Product Design
Subject: Total Amino Acid Content of Tobacco
Abstract
States that the data on total amino acid content in tobacco blends was needed for two projects (E-134 and C-133). Reports that Analytical Biochemistry Labs determined the amino acid content by an analysis of the hydrolysate. Warns that with this method tryptophan is destroyed and possible reduction in amino acids by the slow reaction with carbonyl compounds during hydrolysis can occur. Presents levels of amino acids from the data broken down by cigarette brand; Kent, Marlboro, Vantage, True, and Zack. Includes an analysis for amino acids in burley, Turkish and flue-cured tobacco. States that high aspantic means high burley, high proline means high flue-cured, and high glutamine means high Turkish tobacco. Gives a general ratio of tobacco blend: if burley is increased, both flue-cured and Turkish must go down, unless flue-cured is increased dramatically relative to the Turkish. Finds a contradiction for Vantage and Marlboro brands. States that in both brands the asparlic acid and the proline levels are high, but in Vantage the glutamine is also high. Declares that these results are impossible unless Marlboro and Vantage are adding nitrogen containing additives to their tobacco. Investigates this premise by subtracting protein nitrogen and nicotine nitrogen from total nitrogen. Finds that nitrogen compounds are being added to Marlboro and Vantage, but not to Kent, True or Zack. Reveals that a patent search confirms that Philip Morris uses ammonium phosphate and Reynolds uses a Schiff base. Identifies a Schiff base as an aldehyde combined with a base such as ammonia. Anticipates the need to analyze tobaccos for Schiff bases, and surmises that liquid chromatography would be the most likely method to use.
User-Contributed Notes
Fields
- Author
- LEWIS, CI
- Recipient
- Schultz, Frederick J., Ph.D. (VP of Lorillard, Inc. '89-95)
Modification of tobacco products through use of additives and measuring effects on dependence, behavior, and toxicity.
Modification of tobacco products through changes in tobacco processing and use of blends, and measuring effects on dependence, behavior, and toxicity.
see also: Ammonium bicarbonate, Ammonium carbonate, Ammonium chloride, Ammonium hydroxide, Ammonium sulfide, Diammonium phosphate, and Urea
Philip Morris & Co. Ltd.., was incorporated in New York in April of 1902; half the shares were held by the parent company in London, and the balance by its U.S. distributor and his American associate. Its overall sales in 1903, its first full year of U.S. operation, were a modest seven million cigarettes. Among the brand offered, besides Philip Morris, were Blues, Cambridge, Derby, and a ladies favorite name for the London street where the home companies factory was located - Marlborough.
Cigarette manufacturer (Camel, Winston, Doral)
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MEMORANDUM
TO: Dr. F. J. Schultz
FROM: C. I. Lewis a ) 0--
Total Amino Acid' Content of Tobacco
February 28, 1975
In two of our current projects (E-134 and C-133)
we felt there was a need to know the total amino acid
content of some current cigarette blends. These were
determined by Analytical Biochemistry Labs by an analysis
of the hydrolysate derived from a 4hr sealed tube treatment
of the tobacco with 6N HC1 at 14I5oC. Under this treatment
tryptophan is destroyed and there could be some reduction
in amino. acids by the slow reaction of carbonyl compounds
with the amino acids during, hydrolysis.
Some conclusions can be drawn from this data:
(1) True has a relative very high level of
hydroxyproli;ne.
(2) Marlboro, Vantage and Zack have a very high level
of aspartic acid.
(3) Marlboro and Zack have a lower level of glutamic
acid.
(4) Marlboro, Vantage and Dec. Kent have a lower level
of arginine.
(5) Marlboro and Vantage have a very high level of
proline.
ANALYSIS I2ESULTS
Percent w/w
Lab. No. 14946-1 14946-2
Cust. I.D.
Amino Acid (m.w.) 274K Rel. 274M,
$ Rel.
$
Alanine (89) 0.47- 6.1 0.44 6.0
Valine (117) 0.41 5.3 0.42 5.7
Glycine (75) 0.40 5.2 0.42 5.7
Isoleucin e (131) 0.29 3.8 0.29 4.0
Leucine ( 131) 0.55 7.1 0.48 6.6
Proline ( 115) 0.71 9.2 0.76 10.4
Threonine (119) 0.32 4.2 0.26 3.6
Serine ( 105) 0.40 5.2' 0.31 4.2
Methionin e (149) 0.13 1.7 0.14 1.9
Hydroxyproline (131) 0.05 0.6 0.06 0.8
Phenylalanine (165) 0.39 5.1 0.35 4.8
Aspartic Acid (133) 1.26 16.3 1.39 19.0
Glutamic Acid (147) 1.32 17.1 1.17 16.0
Tyrosine (181)~ 0.20 2.6 0.17 2.3
Lysine (146)~ 0.36 4.7 0.29 4.0
Histidine (1S5) 0.12 1.6 0.12 1.6
Arg,i:nine (174) 0.32 4. 2 0.20 2.7
Cystine/2 (120) 0.01 0.1 0.04 0.6
Ave. M. W. (132)
(1) Total 7.71 7.31
(2) Total N% 2.40 3.02
(3) Total N% due
to Protein~ .82 Y77
(4) Other Nitrogen % 1.58 2.25
'(5) Nicotine, % 1.50 1.73
-(6) N due to .26 .30
, . Nicotine, %
(7) Other Nitrog,en
less Nicotine, % 1.24 1.95
274K
= Nov. Kent 85 O
O
CIO
274M = Oct. Marlboro 85 7~
~+.
~.
N+
ANALYSIS RESULTS
Percent w/w
Lab. No. 14946-3 14946-4
Cust. I'. D~. 274T Rel. 274V
g Rel.
,g g g
Amino Acid (mi.w. )
Alanine (89) 0.48 6.5 0.41 5.8
Valine (117) ' 0.39 5.3 0.37 5.2
Glycine (75) 0.44 5.9 0.37 5.2
Isoleucine (131) 0.27 3.6 0.25 3.5
Leucine (131) 0.50 6.8 0.46 6.5'
Proline (115) 0.65 8.8 0.76 10.7
Threonine (119) 0.30; 4.1 0.27 3.8
Serine (105) 0.37 5.0 0.35 5.0
Methionine (149) 0.13 1.8 0.11 1.6
Hydroxyproline (131) 0.09 1.2 0.05 0.7
Phenylalanine (165) 0.37 5.0 0.36 5.1
Aspartic Acid' (133) 1.24 16.8 1.26 17.9
Glutamic Acid. (147) 1.22 16.5 1.23 17.4
Tyrosine (181) 0:18 2.4 0.19 2.6
Lysine (146) 0.33 4.4 0.29 4.1
Histidine (~155) 0.12 1.6 0.09 1.3
Arginine (174) 0.30 4.1 0.20 2.8
Cystine/2 (120)~ 0.02 0.3 0.03 0.4
Ave. M. W. (132)
-(1) Total . 7.40 7.05
(2) Total N% 2.34 2.63
(3) Total No due
to Protein .78 .75
(4) Other Nitrogen % 1.56 1.88
-(5) Nicotine, % 1.50 2.01
(6) N due to .26 .34
Nicotine, % '
(7) Other Nitrogen
less Nicotine, o 1.30 1.67
74T
Nov. True 85 O
O
G.~
C.' .
274V
= Oct. Vantage 85 T
~
.~
tJ
ANAL1'SIS RESULTS
Percent w/w
Lab. No.
Cust. I.D.
Amino Acid (m.w. ) 15128-1
1375CK
Rel. 15128-2
1375NT
Rel.
Alanine (89). 0.47 5.4 0.47 6.3
Valine (117). 0:47 5.4 0.43 5.7
Glycine (75) 0.49 5.7 0.42 5.6
Isoleucine (131) 0.34 4.0 0.31 4.1
Leucine (131) 0.54 6.2 0.50 6.7
Proline (115) 0.67 7.7 0.71 9.5
Threonine (119)~ 0.26 3.0 0.25 3.3
.Serine (105) 0.66 7.6 0.51 6.8
Methionine (149) 0.23 2.7 0.18 2.~!
Hydroxyproline (131 ) 0.06 0.7 0.06 0.8
Phenylalanine (165) 0.40 4.6 0.36 4.8
Aspartic Acid!(133) 1.72 19.8 1.22 16.2
Glutamic Acid (147) 1.36 15.4 1.25 16.6
Tyrosine (181) 0.19 2.2 0.16 2.1
Lysine (146) 0.35 4.0 0.31 4.1
Histidine (155) 0.16 1.8 0.16 2.1
Arginine (174) 0.27 3.1 0.19 2.5
Cystine/2 (120)~ 0.01 0.1 0.02 0.2
Ave. M. W. (13 2)
(1) Total 8.65 7.51
(2) Total N% 2.39 2.26
(3) Total N% due
-to Protein; .92 .80
(4) Other Nitrogen~ % 1.47 1.46
.(5) Nicotine, % 1.83 1.50
(6) N due to
Nicotine, o .25. .26
(7) Other Nitrogen 1.22 1.20
less Nicotine, ~
1375CK = Dec. Zack 85
1375NT"= Dec. Kent 85
--5-
The following data is a general analysis for free
amino acids in burley, turkish and flue-cured.
Cutters
Burley, ug/g, Lower Primings
Turkish, ug/g Cutters
Flue-cured, uq/q
Alanine 138 671 464
Valine 30 279 146
Glycine 199 194 30
Isoleucine 37 20 20
Leucine 44 , 20 20
Proline 137 1410 4685
Theonine 152 20 157
Serine N.A. 20 200
Methionine N.A. 318 189
Hydsoxyproline
Phenylalanine N.A.
143 N.A.
.446 N.A.
188
Aspartic Acid 1353 583 279
Glutamic Acid 304 866 379
Tyrosine 30 30 30
Lysine 116 30 30
Histidine 402 337 205
Arga.nine
Cystine/2 N.A.
N.A. R.A.
N.A. N.A.
N.A.
Free amino acids should be in equilibrium with total
.amino acid content of the protein.
There appears to be a paradox for both Marlboro and
Vantage, For example:
(1) -high aspartic means high burley
- (2) high proline means high flue-cured
(3) high glutamic means high turkish
If you increase the burley then relatively both the
flue-cured and turkish must go down unless flue-cured is -.
increased dramatically relative to the turkish. But in
Marlboro~ and Vantage the aspartic acid i shig,h and the proline is high. But in the Vantage the
glutamic is high
also. This just can not be unless Marlboro and Vantage
are addi;ng n1troqen containing aca.diitives to~ their tohaccol.
This premise was.investigated by subtracting out
protein nitrogen and nicotine nitrogen from total nitrogeni.
In Table I at the bottom the results are straight forward.
Nitrogen compounds are being added to Marlboro and' Vantage
but not to Kent, True or Zack.
This should'be a patentable item so the Reynolds and
Philip Morris patents were searched for nitrogen compounds.
The results were the patents in Appendix I. Philip Morris
is using ammonium phosphate and Reynolds is using! a Schiff
base and/or pyridine compound. A Schiff base is made by
aombination of an aldehycle with a base like ammonia. Sugars
are aldehydes. It is also known that the quality of tobacco
as defined by Schmuck is soluble carbohydrates/protein.
Therefore one would have to say that what Philip Morris has
done with ammonium phosphate Reynolds is doings with the
Schiff base and pyridine compound and it is all related to
quality of.the tobacco and tobacco smoke.
In the future we are going! 'to have to be able to
analyze tobaccos for Schiff bases. The low volatility of
these materials and'extreme heat sensitivity of these
materials would make liquid chromatography the-most likely
mode of attack.
Attachments
CIL/hsh
Xc: J. H. Bell
A. B. Hudson
11. J. tdinnemeyer
Sa_-McGeady
-Haxty Skladanowski
CIL/hsh
