close

Вход

Забыли?

вход по аккаунту

?

A STUDY OF CERTAIN FACTORS INFLUENCING THE CHEMICAL COMPOSITION OF FLUE-CURED CIGARETTE LEAF TOBACCO

код для вставкиСкачать
The Pennsylvania State College
The Graduate School
Department of Agricultural and Biological Chemistry
A Study of Certain Factors
Influencing the Chemical Composition
of Flue-cured Cigarette Leaf Tobacco
A Dissertation
by
William Lyon Porter
Submitted in partial fulfillment
of the requirements for the degree of
Doctor of Philosophy
August, 1940
Approved:
CjpJlp 3-1
, 194Q>
Professor of Soil and Phytochemistry
7,
Head of the Department
Table of Contents
page
List of T a b l e s .............
iii
List of F i g u r e s .........................
I* Introduction . . . . .
.................
II. Results of Previous Investigations
....
III. Description of Materials and Techniques
Employed ...............................
1. Materials
.....
2. T e c h n i q u e s ...............
1
3
6
6
15
IV. Presentation of the Results Obtained . . .
V. Discussion of the Results Obtained
v
20
. . . . 39
VI. S u m m a r y .........................
VII. Acknowledgments.......................50
VIII. Bibliography........................... 51
47
Hi
List of Tables
page
I. Description of the Hogshead Sanples of
the 1938 C r o p .....................
10
II. Fertilizer Treatments of the Experimental
Plots Stu d i e d...............
11
III. The Effect of Fertilizer Treatments on the
Yield and Value of the 1938 and ly39
Crops Produced on the Experimental
Plots Studied.....................
12
IV. Description of the Pure Culture Organisms
Studied from Commercial Flue-Cured
Tobacco........................
13
V. Description of the Organisms Isolated from
Tobacco Grown on the Chatham Plots. •
14
VI. Analysis of Flue-cured Tobacco Taken from
Hogsheads at Different Intervals of
T i m e ...............
23
VII. The Effect of Fertilizer Treatment on Cer­
tain Constituents of Cigarette Leaf
Tobacco Produced on the Experimental
Plots Studied in 1938. Best Quality.
24
VIII. The Effect of Fertilizer Treatment on Cer­
tain Constituents of Cigarette Leaf
Tobacco Produced on the Experimental
Plots Studied in 1939. Good Quality.
25
IX. The Effect of Fertilizer Treatment on Cer­
tain Constituents of Cigarette Leaf
Tobacco Produced on the Experimental
Plots Studied in 1939. Scrap Quality.
26
X. Milligrams of Steam Volatile Acids (as
Acetic Acid) Produced per 100 ml, of
Culture Solution by Pure Cultures of
Microorganisms Isolated from Cigarette
Leaf Tobacco . . . . . . . . . . . .
27
iv
LIat of Table8 (continued)
page
XI. Milligrams of Formic Acid Produced per
100 ml. of Culture Solution by Pure
Cultures of Microorganisms Isolated
from Cigarette Leaf Tobacco . . . . .
28
V
List of Figures
page
1. The Influence of Fertilizer Treatments on
the Distribution of Total Volatile Acids
in Cigarette Leaf Tobacco from the
1938 and 1939 Crop of the Experimental
Plots Studied* Calculated as mgms.
Acetic Acid per five gms...............
29
2. The Influence of Fertilizer Treatment on
the Distribution of Formic Acid in
Cigarette Leaf Tobacco from the 1938
and 1939 Crops of the Experimental
Plots Studied...................
3-5. The Production of Steam Volatile Acids by
Microorganisms Isolated from Commer­
cial Cigarette Leaf Tobacco. Cal­
culated as mgms. Acetic Acid per
100 m l ............. '.................. 31-33
6-10. Comparison of the Production of Steam
Volatile Acids by Microorganisms
Isolated from Cigarette Leaf Tobacco
from the Experimental Plots Studied.
Calculated as mgms. Acetic Acid per
100 m l ................................. 34-38
30
I. Introduction
Studies on the fermentation of oigar leaf tobacco
have established the fact that microorganisms play a
predominant role in this process and that the number and
kinds of organisms involved are related to the chemical
composition of the leaf.
A number of chemical changes involving the activity
of microorganisms may be followed quite readily during
fermentation, especially when the tobacco has been bulked.
Among th9se changes are those involving ammonia, protein
and non-protein nitrogen, fixed acids, and reducing
materials.
Studies on flue-cured cigarette leaf tobacco in
these laboratories have indicated that bacteria may play a
role in the aging process.
Moreover, as stated before, it
has been found that the chemical composition of the leaf
has a great deal to do with the nature of the organisms
present at any one time.
In order to establish a paral­
lelism between the fermentation of cigar leaf and the
aging of flue-cured cigarette leaf, the present series .of
investigations were undertaken.
These studies involve
(1) the chemical changes occurring in aging flue-cured
cigarette leaf tobaoco; (2) the chemical composition of
flue-cured tobacco as influenced by the field treatment
'it
2
and (3) the effect of certain types of bacteria on the
production of steam volatile acids including formic acid.
t
3
II. Results of Previous Investigations
The studies of tobacco fermentation have been
reported upon by many workers and different conclusions
drawn.
Reviews of these investigations are available in
the literature (20)(21)(23)•
The principal investigations,
however, have been conducted primarily on the fermentation
of air-cured tobaccos, such as cigar leaf.
The results
obtained from the flue-cured tobacco experiments have been
found to differ greatly from the experimental data on aircured tobacco (1)(10)(20).
Certain changes in many of the
chief constituents have been noted (27).
Changes in the
chemical composition of aging cigarette leaf tobaoco have
been reported by Darkis, et al (10)(29).
The aging
process has been ascribed to several causative agents,
among which are microorganisms (11)(20) (23), enzymes (25)
(26) (27), and the reaction between amino compounds and
sugars to form melanoidins (2).
Darkis, et al have
summarized the methods of harvesting and aging in a series
of papers (8 )(9)(10).
The chemical composition of the different flue-cured
types of tobacco vary to a great extent (10)(20).
Due to
this fact it has been reported that the methods of curing
must be varied (12)(15).
The relative quantities of the
different constituents with respect to the position of the
4
sample on the plant have heen discussed (6)(9).
However,
very little experimental data are available on the affect
of fertilization or field treatment on the chemical compo­
sition of cigarette leaf tobacco, although the literature
is voluminous on the work carried out with cigar leaf
types (3)(7)(13)(17)(19)(30).
Reports on the metabolism of steam volatile acids,
including formic acid, are scarce.
Formation of formic
and acetic acids from samples from the hogshead during
aging is mentioned by Darkis, et al but not discussed (10).
MoKinstry (27) investigating the fermentation of
cigar leaf tobacco found that an increase in steam volatile
organic acids is correlated with a rapid proliferation of
microflora, followed by a diminution of these organic
acids and the gradual decrease of the non-protein fraction
of the leaf with the evolution of ammonia.
His work sug­
gests that the formation of steam volatile organic acids
may result from the following chemical reactions:
(1) the deamination of amino acids; (2 ) the fermentation
of “gums" and pectic substances, and (3) the dissimilation
of the non-volatile organic acids of the leaf.
An hypothesis was suggested by McKinstry (27) to ex­
plain decreases in volatile organic acidity.
Since many
of the volatile organic acids are possible energy sources
5
for the microorganisms associated with cigar leaf fermenta­
tion, it is possible that these compounds, produced as
end-products of the fermentation of more complex leaf con­
stituents, may bo in turn utilised for the energy metabolism
of the microorganisms*
III.
Description of
Materials and Techniques Employed
1* Materials
The samples used were obtained from one of the large
tobacco companies*
These inoluded five samples, bought
on the open market and aged in hogsheads in the ordinary
commercial manner.
Descriptions of the samples and
intervals of sampling are recorded in Table I*
At
intervals during the first year of aging, borings of each
hogshead were made, and samples were sent to our labora­
tory*
These samples were dried, milled, and placed In
glass jars to await chemical and bacteriological analysis.
The samples for Section II were obtained from certain
fertilizer plots through the assistance of Mr. E. M,
Matthews, superintendent of the Chatham Tobacco Substation
of the Virginia Agricultural Experiment Station,
The
samples came from a series of eight plots, the treatments
of which were kept constant with the exception of the
potassium which increased from no potash on plot number 1
to 300 pounds per acre on plot number 8 ,
The fertilizer
treatments are summarized in Table II.
The crop produced excellent yields in 1938 and com­
manded a fair price on the market in spite of the fact
that it was grown under conditions of excessive rainfall.
7
For 10 weeks following planting, the rainfall was 50 per
cent greater than normal*
The 1939 crop was grown under
rather dry conditions and, on the whole, produced tobacco
inferior to the 1938 crop, as indicated by the crop value
(Table III).
Harris (18) reported the market value of the
1937 crop of tobacco grown on the Chatham plots, as selling
for a higher price than crops of the following two years.
In 1939, the middle primings were harvested following
a period unfavorable for mineral absorption.
Bottom and
top primings were removed from the stalks during or just
following periods favorable for mineral absorption.
The 1938 crop was grown under a fairly constant,
though high, moisture condition and the mineral absorption
was, therefore, fairly constant.
Representative samples were taken from the bottom,
middle, and top sections of the plants on each plot and
were taken immediately after the flue-curing process.
The samples were dried, milled, and placed in glass jars
for subsequent analysis.
The samples for section III were pure cultures of
organisms isolated from tobacco samples received for
analysis during the two years of the experiment.
Twelve
organisms were ohosen as representative of the commercial
tobacco isolations from the 1938 crop (Table IV).
Seven Isolations from the 1938 Chatham samples were chosen,
not necessarily as representative organisms, "but for com­
parison of the activity of the same type of organisms,
isolated from different qualities of tobacco, in the pro­
duction of steam volatile acidity and formic acid (Table V).
Thirteen organisms were chosen from the 1939 Chatham iso­
lations for comparison of activities (Table V).
These organisms were divided into different types
according to the following classification**
Class A. Typical yellow, Gram-negative organisms
found on most samples of tobacco.
These are subdivided on
their biochemical activities into four types, namely:
Type I. Those organisms that are weakly saccharolytio
(attacking only glucose) and slightly proteolytic
(liquifying gelatin but with slow action on milk casein).
Type II. Those that are intermediate in their re­
actions, being able to produce acids from glucose and
sucrose, attack gelatin more actively, and may or may not
digest casein within 14 days.
Type III. Those that are actively saccharolytic
(producing acid from glucose, sucrose, and lactose, attack
*From unpublished data of Mr. Joseph Naghski and Mr. Walton
Grundy of the Department of Bacteriology at The Pennsyl­
vania State College.
9
gelatin vigorously tout this proteolytic activity is not
apparent in litmus milk due to the sparing action of the
lactose; and,
Type XV. Those that produce acid and gas from car­
bohydrates, liquify gelatin slightly and sometimes pro­
duce sufficient acid in milk to precipitate the casein.
Class B. Includes all other organisms.
This class
is subdivided into three groups, as in the followings
Saccharolytic.
Those organisms showing greater
activity on the carbohydrates than on proteins (gelatin
and milk) •
Proteolytic. Thoso that attack proteins very readily
and may not show activity on the sugars (this includes
most of the spore formers.)
Inert. Those that do not act to any appreciable
extent on either sugars or proteins.
(This last group
Is composed chiefly of Gram-negative red bacteria.)
10
TABLE I
Description of the Hogshead Samples of the 1958 Crop*
i1
Warehouse
Sample Numbers
Georgia
230
235
240
289
South Carolina
231
236
241
290
Eastern N. Carolina
232
237
242
291
West Durham
233
238
243
292
West Reldsville
234
239
244
293
Date Received
Time of Aging (Days)
4/24,39 6/ 6/ 39
153
195
6/24/ 39 9/ 2$3*
213
309
*Tobacco was placed In the Hogsheads on November 20,1938.
11
TABLE II
Fertilizer Treatments
of the Experimental Tobacco Plots Studied*
Plot
No.
Row Treatment
.Side Dressing
of Ko0, Pounds
Pounds
per acre**
per acre Formula
Total
in Treatment,
Pounds per
acre***
1
1000
3-8-0
0
0
2
1000
3-8-3
0
30
3
1000
3-8-6
0
60
4
1000
3—8—6
30
90
5
1000
3-8-6
60
120
6
1000
3-8-6
100
160
7
1000
3-8-6
140
200
8
1000
3—8—6
240
300
Data presented in this table were obtained from Mr*
E, M. Matthews, superintendent of the Chatham Substation
of the Virginia Agricultural Experiment Station* Sup­
plementary treatments involved the application of mag­
nesium limestone at the rate of 200 pounds per acre plus
30 pounds of nitrogen and 80 pounds of P2O5 per acre.
**Side Dressings were applied on either side of tbe row,
about 8 inches from the plants, three weeks after trans­
planting.
***Two per cent of the KgO was applied as muriate, the
remainder as the sulfate.
12
TABLE III
The Effect of Fertilizer Treatments on the
Yield and Value of the 1938 and 1959 Crops Produced
on the Experimental Plots Studied*
1938 CROP
Plot No.
1
2
3
4
5
6
7
8
Yield
per acre
pounds
758
1096
1168
1192
1188
1222
1198
1228
Value
per acre
dollars
.
Price
per pound
cents
140*20
218.08
239.70
254.35
233.63
241.09
222.74
237.53
18.5
19.9
20.5
21.3
19.7
19.7
18.6
19.3
120.40
150.80
155.60
162.00
147.60
156.40
143.60
144.80
13.6
13.8
14.0
14.7
13.6
14.0
12.5
13.5
1939 CROP
1
2
3
4
5
6
7
8
888
1096
1112
1104
1084
1120
1148
1076
*Data presented In this table were obtained from Mr*
E. M. Matthews, superintendent of the Chatham Substation
of the Virginia Agricultural Experiment Station#
13
TABLE IV
Description of the Pure Culture Organlsms Studied
from Commercial Flue-Cured Tobacco
Run No.
Culture No.
------- ---Type and Description
101
84-3
Smooth yellow, liqulfier
102
90-1
Smooth yellow, non-liqulfier
103
50-7-20
Rough yellow
104
54-3-37
B. vulgatus
105
49-8-37
Contaminant from control 7
— ns
Torula (red)
106
107
50-4R-20
White rough
108
50-19R-20
White smooth (gram +,short rod)
109
50-10R-20
White smooth (gram -,short rod)
110
40-S
White smooth (gram +,spore
forming rod)
111
56-A
White smooth (gram -, slow
grower)
112
50-A
E-ll(?)
14
TABLE V
Description of the Organisms Isolated from
Tobacco Grown on the Chatham Plots
Culture
Run No.
No.
115
117
118
121
122
204
208
482
541
616
114
75
222
519
119
120
4
41
209
456
211
7
123
124
113
114
116
201
206
207
210
69
449
196
152
267
10
1
24
137
Type
III
III
III
II
III*
III
III
I
I
Red
Inert
Red
Inert
IV
IV
I
I
I
I
Coccoid
Coccold
III
*Type III hut weak
Gram
Reaction
Leaf
Plot
Year
-
Middle
Middle
Top
Top
Bot tom
Middle
Bottom
8
1
1
2
2
4
1
1939
1939
1939
1939
1939
1939
1939-
-
Bottom
Top
1
.1
1939
1939
-
Bottom
8
1939
Bot tom
1
1939
Bottom
Bottom
2
3
4
8
1
1
1
5
3
1939
1939
1938
1938
1938
1938
we
mm
«*
••
•
mm
-
+
+
-
Middle
Middle
Middle
Bottom
Bottom
Top
Middle
1938
1938
15
2. Techniques
In so far as possible, accepted methods or official
methods of the Association of Official Agricultural
Chemists (4) were followed in this work*
In some cases,
certain modifications were made to increase the accuracy
of the method or to fit the size of sample which had to be
used.
For determinations not in the A, 0. A, C., the
“Standard Methods of Analysis” as used by the research
departments of tobacco companies were used.
For the determination of total volatile acids, a five
gram sample of milled tobacco was suspended In 100 ml. of
carbon dioxide free water and acidified with 5 ml. of 40
per cent tartaric acid.
This mixture was steam distilled
in such a manner that 500 ml* of distillate were obtained
in exactly 50 minutes.
The distillate was then heated to
95°C. and titrated with 0,02 N NaOH using Phenol Red as
the indicator.
The volatile acidity was calculated as
milligrams of acetic acid per five grams of tobacco.
A
blank was necessary and consisted of 100 ml. of carbon
dioxide free water.
The titrated solution from the volatile acid deter­
mination was utilized far the determinations of formic
acid.
The distillate was acidified with 0.1 NHC1 and then
made alkaline with a slight excess of CaCOg.
After evap­
oration to a small volume the excess of CaCOg was filtered
16
off and the filtrate was treated with the following re­
agents ; dilute HCl, sodium acetate solution and mercuric
chloride solution.
This mixture was refluxed for two
hours in a hoiling water hath, the flasks tightly
stoppered and placed in the ice box over night to crys­
tallize the resulting mercurous chloride.
The precipitate
was filtered through a tared Gooch crucible, dried for
30 minutes at 105°C. and weighed.
The weight of Hg2Cl2
was converted to formic acid and calculated as mgms. of
formic acid per five grams of tobacco.
Nicotine was determined according to the semi-micro
method of Avens and Pearce of the New York State
Agricultural Experiment Station (Geneva) (5).
The method for calcium was based on the A. 0. A. C.
method for the determination of this element in plants.
The method for the determination of potassium adopted
by Thomas (17) was used.
Total nitrogen was determined by means of the regular
Kjeldahl method, modified to include nitrogen in the
nitrate form.
Protein nitrogen was precipitated from
another sample with dilute acetic acid, the precipitate
filtered off and used in the regular Kjeldahl method for
protein nitrogen.
The difference between total nitrogen
and protein nitrogen was reported as non-protein nitrogen.
17
Moisture determinations were made according to one
of the methods of the tobacco companies, namely; drying
a five gram san*ple for two hours at 97.5°C. and calculating
the loss in weight as per cent of moisture.
This method
is open to criticism hut is as efficient as most of the
more rapid methods and our laboratory used this method
exclusively.
The reducing materials were determined according to
standard methods.
Five grams of milled tobacco were
placed in a 250 ml. volumetric flask and 175 ml. of water
added.
The suspension was placed in a water bath at 80°C.
for one hour and then allowed to stand over night.
This
suspension was clarified with lead, deleaded and filtered.
25 ml. aliquots of the resulting solution were analyzed
for reducing materials according to the method of
Quisimblng and Thomas (15).
The reducing materials were
calculated as dextrose, even though certain unpublished
data leads us to believe that only a fraction of the re­
ducing power is due to this compound.
All data were calculated as per oent on a moisturefree basis with the exception of total volatile acids,
including formic acid which were calculated as mgms. per
five grams of moisture-free tobacco.
18
In the culture work, 100 ml. of the culture solution
were acidified with tartaric acid and the aforementioned
method followed.
Controls on each set of media were run
and all the data then calculated to mgms. of acetic acid
(HAc) per 100 ml. of solution.
The formic acids were
calculated as mgms. per 100 ml. of solution.
The media used for this work were made according to
the following formula:
1. One pound of yeast was extracted with four liters
of distilled water and steamed in the Arnold for three
hours, the cells allowed to settle, the supernatant liquid
syphoned off, filtered through filter paper with the use
of suction and then through a Berkfeld filter.
2 . 300 grams of tobacco, finely ground, were ex­
tracted in the Arnold for one hour, with six liters of
distilled water, filtered hot through filter paper under
suction and cooled.
3. The following materials were dissolved in water
and made to five liters:
Glucose (tech.)
Asparagin
KgHPO^ (C.P.)
250 gms.
MgSO^
5 gras.
50 gms.
NaCl
5 gms.
12.5 gms.
KN03
25 gms.
4.
Two and one-half liters of yeast extract, five
liters each of the tobacco extract and the salt*-sugar
mixture were mixed and the resulting solution was adjusted
to pH 7.2.
After heating for one-half hour in the Arnold
and filtering, 12.5 liters of distilled water were added
and mixed.
500 ml. of this solution were placed in quart
milk bottles, plugged with cotton and sterilized for 30
minutes in the autoclave at 15 pounds pressure.
Inoculations of the pure cultures were made into 10
ml. nutrient broth tubes and allowed to grow for 48 hours.
At the eid of this period the entire broth suspension was
transferred to the quart bottles and allowed to grow for
the time studied.
One ml. sanples were removed to test
for contamination and 100 ml. samples were withdrawn for
analysis of steam volatile acidity including formic acid.
20
IV. Presentation of the Results Obtained
The samples from the aging experiments resulted In
the data shown in Table VI.
Again the results are re­
ported as moisture-free percentages with the exception of
the steam volatile acids and formic acid which are re­
ported as milligrams per five grams of moisture-free
tobacco.
The results of the chemical analysis of the Chatham
fertilizer plots samples studied, are shown In Table VII,
Table VIII and Table IX.
All results are given in per cent
on a moisture-free basis with the exception of the steam
volatile acids, Including formic acid, which are calcu­
lated as milligrams of the constituent per five grams of
moisture-free tobacco,
steam volatile acids are calcu­
lated as acetic acid.
Protein and non-protein nitrogen,
nicotine, steam volatile acids, formic acid and the fixed
acids were determined for plots 1, 4, and 8 only, because
of shortage of samples.
Fixed acids were run on the 1938
samples only.
Figure 1 presents graphically the trends of the steam
volatile acids due to the field treatments of the tobacco.
Figure 2 represents the same picture for the formic acid.
V
21
The culture work is reported in Tables X and XI.
All
results have been calculated as mgms. of steam volatile
acids, as acetic acid, per 100 ml. of culture solution and
the formic acid as milligrams of formic acid per 100 ml.
of culture solution.
Since the controls on the uninocu-
lated media varied to a slight extent, all the data was
recalculated to a control value of 7.00 mgms* HAc/100 ml.,
since this figure was the average of all the controls.
Figures 3 to 10, inclusive, present graphically the
trend of the production of steam volatile acidity, cal­
culated as acetic acid, at intervals over a period of two
weeks.
In inspecting these graphs, It must be noted that
production of acids was such that with some organisms, a
maximum of 40 mgms. HAc/100 ml. was ample.
However, some
of the results were as high as 84 mgms. HAc/100 ml. and
It was, therefore, necessary to use smaller units.
Samples 115, 117, 118, 122, 204, and 208 show the
comparison of the production of steam volatile acids by
the same type organism (III) Isolated from tobacco having
different chemical composition and, therefore, different
qualities.
Samples 113, 114, 116, and 201 show the same
comparison for Type I organisms.
The samples numbered
115, 121, 119, 123, and 209 show the relative degree of
steam volatile acidity production by organisms of
22
different types.
Samples 101 to 112, inclusive, are
representative types of organisms isolated from commercial
tobaccos; however, all types have not been included.
Table VI.
Sample
No.
%
Total
N
Analysis of Flue-Cured Tobacco Taken from Hogsheads at Different Intervals of Time.
f> NonProtein
N
% NonProtein
N
%
K
%
Ca
1°
Red.
Sugar
Nicotine
%
Nicotine
N
mgms.
T.V.A./
5 gms.
mgms.
hcooe/
n/ k
5 gms.
230
235
240
289
1.53
1.63
1.55
1.58
0.66
0.67
0.68
0.67
0.87
0.96
0.87
0.91
1.38
1.56
1.47
1.78
2.45
2.36
2.36
2.31
24.46
21.15
24.75
20.91
2.29
1.95
2.19
2.04
0.39
0.34
0.38
0.35
7.37
6.60
10.09
9.11
2.39
2.35
2.14
2.61
1.11
1.04
1.05
0.89
231
236
241
-290
1.55
1.59
1.48
1.58
0.66
0.68
0.66
0.69
0.89
0.91
0.82
0.95
2.34
2.41
2.37
2.45
2.08
2.03
2.09
2.03
23.62
22.76
23.75
20.94
1.69
1.61
1.62
1.69
0.29
0.28
0.28
0.29
7.37
7.85
11.06
9.32
2.49
2.36
3.37
2.51
0.68
0.66
0.62
0.64
232
237
242
291
1.53
1.57
1.55
1.63
0.68
0.68
0.73
0.73
0.85
0.89
0.82
0.90
2.69
2.80
2.83
2.65
2.53
2.41
2.49
2.50
23.96
20.59
22.22
19.94
1.50
1.39
1.41
1.55
0.26
0.24
0.24
0.27
7.45
7.33
10.22
9.12
2.70
2.26
2.46
2.56
0.57
0.56
0.55
0.62
233
238
243
292
1.49
1.58
1.46
1.65
0.68
0.70
0.74
0.71
0.81
0.88
0.72
0.94
2.25
2.51
2.55
2.55
2.01
2.13
2.32
2.21
25.07
22.48
23.65
21.08
1.40
1.49
1.33
1.45
0.24
0.26
0.23
0.25
7.35
7.65
9.95
9.44
2.58
2.35
1.90
2.48
0.66
0.63
0.55
0.65
234
239
244
293
1.89
1.85
1.99
1.86
0.75
0.74
0.82
0.75
1.14
0.96
1.17
1.11
2.48
2.81
2.33
2.68
2.46
2.35
2.67
2.23
20.79
21.19
21.09
19.07
2.19
2.04
2.34
2.06
0.38
0.35
0.40
0.36
7.29
7.54
11.05
10.25
2.68
2.33
3.00
2.57
0.76
0.66
0.85
0.69
All results on moisture-free basis
Table VII#
Plot
No.
Leaf
1
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
TOP
Bottom
Middle
Top
2
3
4
5
6
7
8
T.N.
1°
1.97
2.60
3.22
1.79
2.11
2.20
1.71
1.95
2.31
1.59
2.07
2.15
1.63
2.09
2.35
1.71
1.86
2.32
1.77
1.98
2.21
1.80
1.85
2.20
The Effect of Fertilizer Treatment on Certain Constituents of Cigarette' Leaf Tobacco
Produced on the Experimental Plot Studied in 1938. Best Quality#
K
0.91
1.14
1.35
1.18
1.26
1.92
1.91
1.93
2.00
2.20
2.17
2;n
2.46
2.29
2.59
2.68
2.44
2.71
3.18
2.81
2.78
3.24
2.72
2.74
Red.
Mtrls.
1°
22.47
13.48
5.87
23.65
20.06
14.76
24.03
21.83
15.43
27.94
22.18
18.60
27.69
24.45
14.18
28.42
22.63
13.35
24.93
20.89
18.62
22.40
25.29
18.19
P.N.
i
0.96
1.08
1.21
N.P.N.
1°
1.01
1.52
2.01
1.56
3.06
4.70
Nic.
N.
$
0.27
0.53
0.82
0.90
1.03
1.06
0.69
1.04
1.09
0.96
2.01
2.86
0.17
0.35
0.49
Nic.
i
4.68
5.19
5.82
2.70
2.88
3.09
Ca
i
2.76
3.24
3.79
6.19
6.46
7.16
2.39
2.25
2.37
1.88
2.21
2.47
T.V.A.*
H£00H*
Oxalic Citric Malic
Acid
Acid Acid
i
1o
i
0.34
0.31 5.56
0.60 5.23
0.67
1.50
0.87 6.44
0.20
0.24
0.40
n/ k
2.16
2.28
2.38
0.13
0.16
0.33
4.41
2.95
3.31
0.72
0.95
1.02
0.43
0.23
0.43
2.70
1.48
1.79
0.56
0.68
0.80
•
0.99
0.97
1.04
0.81
0.88
1.16
1.01
1.54
2.57
0.17
0.27
0.43
6.38
6.94
6.97
2.40
2.46
2.27
1.84
2.04
2.11
0.22
0.05
0.22
T.N. - Total Nitrogen; K - Potassium; Red. Mtrls. - Reducing Materials; P.N. - Protein Nitrogen;
N.P.N. - Non-protein Nitrogen; Nic. - Nicotine; Nic. N. - Nicotine Nitrogen; T.V.A. - Total Volatile Acids;
HCOOH - Formic Acid; .Ca - Calcium.
Table VIII.
Plot
No.
1
2
3
4
5
6
7
8
Leaf
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
The Effect of Fertilizer Treatment on Certain Constituents of Cigarette Leaf Tobacco
Produced on the Experimental Plots Studied in 1939* Good Quality.
K
%
T.N.
£
—
2.12
2.74
2.10
2.05
2.20
2.06
1.84
2.31
2.21
1.93
2.27
2.05
1.91
2.36
2.09
1.89
2.21
1.95
1.92
2.33
1.88
1.86
2.21
0.73
1.29
1.39
1.46
1.51
1.82
1.75
2.15
2.91
2.43
2.57
3.70
3.05
3.48
3.82
2.99
2.65
3.80
3.50
3.38
4.08
3.44
3.54
Red
liferIs
£
N.P.N.
1°
0.89
1.02
1.23
1.72
o .40
4.73
0.85
0.94
1.36
1.13
1.33
0.78
0.82
0.94
1.10
1.04
1.27
—
— -
14.00
6.54
14.15
17.21
12.46
12.08
17.75
9.69
13.70
22.58
11.02
12.24
20.09
8.93
13.21
21.50
13.60
14.70
20.62
10.17
16.85
20.97
9.59
Nic.
i
P.N,
fo
0 . 0 0
NiCo
N.
fo
—
T.V.A. *
—
HCOOH*
—
0.59
0.82
6.24
5.65
3.22
1.72
2.20
2.69
3.24
0.38
0.46
0.56
7.06
8.35
8.63
3.43
3.70
3.10
1.81
2.42
2.95
0.31
0.42
0.51
6.84
7.35
7.01
2.69
3.40
2.80
n/k
mmrnm
2.95
2.12
1.51
1.40
1.46
1.13
1.05
1.07
0.76
0.79
0.88
0.50
0.63
0.68
0.55
0.63
0.83
0.51
0.55
0.69
0.46
0.54
0.67
T.N. - Total Nitrogen; K - Potassium; Red. Mtrls. - Reducing Materials; P.N. - Protein Nitrogen;
N.P.N. - Non-protein Nitrogen; Nic. - Nicotine; Nic. N. - Nicotine Nitrogen; T.V.A. - Total Volatile
Acids; HCOOH - Formic Acid; Ca - Calcium.
* mgms./5 gms.
t
o1
CJ
Table IX.
Plot
No.
1
2
3
4
5
6
7
8
Leaf
The Effect of Fertilizer Treatment on Certain Constituents of Cigarette Leaf Tobacco
Produced on the Experimental Plots Studied in 1939. Scrap Quality.
T.N.
io
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
Bottom
Middle
Top
2.37
2.13
3.12
2.27
1.93
2.44
2.28
1.92
2.54
2.39
1.72
2.23
2.16
1.98
2.24
2.42
1.90
2.41
2.34
1.96
2.39
2.23
1.72
2.36
Red.
Mtrls.
K
3
. i
0.61
6.70
12.48
1.09
3.70
1.32
7.57
1.27
17.22
1.35
9.12
1.61
5.54
1.97
14.39
1.89
7.49
2.44
5.81
3.08
2.44
21.05
9.86
2.55
3.33
7.83
3.17
16.31
3.44
7.81
4.30
5.07
3.30
18.05
9.46
3.22
4.53
7.72
3.34
16.74
3.50
8.35
4.17
8.03
3.58
17.04
7.08
3.48
.
.
P.N.
_
N.P.N.
Nic.
Nic.
N.
T.V.A.*
HCOOH*
n /k
.. I.....
— J° ....
0.93
0.91
1.27
1.44
1.22
2.05
2.41
3.36
4.51
0.42
0.58
0.79
5.28
5.92
5.57
2.69
3.52
2.95
0.90
0.76
0.91
1.49
0.96
1.32
2.07
2.30
3.28
0.36
0.40
0.57
5.79
8.00
9.50
3.45
2.54
3.02
0.85
0.73
0.92
1.38
0.99
1.44
1.98
2.37
2.14
0.34
0.41
0.54
6.41
6.95
6.93
2.98
3.75
3.01
3.88
1.96
2.36
1.79
1.43
1.52
1.16
1.02
1.04
0.78
0.71
0.88
0.65
0.66
0.65
0.56
0.58
0.75
0.52
0.59
0.68
0.54
0.48
0.68
T.N. - Total Nitrogen; K - Potassium ; Red. Mtrls. - Reducing Materials; P.N. - Protein Nitrogen;
N.P.N. - Non-protein Nitrogen; Nic. - Nicotine; Nic. N. - Nicotine Nitrogen; T.V.A. - Total Volatile
Acids; HCOOH - Formic Acid; Ca - Calcium.
* mgas./5 gms
to
o>
27
ble
me
iDay
Milligrams of Steam Volatile Acids (as Acetic Acid) Produoed
per 100 ml. of Culture Solution by Pure Cultures of Micro­
organisms Isolated from Cigarette-Leaf Tobaooo.
0
1
2
3
4
7
14
am p!
Mgms. Aoetio Acid/100 ml.
iumbe
101
102
112
7.00
7.00
7.00
7.00
7.00
7.00
7.00
7.00
7.00
7.00
7.00
7.00
5.89
5.94
5.76
36.13
30.51
5.62
7.89
12.95
37.99
30.12
7.89
13.49
6.49
5.58
5.73
49.62
36.98
3.81
14.93
22.21
46.09
44.25
10.41
17.57
7.17
6.40
6.74
55.43
41.85
9.26
19.88
27.15
40,0?
81.54
12.45
15.13
7.42
7.14
5.43
58.57
43.54
19.85
15.06
30.84
44,09
27.44
13.59
16.23
16.39
12.17
17.26
55.24
84.67
13.83
31.71
28.04
38.15
22.46
11.41
30.85
3.87
12.50
31.84
49.60
18.26
9.08
26.31
22.49
41,08
19.41
9.66
32.97
115
117
118
121
122
204
205
7.00
7.00
7,00
7.00
7.00
7.00
7.00
5.78
9.86
10.50
10.66
9.72
6.00
5.52
11.30
15.10
12.02
16.90
10.78
13.50
8.90
14.47
18.69
15.70
17.30
10.66
15.75
8.72
16,94
20.10
21.94
18.74
10.20
17.25
13.25
27.91
22.15
30.74
22.70
8.76
23.75
14.95
26.37
25.24
33.22
23.20
8157
28.50
16.25
119
120
7.00
7.00
8.40
7.74
9.52
7.45
8.74
7.62
9.94
7.22
8.39
6.66
7.20
6.98
209
211
7.00
7 .00
7.49
8.02
7.79
6,53
9.44
4.12
9.60
2.92
6.74
2.95
6.50
2.60
123
124
7.00
7.00
36.76
22.36
50.48
35.50
64.98
48.80
56.72
41.76
52.16
89.92
48.01
32.00
113
114
116
201
206
207
7.00
7.00
7.00
7.00
7.00
7.00
7.00
6.86
7.21
5.62
7.74
7.49
9.08
14.48
6.33
5.45
8.58
7.13
6.90
8.34
15.90
5.01
. 5.01
14.02
6.99
7.86
16.32
18.82
5.80
4.13
10.65
7.99
7.76
7.86
16.07
6.20
6.12
7.54
8.43
7.00
12.66
17.32
6.28
6.36
4.42
8.64
6.98
28.58
13.50
103
104
105
106
107
108
109
110
111
no
Table XI.
Time
in Days
Sample
Number
Milligrams of Formic Acid Produced per 100 ml. of Culture
Solution by Pure Cultures of Microorganisms Isolated from
Cigarette-Leaf Tobaoco.
0
1
2
3
4
7
14
Mgms. HCOOH/lOO ml.
101
102
103
104
105
106
107
108
109
110
111
112
4.00
4.00
4.00
4.00
4.00
4.00
4.00
4.00
4.00
4.00
4.00
4.00
4.22
:3.77
3.69
3.02
4.58
2.52
2.97
4.01
2.17
4.01
2.35
0.82
3.62
2.48
2.36
1.90
3.67
1.69
3.07
3.43
2.43
1.22
2.08
0.96
1.77
3.86
2.89
1.72
3.98
2.70
4.11
2.64
2.70
1.82
0.91
1.77
3.13
4.15
2.00
1.42
3.98
3.08
3.64
2.57
2.46
1.73
0.83
1.65
2.99
6.72
1.33
1.34
2.29
6.11
1.69
0.91
2.45
1.55
0.86
1.43
2.65
5.55
1.92
1.66
2.47
5.82
1.42
0.71
2.44
1.33
0.83
1.34
115
117
118
121
122
204
205
4.00
4.00
4.00
4.00
4.00
4.00
4.00
2.30
4.93
2.55
2.61
2.55
2.62
2.50
6.45
8.73
4.39
5.28
1.91
2.69
4.00
8.65
10.44
7.28
7.33
2.55
4.67
6.25
9.13
11.20
11.08
10.25
2.75
3.65
7.15
5.20
12.71
14.28
11.32
2.22
6.65
7.00
4.60
12.77
14.32
11.50
2.24
6.55
6.10
119
120
4.00
4.00
0.62
0.46
0.69
0.46
0.67
0.50 •
0.65
0.55
0.65
0.56
0.55
0.59
209
211
4.00
4.00
0.20
0.18
0.22
0.23
0.21
0.23
0.20
0.24-
0.20
0.21
0.21
0.20
123
4.00
124 . 4.00
2.80
3.23
8.23
8.05
11.27
8.53
7.55
7.45
8.28
5.30
10.12
6.57
113
114
116
201
206
207
210
1.31
1.13
2.90
2.60
0o91
0.85
2.35
1.63
2.28
2.55
2.65
0.90
0.90
6.35
1.39
2.54
1175
1.95
0.85
0.92
7.65
3.74
2.63
0.31
1.82
0.88
0.91
8.00
0.61
2.24
0.30
1.70
0.90
0.85
5.40
0.71
3.16
0.27
1.25
0.93
0.85
4.20
4.00
4.00
4.00
4.00
4.00
4.00
4.00
¥%
oe
*uQ
0e
\u
I
1
<
<
1938
Plot No.
l93Q(good)
CHATHAM
l939(icrop)
SAMPLES
Figure 1
The influence of fertilizer treatments on the
distribution of total volatile acids in cigarette
leaf tobacco from the 1938 and 1939 crop of the
experimental plots studied. Calculated as mgms.
acetic acid per five gms.
5
c
0
0
1E3
\
\
x
E
<n 3
\
in
1
1
O
o
o
X
e2
8U
X
•
* T>
6
O'
2
I
4
P l o t No
1936
8
0
o
o
u
■/
X
*1 •
i
t
a*
2
1 4
8
P l o t No
19 3 9 (good)
CHATHAM
i
4
a
P l o t No
1939 (scro p)
SAM PLES
Figure 2
-The influence of fertiliser treatment on the
distribution of formic acid in cigarette leaf tobacco from the
1938 and 1939 crops of the experimental plots studied
CA
O
I
1
SO)
I
4G
u 40
lOt
5
6
Ti m
in
7
0
in Day*
60
c
o
o
4C
<
I
E
tr
2
103
T»a« in Day*
104
Tim * in Dojrt
, Figure 3
The production of steam volatile acids by microorganisms isolated from commercial cigarette leaf tobacco. Calculated
as mgms. acetic acid per 100 ml.
w
h
n>
aq
t
oO'
6C
106
106
*
* 1 4 -"s— 4— t— i— §—
TtM
*5—
n— a— n v
mi D«ri
40
60
60
c
S
8
* 4C
3
<
I
T
C
9
2
107
>06
Figure 4
The production of steam volatile acids by microorganisms
isolated from commercial cigarette leaf tobacco. (Coat’d.) Calculated
as mgms. acetic acid per 100 ml.
1
(
o
o
4
Z
■
a
to
OQ
«
4
u C
8
N
i
■
a
112
Figure 5
The production of steam volatile acids by microorganisms
isolated from commercial cigarette leaf tobacco. (Cont’d.) Calculated
as mgms. acetic acid per 100 ml.
30
a
o
o
\
4
X
I
a
117
40i
30
5 20
t2>
Ti*« in Ooyi
Figure 6
Comparison of the production of steam volatile acids
by microorganisms isolated from cigarette leaf tobacco from the
experimental plots studied
CA
40i
30
\ 20
206
>4
_Figure 7
Comparison of the production of steam volatile acids
by microorganisms isolated from cigarette leaf tobacco from the
experimental plots studied (Cont’d.)
w
cn
30
g
8
■?0
i.
1
2
Ti m
in Doy»
Figure 8
Comparison of the production of steam volatile acids
hy microorganisms isolated from cigarette leaf tobacco from the
experimental plots studied. (ContM.)
03
o>
40)
30
1
c
8
s
\
\
*
z
i
i
3
3
o
116
114
Ti»» in Day*
40]
20
0
2
3
4
5
6
7
Tina m 0oy»
6
9
10 11 12
13
14
Tin* ir» Day*
Figure 9
Comparison of the production of steam volatile acids
hy microorganisms isolated from cigarette leaf tobacco from the
experimental plots studied. (Cont»d.)
S 30
w 30
20
20
207
210
Tjac in Day*
60
123
T<at in Day*
Figure 10
Comparison of the production of steam volatile acids
by microorganisms isolated from cigarette leaf tobacco from the
experimental plots studied. (Conttd»)
03
oo
39
V. Discussion of the Results obtained
The investigation of flue-cured cigarette leaf aging
processes have been made on samples aged under commercial
conditions and not under controlled conditions in small
bulks.
Satisfactory sampling of hogsheads containing
tobacco obtained from many different lots is quite diffi­
cult; as will be noted later in this discussion the effect
of field treatment on the chemical composition of the
plant is relatively large; therefore, each lot will
probably be different from the others.
For this reason,
certain errors are likely to creep into all results ob­
tained in this manner.
Analytical data which are not er­
ratic could be obtained only if great care was used in
<|
placing the tobacco in the hogsheads and in removing the
samples for analysis.
Although the conventional methods
of sampling were employed in order to attempt to get a
representative sample, the tobacco was not placed in the
hogsheads in any specific manner and samples from several
portions of the hogsheads may not be truly representative
of the whole.
Relatively small changes were noted in the several
constituents, studied over a period of approximately eight
months of aging.
The inorganic materials show little in­
crease or decrease.
The nitrogen fraction remains
40
relatively constant.
However, as in samples 233-292, an
increase in total nitrogen Is made up of an increase in
both protein and non-protein nitrogen, not by an Increase
of one or the other.
Small changes, which may be due to
sampling errors, sometimes are made up of one or the other
nitrogen fractions, as in samples 230-289,
The nicotine content does show a tendency to decrease
but when the results were calculated to nicotine nitrogen,
constant results were obtained.
Since the nitrogen and
potassium contents remain fairly constant, the N/K ratio
shows no change upon aging.
The most significant changes occur in the steam
volatile acid content of the leaf.
During the spring
sweat the volatile acid content remains constant to a
certain extent.
However, upon redrying, this constituent
increases sharply and even though the quantity drops 96
days later, there is still a significant difference In the
quantity at that time and that of the original volatile
acid content.
The formic acid fraction of the steam volatile acid
content remained fairly constant during the eight months
aging period.
With this fact in mind, It was deemed necessary to
study the production of volatile acids by bacteria isolated
41
from cigarette leaf tobacco.
Therefore, the experimental
work described in the third section of this paper was
undertaken.
In order to complete the picture of this phase of the
experiment, our laboratory is planning to continue this
study in a detailed manner.
In discussing the results obtained with the field
treatment samples, it must be remembered that the moisture
relationships for the two years of the study were quite
different, thereby causing differences in the environ­
mental factors affecting the chemical composition of the
leaves.
For this reason the data obtained from the 19-39
crop is not as clear cut as that obtained from the 1938
crop of
tobacco grown on the experimental plots studied.
This is
true not only ofthe chemical data, but also of
the bacteriological data which will be reported at a later
date.
Gribblns (16) has reported upon the nitrogen, potas­
sium, and reducing materials relationships but has not
discussed the interrelationships of these constituents
with other constituents of the plants.
Schweizer (30) has stated that, in Virginia tobacco,
a decreasing sugar content is associated with increasing
nitrogen and nicotine contents.
This relationship is well
42
demonstrated by this data.
It may also he pointed out
that the increase is made up of increases of each of the
several nitrogen fractions and not by one or two of the
components making up the total nitrogen content.
Thus a
ratio between the nitrogen constituents is maintained.
The greater the quantity of potassium absorbed by
the plant, the smaller the amount of nicotine synthesized.
This fact is demonstrated by the relationships between
samples from plots 1, 4, and 8 . Samples from plot 1
contain more than twice as much nicotine nitrogen as do
those from plot 8 .
As the available potash Increases there is a corre­
sponding Increase in the steam volatile acid content of
the lower leaves.
This Increase is true, also, in the
middle and top leaves until the optimum potash avail­
ability is reached and then the quantity begins to level
off or even begins to decrease.
It Is interesting to note, that In each case the
tobacco obtaining the highest market price, contains a
higher quantity of formic acid than the other samples.
These samples received the optimum quantities of potash
and are represented by samples from plot 4.
also of the volatile acid content.
This is true
The leaves obtained from tne middle of the tobacco
plant are considered by the experts to be the best tobacco
and the quality decreases from the middle toward the top
and the bottom of the plant.
Darkis, et al consider the
sugar content to be th© indicator of quality,
however, we
consider the quality to be dependent, not only on the car­
bohydrate content, but on the relationship of the elements
and constituents present, along with the carbohydrates
present.
The position of the leaf on the plant has much
to do with its composition in relation to leaves on other
portions of the same plant.
For example, Tables VII, VIII,
and IX demonstrate the fact that there is a decided dif­
ference between the middle leaves and the top and bottom.
The increase of total nitrogen from the bottom to the top
of the plant is made up of smaller increases of protein
nitrogen and non-protein nitrogen.
Paralleling this in­
crease is, of course, an increase in the quantity of
nicotine in the top over the bottom of the plant.
The oxalic acid and citric acid percentages increase
directly with the height of the leaf on the plant.
This
is true also of malic acid where the potash availability
is low.
However, as the quantity of potassium absorbed
by the plant increases, the opposite fact is true.
The
malic acid content of the bottom leaves Is greater than
that of the higher leaves.
44
The relationships within the volatile acid contents
are identical with the oxalic and citric acids.
There is
an increase in volatile abids from the bottom to the top
of the plant.
It is interesting to note that when the
potash available is excessive, the differences between the
quantity of volatile acids in the middle leaves is almost
identical with that of the top leaves.
Changes in the
formic acid fraction of the volatile acids are very small,
indicating that the fertilizer treatment has little or no
effect on this constituent.
In conjunction with the discussion of the volatile
and non-volatile acids it might be well to note that the
quantities of potassium and calcium Increase in direct
ratio with the fixed and volatile acids.
Since the only definite change which could be detected
significantly in the hogshead aging samples was in the
content of total volatile acids, it was deemed necessary
to study the effect on this production of certain micro­
organisms which were known to be present on tobacco of
this type*
At the beginning of the experiment several organisms
were isolated from commercial cigarette leaf tobacco and
their production or utilization, as the case may be, of
the3e volatile acids were studied.
Figures 3 to 5,
45
inclusive, show that of the twelve organisms studied, we
had approximately three different types of curves.
The
spore forming rods produced large quantities of volatile
acids.
Certain Gram-negative rods produced volatile acids,
but in lesser quantities than the spore formers.
A third
group, which can be classed as inert, had little or no
effect on the volatile acid content of the medium.
With
this in mind, a further study of the production of vola­
tile acids was carried out with organisms, the type of
which were known.
The field treatment samples used in the
fertilizer studies were used in this part of the work*
Exhaustive Isolations from each of the samples from the
plots were made and each organism classed according to
the classification given on page 8 of this paper.
Comparison of Type I and Type III organisms Isolated
from tobaccos of different qualities and chemical composi­
tion show that when certain physiological adaptations were
isolated and their cultural characteristics determined,
the ability to produce steam volatile acids was the same
no matter under what conditions they were grown.
Although
organisms may have different shapes and forms which would
cause them to be classed as types upon isolation, when
studied In culture they are physiologically the same*
A comparison of ths different types of the genus,
Flavobacterium, shows that the production of steam volatile
46
acids is also a criterion of classification.
Each type,
no matter where it is isolated, yields a volatile acid pro­
duction curve of the same type.
However, when the coccoid
forms are studied a slightly different picture is noted.
Coccoids from had or scrap samples do not produce acids
to any great extent.
In this manner they resemble the
inert type of Flavobacterium.
Coccoid forms from fair to
good quality tobacco show good production of steam vola­
tile acids in which they resemble the Flavobacterium.
These facts are illustrated by curves 206 and 207 on
Figures 9 and 10.
Organism 206 was isolated from poor
quality tobacco from plot 1 and organism 207 came from
fair quality tobacco from plot 5.
From this data and from the curves, the conclusions
drawn by the bacteriologists helping in this study are
substantiated; namely, that certain organisms are char­
acteristic of good, medium, and poor quality tobacco.
From this study the following conclusions may be drawn:
(1) Red inert, Type I, and coccoidal forms of bacteria
are characteristic of poor quality tobacco; (2) Type II
and weak Type III organisms are characteristic of fair
quality tobacco; and (3) Type III and Type IV organisms
are characteristic of good quality tobacco.
47
VI. Summary
Studies have been made of the chemical changes
occurring In the aging of flue-cured cigarette leaf to­
bacco, the influence of Held treatment on the chemical
composition of flue-cured cigarette leaf tobacco, and the
production or utilization of steam volatile acids by bac­
teria Isolated from flue-cured cigarette leaf tobacco,
prom the results obtained, it may be concluded:
1. Of the changes occurring during the first eight
months of aging, those involving the steam volatile acid
content appear to be the most significant.
2. The formic acid fraction of the steam volatile
acids is constant during this period.
3. The greater the quantity of potassium absorbed by
the plant, the smaller the quantity of nicotine and other
nitrogen fractions synthesized and the greater the amount
of reducing materials synthesized.
This fact is true up
to the point where the optimum quantity of potassium has
been absorbed and then the relationship is not so close,
in fact the inverse relationship may be found to hold true.
4. Quality of tobacco cannot be measured by the car­
bohydrate content alone.
The relationships of the other
48
constituents to the carbohydrate content must be taken
into consideration,
5. The position of* the leaf on the plant has much to
do with its composition in relation to leaves on other
portions of the plant.
There is an increase in total
nitrogen and each of its fractions, steam volatile acids,
fixed acids, and calcium and a decrease in reducing
materials from the bottom to the top of the plant.
Potas­
sium increases in the same manner until an excess of
available potash is present at which time the bottom
leaves contain the larger quantity.
6 . Three types of organisms are found on commercial
cigarette leaf tobacco with respect to their production of
steam volatile acids, namely: (1) spore forming rods
having a great capacity for the production; (2) certain
Gram-negative rods which produce steam volatile aoids to
a lesser degree; and (3) inert organisms having little or
no effect on steam volatile acids.
7. When certain physiological adaptations of micro­
organisms are isolated and their cultural characteristics
determined, the ability to produce steam volatile acids
is the same no matter from what quality of tobacco they
were isolated.
49
8. Each typo of the genus Flavobacterium has a
characteristic curve for the production of steam volatile
acids and each type is different from the others in the
genus•
/
9, Certain organisms are characteristic of good,
medium, and poor quality tobacco and may be identified by
means of their production of steam volatile acids.
50
711. Acknowledgments
The author wishes to express his appreciation to
Dr. D. E. Haley and Dr. J. J» Reid for their interest
and constructive criticism during the progress of this
investigation and in the preparation of this manuscript.
To Mr. M. P. Grlbbins, many thanks for his help in
several of the determinations used in the data.
The author is also greatly indebted to Mr. Joseph
Naghski and Mr. Walton Grundy for their aid in the bac­
teriological work necessary for the completion of the
experimental work.
51
VIII. Bibliography
(1) Adrian!, P. T e, Ramos, C. G., and Isodro, R. A., (L933)
The chemical composition of cigarettes and
cigarette tobacco. Philippine J. Agr. 4:87-98.
(2) Ambler, J, A., (1929) The reaction between amino
acids and glucose. Ind. Eng. Chem. 21:47-50.
(3) Ames, J. W., and Boltz, G. E., (1915) Tobacco,
Influence of fertilizers on composition and
quality. Ohio Agr. Expt. Sta. Bull. 285.
(4) Association of Official -Agricultural Chemists.
(1935) Methods of Analysis.
(5) Avens, A. W., and Pearce, G. W., (1939) Silicotungstic Acid Determination of Nicotine. Errors
involved and a new technique for steam distilla­
tion of nicotine. Ind. Eng. Chem. Anal. Ed.
11:505-8.
(6) Bailey, C. P., and Petre, A. W., (1937) Modern
cigarette industry. Ind. Eng. Chem. 29:11-19.
(7) Beaumont, A. B*, and Snell, M. E., (1931) Field
experiments with tobacco. Mass. Agr. Expt. Sta.
Bull. 271.
(8) Darkis, P. R., Dixon, L. P., and Gross, P. M., (1935)
Flue-cured Tobacco. Ind. Eng. Chem. 27:1152-57.
(9) Darkis, P. R., Dixon, L. P., Wolf, P. A., and Gross,
P. M., (1936) Flue-cured Tobacco. Ind. Eng.
Chem. 28:1214-23.
(10) Dixon, L. P., Darkis, P. R., Wolf, P. A., Hall, J. A.,
Jones, E, P., and Gross, P. M., (1936) Fluecured tobacco. Ind. Eng. Chem. 28:180-9.
(11) Faitelowitz, A., (1927) The bacterial decomposition
of tobacco as leading to the formation of bases
in the presence of water. Bio. Chem. J, 21:
262-4.
(12) Garner, W. W., (1913) Tobacco curing.
Farmers Bull. 623.
TJ. S. D. A.
52
(13) Garner, W. W., Bacon, C. W., Bowling, J. D., and
Brown, D. E#, (1934) The nitrogen nutrition of
tobacco# U. S. L. A. Tech. Bull. 414:34.
(14) Garner, W. W., Bacon, C. W., and Bowling, J. D.
(1934) Cigar and Cigarette Tobaccos. Relation­
ship of production conditions to chemical and
physical characteristics. Ind. Eng. Chem. 26s
970-4.
(15) Garner, W. W., Bacon, C. W., and Foubert, C# L.
(1914) Research studies on the curing of leaf
tobacco. U. S. D. A. Bull. 79.
(16) Grlbbins, M. P., (1940) The distribution of potas­
sium In cigarette leaf tobacco and its sig­
nificance. Thesis, The Pennsylvania State
College Library.
(17) Haley, D. E., Longenecker, J. B., and Olson, 0.
(1931) Composition and quality of Pennsylvania
cigar leaf tobacco as related to fertilizer
treatment. Plant Phys. 6:177.
(18) Harris, R. G., Haley, D. E., and Reid, J, J. (1938)
The effect of fertilizer treatment on the type
of flora found on decomposing plant tissues.
Soil Scl. Prod., 1938, p. 183-186.
(19) Jenkins, E. H., (1892) Experiments in growing to­
bacco with different fertilizers. Conn, Agr.
Expt. Sta. Ann. Rep. 1-24.
(20) Johnson, J., (1934) Studies on the fermentation of
tobacco# J. Agr. Res. 49:137-60.
(21) Johnson, J., (1935) Investigation of cases of to­
bacco fermentation. Wis. Agr. Expt. Sta. Bull.
430 (Ann. Rept# 1933-34), 33-4.
(22) Jorgensen, A. P. C., (1925) "Microorganisms and
Fermentation." 5th Ed# rev. 1925,
(23) Kraybill, H. R., (1916) Some chemical changes in the
resweating of seedleaf tobacco. Ind. Eng# Chem#
8:336-9.
(24) Loew, 0., (1899) Curing and fermentation of cigar
leaf tobacco. U. S# D. A. Rept. 59.
53
(25) Loew, 0., (1900) Physiological studies of Connec­
ticut leaf tobacco. U. S. D. A, Rept. 65.
(26) Loew, 0., (1901) Catalase. A new enzyme of general
occurrence, with special reference to the
tobacco plant. TJ. S. D. A. Rept. 68.
(27) McKinstry, D. W., (1939) The relation of bacterial
activity to the disappearance of citric and
malic acids in the bulk fermentation of tobacco.
Doctor*s Dissertation. The Pennsylvania State
College Library.
(28) Quislmblng, P. A., and Thomas, A. W., (1921) Condi­
tions affecting the quantitative determination
of reducing sugars by Fehling*s solution.
Elimination of certain errors involved in cur­
rent methods. J. Am. Chem. Soo. 45:1503.
(29) Smirnov, A, I., Erigin, P. S., Drboglov, M. A., and
Mashvovzev, M. Th. (1928) Biochemical charac­
teristics of aging in leaves. Planta. Abt. E.,
Z. wiss, Biol. 6:687-764. (C.A. 23:2463)
(30) Schweizer, J. (1939) The Influence of fertilization
on the chemical and physiological properties of
tobacco. Ann. Rept. Tobacco Expt. Sta.,
Besocki, Java. Abs. in Better Crops with Plant
Pood. March, 1940, p. 36.
(31) Thomas, J. J., (1935) The absorption of nitrogen,
phosphorous, and potassium by Pennsylvania cigar
leaf tobacco as modified by environmental condi­
tions. Doctors Dissertation. The Pennsylvania
State College Library.
Документ
Категория
Без категории
Просмотров
0
Размер файла
2 108 Кб
Теги
sdewsdweddes
1/--страниц
Пожаловаться на содержимое документа