ATOMIC ABSORPTION SPECTROPHOTOMETRY COOKBOOK

Section 6

Foodstuffs Analysis
Fertilizer and Feed Analysis
 Atomic Absorption Analysis Cookbook
Section 6
Table of Contents

10. Foodstuffs Analysis ..................................................................................... 1

10.1 Vegetables, Fruits Analysis Method...................................................................... 1
10.1.1 Sample Pretreatment ...................................................................................... 1
10.1.2 Electrical Heating Atomic Absorption Method ............................................. 2
10.1.3 Flame Atomic Absorption Method ................................................................ 4
10.1.4 Hydride Vapor Generation Atomic Absorption Method ............................... 13
10.1.5 Reduction Vaporization Atomic Absorption Method.................................... 15
10.2 Grains, Beans, Potatoes Analysis Method............................................................. 18
10.3 Meat, Fish Analysis Method.................................................................................. 18
10.4 Fats and Oils, Dairy Products Analysis Method.................................................... 18
10.4.1 Sample Pretreatment ...................................................................................... 18
10.4.2 Electrical Heating Atomic Absorption Method ............................................. 19
10.4.3 Flame Atomic Absorption Method ................................................................ 23
10.4.4 Hydride Vapor Generation Atomic Absorption Method ............................... 23
10.4.5 Reduction Vaporization Atomic Absorption Method.................................... 23
10.5 Spirits, Alcoholic Beverage Analysis .................................................................... 24
10.5.1 Sample Pretreatment ...................................................................................... 24
10.5.2 Electrical Heating Atomic Absorption Method ............................................. 24
10.5.3 Flame Atomic Absorption Method ................................................................ 31
10.5.4 Hydride Vapor Generation Atomic Absorption Method ............................... 31
10.5.5 Reduction Vaporization Atomic Absorption Method.................................... 32
10.6 Processed Foods Analysis Method ........................................................................ 33
10.6.1 Sample Pretreatment ...................................................................................... 33
10.6.2 Electrical Heating Atomic Absorption Method ............................................. 33
10.6.3 Flame Atomic Absorption Method ................................................................ 33
10.6.4 Hydride Vapor Generation Atomic Absorption Method ............................... 33
10.6.5 Reduction Vaporization Atomic Absorption Method.................................... 33

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11. Fertilizer and Feed Analysis ................................................................................. 34
11.1 Fertilizer Analysis Method .................................................................................... 34
11.1.1 Sample Pretreatment ...................................................................................... 34
11.1.2 Flame Atomic Absorption Method ................................................................ 36
11.1.3 Hydride Vapor Generation Atomic Absorption Method ............................... 41
11.1.4 Reduction Vaporization Atomic Absorption Method.................................... 43
11.2 Feed Analysis Method ........................................................................................... 45
11.2.1 Sample Pretreatment ...................................................................................... 45
11.2.2 Flame Atomic Absorption Method ................................................................ 45

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G10. foodstuffs analysis
10.1 Vegetables, Fruits Analysis Method
Reference materials:
Health Testing Methods Commentary, Japan Pharmaceutical Society Publication,
Kanehara Publishing Co.,
Foodstuffs Analysis Methods, Japan Foodstuffs Manufacturing Society,
Foodstuffs Analysis Methods Editorial Commission Publication (Korin Co.)
10.1.1 Sample Pretreatment
a) Nitric acid, perchloric acid decomposition
First, grind and air dry 2 ~ 5g of sample and place in a conical beaker. Wet
with water, add 25 ml of nitric acid, and after mixing, set aside. Next, gently
heat to start a sample reaction. After cooling, add 10 ml of perchloric acid and
gently heat to concentrate. Midway, if the contained material becomes dark, add
2~3 ml portions of nitric acid and continue heating. When the contained
material turns yellowish or colorless, decomposition is complete. After cooling,
add 2 ml of hydrochloric acid, and use water to prepare fixed volumes of
measurement solution.
Remarks:
1) If during the decomposition process the sample becomes carbonized
and hardens, it will certainly explode, so be sure that nitric acid is
always present in the sample while heating.
2) Measurement is suitable for As, Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Na,
Pb, Sb, Sn, Zn, etc., but if halogens are coexistent with As, Sb, Se and
Sn, or if the contained material becomes carbonized, volatilization and
loss will occur.
3) Since there is a possibility of contamination with impurities to
reagents and utensils, prepare processed blank test solutions in the same
way as the samples.
b) Ashing by drying
First, grind and air dry 5~10g of sample and place in a quartz beaker. Then
gently heat on a hot plate. Continue heating until enough water is driven off for
partial carbonization to occur. Then place the beaker in an electrical furnace,
and increase the heat at a rate of 100°C per 1 hour. Then heat at 500°C for

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 several hours to conduct ashing. If ashing is incomplete, wet with 2~5 ml of a
50% magnesium nitrate solution or nitric acid (1+1). After drying, continue
ashing. Add 2~4 ml of water to the ash, and after drying, add 5 ml of
hydrochloric acid to dissolve the salts. Use water to prepare fixed volumes of
measurement solution.
Remarks:
There is a possibility of volitilizing almost all of the elements, with Cd
being volitilized during ashing above 500°C. If halogens are coexistent,
As, Sb, Sn Zn, etc. will be volitilized. Heating above 550°C will greatly
reduce the element collection rate.
10.1.2 Electrical Heating Atomic Absorption Method
a) Target elements
Cd, Pb
b) Measurement procedure
Measurement is conducted using the following procedure. The lamp current
value, and the slit width used are in accordance with Cookbook Section 4, item
7.5 Element Specific Measurement Conditions.
• Cd
Reagents:
1) Cd standard solution (0.01 µg Cd/ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
2) Palladium nitrate (II) solution (10 µg Pd/ml) : Dissolve 0.108 g of
palladium nitrate in 10 ml of nitric acid (1+1), and after bringing up to
500 ml using water, add water to 20 ml of this solution to bring up to
200 ml.
Procedure:
1) Place 15 ml each of pretreated sample (see note) into four 20 ml
volumetric flasks, and add standard solution (0.01 µg Cd/ml) to three of
them in increments in the range of 0.1~2 ml so that one flask contains
no standard solution and the other three have increasing concentrations
of standard solution added to the sample. After adding nitric acid (1+1)
to each of the solutions so they all have the same concentration of acid,
bring up to volume using water. (Note: If the pretreated sample contains

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 more than 0.01 µg of Cd, decrease the sampling volume so that this
becomes less than 0.01 µg.)
2) Place equal volumes of at least 100 µl of each sample prepared in
step 1) into small volumetric flasks, and add equal volumes of the
palladium nitrate (II) solution (10 µg Pd/ml) to each, and mix well for
measurement.
Measurement:
Measurement wavelength: 228.8 nm
Calibration curve concentration range: 0.2~2 ng/ml
Tube: High density graphite tube
Sample injection volume: 10 µl
Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 120 15 R 0.1
2 250 15 R 0.1
3 600 10 R 1.0
4 600 15 S 1.0
5 600 3 S 0.0H
6 2000 3 S 0.0H
7 2500 2 S 1.0
• Pb
Reagents:
1) Pb standard solution (0.2 µg Pb/ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
2) Palladium nitrate (II) solution (10 µg Pd/ml) : Preparation is as
described for Cd reagent, item 2).
Procedure:
1) Place 15 ml each of pretreated sample (see note) into four 20 ml
volumetric flasks, and add standard solution (0.2 µg Pb/ml) to three of
them in increments in the range of 0.1~2 ml so that one flask contains
no standard solution and the other three have increasing concentrations
of standard solution added to the sample. After adding nitric acid (1+1)
to each of the solutions so they all have the same concentration of acid,

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 bring up to volume using water. (Note: If the pretreated sample contains
more than 0.1 µg of Pb, decrease the sampling volume so that this
becomes less than 0.1 µg.)
2) Place equal volumes of at least 100 µl of each sample prepared in
step 1) into small volumetric flasks, and add equal volumes of the
palladium nitrate (II) solution (10 µg Pd/ml) to each, and mix well for
measurement..
Measurement:
Measurement wavelength: 283.3 nm
Calibration curve concentration range: 2~20 ng/ml
Tube: High density graphite tube
Sample injection volume: 20 µl
Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 120 20 R 0.1
2 250 10 R 0.1
3 600 10 R 1.0
4 600 15 S 1.0
5 600 3 S 0.0H
6 2000 3 S 0.0H
7 2500 2 S 1.0
10.1.3 Flame Atomic Absorption Method
a) Target elements
Ca, Cd, Cu, Fe, K, Mg, Mn, Na, Pb, Zn
b) Measurement procedure
Measurement is conducted using the following procedure. The lamp current
value, slit width and flame conditions used are in accordance with Cookbook
Section 3, item 6.4 Element Specific Measurement Conditions. For preparation
of the standard solutions for generating the calibration curve, refer to Cookbook
Section 2, item 3 Preparing Standards.
• Ca
Reagents:
1) Ca standard solution (10 µg Ca/ml) :Refer to Cookbook Section 2,
item 3 Preparing standards

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 2) La solution (50 g/l) : Dissolve 67 g of lanthanum chloride by adding
small quantities of hydrochloric acid (1+1) little by little, and then add
water up to a volume of 500 ml.
Procedure:
1) Place a suitable volume of pretreated sample (containing 5~150 µg
of Ca) in a 50 ml volumetric flask. After adding 3 ml of La solution (50
g/l), bring up to volume with water for measurement. At this time, for a
blank test, perform the same pretreatment as that performed for the
sample, and place the same volume of this solution in a 50 ml
volumetric flask. Then, after adding 3 ml of La solution (50 g/l), bring
up to volume with water. The measurement value obtained with this
solution is used for correction of the value obtained with the actual
sample.
2) For the calibration curve, place stepwise increments of 1~30 ml of
the Ca standard into 50 ml volumetric flasks, add 3 ml of lanthanum
solution (50 g/l) to each flask, and add water to bring all the solutions
up to volume.
Measurement:
Measurement wavelength: 422.7 nm
Calibration curve concentration range: 0.3~6 µg/ml
Measurement conditions: Refer to Cookbook Section 3, item 6.4, 9).
Remarks:
When Na is present in amounts greater that 200 times that of the Ca, and
even the addition of La does not completely control the interference,
conduct measurement using the N2O-C2H2 flame (refer to Cookbook
Section 3, item 6.4, 10).
• Cd
Reagents:
1) Cd standard solution (10 µg of Cd/ml) : refer to Cookbook Section 2,
item 3 Preparing Standards.
2) Ammonium dihydrogen citrate (10 g/l): Dissolve 10 g of ammonium
dihydrogen citrate in approximately 80 ml of water. Add 2~3 drops of
m-Cresol Purple solution (1 g/l), and after adding enough drops of

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 aqueous ammonia (1+1) to adjust the pH to about 9, add water to a
volume of 100 ml. Transfer this to a separating funnel, add 2 ml of
diethyl sodium dithiocarbamate solution (10 g/l) and 10 ml of butyl
acetate. After vigorously vibrating the solution, set it aside. Separate
off the ammonium dihydrogen citrate layer and pass it through dry filter
paper to eliminate the minute butyl acetate bubbles.
3) m-Cresol Purple solution (1 g/l) : Dissolve 0.1 g of m-cresol purple
in 50 ml of ethanol, and add water to volume of 100 ml.
4) diethyl sodium dithiocarbamate solution (10 g/l) : Dissolve 1.3 g of
sodium dithiocarbamate in water, and bring volume to 100 ml.
5) Butyl acetate
Procedure:
1) Place a suitable amount of pretreated sample (containing 0.5~10 µg
of Cd) in a separating funnel, and add 10 ml of ammonium dihydrogen
citrate solution (10 g/l) and 2~3 drops of indicator m-Cresol Purple
solution (1 g/l). Then add aqueous ammonia (1+1) until the solution
becomes slightly purple.
Then add 5 ml of diethyl sodium dithiocarbamate solution (10 g/l), and
after shaking the solution, add 10 ml of butyl acetate and vigorously
shake for about 1 minute before setting the solution down. Separate off
the butyl acetate layer and place it in a 20 ml volumetric flask. Add 5
ml of butyl acetate to the remaining aqueous layer, and repeat the
extraction procedure. Add this extracted butyl acetate layer to the
previous volumetric flask. Keep adding butyl acetate until it is brought
up to the 20 ml volume. This will be used in measurement.
2) For the calibration curve, prepare stepwise standard solutions over
the calibration curve concentration range using the Cd standard solution
(10 µg of Cd/ml), and use the same procedure as that for the sample.
Measurement:
Measurement wavelength: 228.8 nm
Calibration curve concentration range: 0.02~0.5 µg/ml (concentration after
extraction)
Measurement conditions:
Lamp current value : 8 mA
Slit width : 0.5 nm

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 Lamp mode : BGC-D2
Burner height : 7 mm
Support gas : Air
Fuel gas flow rate :
C2H2 0.8 l/min (If the flame turns reddish when the sample is
sprayed, decrease the amount of sample drawn up.)
• Cu I (When concentration is within the quantitation range of 0.2 ~ 4 µg/ml,
and there are no interfering components)
Reagents:
Cu standard solution (10 µg of Cu/ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
Procedure:
1) The pretreated sample may be directly used for measurement, or it
may be diluted with water so that its concentration is within the
quantitation range. At this time, take an appropriate amount of water
and perform the same pretreatment as that used for the sample, and then
measure this solution. The obtained value may be used for correction of
the value obtained in sample measurement.
2) For the calibration curve, place 2~40 ml of Cu standard solution (10
µg of Cu/ml) in stepwise volumes in 100 ml volumetric flasks. After
adding acid epeat the extraction procedure. Add this extracted butyl ac.
Add this extracted butyl acetate layer to the previous volumetric flask.
Keep adding butyl acetate until it is brought up to the 20 ml volume.
This will be used in measurement.
Measurement:
Measurement wavelength: 324.7 nm
Calibration curve concentration range: 0.2~4 µg/ml (concentration in
solution after extraction)
Measurement conditions: Refer to Cookbook Section 3, Item 6.4, 15).
• Cu II (When Cu concentration is below the quantitation range)
Reagents:
1) Cu standard solution (10 µg of Cu/ml) : Same as for Cu I
2) Ammonium dihydrogen citrate (10 g/l)

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 3) m-Cresol Purple solution (1 g/l)
4) diethyl sodium dithiocarbamate solution (10 g/l) :
5) Butyl acetate
Items 2), 3), 4) and 5) are prepared as in 2) ~ 5), respectively, for the
Cd reagents.
Procedure:
1) Place a suitable amount of pretreated sample (containing 2 ~ 40 µg
of Cu) in a separating funnel, and proceed as described in step 1) of the
Cd procedure.
2) For the calibration curve, prepare stepwise standard solutions over
the calibration curve concentration range using the Cu standard solution
(10 µg of Cd/ml). se the same procedure as that for the sample.
Measurement
Measurement wavelength: 324.7 nm
Calibration curve concentration range: 0.1~2 µg/ml (concentration in
solution after extraction)
Measurement conditions:
Lamp current value : 6 mA
Slit width : 0.5 nm
Lamp mode : BGC-D2
Burner height : 7 mm
Support gas : Air
Fuel gas flow rate :
C2H2 0.8 l/min (If the flame turns reddish when the sample is
sprayed, decrease the amount of sample drawn up.)
• Fe
Reagents:
Fe standard solution (10 µg of Fe/ml) : Refer to Cookbook Section 2, item
3 Preparing Standards
Procedure:
1) The pretreated sample may be directly used for measurement, or it
may be diluted with hydrochloric acid (1+50) so that its concentration is
within the quantitation range. At this time, take an appropriate amount

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 of water and perform the same pretreatment as that used for the sample,
and then measure this solution. The obtained value may be used for
correction of the value obtained in sample measurement.
2) For the calibration curve, place 3~60 ml of Fe standard solution (10
µg of Fe/ml) in stepwise volumes in 100 ml volumetric flasks, and after
adding acid using the same conditions as that for the sample, bring up to
volume with water.
Measurement
Measurement wavelength: 248.3 nm
Calibration curve concentration range: 0.3~6 µg/ml
Measurement conditions: Refer to Cookbook Section 3, Item 6.4, 16).
• K
Reagents:
K standard solution (10 µg of K/ml) : Refer to Cookbook Section 2, item
3 Preparing Standards
Procedure:
1) The pretreated sample may be directly used for measurement, or it
may be diluted with hydrochloric acid (1+50) so that its concentration is
within the quantitation range. At this time, take an appropriate amount
of water and perform the same pretreatment as that used for the sample,
and then measure this solution. The obtained value may be used for
correction of the value obtained in sample measurement.
2) For the calibration curve, place 1~10 ml of K standard solution (10
µg of K/ml) in stepwise volumes in 100 ml volumetric flasks, and after
adding acid using the same conditions as that for the sample, bring up to
volume with water.
Measurement:
Measurement wavelength: 766.5 nm
Calibration curve concentration range: 0.1~1 µg/ml
Measurement conditions: Refer to Cookbook Section 3, Item 6.4, 19).
• Mg
Reagents:
1) Mg standard solution (5 µg of Mg/ml) : Refer to Cookbook Section
2, item 3 Preparing Standards
2) La solution (50g/l) : Same as item 2) for Ca reagent

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 Procedure:
1) Place a suitable volume of pretreated sample (containing .2~2.5 µg
of Mg) in a 50 ml volumetric flask. After adding 3 ml of La solution (50
g/l), bring up to volume with water for measurement. At this time, for a
blank test, perform the same pretreatment as that performed for the
sample, and place the same volume of this solution in a 50 ml
volumetric flask. Then add 3 ml of La solution (50 g/l), bring up to
volume with water. The measurement value obtained with this solution
is used for correction of the value obtained with the actual sample.
2) For the calibration curve, place stepwise increments of .5~5 ml of the
Mg standard into 50 ml volumetric flasks, add 3 ml of lanthanum
solution (50 g/l) to each flask, and add water to bring all the solutions
up to volume.
Measurement:
Measurement wavelength: 285.2 nm
Calibration curve concentration range: 0.05~0.5 µg/ml
Measurement conditions: Refer to Cookbook Section 3, item 6.4, 22).
• Mn
Reagents:
Mn standard solution (10 µg of Mn/ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
Procedure:
1) The pretreated sample may be directly used for measurement, or it
may be diluted with hydrochloric acid (1+50) so that its concentration is
within the quantitation range. At this time, take an appropriate amount
of water and perform the same pretreatment as that used for the sample,
and then measure this solution. The obtained value may be used for
correction of the value obtained in sample measurement.
2) For the calibration curve, place 2~30 ml of Mn standard solution (10
µg of Mn/ml) in stepwise volumes in 100 ml volumetric flasks, and
after adding acid using the same conditions as that for the sample, bring
up to volume with water.
Measurement:
Measurement wavelength: 279.5 nm

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 Calibration curve concentration range: 0.2~3 µg/ml
Measurement conditions: Refer to Cookbook Section 3, Item 6.4, 23).
• Na
Reagents:
Na standard solution (5 µg of Na/ml) : Refer to Cookbook Section 2, item
3 Preparing Standards
Procedure:
1) The pretreated sample may be directly used for measurement, or it
may be diluted with hydrochloric acid (1+50) so that its concentration is
within the quantitation range. At this time, take an appropriate amount
of water and perform the same pretreatment as that used for the sample,
and then measure this solution. The obtained value may be used for
correction of the value obtained in sample measurement.
2) For the calibration curve, place 1~10 ml of Na standard solution (5
µg of Na/ml) in stepwise volumes in 100 ml volumetric flasks, and after
adding acid using the same conditions as that for the sample, bring up to
volume with water.
Measurement:
Measurement wavelength: 589.0 nm
Calibration curve concentration range: 0.05~0.5 µg/ml
Measurement conditions: Refer to Cookbook Section 3, Item 6.4, 24).
• Pb
Reagents:
1) Pb standard solution (10 µg of Pb/ml) : Refer to Cookbook Section
2, item 3 Preparing Standards
2) Ammonium dihydrogen citrate (10 g/l)
3) m-Cresol Purple solution (1 g/l)
4) diethyl sodium dithiocarbamate solution (10g/l) :
5) Butyl acetate
Items 2), 3), 4) and 5) are prepared as in 2) ~ 5), respectively, for the
Cd reagents.
Procedure:
1) Place a suitable amount of pretreated sample (containing 6 ~ 60 µg
of Pb) in a separating funnel, and proceed as described in step 1) of the
Cd procedure.

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 2) For the calibration curve, prepare stepwise standard solutions of Pb
standard solution, and use the same procedure as that for the sample.
Measurement:
Measurement wavelength: 217.0 nm
Calibration curve concentration range: 0.3~3 µg/ml (concentration in
solution after extraction)
Measurement conditions:
Lamp current value : 12 mA
Slit width : 0.5 nm
Lamp mode : BGC-D2
Burner height : 7 mm
Support gas : Air
Fuel gas flow rate :
C2H2 0.8 l/min (If the flame turns reddish when the sample is
sprayed, decrease the amount of sample drawn up.)
• Zn I (When concentration is within the quantitation range of 0.05 ~ 1 µg/ml,
and there are no interfering components)
Reagents:
Zn standard solution (10 µg of Zn/ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
Procedure:
1) The pretreated sample may be directly used for measurement, or it
may be diluted with hydrochloric acid (1+50) so that its concentration is
within the quantitation range. At this time, take an appropriate amount
of water and perform the same pretreatment as that used for the sample,
and then measure this solution. The obtained value may be used for
correction of the value obtained in sample measurement.
2) For the calibration curve, place .5~10 ml of Zn standard solution (10
µg of Zn/ml) in stepwise volumes in 100 ml volumetric flasks, and after
adding acid using the same conditions as that for the sample, bring up to
volume with water.
Measurement:
Measurement wavelength: 213.9 nm

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 Calibration curve concentration range: 0.05~1 µg/ml
Measurement conditions: Refer to Cookbook Section 3, Item 6.4, 44).
• Zn II (When Zn concentration is below the quantitation range)
Reagents:
1) Zn standard solution (10 µg of Zn/ml) : Same as for Zn I
2) Ammonium dihydrogen citrate (10 g/l)
3) m-Cresol Purple solution (1 g/l)
4) diethyl sodium dithiocarbamate solution (10 g/l) :
5) Butyl acetate
Items 2), 3), 4) and 5) are prepared as in 2) ~ 5), respectively, for the
Cd reagents.
Procedure:
1) Place a suitable amount of pretreated sample (containing 0.5 ~ 10 µg
of Zn) in a separating funnel, and proceed as described in step 1) of the
Cd procedure.
2) For the calibration curve, prepare stepwise standard solutions over
the calibration curve concentration range using the Zn standard solution
(10 µg of Zn/ml), and use the same procedure as that for the sample.
Measurement:
Measurement wavelength: 213.9 nm
Calibration curve concentration range: 0.02~0.5 µg/ml (concentration in
solution after extraction)
Measurement conditions:
Lamp current value : 8 mA
Slit width : 0.5 nm
Lamp mode : BGC-D2
Burner height : 7 mm
Support gas : Air
Fuel gas flow rate :
C2H2 0.8 l/min (If the flame turns reddish when the sample is
sprayed, decrease the amount of sample drawn up.)
10.1.4 Hydride Vapor Generation Atomic Absorption Method
a) Target element
As

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b) Measurement Procedures
Reagents:
1) As standard solution (1 µg/ml) : Refer to Cookbook Section 2, item 3
Preparing Standards.
2) Potassium iodide solution (200 g/l) : Dissolve 20 g of potassium
iodide in water and bring to 100 ml volume.
3) Sodium tetrahydroborate solution (0.5 w/v%) : Dissolve 2.5g of
sodium tetrahydroborate in 500 ml of sodium hydroxide solution (0.1
mol/l).
4) Hydrochloric acid
Procedure:
1) Add ml of hydrochloric acid (1+1) to 10 ml of pretreated sample, and
2 ml of potassium iodide solution, and then heat, without boiling. After
allowing to cool, add 20 ml of water for measurement.
2) For the calibration curve, use the As standard solution (1 µg/ml) to
prepare standard solutions of incremental concentrations between
10~100 ng, add 4 ml of hydrochloric acid (1+1) and 2 ml of potassium
iodide solution, and perform the same measurement procedure as that
for the sample.
Measurement:
Connect the HVG-1 Hydride Vapor Generator to the atomic absorption
spectrophotometer, and measure the sample. For operation of the HVG-1,
refer to the documentation accompanying the HVG-1.
Measurement wavelength: 193.7 nm
Calibration curve concentration range: 0.5~5.0 ng/ml
Measurement conditions:
Lamp current value: 14 mA
Slit width: 0.5 nm
Lamp mode: BGC-D2

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10.1.5 Reduction Vaporization Atomic Absorption Method
a) Target element
Total Hg
b) Sample pretreatment
Place 5~10 g of sample in a decomposition flask (example of a
decomposition flask is illustrated in the following item), and wet with about 5
ml of water. To this flask, mount a fraction collector and above that a cooling
circulator. Introduce 10~20 ml of nitric acid and 10 ml of sulfuric acid into the
fraction collector (3), and then turn cock A to gradually introduce the contents
to the decomposition flask. (If an extreme amount of bubbles are generated, it is
advisable to let it sit overnight.) With cock B in the open position to allow
condensation to circulate through the decomposition flask, gradually heat up the
flask. Once the intense reaction is completed, close cock B and continue
heating. Periodically open cock B to allow condensation to return from the
fraction collector to the decomposition flask. Rinse the cooling circulator with
a small amount of water, add 5~10 ml of urea solution (urea 200 g/l), boil for a
short time, and after cooling to below 60°C, add 2 ml of potassium
permanganate solution (potassium permanganate 50 g/l), and boil again. At this
time, if the permanganic acid color disappears, repeat the addition of potassium
permanganate solution until the color remains for at least 10 minutes. After
allowing to cool, add hydrochloride hydroxyamine (10 w/v%), reduce the
excess potassium permanganate, and add water to bring solution to 250 ml for
measurement.

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 (1) 2-port round
bottom flask
(2), (3) Cylindrical
fraction funnels
(4) Friedlich cooler

(5) Bent tube

(6) Cock A

(7) Cock B

c) Measurement procedure
Reagents:
1) Hg standard solution (0.1 µg Hg/ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
2) Stannous chloride solution (10 w/v%) : Add 10 g of stannous
chloride to 60 ml of sulfuric acid (sulfuric acid to water ratio 1:20), and
while heating, mix to dissolve. After cooling, bring up to 100 ml using
water.
Procedure:
1) The pretreated sample may be measured as it is.
2) For the calibration curve, prepare standard solutions starting with
incremental volumes between 2~12 ml of the Hg standard solution (0.1
µg Hg/ml), then adding 20 ml of sulfuric acid (1+1), and bringing up to
250 ml using water.
Measurement:
Connect the MVU-1A Mercury Vaporizer Unit to the atomic absorption

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spectrophotometer, and measure the sample. For details on operation of
the MVU-1A, refer to its accompanying documentation.
Measurement wavelength: 253.7 nm
Calibration curve concentration range: 1~5 ng/ml
Measurement conditions:
Lamp current: 4 mA
Slit width: 0.5 nm
Lamp mode: BGC-D2

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10.2 Grains, Beans, and Potatoes Analysis Method
Same as analysis method described in 10.1 Vegetables and Fruits Analysis Method

10.3 Meat, and Fish Analysis Method

Same as analysis method described in 10.1 Vegetables and Fruits Analysis Method

10.4 Fats, Oils, and Dairy Products Analysis Method

Reference materials:
Health Testing Methods Commentary, Japan Pharmaceutical Society Publication,
Kanehara Publishing Co., Foodstuffs Analysis Methods, Japan Foodstuffs
Manufacturing Society, Foodstuffs Analysis Methods Editorial Commission
Publication (Korin Co.)
10.4.1 Sample Pretreatment
a) Nitric acid, perchloric acid decomposition
This is the same as the nitric acid, perchloric acid decomposition of item
10.1.1 a), however, perchloric acid and the sample often do not mix, preventing
successful decomposition. In such cases, use the procedure described in b)
Ashing by drying.
b) Ashing by drying
Place 5~10 g of sample in a quartz beaker, add 1~3 ml of sulfuric acid, and
gently heat on a hot plate at 120°C. After the components, which are volitilized
at low temperatures are driven off, continue heating the sample until
carbonization occurs. (At this time, if intense bubbling in the sample occurs,
add 0.5 ml of nitric acid.) Place this in an electric furnace, increase heat at a rate
of 100°C per hour, and at about 500°C perform ashing over a period of several
hours. If ashing is incomplete, wet with 2~5 ml of 50% magnesium nitrate
solution or nitric acid (1+1). After drying, continue ashing. Add 2~4 ml of
water to the ash, and after drying, add 5 ml of hydrochloric acid to dissolve the
salts. Use water to prepare fixed volumes of the measurement solution.
Remarks:
There is a possibility of volitilizing most of the elements, with Cd being
volitilized during ashing above 500°C. If halogens are coexistent, As, Sb,
Sn Zn, etc. will be volitilized. Heating above 550°C will greatly reduce
the element collection rate.

- 18 -
10.4.2 Electrical Heating Atomic Absorption Method
a) Target elements
Cd, Pb, Cu, Fe, Zn
b) Measurement procedure
Measurement is conducted using the following procedure. The lamp current
value, and the slit width used are in accordance with Cookbook Section 4, item 7.5.
• Cd
Reagents:
Same as those described in item 10.1.2 Electrical Heating Atomic
Absorption Method, Cd.
Procedure:
Using 15 ml pretreated sample volumes, follow the procedure described
in item 10.1.2 Electrical Heating Atomic Absorption Method, Cd.
Measurement:
Same as described in item 10.1.2 Electrical Heating Atomic Absorption
Method, Cd.
• Pb
Reagents:
Same as those described in item 10.1.2 Electrical Heating Atomic
Absorption Method, Pb.
Procedure:
Using 15 ml pretreated sample volumes, follow the procedure described
in item 10.1.2 Electrical Heating Atomic Absorption Method, Pb.
Measurement:
Same as described in item 10.1.2 Electrical Heating Atomic Absorption
Method, Pb.
• Cu (direct quantitation of oil)
Reagents:
Cu standard solution (0.2 µg Cu/ml): Prepare a 10 mg Cu/ml aqueous
solution referring to Cookbook Section 2, item 3 Preparing Standards.
Introduce 1 ml of this solution into a 50 ml volumetric flask, add 10 ml of
methyl alcohol and mix well. Then bring up to volume using 4-methyl-2-
pentanone (MIBK), for the Cu standard solution (0.2 µg Cu/ml). Or,

- 19 -
 measure out 63.257 mg of the oily metallic standard reagent copper
cyclohexylbutyric acid (molecular weight 402.04), add 3 ml of xylene and
5 ml of 2-ethylhexanoic acid. Gently heat to dissolve until it is clear.
After cooling to room temperature, transfer the solution to a 100 ml
volumetric flask and bring up to volume using MIBK (100 µg Cu/ml).
Next, transfer 4 ml of this solution to a 100 ml volumetric flask and bring
up to volume using MIBK (4 µg Cu/ml), and then transfer 5 ml of this
solution to a 100 ml volumetric flask and bring up to volume using MIBK
for the final Cu standard solution (0.2 µg Cu/ml).
Procedure:
1) Using a suitable amount of sample, dilute with MIBK so that the
concentration is within the quantitation range (dilution factor of 5) for
measurement.
2) For the calibration curve, introduce 0.5~5 ml of Cu standard solution
(0.2 µg Cu/ml) in incremental volumes into 50 ml volumetric flasks,
and bring all the solutions up to volume using MIBK.
Measurement:
Measurement wavelength: 324.7 nm
Calibration curve concentration range: 1~20 ng/ml
Tube: pyrolytic graphite tube
Sample injection volume: 10 µl
Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 80 20 R 0.1
2 250 10 R 0.1
3 750 15 R 1.0
4 750 15 S 1.0
5 750 3 S 0.0H
6 2400 4 S 0.0H
7 2700 2 S 1.0
• Fe (direct quantitation of oil)
Reagents:
Fe standard solution : (0.5 µg Fe/ml): Prepare a 10 mg Fe/ml aqueous

- 20 -
 solution referring to Cookbook Section 2, item 3 Preparing Standards.
Introduce 2.5 ml of this solution into a 50 ml volumetric flask. Add 10 ml
of methyl alcohol and mix well. Then bring up to volume using 4-methyl-
2-pentanone (MIBK), for the Fe standard solution (0.5 µg Fe/ml).
Or measure out 60.023 mg of the oily metallic standard reagent ferrous
cyclohexylbutyric acid (molecular weight 670.45). Add 3 ml of xylene
and 5 ml of 2-ethylhexanoic acid. Gently heat to dissolve until it is clear,
and after cooling to room temperature, transfer the solution to a 100 ml
volumetric flask. Then bring up to volume using MIBK (100 µg Fe/ml).
Next, transfer 5 ml of this solution to a 100 ml volumetric flask and bring
up to volume using MIBK (5 µg Fe/ml), and then transfer 10 ml of this
solution to a 100 ml volumetric flask. Then bring up to volume using
MIBK for the final Fe standard solution (0.5 µg Fe/ml).
Procedure:
1) Using a suitable amount of sample, dilute with MIBK so that the
concentration is within the quantitation range (dilution factor of 5) for
measurement.
2) For the calibration curve, introduce 0.5~5 ml of Fe standard solution
(0.5 µg Fe/ml) in incremental volumes into 50 ml volumetric flasks, and
bring all the solutions up to volume using MIBK.
Measurement:
Measurement wavelength: 248.3 nm
Calibration curve concentration range: 5~50 ng/ml
Tube: pyrolytic graphite tube
Sample injection volume: 10 µl
Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 80 20 R 0.1
2 250 10 R 0.1
3 750 15 R 1.0
4 750 15 S 1.0
5 750 3 S 0.0H
6 2400 4 S 0.0H
7 2700 2 S 1.0

- 21 -
• Zn (direct quantitation of oil)
Reagents:
Zn standard solution : (0.1 µg Zn/ml): Prepare a 5 mg Zn/ml aqueous
solution referring to Cookbook Section 2, item 3 Preparing Standards.
Introduce 1 ml of this solution into a 50 ml volumetric flask, add 10 ml of
methyl alcohol and mix well. Then bring up to volume using 4-methyl-2-
pentanone (MIBK), for the Zn standard solution (0.1 µg Zn/ml).
Or, measure out 61.781 mg of the oily metallic standard reagent zinc
cyclohexylbutyric acid (molecular weight 403.86). Add 3 ml of xylene
and 5 ml of 2-ethylhexanoic acid. Gently heat to dissolve until it is clear,
and after cooling to room temperature, transfer the solution to a 100 ml
volumetric flask and bring up to volume using MIBK (100 µg Zn/ml).
Next, transfer 2 ml of this solution to a 100 ml volumetric flask and bring
up to volume using MIBK (2 µg Zn/ml). Then transfer 5 ml of this
solution to a 100 ml volumetric flask and bring up to volume using MIBK
for the final Zn standard solution (0.1 µg Zn/ml).
Procedure:
1) Using a suitable amount of sample, dilute with MIBK so that the
concentration is within the quantitation range (dilution factor of 5) for
measurement.
2) For the calibration curve, introduce 0.5~5 ml of Zn standard solution
(0.1 µg Zn/ml) in incremental volumes into 50 ml volumetric flasks,
and bring all the solutions up to volume using MIBK.
Measurement:
Measurement wavelength: 213.9 nm
Calibration curve concentration range: 1~10 ng/ml
Tube: High density graphite tube
Sample injection volume: 10 µl

- 22 -
 Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 80 20 R 0.1
2 250 10 R 0.1
3 300 15 R 1.0
4 300 20 S 1.0
5 300 3 S 0.0
6 1300 3 S 0.0
7 2600 2 S 0.1
10.4.3 Flame Atomic Absorption Method
a) Target elements
Ca, Cd, Cu, Fe, K, Mg, Mn, Na, Pb, Zn
b) Measurement procedure
Using the pretreated samples described in Item 10.4.1, use the same
procedure as described in item 10.1.3 Flame Atomic Absorption Method
10.4.4 Hydride Vapor Generation Atomic Absorption Method
a) Target elements
As
b) Measurement procedure
Use the same procedure as described in item 10.1.4 Hydride Vapor
Generation Atomic Absorption Method, b) Measurement procedure.
10.4.5 Reduction Vaporization Atomic Absorption Method
a) Target elements
Total Hg
b) Sample pretreatment
Use the same procedure described in item 10.1.5 Reduction Vaporization
Atomic Absorption Method, b) Sample pretreatment.
c) Measurement procedure
Use the same procedure as described in item 10.1.5 Reduction Vaporization
Atomic Absorption Method, c) Measurement procedure.

- 23 -
10.5 Spirits, Alcoholic Beverage Analysis
Reference materials:
Health Testing Methods Commentary, Japan Pharmaceutical Society
Publication, Kanehara Publishing Co.,
Foodstuffs Analysis Methods, Japan Foodstuffs Manufacturing Society,
Foodstuffs Analysis Methods Editorial Commission Publication (Korin Co.)
10.5.1 Sample Pretreatment
a) Nitric acid, perchloric acid decomposition
Introduce 10~20 g of sample into a conical beaker. Add 25 ml of nitric acid,
and then follow the same procedure as that described in item 10.1.1 Sample
Pretreatment, a) Nitric acid, perchloric acid decomposition.
Remark:
When a large part of the sample is sugar, a large amount of acid must be
used and there is a possibility of contamination of the target element with
reagent. In this case, either decrease the sample amount or use the ashing
by drying procedure described next.
b) Ashing by drying
Dissolve 10~20 g of sample in water, and after drying through vaporization,
continue heating until partial carbonization occurs. Place this in an electric
furnace, and follow the procedure described in item 10.1.1 Sample
Pretreatment, b) Ashing by drying.
c) Hydrochloric acid extraction
Add hydrochloric acid (1+1) to 10~20 g of sample at a ratio of 2 ml/10 g of
sample. Set aside for 1 hour, periodically shaking the flask to mix the contents.
Then, either filter and use the filtrate for measurement, or centrifuge at 4,000
rpm for 5 minutes and use the supernatant for measurement.
Remark:
Used for measurement of K, Na, Mg, Cu, Mn, Zn.
10.5.2 Electrical Heating Atomic Absorption Method
a) Target elements
Cd, Cr, Cu, Fe, Mn, Pb, Sn, Zn
b) Measurement procedure
Measurement is conducted using the following procedure. The lamp current

- 24 -
 and slit width are as described in Cookbook Section 4, item 7.5 Element
Specific Conditions.
• Cd
Reagents:
Same as that described in item 10.1.2 Electrical Heating Atomic
Absorption Method, for Cd.
Procedure:
Using 15 ml aliquots of sample pretreated according to a) or b), follow the
same procedure as described in item 10.1.2 Electrical Heating Atomic
Absorption Method, for Cd.
Measurement:
Same as that described in item 10.1.2 Electrical Heating Atomic
Absorption Method, for Cd.
• Cr
Reagents:
Cr standard solution (0.01 µg Cr/ml) : Refer to Cookbook Section 2, item
3 Preparing Standards
Procedure:
1) Samples pretreated using method a) or b) (samples without
cloudiness, and those with low viscosity may be measured without
pretreatment) may be used directly or diluted with nitric acid (1+100) so
that the concentration is within the quantitation range. At this time,
perform the same pretreatment on a suitable volume of water, and
conduct measurement on this solution. The value obtained may be used
to correct the value obtained in actual sample measurement.
2) For the calibration curve, introduce 2~10 ml of Cr standard solution
(0.01 µg Cr/ml) in incremental volumes into 20 ml volumetric flasks.
Add acid according to the same conditions used for the sample, and add
water to bring all the solutions up to volume.
Measurement:
Measurement wavelength: 357.9 nm
Calibration curve concentration range: 1~5 ng/ml
Tube: Pyrolytic graphite tube

- 25 -
 Sample injection volume: 10 µl
Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 120 15 R 0.1
2 250 10 R 0.1
3 700 10 R 1.0
4 700 20 S 1.0
5 700 3 S 0.0H
6 2400 3 S 0.0H
7 2500 2 S 1.0
• Cu
Reagents:
Cu standard solution (0.01 µg Cu/ml) : Refer to Cookbook Section 2, item
3 Preparing Standards
Procedure:
1) Samples pretreated using method a) or b) (samples without
cloudiness, and those with low viscosity may be measured without
pretreatment) may be used directly or diluted with nitric acid (1+100) so
that the concentration is within the quantitation range. At this time,
perform the same pretreatment on a suitable volume of water, and
conduct measurement on this solution. The value obtained may be used
to correct the value obtained in actual sample measurement.
2) For the calibration curve, introduce 2~10 ml of Cu standard solution
(0.01 µg Cu/ml) in incremental volumes into 20 ml volumetric flasks.
Add acid according to the same conditions used for the sample, and add
water to bring all the solutions up to volume.
Measurement:
Measurement wavelength: 324.7 nm
Calibration curve concentration range: 1~5 ng/ml
Tube: Pyrolytic graphite tube
Sample injection volume: 20 µl

- 26 -
 Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 120 20 R 0.1
2 250 10 R 0.1
3 700 10 R 1.0
4 700 20 S 1.0
5 700 3 S 0.0H
6 2300 3 S 0.0H
7 2500 2 S 1.0
• Fe
Reagents:
Fe standard solution (0.02 µg Fe/ml) : Refer to Cookbook Section 2, item
3 Preparing Standards
Procedure:
1) Samples pretreated using method a) or b) (samples without
cloudiness, and those with low viscosity may be measured without
pretreatment) may be used directly or diluted with nitric acid (1+100) so
that the concentration is within the quantitation range. At this time,
perform the same pretreatment on a suitable volume of water, and
conduct measurement on this solution. The value obtained may be used
to correct the value obtained in actual sample measurement.
2) For the calibration curve, introduce 2~10 ml of Fe standard solution
(0.02 µg Fe/ml) in incremental volumes into 20 ml volumetric flasks.
Add acid according to the same conditions used for the sample, and add
water to bring all the solutions up to volume.
Measurement:
Measurement wavelength: 248.3 nm
Calibration curve concentration range: 2~10 ng/ml
Tube: Pyrolytic graphite tube
Sample injection volume: 20 µl

- 27 -
 Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 120 20 R 0.1
2 250 10 R 0.1
3 600 10 R 1.0
4 600 20 S 1.0
5 600 3 S 0.0H
6 2300 3 S 0.0H
7 2500 2 S 1.0
• Mn
Reagents:
Mn standard solution (0.01 µg Mn /ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
Procedure:
1) Samples pretreated using method a) or b) (samples without
cloudiness, and those with low viscosity may be measured without
pretreatment) may be used directly or diluted with nitric acid (1+100) so
that the concentration is within the quantitation range. At this time,
perform the same pretreatment on a suitable volume of water, and
conduct measurement on this solution. The value obtained may be used
to correct the value obtained in actual sample measurement.
2) For the calibration curve, introduce 0.5~4 ml of Mn standard
solution (0.01 µg Mn/ml) in incremental volumes into 20 ml volumetric
flasks. Add acid according to the same conditions used for the sample,
and add water to bring all the solutions up to volume.
Measurement:
Measurement wavelength: 379.5 nm
Calibration curve concentration range: 0.2~2 ng/ml
Tube: Pyrolytic graphite tube
Sample injection volume: 20 µl

- 28 -
 Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 120 20 R 0.1
2 250 10 R 0.1
3 700 10 R 1.0
4 700 20 S 1.0
5 700 3 S 0.0H
6 2300 3 S 0.0H
7 2500 2 S 1.0
• Pb
Reagents:
Same as that described in item 10.1.2 Electrical Heating Atomic
Absorption Method, for Pb.
Procedure:
Using 15 ml aliquots of sample pretreated according to a) or b), follow the
same procedure as described in item 10.1.2 Electrical Heating Atomic
Absorption Method, for Pb.
Measurement:
Same as that described in item 10.1.2 Electrical Heating Atomic
Absorption Method, for Pb.
• Sn
Reagents:
1) Sn standard solution (0.2 µg Sn /ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
2) Palladium nitrate (II) solution (100 µg Pd/ml) : Add 10 ml of nitric
acid (1+1) to 0.108 g of palladium nitrate (II) to dissolve, and bring up
to 500 ml using water.
Procedure:
1) Samples pretreated using method b) (samples without cloudiness,
and those with low viscosity may be measured without pretreatment)
may be used directly or diluted with nitric acid (1+100) so that the
concentration is within the quantitation range. In either case, add the
palladium nitrate (II) solution so that its concentration becomes 10

- 29 -
 µg/ml. At this time, perform the same pretreatment on a suitable
volume of water, and conduct measurement on this solution. The value
obtained may be used to correct the value obtained in actual sample
measurement.
2) For the calibration curve, introduce 1~10 ml of Sn standard solution
(0.2 µg Sn/ml) in incremental volumes into 20 ml volumetric flasks.
Add 2 ml of palladium nitrate (II) solution (100 µg Pd/ml), and add acid
according to the same conditions used for the sample, and add water to
bring all the solutions up to volume.
Measurement:
Measurement wavelength: 386.3 nm
Calibration curve concentration range: 10~100 ng/ml
Tube: Pyrolytic graphite tube
Sample injection volume: 20 µl
Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 120 20 R 0.1
2 250 10 R 0.1
3 800 10 R 1.0
4 800 20 S 1.0
5 800 3 S 0.0H
6 2400 3 S 0.0H
7 2500 2 S 1.0
• Zn
Reagents:
Zn standard solution (0.01 µg Zn /ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
Procedure:
1) Pretreated samples (samples without cloudiness, and those with low
viscosity may be measured without pretreatment) may be used directly
or diluted with nitric acid (1+100) so that the concentration is within the
quantitation range. At this time, perform the same pretreatment on a
suitable volume of water, and conduct measurement on this solution.

- 30 -
 The value obtained may be used to correct the value obtained in actual
sample measurement.
2) For the calibration curve, introduce 2~10 ml of Zn standard solution
(0.01 µg Zn/ml) in incremental volumes into 20 ml volumetric flasks,
add acid according to the same conditions used for the sample, and add
water to bring all the solutions up to volume.
Measurement:
Measurement wavelength: 213.9 nm
Calibration curve concentration range: 1~5 ng/ml
Tube: High density graphite tube
Sample injection volume: 10 µl
Heating conditions:
TEMP (°C) TIME (sec) HEAT GAS (l/min)
Stage 1 120 15 R 0.1
2 250 10 R 0.1
3 450 10 R 1.0
4 450 20 S 1.0
5 450 3 S 0.0
6 1800 3 S 0.0
7 2400 3 S 1.0
10.5.3 Flame Atomic Absorption Method
a) Target elements
Ca, Cd, Cu, Ge, K, Mg, Mn, Na, Pb, Zn
b) Measurement procedure
Using samples pretreated according to item 10.5.1, the procedure is the same
as that described in item 10.1.3 Flame Atomic Absorption Method.
10.5.4 Hydride Vapor Generation Atomic Absorption Method
a) Target elements
As
b) Measurement procedure
Same as procedure described in item 10.1.4 Hydride Vapor Generation
Atomic Absorption Method, b) Measurement procedure

- 31 -
10.5.5 Reduction Vaporization Atomic Absorption Method
a) Target elements
Hg
b) Sample pretreatment
Same as that described in item 10.1.5 Reduction Vaporization Atomic
Absorption Method, b) Sample pretreatment
c) Measurement procedure
Same as that described in item 10.1.5 Reduction Vaporization Atomic
Absorption Method, c) Measurement procedure

- 32 -
10.6 Processed Foods Analysis Method
Reference materials:
Health Testing Methods Commentary, Japan Pharmaceutical Society
Publication, Kanehara Publishing Co.,
Foodstuffs Analysis Methods, Japan Foodstuffs Manufacturing Society,
Foodstuffs Analysis Methods Editorial Commission Publication (Korin Co.)
10.6.1 Sample Pretreatment
Same as item 10.1.1 Sample Pretreatment
10.6.2 Electrical Heating Atomic Absorption Method
a) Target elements
Cd, Cr, Cu, Fe, Mn, Pb, Sn, Zn
b) Measurement procedure
Using pretreated samples, follow the procedure described in item 10.5.2
Electrical Heating Atomic Absorption Method
10.6.3 Flame Atomic Absorption Method
a) Target elements
Ca, Cd, Cu, Fe, Mg, Mn, Na, Pb, Zn
b) Measurement procedure
Using pretreated samples, follow the procedure described in item 10.1.3
Flame Atomic Absorption Method.
10.6.4 Hydride Vapor Generation Atomic Absorption Method
a) Target elements
As
b) Measurement procedure
Follow the procedure described in item 10.1.4 Hydride Vapor Generation
Atomic Absorption Method, b) Measurement procedure.
10.6.5 Reduction Vaporization Atomic Absorption Method
a) Target elements
Total Hg
b) Sample pretreatment
Follow the same procedure described in 10.1.5 Reduction Vaporization
Atomic Absorption Method, b) Sample pretreatment
c) Measurement procedure
Follow the procedure described in item 10.1.5 Reduction Vaporization
Atomic Absorption Method, c) Measurement procedure.

- 33 -
11. Fertilizer and Feed Analysis
11.1 Fertilizer Analysis Method
Reference materials:
Fertilizer Analysis, Agriculture and Forestry Ministry Agricultural Technology
Research Institute
11.1.1 Sample Pretreatment
a) Total analysis
• Ca, Cd, Co, Cu, Fe, Mg, Mo, Ni, Pb, Zn (inorganic fertilizer)
Place 2~5 g of sample in a tall beaker. Add 30 ml of hydrochloric acid and
10 ml of nitric acid. Cover with a watch glass, boil on a sand bath for 30
minutes, and boil over a water bath unydrrochloric acid and 10 ml of nitric acid.
Cover with a watch glass, boil on a sand bath for 30 minutes, and boil over a
water bath until dry. Add a little more nitric acid and boil until dry once again.
After allowing to cool, add about 40 ml of hydrochloric acid (1+20), heat to
dissolve. After cooling, add water to accurately bring up to 200 ml.
Immediately filter through dry filter paper, and use the filtrate for measurement.
• Ca, K, Mg (organic fertilizer, or fertilizer containing organic material)
Place 5~10 g of sample in a platinum dish, ash at about 500°C then transfer
to a tall beaker. Add 10 ml of hydrochloric acid and water to bring up to 30 ml.
Boil for about 30 minutes, and after cooling, add water to accurately bring
volume to 250 ml. Then filter through dry filter paper, and use the filtrate for
measurement.
• K (potassium salt)
Place 2.5 g of sample in a 300 ml tall beaker. Add 200 ml of water and a
small amount of hydrochloric acid to acidify. Cover with a watch glass and boil
for about 15 minutes to dissolve. After cooling, add water to accurately bring
volume up to 250 ml. Then filter through dry filter paper, and use the filtrate
for measurement.
• K (vegetation ash)
Place 5 g of sample in a tall beaker. Wet with a small amount of water,
cover with a watch glass, add 10 ml of hydrochloric acid and water, and boil
for about 30 minutes. Next, boil until dry and after performing silicic acid
separation,, add water to accurately bring volume to 500 ml. Then filter through
dry filter paper and use the filtrate for measurement.

- 34 -
• K (complex fertilizer)
Place 5 g of sample in a 300 ml tall beaker (for samples containing organic
material, place in platinum dish beforehand, and ash at about 500°C), add 10 ml
of hydrochloric acid and bring up to 30 ml with water. Then boil for about 10
minutes. Next add 150 ml of water, boil, and after cooling, add water to
accurately bring volume to 500 ml. Then filter through dry filter paper and use
the filtrate for measurement.
• K (potassium ore)
1)Place 1 g of sample in a platinum dish, and add several drops of water to
wet. 2)Add 5 ml of 46% hydrofluoric acid and 0.5 ml of perchloric acid, and
heat on a hot plate until white fumes of perchloric acid are generated. 3)After
allowing to cool, , add 5 ml of 46% hydrofluoric acid, partially cover with a
poly-4-fluoroethylene watch glass, and heat on a hot plate until almost dry.
4)After allowing to cool, add 5 ml of hydrochloric acid (1+1) and a small
amount of water, and heat on a hot plate to dissolve. At this time, if dissolution
is not complete due to incomplete decomposition, go back to step 2) and repeat
the process to completely dissolve. After allowing to cool, transfer to a 100 ml
volumetric flask, and add water to bring solution up to volume.
b) Water-soluble component analysis
• Cd
Place 50 g of sample in a sample vial having a stopper. Add 10 times that
amount of water (20°C), and shake in an oscillating mixer (200 oscillations per
minute, oscillating width 4~5 cm) for 6 hours, and immediately filter through
glass fiber filter paper having 1 µm pores. If filtering is noticeably difficult to
perform, then centrifuge the previously mixed liquid at 3,000 rpm for 20
minutes, and use the supernatant for measurement.
• Cu, Fe, Mn, Mo, Zn
Place 5 g of sample in a 500 ml volumetric flask, and add approximately 400
ml of water. Shake in an oscillating mixer at 30~40 rotations per minute for 30
minutes. Then bring up to volume with water, and immediately pass through
dry filter paper. Use the filtrate for measurement.
• Mg
Place 1 g of sample in a 500 ml triangular flask and add approximately 400

- 35 -
 ml of water. Using a cooling circulator, boil for 30 minutes, and after allowing
to cool, add water to accurately bring volume to 500 ml. Immediately pass
through dry filter paper, and use the filtrate for measurement.
• K (potassium salts)
Place 2.5 g of sample in a 300 ml tall beaker. Add about 200 ml of water,
and boil for 15 minutes. After cooling, add water to accurately bring volume up
to 250 ml. Then pass through dry filter paper, and use the filtrate for
measurement.
• K (complex fertilizer)
Place 5 g of sample in a small mortar, and add a small amount of water and
grind well. Transfer the supernatant liquid into a 500 ml volumetric flask. After
repeating this procedure 3 more times, transfer all of the remaining material in
the mortar to the flask. Add water to bring up to approximately 400 ml, and
shake at 30~40 rotations per minute in a rotating mixer for 30 minutes. Then
bring up to volume with water. Pass through dry filter paper, and use the filtrate
for measurement.
c) Soluble component analysis
• Ca, Mg
Place 2.5 g of sample in a tall beaker, and add 200 ml of 0.5 mol
hydrochloric acid. Cover with a watch glass, and boil for 5 minutes. After
cooling, add water to bring volume to 250 ml. Then pass through dry filter
paper and use the filtrate for measurement.
d) Citric acid soluble component analysis
• K, Mg, Mn
Place 1 g of sample in a 250 ml volumetric flask. Add 150 ml of citric acid
(citric acid 20 g/l) at 30°C, and shake at 30~40 rotations per minute in a rotating
mixer for one hour (while maintaining 30°C during extraction). Quickly return
to ambient temperature, and add water to bring up to volume. Quickly pass
through dry filter paper. Use the filtrate for measurement.
11.1.2 Flame Atomic Absorption Method
a) Target elements
Ca, Cd, Co, Cu, Fe, K, Mg, Mn, Mo, Ni, Pb, Zn
b) Measurement procedure
Measurement is conducted using the following procedure. The lamp current

- 36 -
 value, slit width and flame conditions used are in accordance with Cookbook
Section 3, item 6.4 Element Specific Measurement Conditions. For preparation
of the standard solutions for generating the calibration curve, refer to Cookbook
Section 2, item 3 Preparing Standards.
• Ca
The analysis method is the same as that described in item 10.1.3 Flame
Atomic Absorption Method, Ca.
• Cd I (When concentration is within the quantitation range of 0.05~ 1 µg/ml,
and there are no interfering components)
Reagents:
Cd standard solution (10 µg of Cd/ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
Procedure:
1) The pretreated sample may be directly used for measurement, or it
may be diluted with hydrochloric acid (1+50) so that its concentration is
within the quantitation range. At this time, take an appropriate amount
of water and perform the same pretreatment as that used for the sample,
and then measure this solution. The obtained value may be used for
correction of the value obtained in sample measurement.
2) For the calibration curve, place 0.5~10 ml of Cd standard solution
(10 µg of Cd/ml) in incremental volumes in 100 ml volumetric flasks.
After adding acid using the same conditions as that for the sample,
bring up to volume with water.
Measurement:
Measurement wavelength: 228.8 nm
Calibration curve concentration range: 0.05~1 mg/ml
Measurement conditions: Refer to Cookbook Section 3, Item 6.4, 11)
• Cd II (When Cd concentration is below the quantitation range, or when
interfering components are present)
The analysis method is the same as that described in item 10.1.3 Flame
Atomic Absorption Method, Cd.
• Co
Reagents:
1) Co standard solution (10 µg of Co/ml) : Refer to Cookbook Section
2, item 3 Preparing Standards

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 2) Cupferron solution (50 g/l) : Dissolve 5 g of cupferron (N-
nitrosophenylhydroxylamine ammonium salt) in water and bring up to
100 ml.
3) Ammonium citrate solution (500 g/l) : Gradually add 250 ml of
aqueous ammonia to 250 g of citric acid. While cooling to dissolve,
add water to bring up to 500 ml.
Remark:
When there is a possibility that the ammonium citrate is contaminated
with Co after dissolving with aqueous ammonia, adjust to pH 9 using
aqueous ammonia and dilute nitric acid. Transfer this to a separating
funnel. Add 25 ml of cupferron solution (50 g/l) and 25 ml of 4-methyl-
2-pentanone (MIBK). After vigorously shaking to mix, set solution aside,
discard the organic layer, and add water to the aqueous phase to bring up
to 500 ml.
4) 4-methyl-2-pentanone (MIBK)
Procedure:
1) Place an appropriate amount of pretreated sample (containing 2.5~15
µg of Co) in a beaker. Add 5 ml of ammonium citrate, and adjust to pH
6~7 using nitric acid and aqueous ammonia. Transfer this liquid to a
separating funnel, and add 10 ml of cupferron solution (50 g/l) and
bring up to 50 ml using water. After accurately adding 5 ml of MIBK,
shake vigorously for several minutes to mix, and then let stand. The
organic phase is used for measurement.
2) For the calibration curve, prepare incremental standard solutions
over the calibration curve concentration range using the Co standard
solution (10 µg of Co/ml), and use the same procedure as that for the
sample.
Measurement:
Measurement wavelength: 240.7 nm
Calibration curve concentration range: 0.5~3 mg/ml (concentration after
extraction)
Measurement conditions:
Lamp current value : 12 mA

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 Slit width : 0.2 nm
Lamp mode : BGC-D2
Burner height : 7 mm
Support gas : Air
Fuel gas flow rate :
C2H2 0.8 l/min (If the flame turns reddish when the sample is
sprayed, decrease the amount of sample drawn up.)
• Cu
The analysis method is the same as that described in item 10.1.3 Flame
Atomic Absorption Method, Cu I.
• Fe
The analysis method is the same as that described in item 10.1.3 Flame
Atomic Absorption Method, Fe.
• K
The analysis method is the same as that described in item 10.1.3 Flame
Atomic Absorption Method, K. Since this result expresses the amount of K,
multiply by 1.205 to express amount of K2O.
• Mg
The analysis method is the same as that described in item 10.1.3 Flame
Atomic Absorption Method, Mg. Since this result expresses the amount of Mg,
multiply by 1.658 to express amount of MgO.
• Mn
The analysis method is the same as that described in item 10.1.3 Flame
Atomic Absorption Method, Mn.
• Mo
Reagents:
1) Mo standard solution (10 µg of Mo/ml) : Refer to Cookbook Section
2, item 3 Preparing Standards
2) 8-quinolinol solution (200 g/l) : Place 20 g of 8-quinolinol (oxine)
in a beaker, and heat on a water bath. Add water to bring volume up to
100 ml.
3) 4-methyl-2-pentanone (MIBK)

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 Procedure:
1) Place an appropriate amount of sample (containing 25~300 µg of
Mo) in a 100 ml volumetric flask. Add 10 ml of 1 mol hydrochloric
acid, 5 ml of 8-quinolinol solution (200 g/l), and water to bring volume
to about 80 ml. Shake vigorously to mix and then let stand. Accurately
add 5 ml of MIBK, shake vigorously for 1~2 minutes and then let stand.
Use the organic phase for measurement.
2) For the calibration curve, prepare incremental standard solutions of
Mo standard solution over the quantitation range, and use the same
procedure as that for the sample.
Measurement:
Measurement wavelength: 313.3 nm
Calibration curve concentration range: 5~30 µg/ml (concentration after
extraction)
Measurement conditions:
Lamp current value : 12 mA
Slit width : 0.5 nm
Lamp mode : BGC-D2
Burner height : 17 mm
Support gas : N2O
Fuel gas flow rate : C2H2 6.5 l/min
• Ni
Reagents:
1) Ni standard solution (10 µg of Ni /ml) : Refer to Cookbook Section
2, item 3 Preparing Standards
2) Ammonium nitrate solution (10 g/l)
3) m-Cresol Purple solution (1 g/l) : Dissolve 0.1 g of m-cresylic
purple in 50 ml of ethanol, and add water to volume of 100 ml.
4) diethyl sodium dithiocarbamate solution (10 g/l) : Dissolve 1.3 g of
sodium dithiocarbamate in water, and bring volume to 100 ml.
5) Butyl acetate
Items 2), 3), 4) and 5) are prepared as in 2) ~ 5), respectively, for the
Cd reagents in item 10.1.3 Flame.Atomic Absorption Analysis Method

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 Procedure:
1) Place a suitable amount of pretreated sample (containing 5 ~ 30 µg
of Ni) in a separating funnel, and proceed as described in step 1) of the
Cd procedure in item 10.1.3 Flame.Atomic Absorption Analysis
Method.
2) For the calibration curve, prepare incremental standard solutions of
Ni standard solution over the calibration curve quantitation range, and
use the same procedure as that for the sample.
Measurement:
Measurement wavelength: 232.0 nm
Calibration curve concentration range: 0.5~3 µg/ml (concentration after
extraction)
Measurement conditions:
Lamp current value : 12 mA
Slit width : 0.2 nm
Lamp mode : BGC-D2
Burner height : 7 mm
Support gas : Air
Fuel gas flow rate :
C2H2 0.8 l/min (If the flame turns reddish when the sample is
sprayed, decrease the amount of sample drawn up.)
• Pb
The analysis method is the same as that described in item 10.1.3 Flame
Atomic Absorption Method, Pb.
• Zn I (when Zn concentration is within the quantitation range of 0.05~1
µg/ml, and there are no interfering components)
The analysis method is the same as that described in item 10.1.3 Flame
Atomic Absorption Method, Zn I.
• Zn II (when the Zn concentration is below the quantitation range)
The analysis method is the same as that described in item 10.1.3 Flame
Atomic Absorption Method, Zn II.
11.1.3 Hydride Vapor Generation Atomic Absorption Method
a) Target elements
As, Se

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b) Sample pretreatment
• Total As (fertilizer)
Place 1~5 g of sample in a beaker. Wet with water, add 2 ml of sulfuric acid,
about 5 ml of nitric acid, and 20 ml of perchloric acid. Heat until white fumes
of perchloric acid are generated, and further, until the sample becomes
concentrated almost to dryness. After cooling, add water to dissolve and after
cooling, transfer to a 100 ml volumetric flask. Add water to bring up to
volume, and pass through dry filter paper. Use the filtrate for measurement.
• Water soluble As
The procedure is the same as that described in item 11.1.1 Sample
Pretreatment, b) Water-soluble component analysis, Cd.
• Total Se (inorganic fertilizer and rock phosphate)
Place 1~5 g of sample in a tall beaker. Wet with a small amount of water,
and add 20 ml of nitric acid. Cover with a watch glass and gradually heat to
decompose. When the liquid volume becomes about 5 ml, add about 10 ml
more of nitric acid and heat again to completely decompose. Continue heating
until the sample is nearly dry. Add approximately 20 ml of water and heat to
dissolve. After allowing to cool, transfer to a 100 ml volumetric flask. Add 10
ml of hydrochloric acid, and add water to bring up to volume. Then pass
through dry filter paper. Use the filtrate for measurement.
• Total Se (sample containing organic material)
Place the sample in a Kjeldahl flask. Add 1~2 glass beads, and add
approximately 10 ml of nitric acid for each 1g of sample. Attach an air cooling
tube and gradually heat. Continue heating so that as the temperature gradually
rises, the nitric acid begins to condense at the bottom of the cooling tube, and
then heat for 10 minutes more. After cooling, carefully add 2~4 ml of
perchloric acid. Attach the cooling tube and gradually heat for 15 minutes.
Detach the cooling tube, heat again until the white fumes of perchloric acid are
generated. Then continue heating for 15 minutes longer. Rinse the insides of
the flask with a small amount of water, and heat again until the white fumes of
perchloric acid are generated, and then allow to cool. Add approximately 20 ml
of water to dissolve, and transfer to a 100 ml volumetric flask. Add 10 ml of
hydrochloric acid , and add water to bring up to volume. Pass through dry filter
paper, and use the filtrate for measurement.

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 c) Measurement procedure
• As
Same procedure as that described in item 10.1.4 Hydride Vapor Generation
Atomic Absorption Method, b) Measurement procedure
• Se
Reagents:
1) Se standard solution (1 µg of Se/ml) : Refer to Cookbook Section 2,
item 3 Preparing Standards
2) Sodium tetrahydroborate solution : Same procedure as that described
in item 10.1.4 Hydride Vapor Generation Atomic Absorption Method,
b) Measurement procedure, Reagents, 3).
Procedure:
1) The pretreated sample may be used for measurement as it is.
2) For the calibration curve, prepare incremental standard solutions
from the Se standard solution (1 µg of Se/ml) with concentrations of Se
ranging from 20~100 ng. Add 4 ml of hydrochloric acid (1+1), and
then add water to bring all solutions up to a volume of 20 ml.
Measurement:
Measurement wavelength: 196.0 nm
Calibration curve concentration range: 1~5 ng/ml
Measurement conditions:
Lamp current value : 23 mA
Slit width : 0.5 nm
Lamp mode : BGC-D2
11.1.4 Reduction Vaporization Atomic Absorption Method
a) Target element
Total Hg
b) Sample pretreatment
• Fertilizer:
Place a fixed amount of sample (2~5 g of inorganic fertilizer, or 5~10 g of
fertilizer containing organic material) in a decomposition flask, and use the
same procedure as that described in item 10.1.5 Reduction Vaporization Atomic
Absorption Method, b) Sample pretreatment , Procedure.

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• Rock phosphate
Place 2 g of sample in a decomposition flask, and wet with a small amount
of water. Add 25 ml of phosphoric acid (1+1) and 2~3 ml of potassium
permanganate (potassium permanganate 50 g/l). Attach a cooling circulator and
heat for about 1 hour to decompose. During this time, if the potassium
permanganate color disappears, cool to below 60°C, and add 2~3 drops of
potassium permanganate solution. Then heat. Repeat this procedure until the
potassium permanganate color remains, and then heat for 10 minutes longer.
After cooling, transfer to a 250 ml volumetric flask. Bring up to volume with
water, and use this solution for measurement.
• Water soluble Hg
Same as that described in item 11.1.1 Sample Pretreatment, B) Water
soluble Components, Cd
c) Measurement procedure
Same as that described in item 10.1.5 Reduction Vaporization Atomic
Absorption Method, C) Measurement procedure.

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11.2 Feed Analysis Method
11.2.1 Sample Pretreatment
Same as that described in item 10.1.1 Sample Pretreatment
11.2.2 Flame Atomic Absorption Method
a) Target Elements
Ca, Cd, Fe, Mg, Mn, Pb, Zn
b) Measurement Procedure
Measurement is conducted using the following procedure. The lamp current,
slit width and flame conditions are as described in Cookbook Section 3, item
6.4 Element Specific Conditions. For preparation of standards for the
calibration curves, refer to Cookbook Section 2, item 3 Preparing Standards.
• Ca
Same as that described in item 10.1.3 Flame Atomic Absorption Method, Ca
• Cd
Same as that described in item 10.1.3 Flame Atomic Absorption Method, Cd
• Cu I (when Cu concentration is within quantitation range of 0.2~4 mg/ml,
and there are no interfering components)
Same as that described in item 10.1.3 Flame Atomic Absorption Method, Cu I
• Cu II (when Cu concentration is below quantitation range)
Same as that described in item 10.1.3 Flame Atomic Absorption Method, Cu II
• Fe
Same as that described in item 10.1.3 Flame Atomic Absorption Method, Fe
• Mg
Same as that described in item 10.1.3 Flame Atomic Absorption Method, Mg
• Mn
Same as that described in item 10.1.3 Flame Atomic Absorption Method, Mn
• Pb
Same as that described in item 10.1.3 Flame Atomic Absorption Method, Pb
• Zn I (when Zn concentration is within quantitation range of 0.05~1 mg/ml,
and there are no interfering components)
Same as that described in item 10.1.3 Flame Atomic Absorption Method, Zn I
• Zn II (when Zn concentration is below quantitation range)
Same as that described in item 10.1.3 Flame Atomic Absorption Method, Zn II

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