NAME: Latiyah Timothy LAB PARTNER: Jasiel Mohammed

ID# 816012983 DATE: 5th November 2019

Course Code: Bioc 2069

Title of Lab: Assay of Tissue Glycogen.
Aim: To quantify glycogen from fed and fasted states of various animal tissues using an
indirect enzymatic/spectrophotometric method
Theory:
Glycogen is a storage form of glucose, that can be readily utilized. Glycogen is a large, branched
polymer of glucose residues. This polymer contains α(1-4) glycosidic bonds while the branches are
made up of α(1-6) glycosidic bonds. (Stryer, 2015)

Fig 1. Structure of Glucose (Stryer, 2015)

Glucose is stored as glycogen because free glucose is soluble and can disturb the osmotic pressure in
the cell and can cause the cell to burst. However, glycogen is insoluble in water and will not disturb
the osmotic pressure of the cell. (Stryer, 2015)
The major sites of storage of glycogen are in the liver and muscle. In the liver, there is a higher
concentration of glycogen of about 10% while skeletal muscle has a concentration of about 2%.
However, more glycogen is stored in the muscle because it has a greater mass. In the liver, the
metabolism of glycogen is used to regulate the glucose levels to meet the need of the whole organism,
while, in the muscle, its metabolism is utilized as energy in the muscle. (Stryer, 2015)
In states where there is high glucose in the blood, glucose is converted to glycogen for storage, this is
termed glycogen synthesis or glycogenesis. Glycogen is synthesised from glucose-6-phosphate.
Glucose 6-phosphate is converted to glucose 1-phosphate by phosphoglucomutase. Then UDP-
glucose attaches glucose residues to the nonreducing end of glycogen via glycogen synthase. And the
branching enzyme produces the (α1→6) linkages at branch points (Mckee, 2016)
 Fig 2. Glycogen Synthesis
(Mckee, 2016)

Glycogen degradation occurs in two parts. First glycogen phosphorylase cleaves nonreducing ends of
glycogen, to produce glucose 1-phosphate. Then the debranching enzyme shifts a block of three
glycogen resides to and expels the residue at the (α1→ 6) branch as free glucose. (Mckee, 2016)
Glycogen is regulated by the hormones, epinephrine, insulin and glucagon as well as allosteric
regulators. Glycogenesis is inhibited when low blood glucose activates glucagon or epinephrine which
then bind to their respective receptors of target cells which triggers glycogenolysis to activate. Insulin
stimulates glycogen synthesis and inhibits glycogen degradation by decreasing the synthesis of cAMP
and activating phosphoprotein phosphatase. (Nelson and Cox, 2017)
In this lab, the glucogen stored in fasted and fed states in various tissues was quantified. This is called
and Assay of Tissue glycogen. The tissue is disintegrated in the boiling solution of KOH. Treatment
with KOH leads to the degradation of proteins and free saccharides. In contrast, glycogen is stable in
the alkali solution. After KOH treatment, glycogen is precipitated with 96% ethanol, washed, diluted
and hydrolyzed in H2SO4. Glycogen hydrolysis produces free glucose, which is determined by an
enzymatic reaction in the presence of glucose oxidase: Glucose oxidase converts glucose to gluconic
acid. The by-product of this reaction is H2O2. The formed hydrogen peroxide reacts under catalysis of
peroxidase with o-dianisidine HCl to form red-violet product suitable for spectrophotometric
determination at 450nm.
APPARATUS AND MATERIALS
1. Heart, liver, muscle and kidney from freshly killed rats/guinea pigs (fed and fasted).
2. KOH (300g/L)
3. Saturated Na2SO4- 0.6 ml
4. Ethanol - 95% (v/v)- 20 ml
5. 2.4M HCl -8 ml
6. 1.0M NaOH
7. Stock glucose solution (0.2 µmol/mL)- 3ml
8. Dissecting instruments
9. Reagents for the estimation of glucose (17.5 ml) :
Glucose oxidase 12.5 mg
Peroxidase 4.0 mg
o-dianisidine HCl - 1% (w/v) in EtOH 0.5 mL
Dissolve all of the above in:
0.5M sodium phosphate buffer, pH 7.2 100mL

PROCEDURE:
Glycogen Assay:
Two millilitres of fed and fasted liver, fed and fasted kidney, fed and fasted heart and fed and
fasted muscle were collected in small test tubes and labelled and placed in an ice bucket. 0.2mL of
saturated Na2SO4 was added to each tube and vortexed. 5mL of ethanol was then added to all the tubes
and left to stand for five minutes. The precipitated glycogen was pelleted by centrifuging the tubes at
3000rpm for 10 minutes. The supernatant liquid was then poured off and discarded. 5mL of distilled
water was added to the pellet to dissolve and the tubes were gently warmed in a water bath at 37°C.
Tube containing glycogen from the liver of a fasted animal: The dissolved glycogen was
poured into a 10mL measuring cylinder and the test tube was rinsed with 2mL of distilled water. All
washings were poured into the measuring cylinder. The volume of solution in the measuring cylinder
was brought up to the 10mL mark by distilled water. The contents of the tube were thoroughly mixed.

Tube containing glycogen from the liver of a fed animal: The dissolved glycogen was poured
into a 100mL measuring cylinder and the test tube was rinsed with 2mL of distilled water. All
washings were poured into the measuring cylinder. The volume of solution in the measuring cylinder
was brought up to the 100mL mark with distilled water. The contents of the tube were thoroughly
mixed.

Tubes containing glycogen from other tissues of both fed and fasted animals: Each tube of
dissolved glycogen was poured into a 25mL measuring cylinder and the test tube was rinsed with
8mL of distilled water. All washings were poured into the measuring cylinders. The volumes of the
solutions in the measuring cylinders were brought up to the 25mL mark with distilled water and both
were thoroughly mixed.

1mL aliquots of prepared glycogen from the measuring cylinders were pipetted into separate
labelled test tubes and 1mL of 2.4M HCl was added to each tube which was then thoroughly mixed.
The tubes were covered with foil and placed in a boiling water bath for 30 minutes.
After boiling, the solutions were neutralised by adding 1M NaOH. The samples were then diluted to
5mL using distilled water. 0.3 and 0.5mL of liver/muscle from fed/fasted animals were pipetted into
separate test tubes and 0.5 and 1.0mL of heart/kidney samples were pipette into other tubes. Distilled
water was added to ensure that the volume of each tube was 1.0mL. 2.5mL of enzyme cocktail was
then added to each tube was foiled and incubated at 37°C for 45minutes. The tubes were cooled to
room temperature and the absorbance read at 450nm
Calibration Curve:
Seven test tubes were labelled 1-7 and stock glucose solution was added to the test tubes in the
following volumes, 0.0mL, 0.2mL, 0.2mL, 0.4mL, 0.6mL, 0.6mL and 1.0mL respectively. The
volume of each tube was made up to 1.0mL using distilled water where necessary. 2.5mL of enzyme
cocktail was added to each tube and incubated at 37°C for 45 minutes. The tubes were then cooled to
room temperature and the absorbance read at 450nm. A calibration curve was then plotted with
Absorbance vs glucose concentration.

RESULTS:
Table 1: Absorbances obtained and the Determined concentration of Glucose for Calibration curve.

Tube # 1 2 3 4 5 6 7

Volume of Stock 0.0 0.2 0.2 0.4 0.6 0.6 1.0

Glucose (ml)
Volume of Distilled 1.0 0.8 0.8 0.6 0.4 0.4 0.0
water (ml)

Volume of Enzyme 2.5 2.5 2.5 2.5 2.5 2.5 2.5

cocktail (ml)

Absorbance450nm 0 0.068 0.068 0.156 0.235 0.236 0.405

mol of glucose 0 0.040 0.040 0.080 0.120 0.120 0.200
Table 2: Results obtained from Assay of Tissue Glycogen from Fed tissue

Fed Tissue HEART KIDNEY MUSCLE LIVER

Mass of Tissue (g) 6.5 11.6 25 25
Final volume (mL) 13 22.8 50 50

Aliquot size mL 0.5 1.0 0.5 1.0 0.3 0.5 0.3 0.5
A450nm Sample i 0.061 0.093 0.106 0.307 0.109 0.217 0.115 0.456
Micromoles of glucose 0.031 0.047 0.054 0.155 0.055 0.109 0.058 0.230
Average glucose content
0.054 0.131 0.201 0.327
(per mL)
Concentration of glucose 6.65 16.33 25.13 163.5
mol (per g of tissue)
Mass of glycogen per 100g 0.12 0.27 0.41 2.67
of tissue
% Mass of glycogen in 1.83 2.32 1.63 10.68
tissue

Table 3 Results obtained from Assay of Tissue Glycogen from Fasted tissue

Fasted Tissue HEART KIDNEY MUSCLE LIVER

Mass of Tissue (g) 4.27 8.8 20.0 24.4
Final volume (mL) 8.5 17.6 40.0 50.8

Aliquot size mL 0.5 1.0 0.5 1.0 0.3 0.5 0.3 0.5
A450nm Sample i 0.005 0.018 0.004 0.005 0.0 0.004 0.006 0.011
Micromoles of glucose 0.0025 0.0091 0.002 0.0025 0.0 0.002 0.003 0.0055
Average glucose content
0.007 0.003 0.002 0.011
(per mL)
Concentration of 0.871 0.375 0.25 0.573
glucose mol (per g of
tissue)
Mass of glycogen per 0.014 0.0061 0.0041 0.0093
100g of tissue
% Mass of glycogen in 0.33 0.069 0.02 0.038
tissue
CALCULATIONS:
Determining μmol of Glucose for Table1:
E.g. using test tube # 2

Stock glucose solution (0.2 μmol/mL)

∴ 0.2mL will contain 0.2 ×0.2 = 0.04 μmol glucose

Determining μmol of Glucose for samples

E.g. using Fed Liver Tissue
From the calibration curve the equation of the line :
𝒚 = 𝟏. 𝟗𝟖𝟐𝟕𝒙

y=mx Y=absorbance x=concentration

Absorbance at 0.3 ml =0.115
0.115
∴ 𝑥 = 1.9827 = 𝟎. 𝟎𝟓𝟖 μmol of glucose
0.058
∴ in 1 ml = × 1 = 0.193
0.3

0.5 ml aliquot of fed liver = 0.230 μmol of glucose

In 1ml= 0.46 μmol of glucose
0.46+0.193
Average amount of glucose in 1ml of fed liver: 2
= 0.327 μmol of glucose
Flow chart 1: Fed Liver

Liver Fed (25 g)

4087.5
= 163.5 μmol/g
25

50 ml (Final volume)
163.5
2
× 50 = 4087.5 μmol

2ml (extract distributed)

= 163.5μmol

5ml (distilled H20 resuspended)

100 ml (Liver fed sample vol. made up)

=100ml x 1.635 μmol= 163.5μmol

1ml (hydrolysis)

5ml (after neutrilisation)

5ml x 0.327=1.635μmol

0.3 ml 0.5 ml

1ml = 0.327μmol
Determining Mass of glycogen (g) per 100 g of Fed Liver tissue
Mr of glycoside = 162
1000μmol= 1mmol
∴ 163.5 μmol = 0.1635 mmol
1mmol =1000 mol
0.1635
∴ 1000
× 162 = 0.027mol/g

∴ per 100 g = 0.027× 100 = 2.67mol/g of glucogen

Percentage mass of glycogen in fed liver tissue:

Mass of glycogen in 100g of fed liver tissue = 2.67g
∴ percentage of glycogen in the fed liver tissue
2.67
= 25
× 100 = 10.68%
Flow chart 2: Fed Muscle

Muscle Fed (25 g)

628.13
= 25.13 μmol/g
25

50 ml (Final volume)
25.13
2
× 50 = 628.13 μmol

2ml (extract distributed)

= 25.13μmol

5ml (distilled H20 resuspended)

25 ml (muscle fed sample vol. made up)

=25ml x 1.005 μmol= 25.13μmol

1ml (hydrolysis)

5ml (after neutrilisation)

5ml x 0.201=1.005μmol

0.3 ml 0.5 ml

1ml = 0.201μmol
Determining Mass of glycogen (g) per 100 g of Fed Muscle tissue
Mr of glycoside = 162
1000μmol= 1mmol
∴ 25.13 μmol = 0.0253 mmol
1mmol =1000 mol
0.0253
∴ 1000
× 162 = 4.1 x 10-3mol/g

∴ per 100 g = 4.1 x 10-3× 100 = 0.41mol/g of glucogen

Percentage mass of glycogen in fed muscle tissue:

Mass of glycogen in 100g of fed muscle tissue = 0.41g
∴ percentage of glycogen in the fed muscle tissue
0.41
= 25
× 100 = 1.63%
Flow chart 3: Fed Kidney

Kidney Fed (11.6 g)

189.53
= 16.33 μmol/g
11.6

22.8 ml (Final volume)

16.63
2
× 22.8 = 189.53 μmol

2ml (extract distributed)

= 16.63μmol

5ml (distilled H20 resuspended)

25 ml (muscle fed sample vol. made up)

=25ml x 0.665 μmol= 16.63μmol

1ml (hydrolysis)

5ml (after neutrilisation)

5ml x 0.131=0.665μmol

0.5 ml 1.0 ml

1ml = 0.131μmol
Determining Mass of glycogen (g) per 100 g of Fed kidney tissue
Mr of glycoside = 162
1000μmol= 1mmol
∴ 16.33 μmol = 0.01633 mmol
1mmol =1000 mol
0.01633
∴ 1000
× 162 = 2.7 x 10-3 mol/g

∴ per 100 g = 2.7 x 10-3 × 100 = 0.27 mol/g of glucogen

Percentage mass of glycogen in fed kidney tissue:

Mass of glycogen in 100g of fed kidney tissue = 0.27g
∴ percentage of glycogen in the fed kidney tissue
0.27
= 11.6 × 100 = 2.32%
Flow Chart 4: Fed Heart Tissue

Heart Fed (6.6 g)

43.9
= 6.65 μmol/g
6.6

13 ml (Final volume)
6.75
2
× 13 = 43.9 μmol

2ml (extract distributed)

= 6.75μmol

5ml (distilled H20 resuspended)

25 ml (heart fed sample vol. made up)

=25ml x 0.27 μmol= 6.75μmol

1ml (hydrolysis)

5ml (after neutrilisation)

5ml x 0.054=0.27μmol

0.5 ml 1.0 ml

1ml = 0.054μmol
Determining Mass of glycogen (g) per 100 g of Fed heart tissue
Mr of glycoside = 162
1000μmol= 1mmol
∴ 6.65 μmol = 0.00665 mmol
1mmol =1000 mol
0.00665
∴ 1000
× 162 = 1.2 x 10-3mol/g

∴ per 100 g = 0.431× 100 = 0.12mol/g of glucogen

Percentage mass of glycogen in fed heart tissue:

Mass of glycogen in 100g of fed heart tissue = 0.12g
∴ percentage of glycogen in the fed heart tissue
0.12
= 6.6
× 100 = 1.83%
Flowchart 5: Fasted Liver Tissue

Liver Fasted (24.4 g)

13.97
= 0.573μmol/g
24.4

50.8 ml (Final volume)

0.55
2
× 50.8 = 13.97 μmol

2ml (extract distributed)

= 0.55μmol

5ml (distilled H20 resuspended)

10 ml (Liver fasted sample vol. made up)

=10ml x 0.055 μmol= 0.55μmol

1ml (hydrolysis)

5ml (after neutrilisation)

5ml x 0.011=0.055μmol

0.3 ml 0.5 ml

1ml = 0.011μmol
Determining Mass of glycogen (g) per 100 g of Fasted liver tissue
Mr of glycoside = 162
1000μmol= 1mmol
∴ 0.573 μmol = 5.73 x10-4 mmol
1mmol =1000 mol
5.73 x10−4
∴ 1000
× 162 = 9.3 x 10-5 mol/g

∴ per 100 g = 9.3 x 10-5 × 100 = 0.0093 mol/g of glucogen

Percentage mass of glycogen in fasted liver tissue:

Mass of glycogen in 100g of fasted liver tissue = 0.0093g
∴ percentage of glycogen in the fasted liver tissue
0.0092
= × 100 = 0.038%
24.4
Flow Chart 6: Fasted Muscle Tissue

Muscle fasted (20.0 g)

5
= 0.25 μmol/g
20.0

40.0 ml (Final volume)

0.25
2
× 40.0 = 5 μmol

2ml (extract distributed)

= 0.25 μmol

5ml (distilled H20 resuspended)

25 ml (muscle fasted sample vol. made

up)
=25ml x 0.01 μmol= 0.25μmol

1ml (hydrolysis)

5ml (after neutrilisation)

5ml x 0.002=0.01μmol

0.3 ml 0.5 ml

1ml = 0.002μmol
Determining Mass of glycogen (g) per 100 g of Fasted muscle tissue
Mr of glycoside = 162
1000μmol= 1mmol
∴ 0.25 μmol = 2.5 x10-4 mmol
1mmol =1000 mol
2.5 x10−4
∴ 1000
× 162 = 4.1 x 10-5 mol/g

∴ per 100 g = 4.1 x 10-5 × 100 = 0.0041 mol/g of glucogen

Percentage mass of glycogen in fasted muscle tissue:

Mass of glycogen in 100g of fasted muscle tissue = 0.0041g
∴ percentage of glycogen in the fasted muscle tissue
0.0041
= 20
× 100 = 0.02%
Flow Chart 7: Fasted Kidney Tissue

Kidney Fasted (8.8 g)

3.3
= 0.375 μmol/g
8.8

17.6 ml (Final volume)

0.375
2
× 17.6 = 3.3 μmol

2ml (extract distributed)

= 0.375μmol

5ml (distilled H20 resuspended)

25ml (kidney sample vol. made up)

=25ml x 0.015 μmol= 0.375μmol

1ml (hydrolysis)

5ml (after neutrilisation)

5ml x 0.003=0.015μmol

0.5 ml 1.0 ml

1ml = 0.003μmol
Determining Mass of glycogen (g) per 100 g of Fasted kidney tissue
Mr of glycoside = 162
1000μmol= 1mmol
∴ 0.375 μmol = 3.75 x10-4 mmol
1mmol =1000 mol
3.75 x10−4
∴ 1000
× 162 = 6.1 x 10-5 mol/g

∴ per 100 g = 6.1 x 10-5 × 100 = 0.0061 mol/g of glucogen

Percentage mass of glycogen in fasted kidney tissue:

Mass of glycogen in 100g of fasted kidney tissue = 0.0061g
∴ percentage of glycogen in the fasted kidney tissue
0.0061
= 8.8
× 100 = 0.069%
Flow Chart 8: Fasted heart Tissue

Heart Fasted (4.27 g)

3.72
= 0.871 μmol/g
4.27

8.5 ml (Final volume)

0.875
2
× 8.5 = 3.72 μmol

2ml (extract distributed)

= 0.875μmol

5ml (distilled H20 resuspended)

25 ml (heart fasted sample vol. made up)

=25ml x 0.035 μmol=0.875μmol

1ml (hydrolysis)

5ml (after neutrilisation)

5ml x 0.007=0.035μmol

0.5 ml 1.0 ml

1ml = 0.007μmol
Determining Mass of glycogen (g) per 100 g of Fasted heart tissue
Mr of glycoside = 162
1000μmol= 1mmol
∴ 0.871μmol = 8.71 x10-4 mmol
1mmol =1000 mol
8.71 x10−4
∴ 1000
× 162 = 1.4 x 10-4 mol/g

∴ per 100 g = 1.4 x 10-4 × 100 = 0.014 mol/g of glucogen

Percentage mass of glycogen in fasted heart tissue:

Mass of glycogen in 100g of fasted heart tissue = 0.014g
∴ percentage of glycogen in the fasted heart tissue
0.014
= 4.27
× 100 = 0.33%
Graph 1: Calibration curve

Graph of Absorbance vs Concentration of Glucose (μmol)

0.45
y = 2.0513x - 0.009
0.4
R² = 0.9987
0.35

0.3
Absorbance (450nm)

0.25

0.2

0.15

0.1

0.05

0
0 0.05 0.1 0.15 0.2 0.25
-0.05
Concentration of Glucose (μmol)

Scale: X-axis Y-axis

DISCUSSION:
Glycogen is a polysaccharide that is made up of glucose residues linked by α(1-4) glycosidic bonds
and α(1-6) glycosidic bonds. Its main role is for the storage of glucose. However, it is also needed to
regulate blood glucose levels in organisms. The main sites for storage are the skeletal muscle and the
liver. The glycogen stored in the liver is used to meet the needs of the organism as a whole while the
glycogen stored in the muscle is used to meet the energy needs of that muscle. As such the liver has a
higher concentration of glycogen that that of muscle, while the muscle stores more glycogen with
regards to mass. (Stryer, 2015)
In the Fed state, there are high levels of glucose in the blood as such excess glucose is converted to
glycogen in a process called glycogenesis. While in the fasted state the blood glucose level is low and
glycogen degradation occurs. (Mckee, 2016)
In this lab, glycogen was quantified using an enzymatic/colourimetric assay of the fed and fasted
states in liver, heart, muscle and kidney tissue of a rat. In the fed state the concentrations of glycogen
found were as follows: 6.65μmol/g, 16.33 μmol/g, 25.13 μmol/g, 163.5 μmol/g for heart, kidney,
muscle and tissues respectively. The results obtained correspond to theory, where it states that
glycogen is mostly stored in the skeletal muscle and the liver. It can also be observed that the liver has
the highest concentration of glycogen of 163.5 μmol/g, this is expected as the liver is stated to have a
higher concentration of glycogen. As for the kidney and heart, the data obtained is expected as these
two tissues are not major stores of glycogen.
Mass per 100 g of tissue as well as percentage mass for the fed tissues was also determined. The
results are 0.12 per 100g, 0.27 per 100g, 0.41 per 100g, 2.67 per 100g for heart, kidney, muscle and
liver respectively. The percentage masses were: 1.83%, 2.32%, 1.63%, 10.68% for heart, kidney,
muscle and liver respectively. These results were also expected as they reflect the theory which states
that liver and skeletal muscle are major stores of glycogen and that liver has a higher concentration as
it is utilized in the whole organism.
In the fed state the concentrations of glycogen in the four tissues were determined as follows:
0.871μmol/g, 0.375 μmol/g, 0.25 μmol/g, 0.573 μmol/g for the heart, kidney, muscle and liver tissue
respectively. As expected the concentration of glycogen in the muscle is low as it needed to provide
energy for the muscle. However, it was expected that the liver concentration would be lower than that
of the tissues as the glycogen stored in the liver used to raise glucose levels in the blood in a fasted
state. This anomaly may have been caused by inaccurate pipetting skills. The high concentrations of
glycogen in the kidney and the heart are expected as these tissues are not major stores of glycogen and
would not have been utilized in a short space of time.
The mass per 100g for the tissues were: 0.014/100g, 0.0061/100g, 0.0041/100g, 0.0093/100g for
heart, kidney, muscle and liver respectively. The percentage masses were: 0.33%, 0.069%, 0.02%,
0.038% for heart, kidney, muscle and liver respectively.
In comparison between the fed and fasted stated it is seen that the levels of glycogen are higher in the
fed state than that of the fasted state. This was expected as in the fed state there are high levels of
glucose in the blood so insulin is activated and triggers the start of glycogen synthesis. While in the
fasted start the blood glucose levels are low and glucagon is activated and triggers the start of
glycogen degradation to raise the glucose levels. (Nelson and Cox)

SOURCES OF ERROR/PRECAUTIONS:

 Ensure that tubes are foiled as the enzyme cocktail is light sensitive
 Inaccutate pipetting may result in incorrect readings and results
ADDITIONAL DISCUSSION:
1. Role of Glycogen in the Liver: The liver is a major storage site of glycogen.The glycogen
stored in the liver as a role in the maintenance the blood glucose levels. The stores in the liver
are also utilized to meet the energetical need of the organism. (Stryer, 2015)

Role of Glycogen in the Muscle: The skeletal muscle is another major storage site of
glycogen, however unlike the liver the glycogen stores are utilized to meet the energetical
needs of the muscle. (Stryer, 2015)

Role of Glycogen in the Kidney: Glycogen stored in the kidney is used for energy, however,
the kidney stores very little glycogen. (Stryer, 2015)

Role of Glycogen in the Heart: The heart uses the stored glycogen for energy, however, it
stores very little amounts. (Nelson and Cox, 2017)

2. Glucose is soluble in water and if stored freely and has a higher osmotic effect on the cell and
cause the cell to burst. However, when stored as glycogen, this form is insoluble in water and
will not disrupt the osmotic pressure in the cell and cause it to burst. (Stryer, 2015)

3. Energy is mostly stored as fats. This is the most concentrated form of energy. It can be stored
anhydrous unlike carbohydrates and protein who which use 4 grams of water for every 1 gram
of carbohydrate/protein. Fats also provide twice the amount of potential energy than that of
carbohydrates and proteins (9 kcal/g vs 4kcal/g). (Nelson and Cox, 2017)
REFERENCES:
Berg, Jeremy M, John L Tymoczko, Lubert Stryer, and Gregory J Gatto. “Glycogen Metabolism.” In
Biochemistry, 8th ed., 618–19. New York: W.H Freeman, 2015.
Mckee, Trudy, and James R Mckee. “Carbohydrate Metabolism.” In Biochemistry. The Molecular
Basis of Life, 6th ed., 33–37. Oxford University Press, 2016.
Nelson, David L., and Michael M. Cox. Lehninger Principles of Biochemistry. New York: W.H.
Freeman, 2017.