Page 75 - Application Notebook - Solution for Food Safety

Application No.L514
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Q Analysis of Melamine and its Related
Substances in Fertilizer Sample 0.50 g Erlenmeyer flask with stopper 200 mL
The analytical sample (fertilizer) was pretreated in Spike
accordance with the test method by adding a standard Hydrochloric acid / water = 1/15 (v/v) 100 mL
quantity of melamine and its related substances. The
pretreatment procedure is indicated in Fig. 4 and the Extraction Ultrasonic extraction for 30 minutes
analytical results are shown in Fig. 5. In this example, five
types of samples were tested, including two types of Centrifugation 2000 × g for 5 minutes
commercially available nitrolime, a synthetic fertilizer that
contains calcium cyanamide, a synthetic fertilizer, and
ammonium sulfate. The quantities of the substances Supernatant
added to the analytical samples, as a percentage of
mass, were equivalent to about 0.035 to 2.8 % Dilution Transfer 5 mL to a 50 mL volumetric flask,
add acetonitrile / phosphate buffer = 4/1 (v/v)
melamine, about 0.035 to 1.6 % ammeline, about up to the marked line
0.035 to 1.1 % ammelide, and about 0.037 to 1.2 %
cyanuric acid. These results demonstrate that the Centrifugation 8000 × g for 5 minutes
Prominence system provides more than adequate
performance for measuring the provisional 0.4 %
melamine limit issued by the Food Safety and Consumer Supernatant
Affairs Bureau, Ministry of Agriculture, Forestry and
Fisheries for calcium cyanamide and fertilizers that
contain calcium cyanamide as an ingredient. Measurement HPLC

Fig. 4 Pretreatment Procedure
(ᶗ) Nitrolime A (ᶘ) Nitrolime B (ᶙ) Synthetic fertilizer containing nitrolime
mAU mAU mAU
30 3 3 10.0 3
7.5
7.5
20
5.0 1
2 5.0
4
10 2 4 2.5
1 4 2.5 2
1
0 0.0 0.0
0.0 2.5 5.0 7.5 10.0 12.5 15.0 0.0 2.5 5.0 7.5 10.0 12.5 15.0 0.0 2.5 5.0 7.5 10.0 12.5 15.0
min min min
(ᶚ) Synthetic fertilizer (ᶛ) Ammonium sulfate
mAU mAU
˙ Peaks
4 1 2 0.75 3 1) Cyanuric acid
2) Ammelide
3 3) Melamine
0.50
3 1 2 4) Ammeline
2 4
0.25
1 4
0.00
0
0.0 2.5 5.0 7.5 10.0 12.5 15.0 0.0 2.5 5.0 7.5 10.0 12.5 15.0
min min
Fig. 5 Chromatograms for (ᶗ) Nitrolime A, (ᶘ) Nitrolime B, (ᶙ) Synthetic Fertilizer Containing Calcium Cyanamide,
( ᶚ ) Synthetic Fertilizer, and ( ᶛ ) Ammonium Sulfate
References
1) Health effects of melamine, etc.: Food Safety Commission of Japan, October 9, 2008, updated April 30, 2009
2) Setting Allowable Limit Values for Melamine in Calcium Cyanamide, Food Safety and Consumer Affairs Bureau, Ministry of Agriculture, Forestry and
Fisheries in Japan, Notice No. 6116, 2012, issued March 25, 2013 and partially revised March 30, 2013
3) Testing Methods for Fertilizers (2016), Food and Agricultural Materials Inspection Center (FAMIC): http://www.famic.go.jp/ffis/fert/obj/
shikenho_2016.pdf (in Japanese)
4) Etsuko Bando and Yuji Shirai: Validation of High Performance Liquid Chromatography (HPLC) for Determination of Melamine and Its Related
Substances in Fertilizer, Research Report of Fertilizer Vol. 6, pp. 27 to 35 (2013)
5) Etsuko Bando and Shigehiro Kai: Determination of Melamine and Its Related Substances in Fertilizer by High Performance Liquid Chromatography
(HPLC): A Collaborative Study, Research Report of Fertilizer Vol. 7 10-21 (2014)

First Edition: Jan. 201

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