Fecht, Johnson, & Subrin 1

Determination of Macroelements, Microelements, Essential Elements, Trace and Toxic Elements

in Fruit Samples
Elizabeth Fechta, Drew Johnsona, and Liz Subrina
a
Department of Chemistry: Analytical Chemistry 1, Students at Butler University, Indianapolis, IN, United States

Abstract
Fruits contain various metals that are essential for life. The metal content of elements in
sample plums were found through by preparing samples through open vessel hot-plate based acid
digestion and closed vessel microwave based acid digestion. Flame Atomic Absorption Spectrum
was then used to determine the macroelements. The samples made using the open method were
found to be a better sample digestion. Through this experiment, it has been determined that
calcium, potassium, sodium, and magnesium elements were found in a plum.
Background/Introduction
Mineral elements provide nutrients and are essential for proper development in
organisms. Various elements including trace and toxic macroelements are found within a sample
of fruit. Some elements are characterized as being macroelements, essential elements and trace
elements. Macroelements include N, P, K, Ca, and Mg and vary in existence based on the type of
fruit. Macroelements are a type of mineral elements that are classified as major elements within
normal life processes because they require amounts greater than 100 mg/dl in the human body.
The human body contains both nonessential and essential elements. Essential elements are
needed in adequate amounts to maintain normally physiological functions of the human body
[1]. Trace elements are also sometimes characterized as essential elements, these include Fe, Mn,
Zn, and Cu. Trace elements are known as inorganic micronutrients many of which are essential
components of enzymes [2]. Trace elements are involved in many vital cellular functions. For
plums, these elements play important roles in metabolic functions, fruit development, and
disease resistance. The amount of exposure to toxic metals through consumption is important due
to the health impacts it can have on humans and the affects it can have on stability of the fruit. A
study at the University of Ibadan, researched the importance of elements for humans in order to
review functions in health and disease conditions so to prevent nutrition-related diseases. These
scientists believed data on mineral contents of human foods and animal feeds are essential for
formulation of feeding regimes and food processing techniques [3].
Soils and environments that fruit plants are grown in impact the mineral elements that are
present in the fruit. These trace elements by plants can occur through the roots (main entry
route) or through leaf stomata after atmospheric deposition over them [4]. The roots can absorb
these elements selectivity and do so in the form of inorganic ions. The uptake of minerals
depends on the soils cation exchange capacity, the soil pH, and the presence of fungi [5]. A lack
of mineral elements can affect the metabolism and tissue structure due to the failed enzyme
system. The most common symptoms of mineral deficiency are stunted growth and discoloured
leaves. The symptoms of a mineral deficiency depend partly on the function of that nutrient in
the plant [3].
In a 1997 study, it was determined through a study utilizing a variety of fruits and
vegetables, that vegetables contain a higher amount of minerals than fruits. Techniques differed
in finding the minerals, but it is wise to note Calcium was detected with O-cresolphthalein

Fecht, Johnson, & Subrin 2

complexone in the presence of 8-hydroxyquinoline, which are not available for this study. This
study took place in Mexico and it must be taken into account the climatic conditions influence
the growth rates of plants and therefore the rates of mineral ion utilization. Genetic factors, soil
and weather conditions, the use of fertilizers, and the state of the plants maturity at harvest all
affect the final level of mineral components in a plant [6].
Through the method of cultural in vitro, macroelements were determined in a plant
material in research conducted by Malgorzata Podwyszynska and Tadeusz Olszewski. Based on
chemical analysis observations indicate that macroelement uptake depends on the gelling agent
and varies between species [7]. There were much higher variations in the contents of Mg and Ca
rose shoots. Additionally, Mg and Ca generally had similar uptake by rose shoots. Results were
enhanced for Ca and Mg uptake when the shoot length or fresh weight was increased in the
experiment.
In a 2015 study, food samples were examined to evaluate elements that are present in
samples and to estimate appropriate dietary intake of elements. Standard solutions of elements
were made, certified reference material was obtained, and food samples were purchased.
Researchers then prepared the food as it would normally be prepared. In some cases this included
cooking. Samples were measured and placed into a Teflon vessel to be digested. Nitric acid was
removed by placing sample vessels on hot plates. The digested solution was filtered through a
0.45 m nitrocellulose membrane filter, transferred to a 50-mL polyethylene flask, and diluted to
the final volume with ultra-pure water, the solutions were analyzed within seven days and
concentrations were measured using an ICAP6300 ICP-OES. Na, K, Mg, and Ca were then
analyzed. Researchers refrained from using glassware and when water was used, they used ultrapure water. Calibration curves were formed for elements at various concentrations and this data
aligns with the reference values of these materials. Contamination tests were also made by
analyzing blank samples [1].
In this work we will determine a variation of elements in plum samples.
Experimental
Preparing the Calcium Stock Solution
0.1233 g of CaCO (Fisher Scientific Company) was transferred into a 100 mL volumetric
flask. About 50 mL of deionized water was added to the flask, and then 1.0 mL of concentrated
HCl was micropipetted (100-1000L micropipette, 6 L, Finnpipette, Fischer Brand) into the
flask. The flask was filled to the mark with deionized water. The resultant solution was 493.7
ppm Ca .
3

2+

Preparing the Calcium Standard Solutions

10.0 mL concentrated HNO and 2 mL Cs were micropipetted (2-10 mL micropipette,
0.02 mL, Finnpipette F2, Thermo Scientific Brand) into eight separate 250 mL volumetric
flasks. To each consecutive flask, volumes of 0 L, 506.4 L, 1012.8 L, 1519.2 L, 2025.6 L,
2532 L, 3038.4 L, and 3544.7 L of the 493.7 ppm Ca stock solution were micropipetted
(100-1000 L micropipette) and filled to the mark with deionized water so that standard
solutions of 0, 1, 2, 3, 4, 5, 6, and 7 ppm Ca , respectively, were prepared.
3

2+

2+

Fecht, Johnson, & Subrin 3

Preparing the Digestates
To six 50 mL volumetric flasks, 5.0 mL of fruit juice was micropipetted (2-10 mL
micropipette). To these 50 mL volumetric flasks and two additional 50 mL volumetric flasks, 2.0
mL of concentrated HNO was micropipetted (2-10 mL micropipette). Each flask was filled to the
mark with deionized water. Four solutions, 3 including fruit juice and 1 blank, were transferred
to digestate vessels - teflon material. These were submitted to a microwave (Mars 6 One Touch
Technology by CEM) in which the temperature was ramped to 150 degrees Celsius over 15
minutes, held for 5 minutes, and cooled for 15 minutes. These digestates were transferred to 50
mL centrifuge tubes for storage. The other four solutions were transferred to 50 mL centrifuge
tubes and digested via hot plate for 30 minutes.
3

Constructing the Calcium Calibration Curve

Standard solutions of 0, 1, 2, 3, 4, 5, 6, and 7 ppm calcium were poured into 8 separate
clean beakers. The absorbance of each solution was determined by running the solution through
the FAAS (Perkin Elmer 2380) and obtaining 10 measurements. The average of the absorbances
were used to obtain a calibration curve yielding an equation y=0.0217x+0.00004 with the yvariable representing absorbance and the x variable representing ppm calcium. The calibration
curve produced an R value of 0.9998.
2

Measuring the Calcium Concentration of Digestates via FAAS

5.0 mL of each digestate (open blank, open sample in triplicate, closed blank, closed
sample in triplicate) was micropipetted (2-10 mL micropipette) into two separate 10 mL
volumetric flasks so that a total of 16 volumetric flasks were prepared. 100 L of Cs was
micropipetted (20-200 L micropipette) into each flask, and 300 L of HNO was micropipetted
(200-1000 L micropipette) into each flask. 60.8 L of a 493.7 ppm calcium stock solution was
micropipetted (20-200 L micropipette) into a single flask of each digestate so that 8 flasks were
spiked with each digestate represented and 8 flasks were unspiked with each digestate
represented. The flasks were filled to the mark with deionized water. The absorbance of each
solution was determined by running the solution through the FAAS. The spike recovery was
calculated, indicating the accuracy of the data.
3

Constructing the Potassium Calibration Curve

Standard solutions of 0, 0.67, 0.96, 1.53, 1.72, and 2.105 ppm potassium were poured
into 6 separate clean beakers. Running the solution through the FAAS and obtaining 10
measurements determined the absorbance of each solution. The average of the absorbances were
used to obtain a calibration curve yielding an equation y=0.0587x+0.0042 with the y-variable
representing absorbance and the x variable representing ppm potassium. The calibration curve
produced an R value of 0.99919.
2

Measuring the Potassium Concentration of Digestates via FAAS

405 L of each digestate (open blank, open sample in triplicate, closed blank, closed
sample in triplicate) was micropipetted (100-1000 L micropipette) into two separate 10 mL
volumetric flasks so that a total of 16 volumetric flasks were prepared. 8 L of Cs was
micropipetted into each flask (5-50 L micropipette), and 24 L of HNO was micropipetted (550 L micropipette) into each flask. 7.45 L of a 972.7 ppm potassium stock solution was
micropipetted (5-50 L micropipette) into a single flask of each digestate so that 8 flasks were
3

Fecht, Johnson, & Subrin 4

spiked with each digestate represented and 8 flasks were unspiked with each digestate
represented. The flasks were filled to the mark with deionized water. The absorbance of each
solution was determined by running the solution through the FAAS. The spike recovery was
calculated, indicating the accuracy of the data.
Constructing the Sodium Calibration Curve
Standard solutions of 0, 0.25, 0.5, 0.75, 1, and 2 ppm sodium were poured into 6 separate
clean beakers. The absorbance of each solution was determined by running the solution through
the FAAS. The data points were used to obtain a calibration curve yielding an equation
y=0.1315x + 0.0284 with the y-variable representing absorbance and the x variable representing
ppm sodium. The calibration curve produced an R value of 0.9924.
2

Measuring the Sodium Concentration of Digestates via FAAS

4 mL of each digestate (open blank, open sample in triplicate, closed blank, closed
sample in triplicate) was micropipetted into two separate 10 mL volumetric flasks so that a total
of 16 volumetric flasks were prepared. 80 L of Cs was micropipetted (20-200 L micropipette)
into each flask, and 240 L of HNO was micropipetted (100-1000 L micropipette) into each
flask. 7.3 L (5-50 L micropipette) of a 1009 ppm sodium stock solution was micropipetted into
a single flask of each digestate so that 8 flasks were spiked with each digestate represented and 8
flasks were unspiked with each digestate represented. The flasks were filled to the mark with
deionized water. The absorbance of each solution was determined by running the solution
through the FAAS. The spike recovery was calculated, indicating the accuracy of the data.
3

Constructing the Magnesium Calibration Curve

Standard solutions of 0, 0.12, 0.24, 0.36, 0.48, and 0.60 ppm magnesium were poured
into 6 separate clean beakers. The absorbance of each solution was determined by running the
solution through the FAAS. The data points were used to obtain a calibration curve yielding an
equation y=0.3541x + 0.0215 with the y-variable representing absorbance and the x variable
representing ppm magnesium. The calibration curve produced an R value of 0.9944
2

Measuring the Magnesium Concentration of Digestates via FAAS

625 L of each digestate (open blank, open sample in triplicate, closed blank, closed
sample in triplicate) was micropipetted (100-1000 L micropipette) into two separate 25 mL
volumetric flasks so that a total of 16 volumetric flasks were prepared. 12.5 L of Cs was
micropipetted (5-50 L micropipette) into each flask, and 37.5 L of HNO was micropipetted
(5-50 L micropipette) into each flask. 0.55 L of a 9020 ppm magnesium stock solution was
micropipetted (0.5-5 L micropipette) into a single flask of each digestate so that 8 flasks were
spiked with each digestate represented and 8 flasks were unspiked with each digestate
represented. The flasks were filled to the mark with deionized water. The absorbance of each
solution was determined by running the solution through the FAAS. The spike recovery was
calculated, indicating the accuracy of the data.
3

Fecht, Johnson, & Subrin 5

Results/Discussion

f(x) = 0.02x + 0
R = 1

Graph 1: Absorbance vs. Concentration (ppm) of Calcium Calibration Curve.

Table 1: Calcium Concentration, Precision, and Spike Recovery of Digestates.
Concentration
(ppm)
SD
%RSD
Spiked Open Blank
3.14101382
0.08864232 2.82209273
Spiked Open Sample
4.83072197
0.20030628 4.14650809
Spiked Closed Blank
3.29769585
0.05254265 1.5933139
Spiked Closed Sample
4.5265745
0.04729587 1.04484894
Unspiked Open Blank
0.05345622
0.02060892 38.5528962
Unspiked Open Sample
1.73394777
0.01407857 0.81193758
Unspiked Closed Blank
0.14562212
0.02060892 14.152329
Unspiked Closed Sample
1.50046083
0.0513158 3.42000268

Spike
Recovery
102.860436
103.167482
105.009759
100.813462

Fecht, Johnson, & Subrin 6

f(x) = 0.06x + 0
R = 1

Graph 2: Absorbance vs. Concentration (ppm) of Potassium Calibration Curve.

Table 2: Potassium Concentration, Precision, and Spike Recovery of Digestates.
Concentration
(ppm)
SD
%RSD
Spiked Open Blank
0.15332198
0.00761863 4.96903995
Spiked Open Sample
2.31232254
0.06377238 2.75793618
Spiked Closed Blank
0.14650767
0.01425315 9.72860496
Spiked Closed Sample
1.78989211
0.01095247 0.61190677
Unspiked Open Blank
0.0174321
0.00933088 53.5269894
Unspiked Open Sample
1.93299262
0.08913038 4.6110045
Unspiked Closed Blank
0.0192344
0.00933088 48.5114083
Unspiked Closed Sample
1.58705281
0.20647444 13.0099286

f(x) = 0.13x + 0.03

R = 0.99

Percent
Recovery
46.858578
130.803423
43.8873331
69.9445848

Fecht, Johnson, & Subrin 7

Graph 3: Absorbance vs. Concentration (ppm) of Sodium Calibration Curve.
Table 3: Sodium Concentration, Precision, and Spike Recovery of Digestates.
Concentration
(ppm)
SD
%RSD
0.0120238 1.9617107
Spiked Open Blank
0.61292776
7
1
0.0277262 3.4635868
Spiked Open Sample
0.80050697
5
1
0.0120238 2.7450326
Spiked Closed Blank
0.43802281
7
9
0.0584381 3.8745966
Spiked Closed Sample
1.50823828
5
4
0.0063624 46.655327
Unspiked Open Blank
0.0136371
3
5
0.0325252 139.46969
Unspiked Open Sample
0.02332066
5
5
0.0063624 41.764146
Unspiked Closed Blank
0.0152342
3
9
0.0191377 2.8318958
Unspiked Closed Sample
0.67579214
3
9

Percent
Recovery
81.204696
105.309798
57.28843
112.797579

f(x) = 0.35x + 0.02

R = 0.99

Graph 4: Absorbance vs. Concentration (ppm) of Magnesium Calibration Curve.

Table 4: Magnesium Concentration, Precision, and Spike Recovery of Digestates.
Concentration
Percent
(ppm)
SD
%RSD
Recovery
Spiked Open Blank
0.46173397
0.00345875 0.74907943
213.061423
Spiked Open Sample
0.52800527
0.03876412 7.34161543
172.842993
Spiked Closed Blank
0.29567919
0.00945112 3.19640889
135.304485

Spiked Closed Sample

Unspiked Open Blank
Unspiked Open Sample
Unspiked Closed Blank
Unspiked Closed Sample

0.47039443
0.0015213
0.15466441
0.0034215
0.11362139

0.00991778
0
0.00423923
0
0.00254688

Fecht, Johnson, & Subrin 8

2.10839579
165.172704
0
2.74092033
0
2.24154917

Table 5: Mass of Metal (g) in a Plum

g/Fruit
SD
199.40399 1.6190355
Calcium
4
5
222.29415 10.249993
Potassium
2
7
77.716096 2.2008389
Sodium
3
6
Magnesiu 17.786407 0.4875114
m
2
5
The construction of calibration curves for all elements yielded curves with R values of
0.9998, 0.9992, 0.9924, and 0.9944 for calcium, potassium, sodium, and magnesium,
respectively. These high R values suggest the presence of a strong linear relationship between
the concentration of each metal and its absorbance.
Between the open and closed methods of acid digestion, the open method via hot plate
analysis tended to yield higher concentrations of the trace metals. In fact, in the unspiked sample
digestates, the open vessel method yielded concentrations that were 0.2335, 0.34594, and
0.04104 ppm higher for calcium, potassium, and magnesium, respectively. For calcium, on the
other hand, the concentration of the unspiked closed sample was 0.6525 ppm higher than that of
the unspiked open sample, suggesting a preference for the closed vessel digestion method.
However, open vessel digestion was favored overall between the four trace metals.
The accuracy of the method could be established by percent recovery, and it was found to
be more accurate for certain metals than other metals. In calcium, for example, the spike
recoveries were 102.9%, 103.2%, 105.0%, and 100.8% for the spiked open blank, spiked open
sample, spiked closed blank, and spiked closed sample, respectively. Other metals yielded less
precise spike recoveries. In potassium, the spike recoveries were 46.9%, 130.8%, 43.9%, and
69.9% for the spiked open blank, spiked open sample, spiked closed blank, and spiked closed
sample, respectively. In sodium, the spike recoveries were 81.2%, 105.3%, 57.3%, and 112.8%
for the spiked open blank, spiked open sample, spiked closed blank, and spiked closed sample,
respectively. In magnesium, the spike recoveries were 213.1%, 172.8%, 135.3%, and 165.2% for
the spiked open blank, spiked open sample, spiked closed blank, and spiked closed sample,
respectively.
A spike recovery of about 100% indicates little matrix effect and little analyst error. In
calcium, the matrix seemed to have little effect on the concentration of calcium in the plum.
However, a spike recovery of less than 100% indicates that there was a significant amount of
matrix effect. In potassium, the spike recovery did seem to indicate that there was a significant
matrix effect for the blanks and the spiked closed sample. However, a spike recovery of greater
than 100% tends to simply indicate error on the part of the analyst. When working with trace
2

Fecht, Johnson, & Subrin 9

elements, errors like this are very common, and are difficult to avoid. The spiked open sample
for potassium seemed to show large error. For sodium, the blanks seemed to experience
significant matrix effect, while the spiked samples seemed to contain a large amount of error on
the analysts part. Finally, in magnesium, large spike recoveries indicated that there was
significant error on the analysts part. Overall, the only sufficiently accurate determination was
that of calcium.
Conclusion
Mineral elements in fruit are important since it plays various roles on the nutritional
contents of the fruit as well as the health of the human digesting it. The mass (g) of each
element in a single plum was found to be 199.4, 222.3, 77.7, and 17.8 for calcium, potassium,
sodium, and magnesium, respectively. Based on the different methods used, the open method
yielded more accurate results.
Acknowledgements
Thank you to Dr. Akinbo for support and help throughout this lab project. Additionally,
thank you to the Analytical Chemistry students at Butler University for the help in preparing
standards to run through the FAAS.

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