Proposition: Assessing bio-activity of pre-popped fortified popcorn during stages of shelf-life.

Justification: Economic and Nutritional

European sales of meat alternatives rose by 450% between 2013 - 2017 (Iannetta P., 2018).
This increase is indicative of a growing market of products tailored for plant-based diets. Graph 1 suggests
rates of EU consumers identified as following 2 of these diets.

Statista, 2017

Graph 1. Percent of self-reported adherence to 2 plant-based diets by 16-24 year olds for different EU
residents during 2017.

Nutritionally, cobalamins (vitamin B12) are commonly obtained through the consumption of other
animals who have stored this vitamin in their body or, through consumption of the animal's lactation or
ovulation. Thus supplementation and/or consumption of fortified foods is advisable if following a strict
herbivorous diet. In response to the apparent increased product demand and to address associated
nutrient deficiency, a simple and potentially viable product may be achieved by covering popcorn in a
fortified yeast extract bound with flax oil -this oil aims to address another potential vegan deficiency of
ALA - however its lipid stability is complex.
Reasons for choosing this yeast extract: first the reported cheese like flavor - for practicing vegans who
may miss this taste. Secondly, to address this target group micro-nutritionally.
The importance of this research is limited specifically to this product. This research only attempts to
scientifically test this fortification during product shelf life, for accurate marketing and consumer safety.
Although the prevalence of B12 deficiency in the general Irish population is unknown.
Research plan: Fortify and assess shelf life of bioactivity.
Tasks / objectives to be carried out:

1. What tests and why? [Why have you chosen these specific methods?]

A review of research on B12 deficiency among vegetarians suggests nutritional yeast fortified with B12
(cyanocobalamin) can be used effectively as a supplement (Pawlak et al. 2013). Testing for specifically for
methylmalonic acid and holo-transcobalamin are advised. As of writing this, other methods are deemed
unreliable. Thus these two tests will be used. As for the frequency, graph 2, showing transcobalamin (TC-
CNClb) levels rising 24hrs after first (of three 9μl doses) of orally consumed cyanocobalamin is suggestive
of a next-day blood as viable timeframe for estimating absorption. (Hardlei et al., 2010)

Graph 2. Effects on transcobalamin levels on 10 individuals during 5 days of oral B12 consumption.

2. What’s the selection criteria?

Include (N=20) Exclude all who

Healthy BMI, both sexes, 18-65 ys., nonsmokers. Take medication,

Meat eaters and plant based.
Both B12 deficient (if present) and adequate at
baseline.
Past eating disorder or current disorder,
Considering changing their diet.
Supplement folic acid*
*This group perhaps is worth excluding as a folate deficiency can mask b12 deficiency (Scaglione &
Panzavolta, 2014). Admittedly, not an ideal exclusion as this issue is nutritionally relevant.

3. Considering this just mentioned relationship, how will folate status be measured?

A Plasma folate assay. Plasma levels of 13.4 - 45.3 nmol/L can be considered within the normal range
when assayed with Lactobacillus casei and converted as 1 ng/mL to 2.265 nmol/L (WHO,2015). A noted
limit: this range is for assessing the general population. Further investigation of range specificity and
potential underlying influencing factors related to folate testing (specifically the accuracy of different
assays) can be considered post review of this proposal.

4. What are the quantities of ingredients?

Popped kernels, 80g.

Fortified yeast extract: 240μg of B12 per serving. Why?

● As adult male/female B12 RDA is suggested at 2.4μg and slightly more for pregnant or lactating
(National Institute of Health , NIH)
● As B12 absorption can be 1-3% passive diffusion and only 2% from gastric intrinsic factor (Pawlak
et al, 2014)
● And as per NIH no upper limit has been identified yet.

Note: As there is only 445ng/g of B12 in the product thought initially to be used up until this point
(Engevita Yeast Flakes). It appears -due to absorption- this product would require additional B12
fortification to address deficiencies.
Considering this, product testing would have to be performed; seeing what concentration can be applied
to flakes/popcorn whilst remaining a desirable edible product. Before adding this reformed product to
the study.

Flax oil:
I assumed it viable to use nitrogen to preserve lipid integrity of ALA against oxidization. Perhaps coconut
oil could be used instead – as ‘Manhattan’ popcorn use this oil method and can obtain shelf life of months.
Protocol:
1. Baseline B12 and Folate status via blood tests.
 Once ~20 subjects have qualified as suitable, arrange time day 0 for blood tests.

2. Fortify and package all required product on Day 0*

3. Give 1 sample product to all subjects and ask to consume before test.

4. Subjects are asked to return on Day 3,7,14,& 30 for blood tests and will be asked to consume the
product 24 hours before test (Day 2,6,13 & 29.)
 Text notify day before test asking to eat sample.

*(Each product is opened on day of consumption. All have been sealed and fortified on Day 0).

Limits
 Diet changes could have occurred that the subject doesn’t notify about.
 Subject may forget to eat popcorn day before and may eat shortly before test.
 Awareness of genetic factors exists yet rate of accounting for this not considered.

Gantt outline of workflow

Duties Weeks

1 4 8

Advertising - -

Research - - - -

Extended research if product still viable. - -

Review data -

Publish -
Dissemination plan
Collect data and organise simply with aim of getting feedback for further testing.

Attempt to communicate data with specialists in B12. Aim to run study again, based on specialist feedback.
Ideally to market a safe prototype of clinical B12 food product. I would not attempt to publish in online
journals- although I’m probably misunderstanding the process, as am expecting that these bodies would
even consider sharing informing me. Perhaps its required that the science is vetted prior to getting the
attention of specialists like the following:

 B12 research institute in (NL)

 Pernicious Anemia Society (UK)
 Nutritional Society (UK)
 European Food Information Council
 European Food Safety Authority

Asking questions like:

Is this product something your Organisation would consider collaborating on?

Would you see the benefit of having lower associated neuro related damage from B12 deficiency in an
emerging high value market?
How could this study be improved slightly?
What is the most significant factor I’ve neglected to consider in the interest of public health?
Bibliography

Iannetta, P., 2018. TRUE-Transition paths to sustainable legume based systems in Europe-H2020. Impact,
2018(6), pp.85-87.

Nils-Gerrit Wunsch, 2017. Vegetarianism and veganism among young adults in selected European
countries. Statistics and facts on Food & Nutrition, Consumer Goods & FMCG. Statistica© 2020.

Pawlak, R., Parrott, S.J., Raj, S., Cullum-Dugan, D. and Lucus, D., 2013. How prevalent is vitamin B12
deficiency among vegetarians?. Nutrition reviews, 71(2), pp.110-117.

Hardlei, T.F., Mørkbak, A.L., Bor, M.V., Bailey, L.B., Hvas, A.M. and Nexo, E., 2010. Assessment of vitamin
B12 absorption based on the accumulation of orally administered cyanocobalamin on transcobalamin.
Clinical chemistry, 56(3), pp.432-436.

Scaglione, F. and Panzavolta, G., 2014. Folate, folic acid and 5-methyltetrahydrofolate are not the same
thing. Xenobiotica, 44(5), pp.480-488.

Pawlak, R., Lester, S.E. and Babatunde, T., 2014. The prevalence of cobalamin deficiency among
vegetarians assessed by serum vitamin B12: a review of literature. European journal of clinical
nutrition, 68(5), p.541.