Original Research Communications--general

Energy content of diets of variable amino acid composition13

Michael E May and James 0 Hill

ABSTRACT variation in the distribution ofdietary nitro- these calculations, and to apply the computerized calculations
gen among the different amino acids is one factor that can mod- to a variety ofmixtures ofamino acids and other foods.
ify the calorie equivalent per weight of amino acid or protein.

Downloaded from ajcn.nutrition.org at UNIVERSITY OF OTAGO SCIENCE LIBRARY on March 3, 2016

This is important to consider when experimental diets with
Methods
different amino acid compositions are compared and when in-
direct calorimetry is used to determine substrate oxidation The heats of combustion and heats of formation of amino
rates. We developed a computer program to compute the en- acids were obtained from standard references (2, 3). Heats of
ergy content, oxygen equivalent, and respiratory quotient for combustion were calculated for lysine, ornithine, proline, and
arbitrary mixtures of amino acids and representative carbohy- histidine from the heats of formation of amino acid, carbon
drates and fats. The calorie content of individual free amino dioxide, and water (see Appendix for step-by-step description
acids was calculated by correction of the heat of combustion of calculations). The biologic energy content was obtained by
for the incomplete oxidation of amino acids characteristic of subtraction ofthe residual heat ofcombustion contained in bi-
humans. Although these computations were presented before, ologic end products from the heat of combustion. The major
we now report the limit ofapplicability ofpublished values and nitrogenous excretory products were assumed to be urea, creat-
the availability of the computer program to do these mine, uric acid, and ammonia. These products were chosen
calculations. Am J C/in Nutr 1990;52:770-6. because of their abundance in urine of normal adults (4). Be-
cause the heats of combustion of these compounds per mole
KEY WORDS Amino acids, branched-chain amino acids, nitrogen are different, the energy content of urinary nitrogen-
energy metabolism, indirect calorimetry containing compounds depends on the distribution of the ni-
trogen among these compounds. The sulfur-containing amino
acids yield a small amount of urinary taurine along with urea,
Introduction
ammonia, creatinine, and uric acid and produce additional
metabolic energy by the oxidation ofthe sulfur to sulfate. These
The energy content ofa foodstuffis determined by multiple
computations were summarized recently (5), but the tedious
factors including the efficiency of intestinal absorption, the
task of computing the urinary metabolite energy content for
fraction ofnutrient or its metabolites lost in the urine, the frac-
all naturally occurring amino acids was simplified in the past
tion of loss into feces of intestinal secretion induced by inges-
by assuming that urine nitrogen is in only one form, such as
tion ofthe foodstuff, and the biochemical pathways of metabo-
urea (5).
lism. The energy content of absorbed nutrients is the heat of
We have written a microcomputer program, METENERG
combustion minus the heat of combustion of products of me-
(ME May, JO Hill, vanderbilt University, Nashville, TN) that
tabolism that are not fully oxidized. These principles are well
computes the metabolizable energy ofthe amino acids after the
known, and the usual factors for caloric content of foods have
user enters the distribution ofurine nitrogen either as a fraction
traditionally been determined from the measured heat of corn-
or as weight of compound excreted per day. Because the uri-
bustion minus the heat of combustion of fecal material and
nary nitrogenous metabolites contain carbon, hydrogen, and
urine (1). Simple sugars, such as glucose, and short-chain fatty
oxygen in addition to nitrogen, the volume of oxygen con-
acids, such as acetic acid, are oxidized nearly completely to car-
sumed in the catabolism ofamino acid sufficient to produce 1
bon dioxide and water after absorption, and the energy content
g ofurine nitrogen (O:N), the metabolic energy equivalent per
is the heat ofcombustion. Amino acids are oxidized to carbon
liter of gaseous oxygen used in oxidation of an amino acid
dioxide, water, and several urinary metabolites containing ni-
trogen; these latter compounds account for the difference be-
tween biological energy content of protein and the heat of com- I From the Departments of Medicine and Pediatrics, Vanderbilt
bustion of protein. It seems intuitive that the biological energy University, Nashville, TN.
2 Supported in part by a pilot and feasibility grant from the Diabetes
content will be greatest when the weight proportion of nitrogen
Research and Training Center, NIH 5 P60 DK20593-lO, and by NIH
is least, and thus each amino acid may have a different caloric
grants DK38088 and DK26657.
value. Address reprint requests to ME May, AS 105 MCN Vanderbilt Uni-
The purpose of this article is to review the chemical and versity, Nashville, TN 37232-2230.
arithmetic bases for computation of the energy value of defined Received September 5. 1989.
food chemicals, to outline the approach to computerization of Accepted for publication December 27, 1989.

770 Am iC/in Nutr l990;52:770-6. Printed in USA. 1990 American Society for Clinical Nutrition
 CALORIE CONTENT OF FREE AMINO ACIDS 771

(H02), and the ratio of gaseous carbon dioxide produced to 8) Sulfur amino acids require additional algebraic terms to
oxygen consumed (RQ) also depend on the distribution of account for energy release and oxygen utilization in oxidation
urine nitrogen among the possible metabolites. METENERG ofsulfur to sulfate and the small change in apparent molecular
computes these quantities for each amino acid after the urine- composition ofunnary nitrogenous compounds occasioned by
nitrogen distribution is specified according to the following the excretion of taurine.
computational algorithm: The above algorithm is not original (5), but the algebraic for-
1) The heat of combustion of urinary nitrogenous corn- mulation of the chemical balance allows the computerization
pounds is computed per mole urine nitrogen by the expression of the calculations, with easy extension of the calculations to
any specified distribution of urinary nitrogenous compounds.
: (F1,).(HJn1) The ionization fraction of organic acids is required to obtain
correct RQ values (5), and the program includes input of the
where F, is the fraction of urine nitrogen in compound i, H,
ionization fraction and the RQ correction for five common
is the heat ofcombustion ofcompound i, and n is the number
short-chain carboxylic acids.
of moles nitrogen per mole i; i represents any urinary corn-

Downloaded from ajcn.nutrition.org at UNIVERSITY OF OTAGO SCIENCE LIBRARY on March 3, 2016

METENERG was written in Turbo Basic 1.0 and compiled
pound containing nitrogen derived from amino acids (includ-
for use on an IBM-PC#{174}--compatible microcomputer under
ing amino acids and proteins).
MSDOS. It requires a minimum of 128 kb RAM memory and
2) The maximum metabolizable energy ofeach amino acid
one disk drive. Printer output uses ASCII codes only and
is computed as heat ofcombustion, amino acid, minus residual
should be compatible with most common printers. The pro-
heat in urinary nitrogenous compounds.
3) The number ofcarbons (ca), hydrogens (he), and oxygens gram files are METENERG.EXE, MOLECULE.FW. MOL-
ECULE.MW, AAFORWT.BAT, and AAMOLWT.BAT. The
(on) in the urine per mole nitrogen are derived from analogous
two DOS batch files allow selection ofeither molecular weights
formulas:
or acyl residue formula weights for the parameter file MOL-
c = (F1,) . (c1/n1) ECULE.CON. The program searches only the default directory
for the latter file and puts output files in the same directory.
h = > (F1,)-(h1/n1) Report files are named RQDI??KA.OUT. The 7? in the name
of the report file is 01 for the first report and then increases
ou = >: (F1,)-(o1/n1) to 99 as additional reports are generated.
This feature allows
where c, h, and o1 are the numbers ofmoles ofcarbon, hydro- multiple reports to be generated in one run of the program.
gen, and oxygen per mole of nitrogen in compound i, respec- When the report counter reaches 99, all subsequent report files
tively. In the calculation ofthe correction for energy content of are named RQDIO1KA.OUT until the report files are cleared.
amino acids, only nitrogenous compounds in the urine will be It is the responsibility of the user to delete old report files by
included rather than all urinary compounds. using appropriate DOS commands.
4) The chemical reaction for biologic oxidation of an amino The program is organized as follows: The first screen displays
acid can be written in general terms as the name, copyright notice, and an option to exit to DOS. The
initialization sections read the chemical formulas and heats of
CcaHhaOoaNna + X02 CCUHhUOOUN + YCO2 + ZH2O combustion of amino acids and other substrates, and the de-
fault distribution of urine nitrogen from the file MOLE-
The molecular compositions of the natural amino acids are
CULE.CON. All numbers are stored as double-precision reals
known and the apparent molecular composition of the urinary
(8 bytes, 14-digit precision) to minimize rounding errors. The
nitrogenous compounds depends on the distribution of urine
main menu allows four choices or exit: enter fractional distri-
nitrogen as detailed in reference 3. The stoichiometric coeffi-
bution of urinary nitrogen, enter mass of urinary metabolites
cients are computed as follows:
with nitrogen, calculate amino acid metabolizable energy and
Y = ca - c#{149}
na O:Nu, and calculate energy parameters ofa food mixture.
Program flow is directed by a SELECT CASE <choice) state-
Z = (ha - hufla)/2
ment, which forces return to the main menu before to exit from
X(2Y+Z+Oufla)/2 the program. There are two functions and 32 procedures that
do the following operations: screen manipulation, 1 1 routines
where Y is the CO2 production in mol/mol amino acid and X to clear specified blocks, print messages, format numbers as
is the 02 utilization in mol/mol amino acid. strings for display, and print successive screens; keyboard input
5) The RQ for a diet is the ratio of carbon dioxide produced routines, 7 routines for entry ofspecified quantities; calculation
to oxygen consumed in complete oxidation of the diet after routines, 10 routines, each for a specific portion ofthe compu-
complete absorption: tations; output routines, 4 routines; and program flow, main
program plus 2 routines comprising groups ofthe input, calcu-
RQ = Z/X
lation, and output routines.
6) The heat equivalent of oxygen (kcal/L 02) is The substrates for diets included in MOLECULE.CON are
22 amino acids, 5 carbohydrates, 3 polyols, 5 short-chain or-
Heq = (Metabolizable energy)/(X . gas constant) ganic acids, ethanol, and 4 fats. Thermodynamic properties for

7) The ratio of oxygen utilization to urine nitrogen produc- the latter 18 substrates were obtained from published literature
(2). The program is supplied with two parameter files; one con-
tion (L 02:g N) is
tains compositions and molecular weights of amino acids
O:N = X.gasconstant/14.0067.n5 (MOLECULE.MW) and the other contains compositions and
772 MAY AND HILL

TABLE 1
Heats ofcombustion and biologic oxidation ofamino acids

Metabolizable energy5

Compound Heat ofcombustion Set 1 Set 2 Set 3 Set 4

kca//g kca//g(kJ/g)

Alanine 4.34 1 3.425 ( 13.70) 3.549 ( 14.20) 3.494 ( 13.98) 4.369 ( I 8.28)
Arginine S. 129 3.254 ( 13.02) 3.508 ( 14.03) 3.396 ( 13.58) 3.784 ( I 5.83)
Asparagine 3.488 2.252 (9.0 1) 2.4 19 (9.68) 2.345 (9.38) 2.7 18 ( 1 1.37)
Aspartic acid 2.875 2.26 1 (9.04) 2.344 (9.38) 2.307 (9.23) 2.689 ( 1 1.25)
Cysteine 3.256 4.3 12 ( 17.24) 4.402 ( 17.6 1 ) 4.362 ( 17.44) 4.792 (20.05)
Cystine 3.0 15 4.093 ( 16.37) 4. 182 ( 16.73) 4. 143 ( 16.57) -

3. 133 ( 12.53) 3.583 (14.99)

Downloaded from ajcn.nutrition.org at UNIVERSITY OF OTAGO SCIENCE LIBRARY on March 3, 2016

Glutamic acid 3.646 3.09 1 ( 12.36) 3. 166 ( 12.66)
Glutamine 4.207 3.089 (12.36) 3.241 (12.96) 3.173 (12.69) 3.61 1 (15.11)
Glycine 3.097 2.01 1 (8.04) 2.158 (8.63) 2.093 (8.37) 2.741 (11.47)
Histidine 4.85 1 3.273 ( 13.09) 3.487 ( 13.94) 3.392 ( 1 3.57) 4.2 16 ( I 7.64)
Isoleucine 6.523 5.902 (23.6 1) 5.986 (23.94) 5.949 (23.80) 6.886(28.81)
Leucine 6.524 5.903 (23.6 1) 5.988 (23.95) 5.950 (23.80) 6.888 (28.82)
Lysine 6.038 4.92 1 ( 19.68) 5.072 (20.29) 5.005 (20.02) 5.686 (23.79)
Methionine 4.456 5.3 13 (2 1 .25) 5.385 (2 1.54) 5.353 (2 1.4 1) 6.06 (25.34)
Ornithine 5.493 4.259(17.04) 4.426(17.71) 4.352(17.41) -

Phenylalanine 6.723 6.229 (24.92) 6.296 (25. 18) 6.266 (25.06) 7.036 (29.44)
Proline 5.681 4.970(19.88) 5.066(20.27) 5.024(20.10) 5.937(24.84)
Serine 3.308 2.532 ( 10. 13) 2.637 ( 10.54) 2.590 ( 10.36) 3. 107 (13.00)
Threonine 4.120 3.434(13.74) 3.527(14.11) 3.486(13.94) 4.101 (17.16)
Tryptophan 6.588 5.787(23.14) 5.896(23.58) 5.848(23.39) 6.419(26.86)
Tyrosine 5.859 5.409 (2 1.64) 5.470 (2 1.88) 5.443 (2 1.77) 6.039 (25.27)
Valine 5.963 5.264 (2 1.06) 5.358 (2 1.43) 5.3 16 (2 1.27) 6.276(26.26)
I 4.781 4.136(16.54) 4.253(17.01) 4.201 (16.80) 4.847 (20.28)
SD 1.324 1.342(5.370) 1.321 (5.283) 1.339(5.321) 1.530(6.402)
CV(%) 27.7 32.4 31.1 31.7

a For four distributions ofurine nitrogen: set 1-90% urea, 3% ammonia, 5% creatinine, 2% uric acid; set 2-100% ammonia; set 3-l00% urea;
and set 4-data from Table 3 ofreference (2) for 100% urea.

formula weights ofthe acyl portion (dehydrated) ofthe amino product. Minor differences would be expected from the use of
acids (MOLECULE.FW). Selection of parameter files is ac- slightly different primary sources, but the large differences in
complished by the DOS batch files. The program as written is our calculations and those previously reported was bother-
appropriate for diets composed of modular components (pro-. some. We replicated the numbers of Livesey and Elia by use of
tein and/or amino acids, carbohydrate, fats), and extension to the compositions ofthe acyl portion for computation of nitro-
natural diets would require modification or assumption
that gen content and formula weights rather than the molecular
the nonprotein nitrogen was negligible in quantity.
corn- The composition for each ofthe amino acids. That is, the previously
putations strictly apply to the portion of the diet that is ab- reported parameter values refer to amino acids in protein when
sorbed or taken parenterally; digestibility is not specifically en- the amino acid data has been expressed as
tered in the program. Because the user is prompted for the diet
components by weight, known digestibility factors (amount ab- Amount ofamino acid = fraction ofprotein nitrogen
sorbed/amount eaten) can be accounted for by entering Weight
in amino acid X acyl formula weight per mole nitrogen
x Digestibility instead ofWeight for each nutritive component
and entering Weight X (1-Digestibility) plus the weight of Ifamino acid composition is expressed in these terms, the sum
dietary minerals as Mineral for the program-this allows cor- of amino acid weights will be equivalent to the protein weight
rect energy computations per weight of the original diet. (The and we can call this mode of expression protein equivalent.
compiled program is available from the authors for $25 (US) Actually, the sum ofamino acid weights obtained by hydrolysis
to cover reproduction and shipping.) of protein will be greater than the weight of the initial protein
by the amount ofwater added to hydrolyze the peptide bonds.
Results and discussion There is no convention stated for expression of amino acid
data in standard tables of food composition (6). Furthermore,
Table 1 shows the heats of combustion and metabolizable the protein-equivalent mode ofexpression ofamino acid corn-
energy for amino acids. Metabolizable energy is presented for position ofproteins is not applicable to diets composed of crys-
four distributions of urine nitrogen. The right-hand column of talline amino acids, as are often used in experiments and in
Table 1 is the metabolizable energy reported by Livesey and parenteral nutrition. The previously published values for me-
Elia (5) under the assumption that urea is the sole urinary end tabolizable energy of amino acids do not apply to diets corn-
 CALORIE CONTENT OF FREE AMINO ACIDS 773

TABLE 2 g protein equivalent and not per g free amino acid (7). This
Heat equivalent ofoxygen for amino acids communication is in agreement with the findings ofthis paper.
These data are ofinterest for two reasons: the design of isoca-
Heat equivalen t ofoxygen consum ed in oxidation5
loric, isonitrogenous research and therapeutic diets requires
Amino acid Set 1 Set 2 Set 3 these computations ifthe distribution ofamino acids is atypi-
cal, and the measurement ofenergy expenditure and substrate
kcal/L (ki/L) oxidation rates by indirect calorimetry or measurement of en-
ergy expenditure in free-living subjects by doubly labeled water
Alanine 4.627 ( I 8.5 1 ) 4.703 ( 18.8 1) 4.630(18.52)
Arginine 4.799 ( I 9. 19) 4.957 ( 19.83) 4.798 (19.19) require proper values of 0:N, H02, and RQ for accuracy.
Asparagine 4.600 (1 8.40) 4.753 (19.0 1) 4.607(18.43) Table 5 shows the effect ofapplying these values to the compu-
Aspartic acid 4.563 ( 18.25) 4.640 ( 18.56) 4.567 (18.27) tation ofthe biologic energy content ofseveral amino acid mix-
Cysteine 5.318 (21.27) 5.359 (21.44) 5.31 1 (21.24) tures. variation in amino acid composition results in different
Cystine 5.308 (2 1.23) 5.350 (2 1.40) .300 (21.20) energy contents despite similar weight percentages of total
Glutamic acid 4.567 ( 18.27) 4.6 18 ( 18.47) 4.70(18.28)

Downloaded from ajcn.nutrition.org at UNIVERSITY OF OTAGO SCIENCE LIBRARY on March 3, 2016

amino acids. For example, solutions enriched in the branched-
Glutamine 4.593 ( 18.37) 4.696 ( 18.78) 4.98(18.39) chain amino acids have a higher caloric value than do formula-
Glycine 3.979 ( 1 5.9 1 ) 4. 1 30 ( 16.52) 4.005 (16.02)
tions enriched in alanine and glycine despite similar or lower
Histidine 4.693 ( 18.77) 4.827 ( 19.3 1) 4.696(18.79)
weight percentages oftotal amino acids. The essential or indis-
Isoleucine 4.64 1 ( 18.56) 4.67 1 ( 18.68) 4.642 (18.57)
Leucine 4.642 (18.57)
pensable amino acids have not only a high metabolizable en-
4.672 (18.69) 4.643 (18.57)
Lysine 4.662 ( 1 8.65) 4.726 ( 18.90) 4.664 ( I 8.65) ergy but also a high oxygen equivalent ofurine nitrogen. Table
Methionine 4.790 ( 19. 16) 4.8 18 ( 19.27) 4.790(19.16) 5 includes several enteral products that contain carbohydrate
Ornithine 4.663 ( 18.65) 4.745 ( 18.98) 4.666(18.66) in addition to amino acids, and the calculated RQ is corre-
Phenylalanine 4.6 17 ( 18.47) 4.640 ( 18.56) 4.618(18.47) spondingly higher for those products. O:N directly reflects the
Proline 4.69 1 ( 18.76) 4.732 ( 18.93) 4.692 (18.77) amino acid composition ofeach diet.
Serine 4.859 ( 19.44) 4.945 ( 19.78) 4.858 (19.43) It can be predicted that it would be difficult to balance both
Threonine 4.629 ( 1 8.52) 4.686 ( 18.74) 4.63 1 (18.52)
calories and nitrogen among test diets of a few amino acids
Tryptophan 4.631 (18.53) 4.671 (18.68) 4.633(18.53)
chosen from the extremes ofcaloric density. The most extreme
Tyrosine 4.63 1 ( 18.52) 4.655 ( 18.62) 4.632 (18.S3)
Valine 4.630(18.52) 4.668(18.67) 4.631 (18.52)
j: 4.688 ( 18.75) 4.757 ( 19.03) 4.690 ( I 8.76)
SD 0.259 ( 1.04) 0.248 (0.99) 0.24(1.01) TABLE 3
CV (%) 5.5 5.2 5.4 Oxygen equivalent ofurine nitrogen

Oxygen used in production

S Sets described in Table 1.
ofurine nitrogen5

Amino acid Set 1 Set 2 Set 3

prising crystalline amino acids. We prepared two parameter
L 02/g N
files containing either molecular weights ofamino acids or acyl
residue formula weights for amino acids, for use with actual Alanine 4.709 4.801 4.801
amino acid weights or protein-equivalent weights of amino Arginine 2.108 2.200 2.200
acids, respectively. There is over twofold variation in the values Asparagine 2.308 2.400 2.400
Aspartic acid 4.709 4.801 4.801
for individual amino acids, and the essential amino acids have
Cysteine 7.015 7.105 7.105
higher metabolizable energy on average than do the nonessen-
Cystine 6.615 6.705 6.705
tial amino acids.
Glutamicacid 7.109 7.201 7.201
Tables 2-4 present the values for free amino acids rather Glutamine 3.509 3.600 3.600
than protein-equivalent amino acids. The heat equivalent of Glycine 2.709 2.800 2.800
oxygen varies little among the amino acids (CV < 6%, Table 2), Histidine 2.575 2.667 2.667
but large variations among amino acids in the values of oxygen Isoleucine I 1.910 12.002 12.002
equivalent of urine nitrogen (CV 57%) and respiratory coeffi- Leucine 11.910 12.002 12.002
Lysine 5.509 5.601 5.601
cient (CV 18%) are shown in Tables 3 and 4. As demonstrated
Methionine 11.816 11.906 11.906
in Table 3, the oxygen equivalent of urine nitrogen is the same
Ornithine 4.309 4.401 4.401
whether the urinary metabolite is urea or ammonia; this calcu- Phenylalanine 15.9 10 16.002 16.002
lated result was verified as a general rule for any monoamino Proline 8.709 8.801 8.801
monocarboxylic acid. The mixture of multiple amino acids Serine 3.909 4.001 4.001
and addition of other caloric sources will tend to minimize Threonine 6.309 6.401 6.401
variation in energy content of natural diets, especially because Tryptophan 9. 109 9.20 1 9.201
amino acids usually contribute only 10-20% of the energy in Tyrosine 15. 1 10 15.202 15.202
Valine 9.509 9.601 9.601
normal human diets.
I 7.154 7.245 7.245
Between the time of first review and publication of this SD 4.117 4.117 4.117
manuscript, an Erratum was published stating that the metabo- CV (%) 57.5 56.8 56.8
lizable energy density of individual amino acids published in
reference 5 of this paper was expressed in units of energy per S Sets described in Table 1.
774 MAY AND HILL

TABLE 4 ERG. The precision ofthe heat equivalent ofoxygen (Table 2)

Respiratory quotient for amino acids supports its use as a measure ofcaloric expenditure. However,
the partitioning of total oxygen consumption into carbohy-
CO2 produced per oxygen
drate and fat oxidation requires correction of the oxygen utili-
consumed in catabolism5
zation by subtracting urine nitrogen X O:N As seen in Table
.

Aminoacid Setl Set2 Set3 3, O:N is highly variable among the natural amino acids. A
problematic but critical question is whether the mix of amino
L/L acids being oxidized is the same under all conditions or among
Alanine 0.835 1.000 0.833 all subjects in a research study. Computation of a correction
Arginine 0.727 1.091 0.727 for urine nitrogen from the dietary amino acid composition
Asparagine 1.011 1.333 1.000 implies the assumption that urine nitrogen is derived from
Asparticacid 1.175 1.333 1.167 amino acids in the same relative proportions as in the diet. This
Cysteine 0.554 0.667 0.556 assumption may be true in a steady state in adult humans or

Downloaded from ajcn.nutrition.org at UNIVERSITY OF OTAGO SCIENCE LIBRARY on March 3, 2016

Cystine 0.588 0.706 0.589 nongrowing animals but is only one ofseveral possible assump-
Glutamic acid 1.004 1. 1 1 1 1.000 tions in states of nitrogen accretion (growth) or negative nitro-
Glutamine 0.893 1. 1 1 1 0.889
gen balance (starvation, postinjury or postoperative catabo-
Glycine 0.862 1.143 0.857
lism). Other assumptions, such as that the mix of amino acids
Histidine 0.906 1.200 0.900
being oxidized is distributed the same as the amino acid distri-
Isoleucine 0.733 0.800 0.733
Leucine 0.733 0.800 0.733
Lysine 0.714 0.857 0.714
Methionine 0.600 0.667 0.601 TABLES
Ornithine 0.727 0.909 0.727 Calorimetric variables ofsome amino acid mixtures5
Phenylalanine 0.85 1 0.900 0.850
Mixture ME RQ Heq 02:N
Proline 0.819 0.909 0.818
Serine 1.006 1.200 1.000 kcal/L
Threonine 0.877 1.000 0.875
Tryptophan 0.871 0.957 0.870 Travasol 8.5%t 495 0.776 4.669 5.126
Tyrosine 0.896 0.947 0.895 AminoSyn 8.S%t 556 0.772 4.687 5.769
Valine 0.750 0.833 0.750 HepatAmine 483 0.772 4.689 6.151
i 0.824 0.976 0.822 FreAmine III 8.S% 344 0.780 4.697 5.876
SD 0.151 0.196 0.148 Nephramine S.4% 282 0.744 4.708 9.372
CV(%) 18.3 20.1 18.0 TrophAmine6% 266 0.780 4.720 6.212
Indispensable-8.5% fl 442 0.787 4.705 8.460
5 Sets described in Table I. Dispensable-8.S%i 27 1 0.864 4.800 4.054
RDA mixture-6.36%55 341 0.761 4.715 9.698
Nutrisource Aminoacidtt 4435ff 0.807 4.694 6.885
Nutrisource AA-HBCtt 47 1 8 0.780 4.706 7.206
imbalance arises for comparisons ofa single amino acid among
HepaticAidII 1077 0.874 4.864 6.740
diets. This is of great interest because the branched-chain TravasorbRenalt 1170 0.945 4.921 6.471
amino acids (leucine, isoleucine, and valine) may have benefi- StandardVivonex 764 0.977 4.950 5.368
cial roles in the modulation of proteolysis in severe catabolic VivonexHighN 758 0.961 4.920 5.636
states (9, 10), and the energy content of the branched-chain Amin-Aid 1769 0.924 4.950 9.534
amino acids is high. For example, the effects of five solutions Traum-Aid HBC 997 0.909 4.917 7.633
on nitrogen balance were studied in traumatized rats ( 1 1): each TravasorbHepatict 1108 0.937 4.964 6.723
solution contained (per L) 50 g glucose and, in addition, the Caseclill 4605ff 0.822 5.397 6.544
first had 1 5 g leucine, the second had 15 g isoleucine, the third Traum-Aid HN 949 0.925 4.912 7.0 14
had 15 g valine, the fourth had 15 g alanine, and the fifth had Isocallill 1073 0.870 4.861 6.755
Ensure11J 1064 0.882 4.878 6.503
an additional I 5 g glucose. The diets were thought to be bal-
Vital HN1 41 16ff 0.940 4.943 6.077
anced for energy (assuming all amino acids and glucose had a
biologic energy content of4 kcal/g), but the actual energy con- S Values calculated for urine nitrogen: 90% urea, 3% ammonia, 5%
tents of the five solutions were 275, 275, 266, 238, and 242 creatinine, 2% uric acid.
kcal/L, respectively. The improvement in nitrogen balance t Travenol, Inc, Deerfield, IL.
seen with the leucine solution compared with the alanine or :t Abbott Laboratories, Inc. North Chicago, IL.
dextrose solutions was as likely to have been because ofthe 14% American McGaw, Inc, Santa Ana, CA.
greater energy content rather than special properties of leucine II His, lie, Leu, Lys, Met, Phe, Thr, Trp, Tyr, Val: 8.5 g/L each.
(10). METENERG will allow such computations to be done Ala, Arg, Asn, Asp, Cys, Glu, GIn, Gly, Pro, Ser: 8.5 g/L each.
#{182}
55 Recommended dietary allowance (8). 3.49 g Cys, 8.38 g Ile, 1 1.17
easily for many other special diets and will allow those compu-
g Leu, 8.38 g Lys, 3.49 g Met, 5.59 g Phe, 5.59 g Thr, 2. 10 g Trp, 5.59 g
tations to be done for different distributions ofnitrogen among
Tyr, and 9.78 g Val in 1 L solution.
urinary metabolites to allow customization for genetic or
tt Sandoz Nutrition, Minneapolis.
pathophysiologic states in which urine nitrogen is shunted if Dry powder (kcal/kg).
away from urea. Norwich Eaton, Inc. Norwich, NY.
The proper interpretation ofthe measurement of respiratory II II Mend Johnson Nutritionals, Evansville, IN.
gases depends on several parameters computed by ME TEN- Ross Laboratories,
#{182}11 Columbus, OH.
 CALORIE CONTENT OF FREE AMINO ACIDS 775

bution ofwhole body protein or that dispensable amino acids mation ofamino acids and related compounds. In: Sober HA, ed.
are oxidized in preference to indispensable amino acids, would Handbook ofbiochemistry. 2nd ed. Cleveland: CRC Press, 1970:
result in computation of different values for 0:N . Errors in B62-4.
3. Dean JA, ed. Langes handbook ofchemistry. 13th ed. New York:
estimating protein oxidation from end products ofamino acid
McGraw-Hill, 1985.
metabolism can affect the estimates ofrates ofoxidation of car-
4. Diem K, Lentner C, eds. Scientific tables. 7th ed. Ardsley: Ciba-
bohydrate and fat and of total energy expenditure obtained
Geigy Corp 1975:663-72.
from indirect calorimetry (5). Determination ofthe oxidation S. Livesey G, Elia M. Estimation ofenergy expenditure, net carbohy-
rates of multiple amino acids simultaneously is technically drate utilization, and net fat oxidation and synthesis by indirect
difficult because of the limited number of tracers for the corn- calorimetry: evaluation of errors with special reference to the de-
mon end product carbon dioxide. METENERG allows corn- tailed composition offuels. Am J Clin Nutr l988;47:608-28.
putation ofbounds ofvalues ofO:N and Heq for differing as- 6. Consumer and Food Economic Institute. Composition of foods:
sumed proportions ofamino acids. dairy and egg products. Agricultural handbook no. 8-1. Washing-
An alternative technique for measurement of energy expen- ton, DC: USGovernment PnntingOffice, 1976.
7. Erratum. Am J Clin Nutr l989;50:l475.

Downloaded from ajcn.nutrition.org at UNIVERSITY OF OTAGO SCIENCE LIBRARY on March 3, 2016

diture in living subjects is determination ofthe decay of specific
8. National Research Council. Recommended dietary allowances.
activity ofdoubly labeled water, which estimates carbon diox-
10th ed. Washington, DC: National Academy Press, 1989.
ide production from the difference in dilution ofenrichment in
9. Brennan MF, Cerra F, Daly JM, et al. Report ofa research work-
water of deuteriurn and oxygen- 18. The difference in dilution shop: branched chain amino acids in stress and injury. JPEN
curves results from the distribution ofoxygen-l 8 into the bicar- 1986; 10:446-52.
bonate pool in addition to the water pool labeled by the deute- 10. May ME,Buse MG. Effects ofbranched chain amino acids on pro-
num. Oxygen consumption is then estimated by assuming that tein turnover. Diabetes Metab Rev l989;S:227-45.
the subjects RQ equals the RQ of the subjects usual diet. Er- 1 1. Freund H, Yoshimura N, Fischer JE. The role of alanine in the
rors in estimation of the diet RQ can lead to errors in estima- nitrogen conserving quality ofthe branched chain amino acids in
tion ofoxygen consumption and thus to error in estimation of the post injury state. J Surg Res 1980; 29:23-30.

total energy expenditure. As detailed in Table 4, there is wide

12. Schoeller DA. Measurement of energy expenditure in free-living
humans by using doubly labeled water. J Nutr 1988; 1 18:1278-89.
variation in RQ among amino acids as well as the known varia-
tion in RQ among fats and carbohydrates (5). The estimation
of energy expenditure by doubly labeled water decay curves APPENDIX
also usually includes the assumption that the subjects are in a
net zero-energy balance during the period under study, which Sample computations for an individual amino acid
may be considered part of the assumption that RQ equals the
1) Urine nitrogen will be assumed to be 90% urea, 5% ammo-
calculated dietary RQ ( 12). The RQ calculated for the diet will
nia, and 5% creatinine. The molecular formulas and heats
be appropriate for the individual subject only ifthe distribution
of combustion of these compounds are
ofoxidized substrates is the same as the distribution of dietary
substrates, and this assumption would be untenable for sub- Urea C1H4O1N2 l5l.Okcal/mol
jects altering weight by deposition or loss ofbody tissue during Ammonia C0H3O0N1 70.6 kcal/mol
the period of study. Creatinine C4H701N3 558.1 kcal/mol
Urine nitrogen can also reflect nucleic acid catabolism and
excretion of many non-amino acid nitrogenous compounds 2) The heat of combustion of urinary nitrogenous corn-
contained in natural diets. Another major assumption of the pounds is (0.9 x 151.0/2) + (0.05 x 70.6/1) + (0.05 X
558. 1/3) = 80.78 kcal/(mol urine N).
above discussion is that urinary nitrogenous compounds are
derived from amino acids. This assumption is valid whenever 3) The C, H, and 0 lost in urinary nitrogenous compounds
the nitrogen content ofdietary amino acids is overwhelmingly arec =(0.9 X l/2)+(0.05 X0/l)+(0.05 X4/3)= 0.5167
(mol/mol N).
large compared with the nitrogen content of non-amino acid
h = (0.9 x 4/2) + (0.05 x 3/ 1 ) + (0.05 x 7/3) = 2.0667
compounds. Foods or experimental diets that contain large
(mol/mol N).
amounts of non-amino acid nitrogen cannot be treated by the
equations or program described above nor can they be allowed
ou = (0.9 X 1/2) + (0.05 x 0/1) + (0.05 x 1/3) = 0.4667
in studies employing indirect calorimetry for computation of (mol/mol N).
4) The remaining calculations will be illustrated with lysine.
substrate oxidation rates.
Finally, we feel that proper design of amino acid-modified
The molecular formula of lysine is C6 14 N 202 The heat .

research diets will require application of the individual energy ofcombustion oflysine is not tabulated but can be calcu-
parameters rather than assumption of constant values for all lated from the heat of formation. The heat offormation is
the enthalpy change for the reaction
amino acids. We are making METENERG available for this
purpose. El
6C+ 7 H2 +02 + N2-C6H14N202

The heat ofcombustion is the enthalpy change for the reac-

References
tion

I. Widdowson EM. Note on the calculation of the energy value of C6H 14N202 + 02 #{248}6 CO2 + 7 H2O + N2
foods and of diets. In: Paul AA, Southgate DA, eds. The composi-
tion of foods. 4th ed. New York: Elsevier/North-Holland Biomedi- The summation reaction
cal Press, 1978:322-6.
2. Hutchens JO. Heat of combustion, enthalpy and free energy of for- 6C+7H2+02+N2-.+6C02+7H2O+N2
776 MAY AND HILL
has an enthalpy that is a sum of heats of formation of the
Because food items are usually specified by weight, we con-
products, ie, vert this result by division by the molecular weight of lysine
(146.19) to 4.933 kcal/g.
(Heat of formation)L, + (heat of combustion)L5
6) The CO2 produced by oxidation of lysine is
= 6 X (heat of formation)co2 + 7 X (heat of formation)H20
(6 - 0.5167 X 2) mol/mol LYS = 4.9667 mol/mol LYS
Substituting the known values for heats of formation, we 7) The H2O produced by oxidation of lysine is
get
(14 - 2.0667 X 2)/2 mol/mol LYS = 4.9333 mol/mol LYS
(Heat of combustion)L, = [6 X 94.38 + 7 X 68.38
8) The oxygen consumed in catabolism of lysine is
- 162.2] kcal/mol = 882.7 kcal/mol
(2 X 4.9667 + 4.9333 + 0.4667 x 2
5) The metabolizable energy is the heat of combustion minus
the heat of combustion of urinary products, ie, - 2)/2 mol/mol LYS = 6.9000 mol/mol LYS.

Downloaded from ajcn.nutrition.org at UNIVERSITY OF OTAGO SCIENCE LIBRARY on March 3, 2016

9) RQ = 4.9667/6.9000 = 0.720 L C02/L 02
882.7 kcal/mol Lys - (80.78 kcal/mol N)
10) Heq = 721.14/6.9000 X 22.414 = 43.663 kcal/L 02
X (2 mol N/mol Lys) = 721.14 kcal/mol Lys 11) O:N = 6.900 X 22.414/14.0067 X 2 = 5.521 LO2/gN