NATURAL TOXINS AOAC OFFICIAL METHODS OF ANALYSIS (2005)

Chapter 49, p. 82

E. Determination A. Acid Titration Method

Inject replicate 20 f..lL portions of domoic acid standard solution Place 10-20 g test portion, ground to pass No. 20 sieve, in 800 mL
into LC system until peaks (measured as height or area) for 3 Kjeldahl flask, add 100 mL HP, and macerate at room temperature
consecutive injections do not vary by more than 3%. Ensure 2 h. Add 100 mL HP and steam distill, collecting distillate in 20 mL
baseline resolution ofL-tryptophan from domoic acid; adjust mobile 0.02M AgN03 acidified with I mL HN03. Before distilling, adjust
phase composition accordingly. apparatus so that tip of condenser dips below surface of liquid in
Make alternate, duplicate injections of test solution from D and receiver. When 150 mL has passed over, filter distillate through
standards. Gooch, wash receiver and Gooch with little H20, and titrate excess
Determine recoveries of domoic acid at 20 f..lg/g level. AgN03in combined filtrate and washings with 0.02M KSCN, using
F. Calculations ferric alum indicator.
Calculate results as follows:
1 mLO,02M AgN03 = 0.54 mg HCN
Domoic acid, I-lg/g = (RIR') x (W'/W) B. Alkaline Titration Method
Place 10-20 g test portion, ground to pass No. 20 sieve, in 800 mL
where Rand R' are average peak heights or areas of test solution and
standard, respectively, and Wand W' are weights injected of test Kjeldahl flask, add ca 200 mL H20 and let stand 2--4 h. (Autolysis
portion, mg, and standard, ng, respectively. should be conducted with apparatus completely connected for
distillation.) Steam distill, collect 150-160 mL distillate in NaOH
Reference: JAOAC74,68(1991). solution (0.5 gin 20 mL HP), and dilute to definite volume.
To 100 mL distillate (it is preferable to dilute to 250 mL and
titrate 100 mL aliquot) add 8 mL 6M NH 40H and 2 mL 5% KI
Subchapter 11 solution and titrate with 0.02M AgN0 3, using microburet. End
PLANT TOXINS point is faint but permanent turbidity and may be easily
recognized, especially against black background.

49.11.01 1 mL 0.02M AgN0 3 =

AOAC Official Method 936.11 1.08 mg HCN (Ag equivalent to 2 CN)
Cyanogenetic Glycosides in Feeds References: J. Am.Chem. Soc. 37,601(1915); JAOAC 4, 151(1920);
Qualitative Test 17,182(1934); 18,347(1935); 19, 589(1936);
First Action 1936 33, 83(1950); 38,96(1955).
Final Action
Prepare sodium picrate paper by dipping strips of filter paper into CAS-74-90-8 (hydrocyanic acid)
__~,&pj<:ricll(;i<Lsolution and drying, then dipping into 10% Na2C03
solution and drying:8torethesepapers in stopperedoo1tle-:---- - ---- - - - .. ----- - --- ---- - ______ _
· I h
Fme II f l ' I d I' 49.11.03
y coP. sma amo.unt 0 p an~ mate~Ia an p ac~ m test AOAC Official Method 997.13
tube. Insert pIece of mOIstened sodIUm pIcrate paper m tube,
taking care that it does not come in contact with test sample. Add Glycoalkaloids (a-Solanine and a-Chaconine)
few drops CHCI 3 and stopper tube tightly. The sodium picrate in Potato Tubers
paper gradually turns orange, then brick red, if plant tissue Liquid Chromatographic Method
contains cyanogenetic glycosides. (Test is delicate, and rapidity First Action 1997
of change in color depends upon amount of free HCN present. (Applicable to quantitative determination of 10-200 mg/kg
This test works well with fresh plant materials, but relatively dry a-solanine and 20-250 mgikg a-chaconine in raw potato tubers .)
substances, particularly seeds of various plants, should be ground
and moistened with H20 and allowed to hydrolyze in stoppered Caution: See Appendix E, safety notes on safe handling of
test tube containing sodium picrate paper. If necessary, small special chemical hazards. a-Solanine and a-chaconine
amount of emulsion may be added.) are toxic. Do not inhale or swallow. Avoid contact with
skin. Handle liquid N carefully. Extremely low
References: JAOAC 19, 94,589(1936); 20,444(1937); temperature (-196°C) can cause skin injury, frost
21,614(1938). injury, or similar burn. Leather gloves and safety
goggles with side shield or face shield should be worn.
Boiling and splashing always occur when filling warm
49.11.02 container or when inserting objects into liquid N.
AOAC Official Method 915.03 Always perform these operations slowly to minimize
Hydrocyanic Acid in Beans boiling and splashing. Store and use liquid N only in
Titrimetric Methods well ventilated place. Due to evaporation ofN gas and
First Action 1915 condensation of 0 gas, the percentage of 0 in a
Final Action confined space can become dangerously low. Due to
(Formed by hydrolysis of glucosides.) the large expansion ratio from liquid to gas (1 to 700),

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