Amendment to the Enforcement Ordina nce of the Food Sanitation Law 
and the Standards and Specificatio ns for Foods and Food Additives 
 
The government of Japan will designate -Apo-8’-carotenal as authorized food 
additives. 
 Summary 
Under Article 10 of the Food Sanitation La w, food additives shall not be used or 
marketed without authorization by the Mi nister of Health, Labour and Welfare 
(hereinafter referred as “the Minister”). In addition, when specifications or standards are 
established for food additives based on Article 11 of the act and stipulated in the Ministry of 
Health, Labour and Welfare Notification (Ministry of Health and Welfare Notification No. 370, 
1959), those additives shall not be used or ma rketed unless they meet the standards or 
specifications.   
In response to a request from the Minister, the Committee on Food Additives of the 
Food Sanitation Council that is establishe d under the Pharmaceutical Affairs and Food 
Sanitation Council has discussed the adequacy of the designation of -Apo-8’-carotenal 
as food additives. The conclusion of the committee is outlined below.  
  Outline of Conclusion  
The Minister of Health, Labour and Welfare, based on Article 10 of the Food 
Sanitation Law, should designate -Apo-8’-carotenal as food additives unlikely to harm 
human health, and establish standard for use and compositional specifications, based 
on Article 11 of the law (see Attachment).      Attachment 
 
 -Apo-8-carotenal 
-アポ-8-カロテナール  
 
CH3 CH3
CH3 CH3CHO
CH3CH3 H3C
 
 
 Standards for use 
Shall not be used in fresh fish/shellfish (including fresh whale meat), konbu 
(kelp)/ wakame  (both Laminariales algae), legumes/pulses, meat, nori (laver), tea, 
and vegetables.  
 
Compositional specifications 
Substance name  -Apo-8-carotenal 
Molecular formula  C30H40O  
Molecular weight   416.64 
Chemical name [CAS number]    
(2E,4E,6E,8E,10E,12E,14E,16E)-2,6,11,15-tetramethyl-17- 
(2,6,6-trimethylcyclohex-1-en-1-yl)h eptadeca-2,4,6,8,10,12,14,16-octaenal  ［1107-26-2 ］ 
Content   -Apo-8-carotenal contains not less than 96.0% of -Apo-8-carotenal 
(C30H40O). 
Description    -Apo-8-carotenal occurs as dark viol et crystals or crystalline powder 
having a metallic luster.   
Identification   (1) A 1 in 20,000 solution of -Apo-8-carotenal in acetone produces an 
orange color. To 5 ml of this solution, add 1 ml of sodium nitrite solution (1 in 20) and 1 
ml of sulfuric acid TS (0.5 mol/L). Th e solution is immediately decolored.  
(2) The test solution prepared in the Assay below exhibits absorption maxima at 
wavelengths of aro und 461 nm and 488 nm. 
Purity  
(1) Lead  Not more than 2.0 g/g as Pb. 
Test Solution  Weigh 2.0 g of -Apo-8-carotenal in a platinum, quartz, or porcelain crucible or a quartz beaker. Heat gradually until the sample melts and becomes viscous. 
Stop heating before the sample starts carbonizing and cool.  Add 1 ml of sulfuric acid, 
and heat by increasing the temperature gradually until white fumes are no longer 
evolved. If necessary, add sulfuric acid ag ain, and heat until the sample is almost 
carbonized. Loosely lid the crucible if necessary, heat in an electric furnace by 
increasing the temperature grad ually, and incinerate at 450–600 C. If carbonized 
matter still remains, crush the residue with a glass rod, moisten with 1 ml of diluted 
hydrochloric acid (1 in 4) and 1 ml of ni tric acid, and heat until white fumes are no 
longer evolved. Then, ignite in the electric furnace to complete incineration. To the residue, add 10 ml of diluted hydrochloric acid  (1 in 4), and evaporate on a water bath to 
dryness. To the residue, add a little amount of  diluted nitric acid (1 in 100), and dissolve 
it while heating. Add diluted nitric acid (1 in 100) again to make exactly 10 ml. When 
the incineration is done at a temperature of 500 C or less, a heat-resistant glass beaker 
may be used. 
Control Solution  Add water to exactly measured 1 ml of Lead Standard Stock 
Solution to make exactly 100 ml. To exactly me asured 4 ml of this solution, add diluted 
nitric acid (1 in 100) to make exactly 10 ml. 
Procedure  Proceed as directed under Method 1 in the Lead Limit Test.     
(2) Arsenic  Not more than 4.0 g/g as As 2O3 (0.50 g, Method 3, Apparatus B). 
(3) Absorption ratio  Measure absorption values (A 1 and A 2) of the test solution 
prepared in Assay at wavelengths of 461 nm and 488 nm. A 2/A1 is 0.80–0.84. 
(4) Subsidiary colors  Not more than 3%. 
Test Solution   To 0.010 g of -Apo-8-carotenal, add tetrahydrofuran with BHT to 
dissolve, and make 100 ml. To 1 mL of this  solution, add ethanol to make 10 mL.  
Procedure  Analyze 10 l portions of the test solution by liquid chromatography 
using the operating conditions below.  Cont inue the chromatography for about 6 times 
the retention time of the main peak. Norm alize the sum of the peak areas of all 
components in the test solution to 100. Cons ider all peaks other than the main peak to 
be derived from subsidiary colors and de termine the peak area percentage.    
Operating Conditions  
Detector: Ultraviolet spectrophotometer (wavelength 463 nm).  
Column: A stainless steel tube of about 4. 6 mm internal diameter and about 25 cm 
length.   
Column packing material: Hexadecylamido propyl silanized silica gel for liquid 
chromatography. 
Column temperature: 30 C.  
Mobile phase: To 20 ml of a 1 in 400 solution of dibutylhydroxy toluene in 2-propanol, add 0.2 ml of N-ethyl- N-(1-methylethyl)propane-2-amine, 25 ml of 
ammonium acetate solution (1 in 500), 455 ml of acetonitrile, and 450 ml of 
methanol. Dilute with ethanol to make 1000 ml . Prepare fresh before use.       
Flow rate: Adjust so that  the retention time of -Apo-8-carotenal is about 7–9 
minutes.  
Residue on Ignition   Not more than 0.10%. 
Assay   
Test Solution   Weigh accurately about 0.04 g of -Apo-8-carotenal, add 10 ml of 
chloroform to dissolve, and add cyclohexane to make exactly 50 ml. To 5 ml of this 
solution, exactly measured, add cyclohexane to make exactly 100 ml. To 5 ml of the 
second solution, add cyclohex ane to make exactly 100 ml.    
Procedure   Measure the absorbance (A) of the te st solution against cyclohexane at a 
maxima absorption wavelength around 461 nm . Calculate the content by the formula:   
Content (%) of -apo-8′-carotenal C 30H40O  
=		ଶ଴଴
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ଶ଺ସ଴ൈ100  
Storage Standards  Store in a hermetic, light-resista nt container under inert gas.  
 
Reagents and Solutions 
N-ethyl- N-(1-methylethyl) propane-2-amine  C 8H19N  A colorless or very slightly 
yellow, clear liquid.  
Content   Not less than 95.0%.   
Density  0.750–0.760.  
Assay  Analyze 1 μl portions of N-ethyl- N-(1-methylethyl)propane-2-amine by gas 
chromatography using operating conditions  below. Measure the areas of all peaks 
obtained to determine the amount from the ra tio of the main peak area using the peak 
area percentage method.  
Operating conditions  
Detector: Thermal conductivity detector  
Column: A fused silica capillary (about 0.53 mm internal diameter and about 15 m 
length) coated with a 1.5- μm thick layer of dimethyl polysiloxane for gas 
chromatography.   
Column temperature: Inject at 50 C, and raise to 150 C at a rate of 10 C/minute.  
Injection port temperature: 200 C. 
Detector temperature: 250 C. Carrier gas: Helium. 
Flow rate: 5 ml/minute.  Injection: Sprit. Split ratio: 1 : 200. Measurement time: 15 minutes 
Dibutylhydroxy Toluene  C
15H24O  White to slightly yellow crystals, powder, or 
granuls.  
Content  Not less than 98.0%. 
Identification   Determine the infrared absorption spectrum of Dibutylhydroxy 
Toluene as directed in the Potassium Bromide Disk Method under Infrared 
Spectrophotometry. It exhibits the absorption at wavenumbers of about 2960 cm-1, 
1430 cm-1, 1360 cm-1, 1230 cm-1, 1150 cm-1, 1120 cm-1, 1030 cm-1, 880 cm-1, 870 cm-1, 
770 cm-1, and 580 cm-1. 
Melting point   69–72 C. 
Clarity of solution  Almost clear (1 g, ethanol (99.5), 20ml) 
Assay  Prepare a test solution by adding acetone to1 g of Dibutylhydroxy Toluene to 
make 10 ml. Analyze 1 μl portions of the test solution by gas chromatography using 
operating conditions below. Continue th e chromatograph for about three times the 
retention time of the main peak. Measure th e areas of all peaks obtained to determine 
the amount from the ratio of the main peak area using the peak area percentage method. 
Separately, perform a blank test to make correction. 
Operating conditions  
Detector: Flame-ionization detector .  
Column: A fused silica capillary (about 0.25 mm internal diameter and about 30 m 
length) coated with a 0.25- μm thick layer of dimethyl polysiloxane for gas 
chromatography.   
Column temperature: 190 C.  
Injection port temperature: 240 C. 
Detector temperature: 250 C. 
Carrier gas: Helium. 
Flow rate: 1.33 ml/minute.  Injection: Sprit. Split ratio: 1 : 100.  
Tetrahydrofuran with BHT   [K9705] Use tetrahydrofuran containing 0.025% of 
dibutylhydroxy toluene.