Aoac 981.16

45.1.12 (c) Sulfanilic acid solution.—10%.
Add 100 g sulfanilic acid to

AOAC Official Method 981.16 ca 500 mL H2O. Add NH4OH with mixing until dissolved (ca
Niacin and Niacinamide 40 mL). Adjust to pH 7.0 with HCl (1 + 3) and dilute to 1 L with H2O.
in Foods, Drugs, and Feeds Filter, and store in cool, dark place. Prepare fresh every 2 weeks.
Automated Method
(d) Cyanogen bromide solution.—5%. Weigh ca 50 g CNBr into
First Action 1981
Final Action 1982 beaker containing 300 mL H2O at ca 40°C, and stir to dissolve. Cool,
and di lute to 1 L with H2O. Prepare fresh every 2 weeks.
AACC–AOAC Method (Caution: CNBr is very poisonous. Weigh compound and prepare
[Applicable to products for which manual method, 961.14 (see solution in hood.)
45.1.10), is applicable and that contain ≥2 µg/g.] (e) Sampler wash solution.—Dilute 3.0 mL wetting agent to 2 L
A. Apparatus with H2O and filter through Whatman 2V paper.
(a) Automatic analyzer.—Technicon AutoAnalyzer II system (f) Calcium hydroxide slurry.—Add 22 g Ca(OH)2 to 200 mL
with flow scheme as in Figure 981.16 (Technicon Instruments volumetric flask and add ca 100 mL H2O. Shake to disperse and
Corp., man i fold No. 116-D137), or equiv a lent (Pulse dilute to volume with H2O. To use, transfer to 250 mL beaker on
Instrumentation (1992) Ltd., 433 Birch Cresent, Saskatoon SK magnetic stirrer and stir at rate to ensure homogeneity.
Canada S7N 2K2; www.autoanalyzer.com).
(g) Basic solution for waste container.—Dissolve 150 g NaOH in
(b) Collection funnels.—Disposable funnels, 20 mL, are convenient.
300 mL H2O in 4 L reagent bottle. Place bottle in hood and pump
(c) Pipets.—Rapid dispensing pipets are convenient in multiple
waste into this bottle.
analyses.
B. Reagents (h) Niacin standard solutions.—(1) Stock solution.—10 µg/mL.
Weigh 50.0 mg nicotinic acid (stored in desiccator) into 500 mL
( a ) We t t i n g a g e n t . — 3 0 % a q u e o u s B r i j 3 5 s o l u t i o n
volumetric flask, dissolve and dilute to volume with H2O. Daily,
(polyoxyethylene(23)lauryl ether; CAS-9002-92-0).
dilute 25.0 mL to 250 mL with H2O. (2) Working standard
(b) Phosphate buffer solutions.—(1) Stock solution.—Dissolve solutions.—Pipet 30, 20, 10, and 5 mL stock solution into 100 mL
130 g Na2HPO4 and 71 g KH2PO4 in ca 900 mL warm H2O. Cool to volumetric flasks containing 5 mL Ca(OH)2 slurry. Add H2O to ca
room temperature and dilute to 1 L with H2O. (2) Working 55 mL, autoclave, and treat test portions and standards alike. Final
solution.—pH 6.7. Dilute 150 mL stock solution to 1 L with H2O and
solutions will contain 3.0, 2.0, 1.0, and 0.5 µg niacin/mL.
add 1.5 mL wetting agent. Filter through Whatman 2V paper before
use. (3) Test portion buffer solution.—pH 7.6. Dissolve 272 g
C. Preparation of Test Sample
Na2HPO4 and 48 g KH2PO4 in ca 1.8 L warm water. Cool to room
temperature and dilute to 2 L with H2O. Grind representative test sample to pass No. 40 sieve.
Figure 981.16. Flow diagram for automated determination of niacin and niacinamide.
 2005 AOAC INTERNATIONAL

D. Determination and pump set of standards and test solutions through system. Use
Transfer accurately weighed test portion (1.5 g maximum) one standard with every series of 20 test solutions to correct for any
containing ca 0.2 mg niacin to 100 mL volumetric flask. Add 5 mL drift. If test solution is more concentrated than highest standard,
Ca(OH)2 slurry, using Rainin pipet, or weigh 0.55 g Ca(OH)2 into dilute test solution with test portion buffer solution to bring peak
flask. Add ca 50 mL H2O, cover with foil, swirl, and autoclave 2 h at height into range of standards. After all test solutions have been run,
121°C. For products which spatter during hydrolysis (high oil replace CNBr line with H2O. Let pump ca 15 min and resample
content), use 125 mL Erlenmeyer and transfer to 100 mL volumetric filtrates to obtain corresponding blank values. Alternatively, dual
flask after autoclaving. channel instrument may be used for simultaneous blank corrections.
While all solutions are still hot, add 10 mL 1.5M HCl (1 + 7) with Determine concentration, C, (µg/mL) from standard curve:
auto pipettor, and swirl to dissolve remaining Ca(OH)2. Be sure all
Ca(OH)2 is dissolved. Let cool to room temperature. (Standard and
test solutions may be stored at this point.) Niacin or niacinamide/g, mg = C × 10/W
To test and standard solutions, add 25 mL test portion buffer
solution from pipettor, 2 drops wetting agent, and dilute to volume
where W = g test portion.
with H2O. (Precipitate forms and final pH will be ca 6.7.) Shake, and
filter through Whatman 2V paper (disposable collection funnels are Reference: JAOAC 62, 1027(1979).
convenient).
Pump high standard solution (3 µg/mL) through system and set CAS-59-67-6 (niacin)
recorder pen at 100% with standard calibration adjustment. Aspirate CAS-98-92-0 (niacinamide)
 2005 AOAC INTERNATIONAL

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45.1.12 (c) Sulfanilic acid solution.—10%.

Add 100 g sulfanilic acid to

 2005 AOAC INTERNATIONAL

 2005 AOAC INTERNATIONAL

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