AOAC-RI ERP Book - Gluten.pdf

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Table 2. Comparison to existing methods
Samples
N¡ean values (Þpm)
Neogen Offic¡al Method 2012.0 1
Soy fìour
Rice c€real
Rice cnsp
Granola bars
R¡ce cereal
Rice mix
Soy crisp
G¡nger c€ke m¡x
C)at cereal
Chocolate wafers
R¡ce cocoa cereal
Ivlult¡grain cereal
Corn flakes cereal
Chocolate oat cereal
14.20
49.50
73.7 0
33.50
57.80
63.1 0
30.00
46.60
29 60
40,30
38.50
85.20
9.00
14.10
16.20
62.70
71.80
43_40
64.00
61.60
30.90
48.60
29 10
40.1 0
42.20
84.00
10.60
15.30
¿¡nd concentralions (ppni) of'lhe controls f'onìt a standard curve.
aud the sample ppm values olgliadin are intcrpolated fì'onr the
culve based on their o¡rtical densit¡, values.
Test Kit lnformation
(a) Kir name.-Veratoxd R5 Gliaclin resr kil.
(b) Cat. ,M.-85 10.
(c) Ordering inforntaÍion.-ln the United S/ates.-Neogen
Corp.. 620 Lesher Pl. I-ansing. MI 489t2. phone: 517-372-
9200, lax: 517-372-0108. \\/w\\,.neogen.cotn. Outside the
1..' ttitL,¿ St¿ttes.-lonracr abo\ !: ao; ia¡ii ¿iii''iUurur intblrnal ion,
Test Kit Reagenfs
(a),4nfibodt,-cooÍed nticrou'ells GB).
(b) Red-ntarked n ix ing ve I l:¡ (18).
(c) Yellou-labeled lrottles of 1.2 rnl. each 0.0. 2.5. 10.0.
20.0. and 40.0 ppnr gliadin standards.
(d) Trvo blue-labeled botlles of' 4 mL enz¡me-labeled
antibod¡ conjugate soluliou.
(e) One green-labeled bottle ol' 12.5 mL K-Blue substrate
solution (tetlamethl,'lbcnzidine).
(f) One red-labeled bottle of 12.5 ml- Red Stop solution
(0.00069'" NaF + cresol red).
(g) One bottle of 40 mL l0 mM PBS-'I\r'een u,ashing re¿ìgent
in a rvidc mouth botlle. Llach bottle can prepâre I l- in distillcd
or deíonized uater (pLI 7.4).
(h) One fbil pouch sanrplc diluent corìcenlrate ol' l0 ml\,f
PBS dr¡' pou,der containing cnouglr pou'cler 1o prepare I I- of'
dilution bufìbr.
(i) 'l\r'o cups ol50 g cxiraction additive.
0) Plastic scoop 10 rneasurc extÍactioll additive.
I-|p( ..: JoiJRNAI-()t.AOACINÌr,.rìNÄi' . l.2013 123
Additional Supp/res and Reagents
(Rcquircd but nol includcd in the test kit).
(a) Gliadin renaturing cocktail solution lol lieat-processed
samples (Neogcn item No. 8515).
(b) 'l\ro I I- bottles to prepare rvashill,q solr¡tion ancl sarnplc
extraot dihrtiou solution.
(c) 'lèst tubes to perlbrnr sanr¡:rle extrirct dilution.
(d) I'irncr.
(e) l-hree reagerlt boats f'or l2-channel pipettor.
(Ð Wash bottle.
(g) Paper torvcls or equi'''alcnt absorbent rnateriâ1,
(h) \\¡alerprool r¡rarkcr.
(i) Distilled or deionizcd u¿rter.
(j) l-aborator¡ gladc ethanol (190 proof).
Apparatus
(a) Scale capable of u,eighrne 0.2,5 + 0.01 g.
(b) Microrvell reâder or strip reader rvith a 650 nrn fìlter or
equivalent rvith Neogen Veratox sofiu,are.
(c) 5-300 pL l2-channel acl.jrÌstable pipettor.
ttll 5-200 pl- aJju:taLrle lri¡ruttur.
(c) Cornpatiblc li¡rs l-or pipettors.
(f) Orbital rotâtor ol shakel' to hold 50 cc centrifìge tutres
during extraction.
(g) Oven or u,a1er bath adjustable to 50"C ifanalyzing heat-
processed samples.
Reference Material Used in Standard Preparation and
Spiking
Material used ftrr preparation of kit standards and spikin_e
erperirnenls \\:as Gliadin G3375 fiorn Sigma-Aldrich
(St. l,ouis, MO) calibrated a-sainst the Prolamine Working Group
standarcl: Zu'ickau, Gennanr'(4). Spiking solution s'as prepared
by l'eighing 25 mg of the above material usiug an analytical
balance uith 0.001 g precision and quantitativell' tlanslèrring
to a 50 mL pol¡,prop¡.'lene centrilìrge tube. ]-he rnaterial rvas
then dissolved in 25 mL deionized rvater and vortexed fòr 2 rrin
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Scatter plot of methods comparison. Fígure 1.