
(U
CO
CO
<u
o
u
<u
a.
[5
00
CO
CO
m
iH
^3
CI
CO
O
CO
M
iH
4-1
3S
CO
X
(U
o
H
4J 0)
O 4J
CO CO
U Q
o
00
o
00
4J
00
00
M
J3
so
vO
CN
r>.
H
O
00
4J - H
4J
c r <u CO
CO rH
<U i H
(X3
00
CO
ro
o
00
S o
CO
u
C
.a
CO
Xi
V4
0)
a.
X
13
4-1
0)
>
iH
CO
CO
-i
0)
O.
X
Cd
O
>
c
H
liO
c:
CO
5
-o
(U
iH
a,
S
CO
CO
O
CO H
c U
o (U
H a.
4J
4.1
}-i
4-i
X
<u
O
CO
a>
u
o c
CO o
V4 H
(U
fH
<
u
N
H
a
<u o
3
TJ
CO rH
0)
3
(U
rH
rH
<
CO
a
o
4-1
CO
4J
0)
X
(U
.c
H
25
in Table 4). According to this result, it appears that under the
conditions of extended storage, the organic chemicals in the chipped
heartwood are either lost by evaporation or converted into unextractable forms (possibly by air oxidation or decomposition).
Extraction periods were also compared.
26
(PMR-1, IR-1).
A rather
showed several peaks in the region 6.0-7.0 ppm which suggests the
presence of aromatic hydrogens.
were obtained from mesquite heartwood samples for which the mesquite
was harvested in the fall (Experiment 1) or spring (Experiment 8)
compared with the winter (Experiments 5-7).
Taking into account the seasonal variations in the amount of
27
material which may be extracted from the heartwood, extraction
yields were found to be reproducible.
tion of two different heartwood samples gave yields of 4.0% and 4.1%
(Experiments 1 and 8). Similarly, extraction of two different
heartwood samples with benzene-ethanol (1:2) gave yields of 10.6%
and 11.7% (Experiments 9 and 11).
Heartwood from Chemically
Defoliated Mesquite
The heartwood samples used for Experiments 14 and 15 were
obtained from the same chemically-defoliated mesquite plant. The
extraction sample for Experiment 14 was heartwood that came from
the portion of the plant that was above the ground, while the extraction sample for Experiment 15 was the heartwood that was in the
portion of the plant that was below the ground.
The extraction yields were 7.9% and 12.5%, indicating that the
extractable organic chemicals in chemically-defoliated mesquite
heartwood were not affected greatly by the chemical spray (compare
the Experiments 14, 15 and 10 in Table 4).
Although 6 months
elapsed between the time the chemical was sprayed and the mesquite
was harvested, the yield of extracted material in Experiment 15 was
found to be similar to the yield of extracted material in Experiment
10.
result from some decomposition of the above ground mesquite heartwood components after chemical defoliation.
PMR spectroscopic analysis of the extracts showed that both
extracts from the chemically-defoliated mesquite were quite similar
28
to the various extracts from freshly harvested mesquites.
Other Parts of Mesquite Plant
Limited data were also obtained from the extraction of chipped
sapwood, chipped bark, and shredded leaves with diethyl ether in a
Soxhlet extraction apparatus. Results are recorded in Table 5.
Comparison of extraction data for the various components of
mesquite obtained from the same harvesting reveals that with diethyl
ether, more material can be extracted from the heartwood (Experiment
1 in Table 4) than from the sapwood, bark, or leaves (Experiments 16,
18, and ,9 in Table 5).
29
u^
o
'O
r-i
0)
H
SH
^s
'X3
0)
4-) i H
a
CO
u
p
CO
H
iH
>.i
7-i
4-1
CO
CN
CM
CO
CM
CN
CO
(U
Cd
s
C
H u
o Si
4->
O
CO 0\
u 0)
4-1
^3
O
O
:5
CN
r^
CM
r^
r^
iH
vO
X H
Cd H
CO CO
CO 0)
>
CU CO
u (U
H hJ
cr ^3
CO a)
(U T3
S T3
<U
T3 U
(U j a
Q. CO
U-t GO
4-1 0)
Xi . H
00 O.
H
e
( U CO
Csj
OS
CM
CO
IS CO
(U
CO
3
a.
H X )
j::
CJ
CO
14-1
o M
u
CO
c PQ
o
H
4-1 T3
OJ
a
CO cn.
u O4J H
X ^
Cd CJ
LO
0)
o
H
4J 0)
O 4-1
CO CO
4-1
cn
ON
X
CM
vO
CN
in
-H
O
C3N
ON
CJN
S-i
0)
C
d) o
CT\
<3^
4-1 ' H
H 4J
cu
3 O
cr (U CO
CO rH
Q
(U
2 o
H
T3
O
rH
ON
CT
<T>
o%
CJ
CO
O
O
O
u
CO
0^
4J
c
CO
y-i
(Xl
X)
13
O
0
o
o
5
a
CO
a
CO
CO
CO
:s
CO
(U
M
CO
pq
CO
(U
>
1-1
4J
J
CU
M
4-1
X
0)
CO
CS
e
H
M
a
vO
00
OS
o
X
Cd
ClH
CO
30
Chromatography and Spectral Analysis
of the Separated Material
methanol eluted the polar fraction (about 90% of the original heartwood extract) from the column.
A proton magnetic resonance spectrum (PMR-2) of the non-polar
heartwood component showed only aliphatic proton absorptions at
0.9-2.2 ppm.
The car-
31
of interest.
heartwood component in ethanolic solution shifted to longer wavelength upon the addition of NaOH solution).
Separation of the heartwood extract polar components by preparative thin layer chromatography was also attempted.
Although
32
graphy would offer the best possibility for separating the polar
fraction of the heartwood extract into the individual components
in sufficient quantities to allow for their identification.
However,
an appropriate analytical technique was needed to monitor the effectiveness of the column chromatography in separating the individual
components of the heartwood extract polar fraction.
Using this
33
into serveral different fractions (Fractions A, B, C, D in Figure 5).
WEIGHT
(mg)
200
100
200
VIAL NUMBER)
Figure 5.
-1
compounds with IR absorptions at 1610 cm
-1
and 1510 cm
''
. The
96-108, HPLC 4) (vide supra) was combined and applied to the column.
j^
tical methods (the TLC plate showed a single spot after development
and the HPLC chromatogram showed only a single peak, HPLC-5).
The major component (Compound 1) in the heartwood polar fraction was examined by infrared spectroscopy (IR-4), proton magnetic
resonance spectroscopy (PMR-4), and ultraviolet spectroscopy (UV-1).
The IR spectrum exhibits broad OH stretching absorption at 3750 to
3000 cm"
II
35
indicating the presence of OH groups. Integration of this broad
absorption seemed to indicate a multiplicity of OH groups. A doublet
at 6.8 ppm and sharp singlet at 6.4 ppm in a ratio of 3:2 lead to an
assumption of 5 (or a multiple of 5) aromatic hydrogens being present
in Compound 1_, The peaks around 2.9 ppm might be attributable to
methylene hydrogens adjacent to a carbonyl group (-CH^-COOH) or an
alcohol group (-CH2-OH).
Compound 1_ is
MJ.
(methanol) = -51.0
Decomposition of Compound 1^
It was noted that when a methanolic solution of Compound 1
36
Table 6.
Solvent
Solubility of Compound _1
in Various Solvents
Insoluble
Partially Soluble
Soluble
CCl
CH2CI2
CHCl.
Et^O
Mi
CH^COCH^
CH^OH
H^O
w e r e examined b y h i g h p r e s s u r e liquid chromatography
immediately
II
(HPLC-6)
(When the r e t e n t i o n
A l l the H P L C
37
peak gradually decreasing and new peaks gradually increasing.
(Quantitative data could not be obtained due to a slow evaporation
of solvent from the sample which produced changes in concentration.
To determine whether the peaks areas were increasing or decreasing,
the relative intensities of peaks in a chromatogram were compared.)
Since the possibility that these changes arose from some type of
contamination appears to be small, it seems that a photodecomposition of this sample might have taken place.
A more detailed study of the photodecomposition of Compound 1_
was conducted following Flow Scheme A in Figure 2.
Before irradiation with the light source (a 60 W light bulb) ,
**
"^
After
si
these samples had been irradiated by the light source for 24 hours
41
!
II
;
above.)
38
a possible source of the observed changes. A direct comparison of
photodecomposition and acid-catalyzed decomposition was performed
according to Flow Scheme B in Figure 3.
Although the sample (Sample 3) used for study had slightly
decomposed (HPLC-15), it could still be used as a standard.
Sample
"'I
-^
m
numbered 2, 5, 8, 10, and 11 in HPLC-15 and HPLC-17).
A third
portion (1/3) of the original solution was examined by HPLC immediately after adding one drop of concentrated HCl.
If acid-catalyzed decomposition had been major factor in the
composition changes, the chromatogram should have changed greatly
upon the addition of concentrated acid.
It
II
39
the Compound 1^ are apparently very sensitive to light. When subjected to light irradiation, decomposition occurs.
Treatment of Compound 1 with Base
The PMR and IR spectra of Compound 1^ indicate that it might be
a phenolic compound.
is a phenolic compound.
It was found that the UV spectra of a methanolic solution of
Compound 1 changes markedly when subjected to high concentrations
^
^
H
fi\
"I
.'I
of base. After the sample solution was treated with base, it appears
that the composition of the sample might have changed (curve F in
UV-5).
2.9 x 10
did not reconvert the absorption of the base treated sample solution completely to the original absorption which indicates a possible irreversible reaction with base.
A significant change was observed upon addition of excess
_2
(more than 20 drops) of 2.9 x 10
M NaOH solution (curve J in
UV-7, where curve A' was the untreated sample absorption and I was
_2
treated with 3 drops of 2.9 x 10
M NaOH solution).
II
40
Structural Determination of
Compound 1^
Silylation of Compound 1^
The mass spectrum of Compound 1_ did not provide an identifiable
parent peak.
M)
used for a model study in order to determine the best reaction conditions for silylation.
m
I
^
II
o f exp er iment s.
Using hexamethyldisilazane (HMDS) and trimethylchlorosilane
(TMCS, as a catalyst) and phenol,refluxing for two hours gave
complete conversion to the phenyl trimethylsilyl ether.
Completion
41
reagent mixture, Compound 1^ was dissolved in a small amount of pyridine prior to silylation.
immediately after the HMDS and TMCS were added. During the reflux
period, all of the precipitate (probably NH.Cl) sublimed out of the
reaction flask onto the inner condenser surface leaving a clear solution.
ed to cool.
"M
pletely.
N e w absorptions at 1260 cm
1 0 8 0 cm
^
2
3
SI
i
II
42
very different chemical environment from the others (the one that
shows a -0,25 ppm signal after been silylated).
Owing to technical problems, the mass spectrum of this trimethylsilyl ether derivative of Compound 1^ was not reproducible.
Therefore the actual molecular weight is still doubtful.
According
nolic hydrogen would increase the molecular weight of original compound by 72 mass units and there are five trimethylsilyl groups.
Compound 1^ should have a molecular weight of 290.
III
Esterification of Compound 1
An acetate derivative of Compound 1_ was prepared by reaction
^
A]
Si
I
esterification procedure.
phenol was refluxed with acetic anhydride and pyridine (which serves
as solvent and a proton acceptor) for 0.5 hr, work-up of the reaction
mixture revealed that all of the phenol had been converted to phenyl
acetate.
'
!i
l\
n
43
mixture was cooled and transferred to a separatory funnel for the
extraction.
material.
After separation and drying, the organic layer was transferred
to a round bottomed flask and the solvents were reomoved in vacuo.
The residue was a light brown solid with melting point of 145-147C.
The IR spectrum (IR-7) of this ester derivative shows the completed disappearance of the OH absorption of Compound 1^ at 3750 to
'^
-1
3000 cm
which indicates that the reaction had gone to completion.
-1
*
Ml
"^
T|
The
a)
?i
,|
44
2.23 ppm and A^ : A^ = 4 : 1) .
b) The integration ratio of PMR-7 shows a total hydrogen number
of 24 (or a multiple thereof) in the acetate derivative.
Since each
This result
jj[
tot
is to be expected since substituting the hydroxyl groups with acetoxy groups should cause a strong deshielding effect to neighboring
hydrogens.
*^'
"I
'^
l
||
(
absorption changes could also have resulted from some side reactions
which might have decomposed Compound 1_ during the esterification.
In the shift reagent experiment, the acetate derivative of
Compound 1_ was dissolved in CDC1 and analyzed by PMR spectroscopy
(PMR-7).
were then added to the PMR sample tube. After each addition, the
45
PMR sample tube was weight and the PMR spectrum was taken (PMR-8 to
13).
Table 7.
As more shift
^.
^
m
tions with the integration ratio: B^ : B^: B^: C: G^ : G^: F: A": A'' =
1 Z 3
i Z
z l
1:1:1:2:1:1:2:3:12 This ratio is again consistent with the presence
^
;?l
tive to the shift reagent (compare PMR-8 and 13) . Relative sensitivity to shift reagent for all the absorptions was:
G
> B
dl
)i
,!
46
Table 7. Effect of Shift Reagent upon the Chemical
Shift of Absorption in the PMR Spectrum
of Acetylated 1
Shift
Reagent
Added(mg)
Before
.," ,.
Addition
First
Addition^
(11.9 mg)
Second
Addition
a
h
^2
^3
^
h
"l
^
^1
^^2
(ppm) (ppm) (ppm) (ppm) (ppm) (ppm) (ppm) (ppm) (ppm)
(8.6 mg)
Third
Addition
(20.1 mg)
5}
i^j
g|
8.37 7.86 7.57 7.08 6.82 6.18 3.47
2.63 3.14
Fourth
Addition
(12.9 mg)
8.83
Fifth
Addition
(8.8 mg)
Sixth
Addition
(16.0 mg)
8.10
7.70
7.17
7.37
6.55
3.67
2.78
3.58
^ See PMR-4.
^ After each addition of shift reagent, the PMR sample tube was
shaken vigorously and the PMR spectrum was taken after all of the
shift reagent had dissolved.
?!
l
'w
J[
J,
II
"
47
Table 8.
Shift
Reagent
Added(mg)
Before
Addition
First
Addition
(11.9 mg)
Second
Addition
h
^2
^3
^
S
S
^
^1
^2
(ppm) (ppm) (ppm) (ppm) (ppm) (ppm) (ppm) (ppm) (ppm)
0
0.32
0.30
0
0.18
0.12
0
0.12
0.08
^
0
0.09
0.07
^
0
0.46
0.39
0
0.25
0.25
0
0.19
0.16
0
0.10
0.09
0
0.37
0.25
(8.6 mg)
.^
"!
j2
Third
Addition
(20.1 mg)
0.55
Fourth
Addition
(12.9 mg)
0.46
0,36
0.17
0.12
0.70
0.41
0.24
0.19
0.54
vi
:?l
0.24
0.14
0.09
0.55
0.37
0.20
0.15
0.44
: I
CI
ii
II
Fifth
Addition
(8.8 mg)
.1
0.28
0.10
0.10
0.07
0.36
0.23
0.17
0.13
0.26
Sixth
Addition
(16.0 mg)
0.54
0.22
0.14
0.09
0.53
0.32
0.20
0.17
0.46
48
tical with that of Compound 1^ some IR spectra of flavonoid compounds
showed types of absorption in the regions of 3750 to 2800 cm
1800 to 1200 cm
and
Compound ]^,
A number of literature references regarding flavonoid compounds
were then searched.
In this
^J
5l
"H
n
of the trimethylsilyl ether derivative of Compound 1^ (PMR-6) .
Jlj
ai
II
()
at 6.1 ppm and 5.9 ppm shown in the PMR spectrum of the catechin
derivative are not present in the PMR spectrum of the derivative of
Compound 1^. Instead the latter shows a singlet at 6.33 ppm.
This
49
^.Sppm
^.8 ppm
f\
5.9 ppm
OR
H H ^ ' ) "Q'2pDm
V/
3.9 ppm
'^{
CH-
2.8 ppm
n
m
R = -Si-CH3
CH.
'!
-'i
<l
^1
II
I
25:6'
8 6
Jjiiiu
10
Figure 6.
IV
0
ppm
50
COMPOUND 1:
R=H
DERIVATIVES:
R'= -C-CH
or
R'= -Si-CHq
I
CHo
n
Figure 7.
:%l
ill
strongly affected by the acetoxy group than that at position 5.
II
C, 62.07; H, 4.83.
Calculation
Since
15 14 D
51
of the Compound 1_ was also made by measuring the coupling constant
for the hydrogens attached to these positions in the Compound 1_
(PMR-4) and trimethylsilyl ether derivative of Compound 1^ (PMR-6).
It was found that J^^ = 8 Hz.
13
CI
OH
II
!
B
Figure 8.
It is expected that in trans-2,3-flavan derivatives, the conformation in which both the 2- and the 3-substituent are quasi-equatorial
(structure A^) will be highly favored at room temperature.
14 15
'
52
CHAPTER IV
SUMMARY AND SUGGESTIONS
FOR FURTHER RESERACH
Summary
During the research period, attention has been focused primarily upon the reddish-brown heartwood of mesquite, Prosopis
juliflora.
*!
the amount of material extracted from the heartwood and the overall
^i
The amounts
,^
the heartwood of mesquite were found to depend upon the solvent and
the season of the year when the mesquite was harvested.
['
ii
In order to separate the heartwood extract of mesquite into
individual components or fractions of components, portions of the
heartwood extract were submitted to
methods, such as:
various chromatographic
53
at least twelve components.
phenolic compounds,
A high pressure liquid chromatographic system was developed for
analyzing the individual components of the polar fraction of the
mesquite heartwood extract.
of the polar extract. Compound 1_, was isolated from one of these
fractions.
Jj
41
'H
1i
;||
spectroscopic methods.
.,
il
upon the acetate derivative of Compound 1^.
From the accumulated data, the structure of Compound 1^ is proposed to be:
(MONOHYDRATE)
Figure 9.
54
Suggestion for Future Research
^J
proposed structure.
li
%l
z\
-.i
It would be useful if
More
'\\
ji
II
LIST OF REFERENCES
1.
2.
3.
4.
Browning, B. L., "Methods of Wood Chemistry," Vol. 1, Interscience, New York, 1967, p. 79-82, 87.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
Weinges, K., and Paulus, E., Liebigs Ann. Chem., 1965, 681,
p. 154.
55
Jf
^t
J[
J!j
^
SI
2
'^l
-II
APPENDIX A
*l
m
High Pressure Liquid Chromatograms
II
II
56
57
::::=:5g:=l3i||sS:::::M||^
:i::::x::::::W:::::::Sffi:::::::::ffi+!i:
:::t--::::::::x::-V--+i^-iiif+i tX
'
J _
1_
tiix-t
J_L
'
'
y-r-4-r--it!
' i ' it^r-nH
| | jn | | / ; IT)
|--L-4--U- ll IIIM 11 -j^JLL
^4(1-13ol4i--i42(J4-4-|-4i>|fr4+r^o
o
(0
u
u
^!ii||lilllllllllllll|j:ttltltt^^
-+ - ^ - r t - - r ^ ^ ^ ^ ^ ^ : ^ = ^ = g ^
1 II -l-Ll-L-L4-L.J-^
u^
O
O
1
,
' [
::::::::::--t-^++4
4 i t t l r 'ill' 1i i^ 1' 1l^' ^' l1^1 t' ^
.
t
1 1 r !' i
' I
-
|_j
'1
1 1
.,
u:
^
2
''
! i
1 '
' 1
i
t -
X :::-: ::::
:T::
-4T
^,L
.-i...
1
'
' '
1 i : 1
-H
Uj. ^ ^ ^ ^ ^ ^
r
: '
1 '1
1
11 .,
1
1 1
_' _ i1!
I ! 1
Il's
'i'
M
f
1 1 IT-I
1 1
'
L-+.4--i
'
-i
'
'
1!
'1
1'
100-
i - t -go - 1
i 1 ' 80 1
'
-^-t
i i 1
11
1i 1
'L
j-
: ' '
1 1' 1
""Tt"4'"T'i
t 1
u-aJ
M
'
iJ
'
1 ' 1 M
1
1
, '
i 1
'
) 1 1
'
tI
i1
'
1
II
' M
11
I I
M
^zx-ix
i
IrrJ
i/u -
<
' . 11
: ' 11
11i
1 1
1 .11 M i l t
1 1 '
1 >
'
1 I : !'
''
; ! 11
1'
i i 1 I 1 ' i 1 1
'
rUCn
111 !
'
111
I
I
1
'
1
1I
1
'
'
1
'
1 1!
1 I
, 1
; i':
0)
at
cr
' H -
'
1' 1i
1
CO
(U
-I-;!
^
S
u
'-:
: ' 1
' '
T1
CO
iH
I; '' , :
'1 ;
1 ;
1
!
!
;
i
'
i 1 ;
' !;1 1 1 !
1
1
1
1
'
I
CJ
1-3
PU
in
'
' l ! Pl
,', '.
1 1 1 i1 i
1
1
1
1
" r
111
1 '
111
III
'
, 1
1 1 1 1
M'
H
'
1
I'll
1 1 '
1 '
'
'
1
1.
,'
II
/{J
1 1
.
I I '
1
! !'
1 ' 1
1' 1t
1 i
'
I
11
. ' .11
!i /
!1 /
' : / ,
/
1i 1i 111 i1 ;11 1
: 1 1 I ; 1 H 1 1 i I j 1 i :1 !/ ' '1 1 ' .
1 11 11 i i !1 I 1' 1
otJ 1
i l l '
1 1 AU
i II 1 I h
'
i 1
'
' ' t
I ' M
' du
i,|i i l i l
)..!, . -
'
*o
^ ij
'/ :
' : ' \j
. . . ; i . . . ^ i f r r - L . Ja-i-^
-: rT--i^^^^-4k=:t4=^r^7lT-t[H-iTtT
- - H ill,Ml ' h ' ! "\i"''i
' r H 1 'MiM'^' ' ' 'U
t . nM x' . . i tl Xl 1
;'
11
M i 1U
i1 1 1
111
1i 1
1 it t :1 ,
M '
'
I I !
""- t
^
1 ' 1
1L
'
it"T;
~r -"-rr
i 1'
'
1
1
1
1
Ml
til
I ' l l
t '1 i i
1
1' ,
] . ii 1 7 '
! 1 !
lilt
1. ,
' i 1
1 1 1 1 ' 1
I1 l 1 1 .i :
1 i
' 1 1
< '
3
CO
H
, *- ^ 1
l i t 4J^_ i..,
'
( 1 ' 1' i ! M 1
Tj^t\
TT__
ii-Li^-[|t-|-l-V4!4-T-+4i4J5--4
I f f -Mt-^1 111
1' II 11 1;1 III
i 11i I I
. 1 . T ' 1 1 1 1 1 1 ii>ii
II
't !
(U
3
! ' ,
1 1i
>
1 1 1 .
1 1 ; : i ,' ;
1;'
^^vi,!;!
'
;:
'11
'
11;
1 1 i
11
'
I i1
' j1 1 ' ^^--r
T 1t - i "T 1
^ 1i 1i
1
i i
(d
11
'
'
1 ! i 1
i:4::::4.,..-^fr.xlr|-Xlt_4^_4.
t x Ml nn ; M T !^
f f - ' -1 " 1 n tT ' T M
-r
_.,
'
M'
it
! i
1 1 1
1 1
j:-i---^j:
r|r
nx'riol xd:
i
1
'nn
' \
11
i Inn
inn
1:
1
1 11
'' |
M . i. i i
. ..!, .l i U .
.-i.,..L..
i I 1 1
1 1 1 i
1 ' !
'1
'
1
1
I I I
L'
u--i-.-J
M ,=F
Hi
^! rH
' i ^
i1 i1 t.,
, : 1 ; 1 1 1 I i 1 1
' '
M
r
^ 1i' r '1 l l ! i 1M!
'
Tn
;!
,1
1 1
_p
1
.--L-lrn
! ' 1 1 !i4--.-J
1 1
1 !
'
1 1
M i l1
11
^_ _H
'
..H-4--u-U.Li
- ti' l! ' , U
1
1 '
1'
'
jJJ^jj^.^-tii-L-^i-!^
S .-_ 1X4-T--t-4-4-t-t-.--i
-iTLLj ..-X - . ; t -titt - _ 4 :i- - -.,
j \i --\-^
4_i ilL.XUiz^-f-1-
:::::::+::5t:i::::::::::::::::::::::
^ r r rt-t- 1-+rL i + f t
i-a-.-TUxi-^^
4;-_^_.._ln:44:_,.,T._.
^-4-11
._ 1
11-,
u
u
II
It
II
58
4-t
c
>
o
CO
u
0)
H
no
cd
o
s
ll
PQ
o
4J
o
cd
u
cd
00
CI
cd
s
o
u
i
*i
II
CI
CM
II
a<
PC CL>
00
H
59
c
0)
>
rH
O
cn
a
a;
H
Cd
O
s
o
**A Si
d
o
Cd
jj
cd
UL*
Si
CO
SI
II
(l4
o
cd
u
60
O
S
o
u
CI
CM
0)
3
t)0
H
60
o
u
^1
>%
s:
a
cd
o M
a 00
o o
H 4-1
U
O
Cd
cd
o
u
u
J2
p^
M-l
CJ
o c
G
a i 3H
o O
H
P
U
O u
Pk G
0)
V4 >
i
O H
f-) O
Cd cn
- ^ <u
1 H
TS
o
HJ Cd
(Xi ^
s
m
M
3
00
H
l
SI
I
SI
CI
>l
II
61
i
i
o
O
CJ
14-4
o
B
cd
u
00
o
o
u
Si
CJ
CJ
Pui
9S
n
ll
I
CI
II
It
CI
I
CJ
(U
00
H
FS4
II
62
i
1
\
[
1
!
;
'
'
I
1
'
"
'
1
I
1
(
'
~ '
-'
,i .
..
...
.,j
'
!
'
9\^
1
I
1
1
i
'
;
;
/'J
' 1
i
'
i
1
1
'
1 1
I
'
1
1
1'
.
M ! '
'
j .
... . j .
' .
M M
' , 1
'
__!;
1 1 1 o u
'
1
" -
x;
-N
1 .
'
'
'M
l
Ml,
' 1
'"I
1
B
u
' '
'
'
i t1 ,
1 '
11:
hm
--
1 M
L -
.
pL,
-1
LO
-i
a)
u
3
00
'
1
"
] - ^
'
'
-J"
i
(
^ ; K
M ~ = !
1,'
o
s:
j
i
l l "
f-rrt-
CI
CI
I
I
4-1
33
i-r
TTTT
! 1 1 11 i 1 1
-i-\
Ml!
.
'
T T
(U
i
1
il
11 i n
1 ^
CI
a.
B
o
o
M , ^
, 1 .. , ,T,..
i
J . .
i .
C ..
1
I
1 .
--I
ll
4-1
1 ,
H
CO
O
!
:
'
1i
U-l
D
8
t L
' \\ -I*-*
^'J
1 [
'
1 1
CJ
MM
MM'
MM I ' l l
'''' '
1
1
'
i M
11
"M,,!;
! 1 1 1J i ;
1 '
'
i
. ' ' 1
'i
;I' I
'
1
1
e
o
'
'
=d
i.
.1 **u '
T3
C
3
O
cx
1
\
'_J^^ "^
1
1
'
'
(rs
1
1
;
' I
'
'
uij
'
i n
IX)
'
'
-j-j.i.
;
1
uu
1
1
I I I '
1 '
'
ou
'
,
! '
'
1
1
'
' 1
> ,
auc:,i_
1 .
1..
; 1
i
' '
'
'
'
; fli<i
'
'
/
/
'
jfSJ
'
I
:
i r-r :
1 3 1
\i
'
* 1
. . .,. i
.
c
r-
..
1'
^^^-^^-^
'
..,,
'
., ,
' 11 1
'
, . , ...i
'
1() 0 '
,
'
' ! '
^1
1 .
, .
'
" ; - 1 "
'
. . .
i
:
I
'
1
1
. 1. . .
- 1
,
'
i1
'
,
'
'11
i
1
)l
II
63
CN
3
O
100
1
1
'
,
... \i 1 1
j i 1
',
11
,
1
!
i
1 ,
. t3
t * 11 i -
1 ,
'
' ! ' i
.
' i 1
..m
t
',' 1
; '
1
1
!
;
I
!
'
'I
'1
1
*
'
'
. .;
1
1
'^
\
i
1
i
1
1
1
1
1
'1
! ' '
'
i.
1
I
1
1
M:
I '
'
'
;
^
:
'
\
\ '
'
90
8D
'
'
1
j
-^^
- . , . i .
.1
i
'
L.-
U
c-
M
3
00
.^
~-'-
^
_
o
.
^
r
^o
Un
1
30
:-
^
1.
M-^^
" ^ , .
.
1
1
\
'
--..
1
' ' :
1 1 1
t
7 :
'
l . l
'
'
I
Ml
I
I
f1
'^1
VI
1in
1
1
, .... 1
'
,
.
. .
i
-
...._... 4
^
i
1.
1
,\
1
!
. M
j1
..
.\
1 '
, ,, M,
'
ij
J1
. . .
1(DO
"I
;i
o
o
s:
e
o
dj
j'' '
S^
\ ^ '
-Tr \ '
I
\
rH
1
1
1
ll
'
!1
t'^
''
[
1
,
1
'!
Cu
\\
hH^
i
>
.... . i , . ,
M
'
11
._ ._.,1
1
1, ; '
o
CO
\'
"
'
'
l.l-i
^j...-'"'
-"^
' ^ l
^'
~^
1 '
1
..
t '
1' '
SI
$:
64
11
U.. . .
^ 1
-U.U
i-^
;
1
-4
'
1-
'
i
11
... 1
-4-^1
.....M
TT"
1 '
-1-^
..
) <
'
1
M
'
'
I-
-.. . .
, ^
.-
'
'
1 M 1
^ ' I '
'
1
'
'
11
j
:
'
'
1
I
"
'
i1
'
i
1
'
'
' ._J_
'
'1
'
'
'
' M
' 11 '
M
;
'
-M-J
S.I
'
11
1
I
'
1
'
'
:
1
>
'
1
1
'
: . . ! . .
1
1
'
1 .
1.
pL.
it
81
PLI
'
33
'
1
!
'
IT' ^
'
'
1
J !
*
1;
)
>
i
o
o
s:
'
'~
W-.
CU
'
3
00
H
'
4-1
00
I
CJ
'
1
1
I
I
:i
:i
M
1
'
rn
[
ll
o
e
o
o
'
1
1
i
i
1
I
i
t
H
CO
in
1
1
o
c
o
j
j
, . , I' ,
X1
'
,
[
:
!
1
1
1
1
1
1 , .
'
CJ 1
11
\
l4-(
'1
^i
'
'
fj
.,..-..
. , 1. +.,.
'T
1 '
>r
11 0
. 1
1
'.
1,
i ,
~~^
1
' '
..
i1
...
2 ib/
'
o
CJ
'
1
. M
. , 1
'l
' 1
-9
o.
'1 A
; 1 M
13
C
3
O
-1
1 ,
ro
'
.'ao-i
1
1 in
-y-^
-4^1
.: . . .
^
'.
-==<_4'1
1 ^
-^
- - O :
'
1
1,
1^^
. ..
'
p-^
TH
t..
'
L: i . >
.1
'~r1
1
i
c
E
;i
65
*l
^t
It
'.I
I
'I
:i
SI
.1
I
f
66
in
t3
C
3
O
o.
B
CJ
c
o
r
:i
CO
o
cu
e
o
o
(U
T3
O
I
*i
:i
:i
4-1
!l
;i
VI
o
PI
- ^ 0 '
70
80
'
I
1
1
1
'
1
1
^40
1 .
30
'
I
1
)
1
u. i
<
;
1
1
1
1
i
, .
1
1.
TO
1
fM
lOoT
i
'
20-
, .
''
;
1
1
1
'
1
I
'
1 1 ,g
'
>
60
>
.y
""^
!=i:^
14
<
. .
I
i_,. _.^. . , . ; . .
1
1
1
i
1
1i,
-.
'
'
0)
U
3
00
SI
f
67
' 1
M'
T\
1
1,1
t"
iM'IM !^i|i
r';
1 M''
: i
IMI
tIM
i ;
IM;
il'i
1 1M M :
I'M
Mi ii! 1 ! M M
llll
, M | !M I M ' I liH; 'ii' i4
iM l!|M i l 1! '! iIlI lM' liii
i; MM
Mi'
'1 i|! Ii i i ! Ml illMi
i M ! 1 li i l l i iM
ii IM MM
i | i i i l l ! II
Ml
''
ij 'Ml
'1 MTM M M iMJ MM;ii Ml i
i! iiii Mi !i!l III!
M Mj! IM
M ! ii M
1 M'
1 ii 1r\ll MTTM- M'lil liiHi liii IM
Mi ;Mi ' 'MiM 'M^
'\ H
'j
M i l
M1
1 M1 ;1 1M
1 "
M' i
MM1
1i
:'
M 1
!|,:
!!
l : ']
liii
1 l'
Ml
''
l i i i ii
M li,.
1'
Ii:i iMM
j i ! ii!' 1M
' MM
IM;
1''
liii
IjM 1
M l i l i i IMiiil!!
1 I!
i il1
ll
i .^
1 M ;
;!M M
ill!
i Hi!
j IH
ill
Ij
:M.
M|
|M! H^^
^ Mil j ; ii
1 i ! 1 : '
M l '
, 1 M M -!-*
i 00 ' M M i l
90 M '
1 1
i ' i
M ' '
! i
''
'
'
l . l .
'
: M
Cd
; <!'
0)
iiii
liii lij!
iii'
'Ml
i lMM,
''
'
'
M'
. . M
11'
i l l I.
'Ml
1' '
' ' i
'
1 i
'
'
'
Mil '
i'li ii I
'
'
'
!''!
''^
'
(U
X 0)
Mi
ill'
'
'
'
IMi MM i i j i
MM
100 ;M
: ' 40
s0 ; '
'
pL.
30
ii - H '
M ; :
i ;,
1'
'
'
'
i
1
20
.M
. 1
I
:
'
1
1
1
1
-'1^^
10
1
i
1
1
i 11
'
1 :
M;
o
CM
0)
*v
1
I t '
-M
M 1
"H
HJ
(U
Si
4J
o Si
1
Ml'
1
1
O
CJ
rH
H
I
1
1
Q.Si
B 00
Il'i
Ml!
( 1 '
'1
II
U
M
il
o -u
\ ' \ '
in
; M
'
I M M
:
M : i !M! i
iiMi
1 i 1
i M i
MM
'' M
fH| U
3
O O
C CO
3
: 1 M
M '
u
o
M:
M!|i l l liii Mi
M.'
1 ill
1
MJM';
1M ;
1M
iii i i i
i'l
M i 1M l
'
! ' ' i
iiii
liii
;' 1
Hi
1 M i
1
,
M ' '
Mil
MM
u
u
M M
i 1 M i 1 i!
. M ;
. 1 '
if '
'i
MM
'
I : )
1
M
'
1% :
1 '1 '
o
H
4J
cd
H
1 '
! M
' i M
' ii
iiii
M M
1 ' '
1 i (
, : 1
jMi
III!
ill'
|t^
i t
1.
1
ii ij_i;l
'1
:1
;!
11
M
->n'
M
. t
IMJlM!:: MM
1 iilii!!! li
i M t ^ ; ii iH- ' M ;
M.
ii M
i 1
MM
i1 1
1 Mi^'
' :,
'
II i!i |
1 i l i ' l M l h lih
iM M
i'
Ml
1 '
illi
<\ I *
iiii!
MM
i l ! ' M ii Mi'
!!' M iTi! 1 ii i
| M MM
Mil
'M
i i.M
in
HI
TMT
Mil
iiii
MM
MM
'
'
j M ; 1
'
Il-j
11
r.i\
' '
i'l!
M:i
11
1 ;
l . l
M'' MM
iij!!
Mi!
1i '
ll Ml
3
00
>
;i
.'I
68
'ii
i J! i i
i
1 1
M
!
T T
1
'
i
TT
!
i
ll
i i i i
1 '
'
lill
' 1 1 :
illi
' I ' M
i ' ii
1 1
M!
1 Il'i
i'l"
! MM 111
'
1 M
iJi'
ii 'Iii
'M' IMI
M M
iii!
Ml
ii iM
' ii 1
MM
Mj
i t !
iiii
i!M Mi
i i ' ' l i i i M'^ ' M ' M ' i l i M l i i i i
i l l ; I'M
i l l ! JMi - M M i l M i i l M 1
' E^
1 ., 1
i j i
liii
M i l
! MM i
: MMiii i i ^ i i l i
il Mil
'
|l
1 ii|M|i
^ Mi:
1i
r r iTi 11ii
1
i l l !
i M ^'
1
! ll i
t ij MM i i i i
M
'1
MM i i i i
MM i . "
iiJI
1 1 1 M |l
M M
1 { ' [
'.''l
M!'
M'l
'
'
Ml
ij'il^'ll
i i i i M ! 1 !i;i
1 'i^i 1iiii i'li ill! Mi
Ml
l! i l i l ll Mli MM ; i i i
Mi
'' Hli ilil
,j.,' '
' 90 ' M . i 1
r i .1
r-f- -
' 1 i!
i: i II !h
ll
ii
Miii
ii Mi
i MM
jiM
11
11
'III
;iM
< Mil
1 MJI
ii iiM
!' M'
M'
11^
'
1 1
M ' '
i , . '
1 : M
;,i0 0
' 1 . :
'
il
Tni ! M. 'Ml
Mil
1 1 ,
M'l
M
1
I ' M
'
"
' ' M
; 1 M
i. i M
' M '
' I '
5 0 '" "I
' 40
:i"
MM
M M
MM
iM!
1
.'
1
:' *
i M1
'
i M ,
MM.
.iiii
i: M
U-i
'O
c3
a
B
Mi'M
Ml liii
iiii M
'
'
MM M
' j ' :,
0
CJ
M.
.iii
i 1 1
: i
in
'
MM
iiii
i j i ' ' M:
0k
:'
i ''' .
' :
ii'i
CM
i-rf
'\]
~ . \ ' ,
30
i"
MM2B ' i
1
^ 1
^i
; 1
,'' 1 '
1
1
;:
' ,
1
t
0)
10
:
3
00
H
{4
I
't
l{
II
I
1
!l
ii
CO
X u
! i ,.
<U
0
M
3
0
X
CM 00
rH H
1 HJ
0
HJ 0
PL4 Si
>
3
0
U X
0)
u ^
H CM
cd
U
^\ 0
; .
1 M 1 :
1',
0
H
U
Cd
H
TJ
Cd
u CO
u M
IMi M i f i i '
'
iiil liii M i i i
M
M
MM
M'
iiii M M ' i i i
MM
ii iMi i ; :M
M1
M ^
'' '
iMi i l i l
M
! j
'M
li ii Mi
in
iM'
1 , . ' 1 I M M M
M i ' . 1 M i 1IiM
M ' . i I , ' 1 1 ' .
'Mi M i '
^ ^
_' T ^^^^^^^,
i M 11 ' j :
Mil Il'i
fc 5
b
M ii'l
^TM
Ij IMiii
'li
1 . ! 7i
\
i :
: : i i | j | ':
iiiiM''
i 1 ^
iii i
I'M
1 ii Ml
IMMM'
11 i
till
:' i
i| 1
1
Mil
M l ,
1 iM M i i
Si
c *J
"Z. -H
'
IM^
'1
'
JO j ' 1 ' M 70 i M
;|"MTM';
M M 1 ; ,' ' ;i 1 ! '
Mil
1 1 1 ;
l i i i iiM i i i '
Mil
!
I'i'
iiii
ill! W il'i
1ilM M' iiii
iilii
1'
Miy
!!l M
! i ; ' t 1
ill!
^ ^ ^ 1
Ml
i
Iii
il
.
IM'
MM ' '
M::M'E
T . MiMT
i|M
1
M MM
'
M
iii; i'
M!
Mil i i i '
i !
ill!
iM 1
M:i
i iM
Ml
i ii'!
ij i i ^ ! II M l ! i i i i j M i
MM
Mil
iM I i i '
1 .
'Ml
iMi
': 1
iiii
il
1 M i
I ii i
Ml l i i i
1;
M il
1 ' 1
lMJ!
i | i |MM!i| M.!;
l ! IM 11 j l iMi
1 1 1
Ml!
III!
Ml
MM
Mi!
Mil
in
'
Mil
II
: TTTTi
' 1.
1 1 I
1 ' '
i'ii
M'
I ;
Ml:
i l l ' li
ii'i l i !
M
ii
1 M Mj
MI
! i i ! i II
1 |Mi IM
ll
^1
If i
:i
;i
ll
69
6
a
o
H
(0
Cd M
H 3
O
o
Cd aa
4J
VI
u 00
**
V4 u
(U o
4 J 4^
(4-1
Cd 0)
u
Hi u
3
d o
CO
c
3
4J
a^00
e H
o
CJ
Ok
CO
fH
I
I
4
a
1
il
il
HJ
OJ
X
4->
:i
I
X
o
K-3 4J
PU H
PC ?
*
CM
CM
0)
u
3
00
II
f
70
' 1
i
MI
I ' lM
M
1 : i 1 i1 1' ll III!
M ill!
Ml! i: M l 1i l i i i i i i i i! I i i l
iiil i
'
1 '
. M
i
1
Mi i | i i
|M iiii i
:ii i i i i
iM i l M 1
! i i i i '
Ml
MM 1
iii
(
i 1
M l
'
ill
'
1
1
i liii
1 i
il iiii
li
|1
11T
Hli
MM
1 M ;!
M
M
' 1 M
,1
i 1
' M
ii:
1 i M \M i
j Iiil
"T
i
1 .
'
; : .
'
'
I i
iiii
i|
MM
1 !
iii:
I l M
'
iiii
iiii
i M 1
ilil
i! M
'
i l l ' iMi
iii 1
M '
:'^U
i i
1 M
M '
M i l
i'l iiii ^
,1M
1' '
i M 1
i ! !
iiii
'Ml
HPl
i ' 60
M ' '
' i 1
III
MM
j!
!i
MM
ii i i i i
il
|ii
!i|! 1 l i l l !
1
1
1
1 !
Mli li
ii!!
M
1
' . 1
u
<u
: ' M
liii
4J
, 1
'
ilili!
iiii
1 ,
i ' i
iii^
IM
iiii
'ii
<
M l
>
'I tM '
li!
i!il
< I 1
; M i
M -
1 1
IiM
M I ! i M 1
\ ' '. t
i '
'
<
M ' ''
'\
;.
' 1
, 1
, i i
i Mi
CN
1 ^
_ ^
'
'1
30
J 1
3
00
H
' 1
10
20
"
(U
fa
t
'I
1
:i
:i
hJ
1
CJ
h J 4-1
P-I H
-I-"
M. 1
; 1
TH
Xi
<
r H 4-1
fs
*0
li 1
CJ
i'
' i
0)
CJ
;;
M
M' 1
in
']
50
U
O
14-1
I
1 i i i 1
1
1
i ;
1 ;i
CN
r'.
3
13
O
C CO
3
O 4J
C U ^
E 00
1 M
M l
i M 1
MM
MM
M !
M
liii
H
4J CO
CO u
H
3
TS o
Cd DC
u
u
i '
- - V
'MMi li
1 1
1
M
i l ' i
lO
MM
. M
; i 7 0 1
1
i i
iiji li ii iiii
ill! ! 1
iiii
Mi'
M . I
i;M ;; i j i M i
' i i ; ; ;' i; I'M M M
80
MM
MM
90
11
1
iiii
ill!
ii! Ilil
Mdiih i M i i l | i i 1. ^ 1IiM
i! i
iiil
MM
: ii'l ii i 1
in
<4-l
1 M I1 ! '
-'
1 , :
iM
M i i ill;
In
^ * ^
i N i ii
MiM
1 1
:i
'^
FM
M
ii:
i M
M 1
1 , '
i , ,
M M
i 1 1 1 1Mil
:
!i M l
Ml'
i ll h
MMi;
;M :
iii.
i ''
MM 1 M
M 11 M ' 11
III |i
i M | Mii
M
", IM!
IM h !M ; i 1l :i 1l
1 !' 1'
M i l l i iiii jlM
I'll
' 1 M
'
'
iiii
iii'
I'll i ! ! 1
I'll
M'i
i 1 i 1M i i | i
1 i i l i i i ii ill'
Mil
liii
Mil
'M i 1! 1iTTTh
il iii i i: i I ; ;
'Ml i i
1 1 1 '
1 1 Illi 1 ii ! ' ! ! Ml Mi iiM
i 1
iii! ' i ' l - 1 l i i i i i iiM 1! MM ! iiI 1 i!
M MM
111 M! ii , M ! ii'l 1 , 1 :
iiii
:
iiii
ii|
1 ! 1
ii11 iii Mi
II ilM liii
! MM
iii'
'iM
M i"
Ml MM M ll M l
Ml MM ii : - i M ;
M i l
i Ml
ijli
i ! 1 MM
MM
! 11 i
Ml
i li!
1 i|i i
'
71
fO
OJ
rH
a.
e
Cd
CO
<4-l
o
B
cd
u
00
o
.1-1
cd
o
u
Si
CJ
in
rH
I
CJ
HJ
Pu
CN
u
3
00
H
fa
I
1!
>i
72
.u
a,
a
oO
c
H
cu
CQ
^
(U
4-1
*4-l
CO
3
O
- l l
CO
(1) CN
rH r^
a
B
U
CO o
U-l
CO
M
vO
rH
CO
1 O
CJ
hJ
CD
PL.
_^
4-1
CN
0)
u
3
oc
H
fa
73
3
O
H
4.J
CO
H
13 CO
CO u
)-i
3
u o
(1
}-l CN
(U
4J
14H
fO
r^
VJ
O
'-^
(U
(U a
rH V4
CL 3
B O
CO CO
CO
4-1
^X
r^ c:0
H
1 iJ
CJ
QJ
HJ
PI ^
33 4-1
iH
CN
(U
}-i
3
00
I
I
E
I
I
I
I
I
I
74
(U
3
O
00
Xi
-3
<
rH
U CJ
OJ X
4-1
m
d
^3
OJ
4-1
ro
CO
U
CD 4-1
rH 3
a O(1)
E
CO 3
CO O
CJ
M-l
00
o
a.
HJ
o
pLi
u
Q
B
iH
CJ
CN
<U
U
3
00
fa
APPENDIX B
I
I
75
76
CL
Q.
--CM
a
CO
u
4-)
-i-ro
X
Cd
-3
O
O
:s
4J
i-i
^-^
H
3
cr
0)
-in
0)
-3
3
>-i
CJ
J-UD
p-i
00
(1)
3
00
H
fa
I
oO
a:
-1
ICPi
:i
.
^1
77
E
CL
-KNI
-<^
O
4J
3 O
O CO
H M
4-1 4J
O >4
CO fa
'
*^
-^-J fa - 3
O
U
O
CO
-u")
iH
4J
0
Pi
1
5-1
CO
OJ
3 X
O
2: a)
4.)
> -H
CN 3
cr
Oi
CO
S a;
-<D
CJN
CN
(U
M
3
'C^
00
fa
cr.
-OO
Q.
-cn
lo
78
'I
I Q.
(f) >
^CNI
OJ
4-)
cr
CO
(U
2
U-l
o
3
4-
O
H
4-)
4-t
cd o
)-i
CO
fa
}-i
4J
u X
in
CO cia
rH
O -3
Pui o
Ok
o
5
CO 4J
(ai CO
LOD
PLI
ro
i)
>-i
-r^
rO
I
CO
-a
.o
3
OC
0)
33
79
-Q.
a.
-U_
-1
3
3
O
a
B
o
U.
CJ
E
3
U
4-1
o
ex
CO
on
P-.
1
Pi
Pu
CO
0)
V4
3
00
X
o
o
I
I _^
80
E
Q.
rCM
cu
u
*oo
cr
CO
0)
-i
4-)
O
4-
(U a
cu CO
CO
4J
.in
Pi X
S w
*
in
I
Di
2
-CD
fa CO
CM
CO
o
}-l
;c^
3
00
H
fa
in
cr
-co
QL
CJ>
-3
o
o
5
CU
CO
81
i-E
-CL
U
d)
5 .Hi
fa]
'3
rH 3
>^ 3
rH O
H cu
cn B
r-\
Si
4-> M-l
0) o
B
H CU
V4 >
H -H
4->
> CO
v>
>
-H
'-'
2
0)
Cu Q
CO
ro
(U
M
3
00
H
fa
82
:E
Q.
CL
i A
1
'm
_i
: J
1 -1
T3
3
3
O
CU
B
o
CJ
; -\
14-1
;^ -1
1 ~
(U
>
4-1
CO
-(
i]
di
i M
1 tn
>
tU
4J
CO
1_
11 1
1
1 o
' "
: -
0)
a
<
M
r^
1
Pu
r*
'
4J
<r
fO
a
p
3
00
H
, ~
"
fa
OO
I -
X
o
83
:E
Q.
-U.
(Nl
ir><i
'
1
]
- 1
4-)
_i
(U
00
CO
-J
1H
(NLL-
t rn
*
J
,
-^
-1
1
i
4-1
~i
-1
Pd
4J
M-l
H
J3
CO
3
3
O
CU
B
M-l
o a
3 <4-l
O O
0)
>
T3 H
- 3 4-1
<
4-1
CD
U
rvi
cd
>
H
}-l
0)
Q
>
CO
I
Pi
(U
4J
cd
4->
0)
Pi
<
CM O
m
ro
fSl
(U
>u
3
CM CD
00
84
E
CL
(U
00
CO rH
0)
&i TJ
3
4-1 3
M-l O
H CU
CO
e
o
CJ
M-l
o
3
O
H
4-1
H
-3
"3
<
T)
M-4
O
0)
>
H
4-1
CO
>
H
V^
OJ
C5
O
O (U
QJ 4J
CO CO
4-1
A
OJ
CJN u
1 <
pli
2 o
Pu 4->
r>.
v ^
OJ
u
3
00
H
fa
CL
i:
' X
''
85
-CL
_,
! J
1 H
CL
X
o
86
_i
-1
iJ
0)
00
(U rHl
3
3
O
4J
M-l
H
-^
CO
CJ
M-l
O
(4-1
3
O
H
O
<U
4J
>
H
^3
-3
-H
4-
CO
Si
}-l
<
>
4J
U Q
3
O
fa
0)
4->
CO
rH
rH
I
OCJ
OJ
O
<
Pu
iJ
2 o
CO
ro
Qi
U
3
00
fa
cn
X
o
- o
87
i:|
4-1
OJ
00
CO
(U
rHl
ttS T )
3
4J 3
O
CU
M-l
CO
o
CJ
uu
MU
O
H
(U
>
4-1 1
H iJ
CO
T3
-3
<:
>
r^
H
M
0)
u Q
uu
H
OJ
r~
4-1
CO
9k
4->
0)
rg
rH
1 <
oi
Pi
OS
ro
<U
U
3
Ot
X
O
4J
88
;E
:cL
1 n
gen
4-1
' H
1 r^
, ^
' -1
1 _j
-4
1 -;
1 ~
i
_
,
,
B
o
CJ
M-l
O M-l
O
H
4J
H
-O
tn
3
O
CU
- 1
-7
4J
M-i
H
CO
1
- .
CO r H |
(U
Pi - 3
1
-I
OJ
>
-H
4-1
CO
-3
<
x: (u
4J
><;
_
' _,
.S3
, -~
-^
CU
CO
4-1
CO
^
I
o
o
o
cr
a.
4-1
ro
0)
rH
1
O
<
o
Pu
o
ro
1
4
>
-H
M
Su
3
00
H
fa
*J
APPENDIX C
Infrared Spectra
89
90
-oJ
lir!
Cd 0)
! -
4J -H
*J
>-i
"i ^
i :
X
w
M5
1^2
3
3*
CO
(y
M i
1
O
o
> "^H
j ' 5 o
X
- ' ? .1 u
cd ^3
1
i
1 0) o
' 3S -H
*J
1 1 0) o
' I 4-> Cd
M
-H M
M
: '
3 fi< 4 J
U
cr
h
Cd
<u td
CO
u
M 1
i 2 -
O X
0) Pi Cd
TJ 1
'
i t 3 CS 3
1 1
MO O
'I 11 CJ Z O
>
5 1
3
,
i
-:
1
l
1
0)
V4
00
.1
r ) r"vu3-.Yjii
4J
iH CM -l
Cd
l
pel pd 0)
M M 3C
91
(U
cr
CO
0)
2
3
o
H
4-1 4J
o o
cd cd
cd
M -3
O O
PU O
!S
4J
PO >-l
I
Cd
Pi
tt)
M X
CM
<r
(U
M
3
00
H
fa
iUvr:^v.M.^n
92
NOISSIWSNVlil iN33a3d
IT.
TJ
3
3
O
CU
o
CJ
u
u
o
d)
cu
CO
ai
I
Pi
<r
<u
u
3
00
H
fa
93
1-
J 3
-.-r.-
ma*
.-
4J
H
3
cr
CO
<u
2
__ - ~ 1
I
i
- -i
..
.::" i "
3
u u
u u
0
tt)
cd
M
CO
a u
pc5
M
73
O
* O
m >
1 a
CKi Cd
M
CO
0)
M
00
( I .', ..ur.'.NYii
94
0153''''; *''liJ. I N s - ^ i o
-;
0)
>
CO
>
H
M
0)
C3
u
QJ
4J
fa
CO
rH
X rH|
4J
(U ^ 3
e 3
H
U
H
3
O
^
> o
vD CJ
I
ai M-l
M
O
m
<r
cu
M
3
00
H
fa
95
'3
3
3
O
a
B
o
u
(U
>
H
4J
>
H
0)
Q
(U
4-1
CO
4J
(U
<
0)
}-i
3
00
APPENDIX D
Ultraviolet Spectra
96
97
-3
6
3
O
4J
0)
3.
CO
tt)
M
3
00
98
3
O
H
U
CO
H
-3
CO
U
cn
u
3
O
33
>3CM
M-l
d)
4-1
M-l
<u
o
VJ
CO 3
rH CO
-3
3
3
4.J
l^-t
00
o H
CU H J
g
o CU
CJ
X
4J
CM
>
D
00
-a<u
u
3
00
X
4-t
99
CO
Cd
H
pa
O )-i
H 3
4J O
- 3 00
-*
Cd
>-l
u u
o
uu
u
tt) tt)
4J
(4-1
u
ctl 3
o
^\ CO
4J
3 Si
3 00
O H
CUhJ
o tt)
CJ j r
Ok
CO <-.
1 u
> H
X 5
CJ>
-.*
tt)
u
3
00
100
CO
o u
H
3
4J O
cd 33
H
" 3 CM
cd r*
u
U M
M O
(4-1
tt) tt)
*J
o
u-l M
CO
3
, O
M|CO
^3 4-
3 JS
3 00
O -H
CU H-)
g
0 tt)
CJ J3
4-)
sf J3
1 4J
> -H
D 5
O
uo
tt)
3
00
101
4-)
3
(U
4-1
CO
a;
}U
H
TS
H
O
<
-3
3
CO
0)
cn 0)
CO r H
pQ
cu
B
m
> CO
CO
I
> uu
s o
m
tt)
>u
3
00
H
fa
102
tt)
M
CU
CO
4J
3
tt)
E
4J
CO
tt)
V4
H
<
I
>
3
CM
m
tt)
3
00
103
tt)
M
I
CO
14-4
o
4-1
3
tt>
B
u
cd
tt)
u
H
tt)
CO
CO
PQ
I
>
X
ro
m
tt)
3
00