1 FST 202 Food Analysis

Measurement of Respiration Rate of

Banana through Gas Chromatography
Mary Jane T. Coloma, MSFSc, University of the Philippines, Los Baños, Laguna, Philippines

ABSTRACT
Accurate determination of respiration is important for assessment of metabolic processes in plant tissues. In this
study, gas chromatography was used to measure the respiration rate of banana, a climacteric fruit that has a short shelf life.
Unripe and ripe banana fruits were stored in an aerobic jar and subjected to Shimadzu gas chromatography. Results
indicated that the concentrations of CO2 and O2, which were determined based on the height of their peaks in the
chromatogram, increased after 2 hours. However, based on theoretical principle, the level of O 2 should decrease as it is
consumed during respiration to break down glucose into simpler molecules, such as CO2 and H2O. Respiration rate was
calculated and found to be higher in the ripe banana than in the unripe banana. The amount of heat produced was also
determined and could be correlated with the respiration rate. Respiration quotient, which is the ratio of CO2 produced to O2
consumed, were determined, and the values were outside the reported range for fruits. For future works, collection of gas
samples should be as short as needed to accumulate sufficient gas for accurate measurement. Inspection of possible leak in
gas-tight syringes should also be conducted.

Keywords: respiration, gas chromatography, banana

INTRODUCTION column, detector, and electronic recording/data

handling system (Grob and Barry, 2004).
Similar to other types of chromatography, GC
Gas chromatography (GC) is an
employs a mobile phase, that is, carrier gas of
analytical technique that can be used to
high purity, and a stationary phase, either
determine the amount of gases consumed and
packed or capillary columns. The packed
produced by fruits (Lyons et al., 1962). GC
column consists of a liquid coated on an inert
was first used by James and Martin (1952) to
support, whereas the capillary column is made
separate and quantify fatty acids. Since then,
of a hollow silica glass (Nielsen, 1998).
GC technique has greatly improved and has
Separation of volatile compounds occurs when
wide applications. GC can be used to
the carrier gas flows through the injector and
determine a variety of compounds, such as
the sample is pushed into the GC column; the
fatty acids, triglycerides, cholesterols, gases,
sample is then delivered into a detector
solvents, alcohols, sugars, amino acids,
(Yokotani et al., 2009). Commonly used
peptides, vitamins, and food additives
detectors are thermal conductive detector,
(Nielsen, 1998). Ideally, GC is suitable for
flame ionization detector, and electron capture
evaluation of thermally stable volatile
detector (Nielsen, 1998). During partitioning
substances. In GC, compounds are identified
in the mobile and stationary phase, the
and analyzed through separation, vaporization,
different components of the sample will reach
and purification without requiring
the detector at varied times; the detector has a
decomposition (Harris, 1999). Prior to GC,
sensor that then sends signal to the recorder,
different methods of sample preparation are
and the results are displayed on graphs or
conducted. Such methods include isolation of
tables (Yokotani et al., 2009).
solutes from food, headspace method,
distillation, solvent extraction, solid-phase
Respiration provides energy for
extraction, and direct injection (Nielsen,
biochemical processes (Fonseca et al., 2002).
1998). The major components of GC are gas
In this metabolic process, a variety of
source, flow controller, injection port, oven,
substances vital for metabolic pathways are
2 FST 202 Food Analysis
produced. Respiration is a vital factor
considered in prolonging the shelf life of fruits
and vegetables. In aerobic respiration, organic
reserves are broken down to simple molecules,
such as CO2 and H2O, and energy is released.
Moreover, O2 is consumed in a series of
reactions. Factors affecting respiration include
the nature of the commodity, such as its type
and maturity, and temperature (Song et al.,
1992). Different metabolic activities occur at
different temperatures and lead to different
respiration rates. In climacteric commodities,
such as banana, the respiration rate is high
during the early development phase, and then
decreases until a rise, which coincides with
ripening, occurs (Lopez–Galvez et al., 1992).
Banana, one of the most popular tropic
fruits, has a climacteric nature, which shows
peaks of ethylene production and cell
respiration associated with ripening after
harvest (Knee, 2002). Banana has a relatively
short shelf life and leads to economic losses; in
this regard, post-harvest technologies are
applied to reduce such loss (Heydari et al.,
2010). One of these technologies is modified
atmospheric packaging (MAP), in which the
O2 content within the packaging is reduced to
approximately 5% to delay the ripening of
banana. The design of MAP relies on
respiration rate, which is measured as a
function of O2 and CO2 in a closed system
(Ullah et al., 2006).
This study aims to employ GC for
measurement of respiration rate of banana.
Results could be vital for designing MAP to
increase the shelf life of this commodity.
MATERIALS AND METHODS
Unripe (green) and ripe (yellow)
bananas were purchased from a local market.
Each fruit was weighed and placed in a
respiration jar (Figure 1). The jar was sealed
with rubber stoppers fitted with a sampling
port and flushed with a gentle stream of air.
Samples (1 mL) were collected from the
headspace by using a gas-tight syringe. The
samples were then injected into the GC
(Shimadzu Figure 1) equipped with thermal
conductivity detector to determine CO2 and
O2 levels at time zero. Standard gas
containing 1% O2 and 15% CO2 was also run
in the GC. The jar together with its content
was allowed to stand undisturbed for 2 hours.
After which, 1 mL of samples were collected
again from the headspace to give
measurement at time 2 hours. Data displayed
by the computer connected to the GC were
recorded.
Percent CO2 and O2 were calculated
using their corresponding peak heights with
the following equations:
where GC response is the height of peaks
shown in the gas chromatograph. The
determined values were then used to
determine respiration rate (mgCO2/kg hour of
the sample) as follows:
( ) ( )
where (%CO2)t is the percent CO2 at time
interval (2 hours), (%CO2)0 is the percent
CO2 at time zero, w is the fresh weight of the
sample (kg), and Vf is the volume of free
space (volume of jar-volume of fruit).
Heat production in kcal and BTU as well as
3 FST 202 Food Analysis

respiration quotient (RQ) were also

calculated with the following:

( ) where heat produced is presented in kcal/kg

hour and BTU/ton day.
( )

Figure 1. Set up and GC system. Unripe and ripe banana fruits were placed in respiration jars for 2
hours (left). Samples were withdrawn from the headspace of the jars and inserted into GC (GC-2014,
Shimadzu; right).

respiration. The loss of O2 can be used to

RESULTS AND DISCUSSION measure respiration rate. However, the
change is usually small and thus difficult to
Figure 2 and 3 show the
measure. Meanwhile, the small change, that
chromatograms of the standard, unripe
is, increase, in the CO2 level can be easily
banana, and ripe banana at time zero and
measured because of its low atmospheric
after 2 hours, respectively. The baseline
concentration (0.3%). In this regard, the
peaks of CO2 (at time zero) showed evident
change in CO2 is typically used to measure
increase after 2 hours, and the peaks of O2
respiration rate (Saltveit, 2019). The balanced
also increased but at a lower degree. In
equation for aerobic respiration is as follows:
principle, the level O2 decreases as it is
consumed by the respiring tissue and that of C6H12O6 + 6O2 + 6H2O  6CO2 + 12
CO2 increases as it is produced during H2O + 673 kcal (38 ATP)

(a) (b) (c)

Figure 2. Chromatograms at time zero. (a) Standard, (b) unripe banana, and (c) ripe banana placed
in a closed chamber at room temperature. About 1 mL of headspace gas was drawn by the syringe
and injected into the GC. The charts showed the peaks of CO2 and O2 from the climacteric and
respiration process.
4 FST 202 Food Analysis

(a) (b) (c)

Figure 3. Chromatograms after 2 hours. (a) Standard, (b) unripe banana, and (c) ripe banana
placed in a closed chamber at room temperature after 2 hours. About 1 mL of headspace gas was
drawn by the syringe and injected into the GC. The charts showed the peaks of CO2 and O2 from the
climacteric and respiration process.
continue to respire, a large amount of
Table 1 shows the concentrations of O2 and chemical energy is released as heat (Sietvalt,
CO2 in the standard, unripe banana, and ripe 2019). In terms of RQ, Fonseca (2002)
banana at baseline and after 2 hours. The reported that its different values correspond
calculated respiration rate, heat production, to the type of substrate utilized. If RQ is
and RQ are also presented. The respiration equal to 1, lipid is consumed; if it is higher
rate and heat production are higher in the ripe than 1, organic acids are used up; and if it is
banana than those in the unripe fruit. After lower than 1, lipids are utilized. Fagundes
reaching the climacteric peak, the respiration (2013) reported that RQ varies at different
rate increases, reaching its maximum value of temperatures and is affected by CO2
125 mg CO2/kg hour, then declines slightly production and O2 consumption. The normal
but is still at a high level (Seymour, 1993). RQ value in literature ranges from 0.7 to 1.3
Consistent with this report, the present study (Beaudry et al., 1992). The obtained values in
reported increasing respiration rate, with a the present work are outside this range,
value of 114.12 mg CO2/kg hour, which is which could be attributed to increased O2
within the maximum value. Meanwhile, the concentration due to possible errors during
heat production, both in kcal and BTU, are the GC operation.
higher in the ripe banana. As the fruit ages or

Table 1 Concentration and production rate of O2 and CO2 measured from unripe and ripe banana
fruits.
Unripe Ripe
%O2 at t=0 17.04% 15.26%
%O2 at t=120 17.15% 17.87%
%CO2 at t=0 0.09% 0.09%
%CO2 at t=120 0.70% 0.93%
Respiration rate 64.05 mg CO2/kg hr 114.12 mg CO2/kg hr
Heat production (Kcal) 0.16 Kcal/kg hr 0.29 Kcal/kg hr
Heat production (BTU) 14090.46 BTU/ton d 25106.37 BTU/ton d
Respiration quotient 5.65 0.32
Values are means of duplicate trials.
5 FST 202 Food Analysis
CONCLUSIONS AND
RECOMMENDATION
This study successfully applied GC to
determine the respiration rate of unripe and
ripe banana fruits. The chromatographs
showed the peaks of O2 and CO2. The height
of the peaks was then used to determine the
concentrations of both gases, which increased
after 2 hours. However, based on theoretical
principle, the level of O2 should decrease as it
is consumed during respiration to break down
glucose into simpler molecules such as CO2
and H2O. The values were then inputted in
equations of respiration rate. The calculated
respiration was higher in the ripe banana than
in the unripe banana. This finding is consistent
with the fact that respiration increases during
ripening. The amount of heat produced was
also determined and could be correlated with
the respiration rate. As fruits continue to aged
and respire, a significant amount of heat is
generated in the process. The RQ, which is the
ratio of CO2 produced to O2 consumed, had
values that were outside the reported range for
fruits. The results should be interpreted with
caution as possible errors were incurred during
the GC operation. Collection of gas samples
should be as short as needed to accumulate
sufficient gas for accurate measurement.
Inspection of possible leak in gas-tight
syringes should also be conducted.
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