Draft Guideline for Food Safety Assessment of Genetically 
Modified Foods Derived from Recombinant-DNA Organisms  
 
 
CHAPTER ONE: GENERAL PROVISIONS  
I. BASIS OF THE RULE  
II. OBJECTIVE  
III. DEFINITIONS  
IV. SCOPE  
CHAPTER TWO: FOOD SAFETY ASSESSMENT OF GM FOODS 
CHAPTER THREE: FOOD SAFETY A SSESSMENT OF GM FOODS DERIVED 
FROM RECOMBINANT-DNA PLANTS  
CHAPTER FOUR: FOOD SAFETY ASSESSMENT OF GM FOODS DERIVED 
FROM RECOMBINANT-DNA MICROORGANISMS  
ANNEX I - INFORMATION REQUIRED TO APPLY FOR THE SAFETY 
ASSESSMENT OF GM FOODS DERIVED FROM RECOMBINANT-DNA 
PLANTS  
I. APPLICATION  MATERIALS  MUST  INCLUDE  THE  FOLLOWING  
INFORMATION:  
II. LITERATURES  AND  REFERENCES  
ANNEX I - INFORMATION REQUIRED TO APPLY FOR THE SAFETY 
ASSESSMENT OF GM FOODS DERIVED FROM RECOMBINANT-DNA 
MICROORGANISMS  
I. APPLICATION  MATERIALS  MUST  INCLUDE  THE  FOLLOWING  
INFORMATION:  
II. LITERATURES  AND  REFERENCES  
ANNEX III - SAFETY ASSESSMENT OF POSSIBLE TOXICITY  
ANNEX IV - SAFETY ASSESSMENT OF POSSIBLE ALLERGENICITY  
ANNEX V - SAFETY ASSESSMEN T OF UNINTENDED EFFECTS  
 1Chapter One: General Provisions  
 
 
I. Basis of the rule  
This guideline is prescribed in accordance with the provision of paragra ph 1 of Article 14 of 
the Law Governing Food Sanitation.  
  
II. Objective  
This guideline explains the information required for food safety assessment of genetically modified foods (hereafter as GM foods) de rived from recombinant-DNA organisms. The 
original developers are required to submit necessa ry information to register their genetically 
modified organisms (hereafter as GMO) to the  Department of Health (DOH) before 
introduce the GM foods into market. This do cument also provides guideline for food 
manufacturer, food trader, and health authoritie s to evaluate food safety of the GM foods.  
 
 
III. Definitions  
 
1. Gene modification techniques  
Techniques that apply genetic engineering or modern biotechnology to transfer or insert 
genetic material into a living cell or organism to result in genetic modification of the cell or 
organism. The technique does not include conventional breeding, cell fusion within 
taxonomic family, protoplast fusion within taxonomic family, hybridization, induced 
mutagenesis, ex vivo fertilization, somatic mutation, and polyploidy induction.    
2. Genetically modified foods (GM foods)  
Foods and food additives that are produced or  manufactured from raw materials consisting 
of or containing genetically modified organisms.  
  
3. Conventional counterpart  
A related variety/species, its components and/or products for which there is experience of 
establishing safety based on common use as food.  
  
4. Genetically modified organism (GMO)  
An organism with genetic material that has been altered by gene modification techniques.  
  
5. Host  
 2An organism to which gene(s) or DNA fragme nt(s) are inserted or transferred by gene 
modification techniques  
  
6. Vector  
An agent that is used to deliver selected foreign DNA into host for proliferation and 
expression.  
  
7. Transferred or introduced or inserted gene (or DNA fragment)  
Any foreign gene (or DNA fragment) that is transf erred or introduced or inserted into host.  
 
 
8. Gene products  
Any product derived from the transferred or  introduced or inserted gene(s) (or DNA 
fragment(s)).  
  
9. Gene (or DNA fragment) donor  
An organism that provides gene(s) (or DNA fragment(s)) for gene modification process.  
 
 
10. Unintended effects  
By introducing a specific trait into an organism through the insertion of DNA sequences, 
additional traits may, in some cases, be acqui red or existing traits be lost or modified. 
 
 
 11. Nutrient
l  
Nutrient means any substance normally consumed as a constituent of food that provides 
energy; or that is needed for growth and development and maintenance of healthy life; or a deficit of which will cause characteristic bioc hemical or physiological changes to occur.  
 
12. Recombinant-DNA plants modified for nutritional or health benefits The recombinant-DNA plant exhibits a particular  trait in portion(s) of the plant intended for 
food use; and the trait is a result of: (i) in troduction of a new nutrient(s) or related 
substance(s), (ii) alteration of either the quantit y or bioavailability of a nutrient(s) or related 
substance(s), (iii) removal or reduction of undesirable substance(s) (e.g. allergens or toxicants), or (iv) alteration of the interaction(s)  of nutritional or health relevance of these 
substances.   
13. Tolerable upper intake levels (or upper levels or UL) 
                                                
 
l Please refer to the“General Princi ples for the Addition of  Essential Nutrients to  Foods (CAC/GL 09-1987)”. 
 3The highest average daily nutrient intake level that is likely to pose no risk of adverse health 
effects to almost all (97-98%) individuals in the general population.  
 
 
14. Bioavailability 
Bioavailability is the degree to which food nutrients are available for absorption and 
utilization in the body. 
  
15. Acceptable daily intake (ADI) 
Acceptable daily intake is a measure of the amount of a specific substance in food or 
drinking water  that can be ingested over a lifetime without an appreciable health risk. An 
ADI value is based on current research, with long-term studies on animals and observations 
of humans. First, a No Observable (Adverse) Effect Level, the amount of a substance that 
shows no toxic effects, is determined on the basi s of animal studies intended to measure an 
effect at several doses. Usually the studies ar e performed with several doses including high 
doses. Then, the NOEL (or NOAEL ) is scaled by a safety factor, conventionally 100, to 
account for the differences between test anim als and humans (factor of 10) and possible 
differences in sensitivity between humans (anot her factor of 10). The ADI is usually given in 
mg per kg body weight per day. 
 
 
16. Dietary exposure assessment 
Exposure assessment means the qualitative and/or quantitative evaluation of the likely intake 
of biological, chemical, and physical agents via food as well as exposures from other sources 
if relevant. Dietary exposure to (or intake of) food chemicals is estimated by combining food 
consumption data with food chemical concentration data. The general equation is:       
 
 ＝    Dietary exposure  
(mg/kg body weight/day)  Σ(                                                                                       )  
Body weight (kg/person)Concentration of chemical          Food consumption  
in food [mg/kg]                    [kg/person/day]  ×
 4Chapter Two: Food Safety Assessment of GM Foods  
 
 
This chapter describes food safety assessment of GM foods derived from recombinant-
DNA organisms. Information required varies according to the sources of the GM foods 
(plants, animals, or microorganisms).  
 
Food safety assessment of GM food s derived from recombinant-DNA organisms  is 
conducted in three phases as follows.  
 I. Phase I assessment is conducted according to the following information:  
Information of the recombinant-DNA organisms   
 
II. When the results of the phase I assessmen t indicate the presence of possible toxin(s)  
and/or allergen(s) in the GM foods derived fr om recombinant-DNA plants, it is necessary 
to carry out phase II assessment. Phase II assessment is conducted according to the 
following information.  
A. Safety assessment of possible toxicity   B. Safety assessment of possible allergenicity   
 
III. When phase I and phase II assessments of the GM foods derived from recombinant-DNA plants indicate that the available data ar e insufficient for a thorough safety assessment, 
properly designed animal studies are requested on the whole foods
2.  
 
                                                 
2 Whole food is defined as the general eatable parts of  the GM foods, of which the component has not been 
refined, processed, conc entrated, or purified.  
 5Chapter Three: Food Safety Assessment of GM Foods Derived 
from Recombinant-DNA Plants 
 
 
Food safety assessment of GM foods derived from recombinant-DNA plants  is conducted 
in three phases as follows.  
 I. Phase I assessment is conducted according to the following information:  
Information of the recombinant-DNA plants (for details see Annex I)  
 
II. When the results of the phase I assessmen t indicate the presence of possible toxin(s) 
and/or allergen(s) in the GM foods derived fr om recombinant-DNA plants, it is necessary 
to carry out phase II assessment. Phase II assessment is conducted according to the 
following information.  A. Safety assessment of possible toxicity  (for details see Annex III) 
B. Safety assessment of possible allergenicity  (for details see Annex IV) 
 
III. When phase I and phase II assessments of the GM foods derived from recombinant-DNA plants indicate that the available data ar e insufficient for a thorough safety assessment, 
properly designed animal studies are requested on the whole foods.   
IV. Scope  
This Guideline addresses safety and nutritional aspects of foods consisting of, or derived 
from, organisms that have a history of safe use as sources of food, and that have been 
modified by modern biotechnology to exhibit new or altered expression of traits. This 
document does not address GM animal feed or animals fed with GM feed, the beneficial or 
health claims of nutritionally enhanced GM plants. This document also does not address 
environmental risks of GM plants.  
 6Chapter Four: Food Safety Assessment of GM Foods Derived 
from Recombinant-DNA Microorganisms  
 
Food safety assessment of GM food s derived from recombinant-DNA microorganisms  is 
conducted in three phases as follows.  
 I. Phase I assessment is conducted according to the following information:  
Information of the recombinant-DNA microorganisms  (for details see Annex II)  
 
II. When the results of the phase I assessmen t indicate the presence of possible toxin(s) 
and/or allergen(s) in the GM foods derived from recombinant-DNA microorganisms, it is 
necessary to carry out phase II assessment. Phase II assessment is conducted according to the following information.  
A. Safety assessment of possible toxicity  (for details see Annex III) 
B. Safety assessment of possible allergenicity  (for details see Annex IV) 
 
III. When phase I and phase II assessments of the GM foods derived from recombinant-DNA microorganisms indicate that the availabl e data are insufficient for a thorough safety 
assessment, properly designed animal studies are requested on the whole foods.   
IV. Scope  
This Guideline addresses safety and nutritional aspects of foods consisting of, or derived 
from, microorganisms that have a history of safe use as sources of food, which have been 
modified by modern biotechnology to exhibit n ew or altered traits. This document does not 
address safety of microorganisms used in agricult ure (for plant protection, biofertilizers, in 
animal feed or food derived from animals fed the feed etc.), risks related to environmental 
releases of recombinant-DNA microorganisms use d in food production, safety of substances 
produced by microorganisms that are used as a dditives or processing aids, including enzymes 
for use in food production, specific purported health benefits or probiotic effects that may be attributed to the use of microorganisms in food, or issues relating to the safety of food production workers handling recombinant-DNA microorganisms.  
  
 7ANNEX I - INFORMATION REQUIRED TO APPLY FOR 
THE SAFETY ASSESSMENT OF GM FOODS DERIVED FROM RECOMBINANT-DNA PLANTS  
 
 
I. APPLICATION MATERIALS MUST INCLUDE THE FOLLOWING 
INFORMATION:  
 
A. DESCRIPTION OF THE RECOMBINANT-DNA PLANT  
A) Identity of the crop and the transformation event(s) to be reviewed;  
B) type and purpose of the genetic modification.  
 
B. DESCRIPTION OF THE HOST PLANT AND ITS USE AS FOOD  
A) Common or usual name; scientific name; and, taxonomic classification;  
B) history of cultivation and development through breeding, in particular identifying 
traits that may adversely impact on human health;  
C) information on the host plant’s genotype and phenotype relevant to its safety, including any known toxicity or allergenicity; and  
D) history of safe use for consumption as food.  
 
C. DESCRIPTION OF THE DONOR ORGANISM(S)  
A) Its usual or common name, scientific name and taxonomic classification;  
B) information about the natural history as concerns food safety  a. Information on naturally occurring toxins, anti-nutrients and allergens; for microorganisms, additional information on pathogenicity and the relationship to known pathogens; and  
b.  information on the past and present use, if any, in the food supply and exposure 
route(s) other than intended food use (e.g.  possible presence as contaminants).  
 
D. DESCRIPTION OF THE GENETIC MODIFICATION(S)  
A) The description of the transformation process should include:  a. information on the specific method used for the transformation (e.g. Agrobacterium-mediated transformation);  
b. information, if applicable, on the DNA used to modify the plant (e.g. helper plasmids), including the source (e.g. plant, microbial, viral, synthetic), identity and expected function in the plant; and  
c. intermediate host organisms
3.  
B) Information should be provided on the DNA to be introduced, including:  
a. the characterization of all the genetic components including marker genes,  
b. regulatory and other elements affecting the function of the DNA;  
c. the size and identity;  
                                                 
3 Intermediate recipient organisms mean  the organisms (e.g. other bacteria or  fungi) used to produce or process 
DNA prior to introduction into the final recipient organism.  
 8d. the location and orientation of the sequen ce in the final vector/construct; and  
e. the function.  
 
E. CHARACTERIZATION OF THE GENETIC MODIFICATION(S)  
A) Information should be provided on the DNA insertions into the plant genome; this 
should include:  a. the characterization and description of  the inserted genetic materials;  
b. the number of insertion sites;  
c. the organisation
4 of the inserted genetic material at each insertion site including 
copy number and sequence data of the inserted material and of the surrounding region, sufficient to identify any substan ces expressed as a consequence of the 
inserted material, or, where more appropriate, other information
5 such as 
analysis of transcripts or expression products to identify any new substances that may be present in the food; and  
                                                d. identification of any open reading frames
6 within the inserted DNA or created 
by the insertions with contiguous plant genomic DNA including those that could result in fusion proteins.  
B) Information should be provided on any newly expressed substance(s)
7 in the 
recombinant-DNA plant.  
This should include information listed from a. to e. If the newly expressed substance(s) in the recombinant-DNA plant are evaluated to be substantially equivalent to existing substances with known history of safe use as foods, no further assessment is required. Otherwise, assessment as listed in B) of F should be conducted.  
a. The gene product(s) (e.g. a protein or an untranslated RNA);  
b. the gene product(s)’ function;  
c. the phenotypic description of the new trait(s);  
d. the level and site of expression in the plant of the expressed gene product(s), and the levels of its metabolites in the plant, particularly in the edible portions; and  
e. where possible, the amount of the target gene product(s) if the function of the 
expressed sequence(s)/gene(s) is to al ter the accumulation of a specific 
endogenous mRNA or protein.  
C) In addition, information should be provided:  
a. to demonstrate whether the arrangement of the genetic material used for insertion has been conserved or whether significant rearrangements have occurred upon integration;  
 
4 The information should be described comprehensively so  that the identification of  any substanc es expressed 
as a consequence of the inserted, modi fied or deleted material is possible.  
5 The information should allow the identification of an y possible newly expressed materials presented in foods 
being made.  
6 Including those that can result in fusion proteins.  
7 Newly expressed substance(s) are those expressed materials, such as proteins, untranslated RNA, or 
metabolites, etc., that are expressed by the foreign DNA  inserted through genetic modification, and are not 
natively in the host strain.  
 9b. to demonstrate whether deliberate modifications made to the amino acid 
sequence of the expressed protein result in changes in its post-translational 
modification or affect sites critical for its structure or function;  
c. to demonstrate whether the intended effect of the modification has been achieved and that all expressed traits ar e expressed and inherited in a manner that 
is stable through several generations cons istent with laws of inheritance. It may 
be necessary to examine the inheritan ce of the DNA insert itself or the 
expression of the corresponding RNA if the phenotypic characteristics cannot be measured directly;  
d. to demonstrate whether the newly expressed trait(s) are expressed as expected in 
the appropriate tissues in a manner and at levels that are consistent with the associated regulatory sequences driving the expression of the corresponding gene;  
e. to indicate whether there is any evidence to  suggest that one or several genes in 
the host plant has been affected by the transformation process; and  
f. to confirm the identity and expression pattern of any new fusion proteins.  
 
F. SAFETY ASSESSMENT  
A) Compositional analyses of key components  a. Key components are those components in a particular food that may have a substantial impact in the overall diet. They may be  a) major constituents (e.g. fats, proteins, carbohydrates);  
b) minor compounds (e.g. minerals, vitamins);  
c) anti-nutrients (e.g. enzyme inhibitors); and  
d) key toxicants that are those toxicologically significant compounds known to be inherently present in the plant, such as those compounds whose toxic potency and level may be significant to he alth (e.g. solanine in potatoes if the 
level is increased, selenium in wheat) and allergens.  
b. Crop composition databases published by international organizations
8.  
c. Analyses of concentrations of key components of the recombinant-DNA plant, especially those typical of the food, should be compared with an equivalent analyses of a conventional counterpart  grown and harvested under the same 
conditions. The comparison should meet the following requirements:  a) The comparator(s) used in this assessment should ideally be the near isogenic parental line. In practice, this may not be  feasible at all times, in which case a 
line as close as possible should be chosen.  
b) In some cases, a further comparison with the recombinant-DNA plant grown under its expected agronomic conditions may need to be considered (e.g. application of an herbicide).  
c) The statistical significance of any observed differences should be assessed in the context of the range of natural vari ations for that parameter to determine 
its biological significance.  
d. Experimental design  
                                                
 
8 Such as OECD and ILSI published documents that co uld be used as reference materials in conducting 
comparative compositional analyses of key component s, e.g. ‘Consensus Documents for the Work on the 
Safety of Novel Foods and Feeds’ from OECD, and ‘Crop Composition Database’ from ILSI.  
 10a) The location of trial sites should be representative of the range of 
environmental conditions under which the plant varieties would be expected 
to be grown.  
b) The number of trial sites should be sufficient9 to allow accurate assessment 
of compositional characteristics over this range.  
c) Trials should be conducted over a sufficient number of generations10 to 
allow adequate exposure to the variety of conditions met in nature.  
                                                d) To minimise environmental effects, and to reduce any effect from naturally occurring genotypic variation within a crop variety, each trial site plots should be replicated and arranged randomly.  
e) An adequate number of plants should be sampled and the methods of analysis should be sufficiently sensitive and specific to detect variations in key components.  
 
B) Initial assessment of possible toxicity and/or allergenicity of newly expressed substances in recombinant-DNA plants.  a. The following information should be provided for initial assessment of possible toxicity of the newly expressed substances in recombinant-DNA plants.  a) The concentration of the substance in the edible parts of the recombinant-DNA plant, including variations and mean values;  
b) the chemical nature and function of the newly expressed substance; and  
c) in the case of proteins, the assessment of potential toxicity should focus on  
1. amino acid sequence similarity betw een the protein and known protein 
toxins and anti-nutrients (e.g. protease inhibitors, lectins);  
2. stability to heat or processing; and  
3. stability to degradation in appropriate representative gastric and intestinal model systems.  
b. The following information should be provided for initial assessment of possible allergenicity of the newly expressed proteins in recombinant-DNA plants.  a) The source of the introduced protein  
1. Information should describe any reports of allergenicity associated with the donor organism.  
2. When reasonable evidence of IgE medi ated oral, respiratory or contact 
allergy of the donor organism is available, the following information may be provided. 
1) Availability of sera for screening purposes;  
2) documented type, severity and frequency of allergic reactions;  
3) structural characteristics and amino acid sequence; and  
4) physicochemical and immunological properties (when available) of known allergenic proteins from that source.  
 
9 To assess the characteristics of th e compositions of the GM plants, th e “Guidance documents on field trails 
of the Genetically modified plants” is sued by the Council of Agriculture can be used as a reference guideline.  
10 Sufficient number of generations me ans that plants should have a cert ain number of generations by which 
the plants have sufficiently experienced all kinds of natural environmental variations. Please refer to the 
“Guidance documents on field trails of the Genetically modified plants” issued by the Council of Agriculture 
for more details.  
 113. Genes derived from known allergenic sources should be assumed to 
encode an allergen unless scientific evidence demonstrates otherwise.  
4. When genes are derived from non-allergenic sources, the following information should be provided.  
1) Level and location of the newly expressed protein; and  
2) function of the newly expressed protein.  
b) Amino acid sequence homology  
1. Sequence homology searches comparing the structure of all newly expressed proteins with all known allergens should be done. Searches should be conducted using various algorithms
11 to predict overall 
structural similarities.  
2. Strategies such as stepwise contiguous identical amino acid segment searches
12 may also be performed for identifying sequences that may 
represent linear epitopes. 
3. IgE cross-reactivity between the n ewly expressed protein and a known 
allergen should be considered a possibility when 
1) there is more than 35% identity in a segment of 80 or more amino acids (FAO/WHO 2001). 
2) Linear epitope(s) is identified or there are other scientifically justified criteria. 
4. A positive sequence homology result
13 indicates that the newly 
expressed protein is likely to be a llergenic. If the product is to be 
considered further, it should be assessed using serum, if available, from individuals allergic to the identified  allergenic source. If there is no 
available serum, other scientifically sound methods and tools
14 may be 
used to assess the allergenicity potential of the newly expressed protein(s).  
c) Pepsin resistance  
1. Resistance to pepsin digestion should be conducted with a consistent and well-validated pepsin degradation protocol.  
2. Further analyses are required for a newly expressed protein with a pepsin digestion fragment larger than 3.5 kDa.  
c. Alternative protocols may be used wher e adequate justification is provided. 
Initial assessment of possible toxicity and/or allergenicity of the newly expressed substances in recombinant-DNA plants may require the isolation of the new substance from the recombinant-DNA plant,  or the synthesis or production of 
the substance from an alternative source,  in which case the material should be 
                                                
 
11 Such as FASTA or BLASTP, etc.. 
12 The size of the contiguous amino acid search should be based on a scientific ally justified rationale in order to 
minimize the potential for false ne gative or false positive results. 
13 A negative sequence homology result  indicates that a newly expressed pr otein should be considered along 
with the other data outlined under this strategy in asse ssing the allergenic potentia l of these prot eins. Further 
studies should be conduc ted as appropriate.  
14 Methods including targeted-IgE serum screening, the construction of the international sera databank, the 
application of the animal models, screening for the T- cell epitopes on the newly expressed proteins, and/or the 
antigen related structural modules etc..  
 12shown to be biochemically, structurally, and functionally equivalent to that 
produced in the recombinant-DNA plant.  
d. When the results of the initial assessment indicate that the newly expressed 
substance is possible toxin(s) and/or allergen(s), further assessment should be 
conducted according to Annex III and/or IV .  
 
C) Evaluation of metabolites  
Some recombinant-DNA plants may have been modified in a manner that could 
result in new or altered levels of various metabolites in the food.  
a. Consideration should be given to the potential for the accumulation of metabolites in the food that would adversely affect human health.  
b. Safety assessment of such plants requires  investigation of residue and metabolite 
levels in the food and assessment of any alterations in nutrient profile.  
c. Where altered residue or metabolite levels are identified in foods, consideration should be given to the potential impacts on human health  using conventional 
procedures for establishing the safety of such metabolites
15.  
 
D) Nutritional modification  
The following information should be provided for the assessment of foods derived 
from recombinant-DNA plants modified for nutritional or health benefits.  
a. The likely intake of the food derived from the recombinant-DNA plant should 
be estimated. The expected intake of the food should be used to assess the nutritional implications of the altered nutrient profile both at customary and 
maximal levels of consumption.  a) Upper levels of intake  for many nutrients
16 that have been set out by DOH 
may be considered, as appropriate. The basis for their derivation should also be considered in order to assess the public health implications of exceeding these levels. The safety assessment of related substances should follow a case-by-case approach, taking into account upper levels as well as other values, where appropriate. b) Although it is preferable to use a scientifically determined upper level of 
intake of a specific nutrient or related substance, when no such value has been determined, consideration may be given to an established history of safe use for 
nutrients or related substances that are c onsumed in the diet if the expected or 
foreseeable exposure would be consistent with those historical safe levels. 
b. Although the recombinant-DNA plant components may be individually assessed as safe, the impact of the change on the overall nutrient profile should be determined.  
c. When the modification results in a food product, such as vegetable oil, with a composition that is significantly different from its conventional counterpart, it may be appropriate to use additional conv entional foods or food components (i.e. 
foods or food components whose nutritional composition is closer to that of the 
                                                
 
15 E.g. procedures for assessing the human safety of chemicals in foods.  
16 Please refer to the “Tolerable Upper I ntake Levels” ( http://www.fda.gov.tw/gra dation.aspx?site_content_  
sn=271 ) issued by DOH for more details.  
 13food derived from recombinant-DNA pl ant) as appropriate comparators to 
assess the nutritional impact of the food.  
d. It is also important to ascertain to what extent the modified nutrient is 
bioavailable and remains stable with time, processing and storage.  a) Where appropriate, information may be needed on the different chemical 
forms of the nutrient(s) or related substance(s) expressed in the portion of the plant intended for food use and their respective levels.  
b) Bioavailability of the nutrient(s), related substance(s) or undesirable 
substance(s) in the food that were the subject of the modification in the recombinant-DNA plant should be established, where appropriate. If more than one chemical form of the nutrient(s) or  related substance(s) is present, their 
combined bioavailability should be established, where appropriate.  
e. Animal feeding studies may be warranted for foods derived from recombinant-DNA plants if changes in the bioavailab ility of nutrients are expected or if the 
composition is not comparable to conventional foods.  
f. Dietary exposure to a nutrient(s) or related substance(s) should be evaluated in 
the context of the total diet and the assessment should be carried out based on the customary dietary consumption by the relevant population(s) of the corresponding food that is likely to be displaced. When evaluating the exposure, 
it is appropriate to consider information on whether the consumption of the modified food could lead to adverse nutritional effects as compared with consumption of the food that it is intended to replace.  a) The concentration of the nutrient(s) or related substance(s)
17 expressed in the 
portion of the plant intended for food use should be determined.  
b) Consumption estimates are based on food consumption data of Taiwan18 
when available, using existing guidance19 on estimation of exposure in a given 
population(s). When the food consumpti on data are unavailable, the Food 
Balance Sheets issued by the Council of Agriculture (COA) may provide a useful 
resource.  
c) To assess the safety of a food derived from a recombinant-DNA plant 
modified for a nutritional or health benefi t, the estimated intake of the nutrient 
or related substance in the population(s) is compared with the nutritional or 
toxicological reference values, such as u pper levels of intake, acceptable daily 
intakes (ADIs) for that nutrient or rela ted substance, where these values exist. 
This may involve assessments of different consumption scenarios against the relevant nutritional reference value, ta king into account possible changes in 
bioavailability, or extend to probabilistic methods that characterize the distribution of exposures within the relevant population(s). 
                                                
 
17 Please refer to the “Compositional analyses of key components” of the “Guideline for Food Safety 
Assessment of Genetically modifi ed Foods Derived from Recombinant-DNA Organisms” in Annex I for 
more details.  
18 Please refer to the “Nutrition Survey in Taiwan” and “Food Composition Tables ” ( http://www.fda.gov.tw/  
gradation.aspx?site_content_sn=271 ) issued by DOH for more details.  
19 A model for establishing upper levels of intake for nutrients and related substa nces. Report of the Joint 
FAO/WHO Technical Workshop on nutrie nt risk assessment. WHO Headqua rters, Geneva, Switzerland, 2 –
6 May 2005.  
 14g. In consideration of geographical and cultural variation in food consumption 
patterns, the nutrient and the populations affected should be identified for nutritional changes to a specific food.  
h. Attention should be paid to the partic ular physiological characteristics and 
metabolic requirements of specific populati on groups such as infants, children, 
pregnant and lactating women, the elderl y and those with chronic diseases or 
compromised immune systems.  
i. If the characterization of the food indicates  that the available data are insufficient 
for a thorough safety assessment, properly designed animal studies could be requested on the whole foods.  
 
E) Food processing  Information should be provided describing  the processing conditions used in the 
production of a food ingredient from the recombinant-DNA plant.  
F)  Antibiotic resistance marker genes  The following information should be provided  for safety assessment of the antibiotic 
resistance marker genes. a. Describe the clinical and veterinary use  and importance of the antibiotic in 
question; 
b. whether the presence in food of the enzyme  or protein encoded by the antibiotic 
resistance marker gene would compromise the therapeutic efficacy of the orally 
administered antibiotic; 
c. safety of the gene product; and 
d. documentation, if any, regarding the approval and consumption of the antibiotic resistance marker genes in other countrie s. Any other information available to 
demonstrate the safety of the anti biotic resistance marker genes. 
 
G) Other considerations  a. Some recombinant-DNA plants may exhibi t traits (e.g., herbicide tolerance) 
which may indirectly result in the potential for accumulation of pesticide residues, altered metabolites of such residues, to xic metabolites, contaminants, or other 
substances which may be relevant to human health. The safety assessment 
should take this potential for accumulation into account. Conventional procedures for establishing the safety of such compounds (e.g., procedures for 
assessing the human safety of chemicals) should be applied.  
b. The safety assessment should be reviewed  in the light of new scientific 
information that calls into question the conclusions of the original safety assessment.  
 
 
II. LITERATURES AND REFERENCES  
 
A. Documentation regarding the approval and con sumption of the GMO in other countries.  
B. Any other information to demonstrate safety of the GMO as food.  
C. For safety assessment of unintende d effects, refer to Annex V.  
 
 15ANNEX II - INFORMATION REQUIRED FOR THE SAFETY 
ASSESSMENT OF PRODUCED USING RECOMBINANT-DNA 
MICROORGANISMS 
 
I. APPLICATION MATERIALS MUST  INCLUDE THE FOLLOWING 
INFORMATION 
 
A. DESCRIPTION OF THE RECOMBINANT-DNA MICROORGANISM  
A) Description of the bacterial, yeast, or fungal strain; 
B) Description of the food produced using  the recombinant-DNA microorganisms; 
C) Information about the preservation and identification of the recombinant-DNA 
microorganisms used in food production or contained in food  
 B. DESCRIPTION OF THE HOST MICROORGANISM AND ITS USE AS 
FOOD  
The data and information should include, but need not be restricted to:  A) Identity  
a. scientific name, common name or ot her name(s) used to reference the 
microorganism; 
b. strain designation;  
c. information about the strain and its source, or accession numbers or other information from a recognized culture repository from which the organism or its antecedents may be obtained; and  
d. if applicable, information supporting its taxonomical assignment. 
B) History of use and cultivation, known information about strain development 
(including isolation of mutations or antecedent strains used in strain construction);  
C) Information on the recipient microorganism’s and related species
’ genotype and 
phenotype relevant to safety, including  
a. any known toxins, antibiotics, antibiotic resistance factors or other factors related to pathogenicity, or immunological impact;  
b. the genetic stability of the microorgani sm including, as appropriate, the 
presence of mobile DNA elements, i.e. insertion sequences, transposons, plasmids, and prophages. 
D) History of safe use in food production or safe consumption in food, which may include information on 
a. how the recipient microorganism is typically grown;  
b. how the recipient microorganism is typically transported;  
c. how the recipient microorganism is typically stored; 
d. quality assurance, including those to verify strain identity and production specifications for microorganisms and foods; and 
e. whether these organisms remain viable in the processed food or are removed or rendered non-viable as  a consequence of processing. 
E) Information on the relevant production parameters used to culture the recipient microorganism. 
 16 
C. DESCRIPTION OF THE DONOR ORGANISM(S)  
A) Identity.  
a. Scientific name, common name or other na me(s) used to reference the organism;  
b. strain designation;  
c. information about the strain and its source, or accession numbers or other 
information from a recognized culture repository from which the organism or its antecedents may be obtained; and 
d. if applicable, information supporting it s taxonomic assignment; its usual or 
common name, scientific name and taxonomic classification; 
B)  information about the organism or related organisms that concerns food safety; 
C)  information on the organism’s genotype and phenotype relevant  to its safety 
including any known toxins, antibiotics, anti biotic resistance factors or other factors 
related to pathogenicity, or immunological impact; and 
D) information on the past and present use, if any, in the food supply and exposure route(s) other than intended food use ( e.g., possible presence as contaminants). 
 D. DESCRIPTION OF THE GENETIC MODIFICATION(S)  Including vector and construct  
A) The description of the strain construction process should include:  
a. information on the specific method(s) used for genetic modification; 
b. information on the DNA used to modify the microorganism, including  a) the source (e.g.  plant, microbial, viral, synthetic); 
b) identity;   c) expected function in the recombinant-DNA microorganism; and  d) copy number for plasmids; 
c. intermediate recipient organisms.  
B) Information should be provided on the DNA  added, inserted, deleted, or modified, 
including:  
a. the characterization of all genetic components, including  a) marker genes;  
b) vector genes; 
c) regulatory elements; and 
d) other elements affecting the function of the DNA; 
b. the size and identity; 
c. the location and orientation of the sequen ce in the final vector/construct; and  
d. the function. 
 E. CHARACTERIZATION OF THE GENETIC MODIFICATION(S) 
A) Information should be provided on the DNA modifications in the recombinant DNA microorganism; this should include:  
a. the characterization and description of th e added, inserted, deleted, or otherwise   
modified genetic materials, including plasmi ds or other carrier DNA used to transfer 
desired genetic sequences. This should include  
a) an analysis of the potential for mobilization of any plasmids or other genetic elements used;  
b) the locations of the added, inserted, deleted, or otherwise modified genetic materials (site on a chromosomal or extrachromosomal location); and  
 17c) if located on a multicopy plasmid, the copy number of the plasmid; 
b. the number of insertion sites;  
c. the organization of the modified genetic material at each insertion site including 
b) the copy number and sequence data of the inserted, modified, or deleted 
material;  
c) plasmids or carrier DNA used to transf er the desired genetic sequences; and  
d) the surrounding sequences; 
d. identification of any open reading frames within inserted DNA, or created by the 
modifications to contiguous DNA in the chromosome or in a plasmid; and 
e. particular reference to any sequences known to encode, or to influence the expression of, potentially harmful functions. 
C) Information should be provided on any newly expressed substance(s) in the recombinant-DNA microorganism.  This should include information listed in a to f . If  the newly expressed substance(s) 
in the recombinant-DNA microorganism were evaluated to be substantially 
equivalent to existing substances with know n history of  safe use as foods, no further 
assessment would be required. Othe rwise, assessments listed in B)  of F  should be 
conducted. 
a. the gene product(s) (e.g.  a protein or an untranslated RNA);  
b. the gene product(s)’ function;  
c. the phenotypic description of the new trait(s);  
d. the level and site of expression (intracellular, periplasmic - for Gram-negative bacteria, organellar - in eukaryotic microorganisms, secreted) in the microorganism of the expressed gene product(s), and, when applicable, the levels of its metabolites in the organism;   
e. the amount of the inserted gene produ ct(s) if the function of the expressed 
sequence(s)/gene(s) is to alter the level of a specific endogenous mRNA or protein; and 
f. the absence of a gene product, or alterations in metabolites related to gene products, if applicable to the intended fu nction(s) of the genetic modification(s). 
  C) In addition, information should be provided:  
a. to demonstrate whether the arrangement of the modified genetic material has been conserved or whether significant rearrangements have occurred after 
introduction to the cell and propagation of the recombinant strain to the extent needed for its use(s) in food production, including those that may occur during its storage; 
b. to demonstrate whether deliberate modifications made to the amino acid sequence of the expressed protein result in changes in its post-translational modification or 
affect sites critical for its structure or function; 
c. to demonstrate whether the intended effect of the modification has been achieved 
and that all expressed traits are expressed and inherited in a manner that is stable 
for the extent of propagation needed for its use(s) in food production and is consistent with laws of inheritance. It may be necessary to examine the inheritance of the inserted or modified DNA or th e expression of the corresponding RNA if 
the phenotypic characteristics cannot be measured directly; 
d. to demonstrate whether the newly expressed trait(s) is expressed as expected and 
targeted to the appropriate cellular location or  is secreted in a manner and at levels 
 18that is consistent with the associated re gulatory sequences driving the expression 
of the corresponding gene; 
e. to indicate whether there is any evidence to  suggest that one or more genes in the 
recipient microorganism has been affected  by the modifications or the genetic 
exchange process; and 
f. to confirm the identity and expression pattern of any new fusion proteins. 
 
F. SAFETY ASSESSMENT  
A) Compositional analyses of key components  
a. Analyses should be performed by comparing the concentrations of key 
components of foods produced by recombinant-DNA microorganisms to a conventional counterpart produced under th e same conditions using an equivalent 
analysis. 
b. Ideally, the comparator(s) used in this a ssessment should be food produced using 
the near isogenic parent strain. 
 
B) Initial assessment of possible toxicity and allergenicity of newly expressed substances 
in recombinant-DNA microorganisms 
a. For initial assessment of possible toxicity of the newly expressed substances, the 
following information should be provided. a) Function and concentration in the food; 
b) the expected exposure, possible intake, and possible effects on diets; 
c) history of safe use as food; 
d) information should be provided to ensure that genes coding for known toxins or anti-nutrients present in the donor organisms are not transferred to recombinant-DNA microorganisms; 
e) in the case of proteins, the assessment of potential toxicity should focus on  
1. amino acid sequence similarity between the protein and known protein toxins 
and anti-nutrients (e.g. protease inhibitors, siderophores);  
2. stability to heat or processing; and 
3. stability to degradation in appropriate representative gastrointestinal model systems.
 
4. Appropriate oral toxicity studies (Annex III) may be carried out in cases 
where the proteins present in the food show no close similarity to proteins that have been safely consumed in food, nor has been consumed safely in food previously, taking into account its biological function in microorganisms where known.  
b. For initial assessment of possible allergenicity of the newly expressed proteins, the 
following information should be provided. 
a) The source of the introduced protein  
1. Information should describe any report s of allergenicity associated with 
the donor organism.  
2. When evidence of IgE mediated oral, re spiratory or contact allergy of the 
donor organism is available, the following information may be provided.  
1) The availability of sera for screening purposes;  
2) documented type, severity and frequency of allergic reactions;  
 193) structural characteristics and amino acid sequence of the known 
allergenic proteins from that source; and  
4) if available, physicochemical and immunological properties of the 
known allergenic proteins from the source.  
3. Genes derived from known allergenic  sources should be assumed to 
encode allergens and be avoided unle ss scientific evidence demonstrates 
otherwise. The transfer of genes from  organisms known to elicit gluten-
sensitive enteropathy in sensitive individuals should be avoided unless it 
is documented that the transferred gene  does not code for an allergen or 
for a protein involved in gluten-sensitive enteropathy.  
4. When genes are derived from non-allergenic sources, the following 
information should be provided.  
1) Level and location of the newly expressed protein; and 2) function of the newly expressed protein. 
b) Amino acid sequence homology
  
1. Result on sequence homology searches comparing the structure of all 
newly expressed proteins with all known allergens must be provided. Searches should be conducted using various algorithms  to predict overall 
structural similarities. 
2. Strategies such as stepwise contiguous identical amino acid segment searches must be performed for identifying sequences that may represent linear epitopes.
  
3. IgE cross-reactivity between the n ewly expressed protein and a known 
allergen should be considered a possibility when 
i. There is more than 35% identity in a segment of 80 or more amino 
acids (FAO/WHO 2001). 
ii. Linear epitope(s) is identified or th ere are other scientifically justified 
criteria. 
4. A positive sequence homology result20 indicates that the newly expressed 
protein is likely to be allergenic. If the product is to be considered further, it should be assessed using serum, if av ailable, from individuals allergic to 
the identified allergenic source. If there is no available serum, other scientifically sound methods and tools may be used to assess the allergenicity potential of the newly expressed protein(s).  
c) Pepsin resistance 
1. The assessment should be conducted with a consistent and well-validated pepsin degradation protocol. 
2. Further analyses are required for a newly expressed protein with a pepsin digestion fragment larger than 3.5 kDa.
 
3. Alternative protocols may be used where adequate justification is 
provided.  
c. For proteins from sources not known to be allergenic, and which do not exhibit 
sequence homology to a known allergen, ta rgeted serum screening or other related 
IgE serum screenings and immunological tests might be considered. 
                                                 
20 A negative sequence homology result indicates that a newly expressed protein shou ld be considered along 
with the other data outlined under this strategy in asse ssing the allergenic potentia l of these prot eins. Further 
studies should be conduc ted as appropriate.  
 20d. Initial assessment of possible toxicity and/or allergenicity of the newly expressed 
substances in recombinant-DNA microorganisms may require the isolation of the new substance from the recombinant-DNA microorganism, or the synthesis or production of the substance from an alternative source, in which case the material should be shown to be biochemically, st ructurally, and functionally equivalent to 
that produced in the recombinant-DNA microorganism.
 
e. When the results of the initial assessm ent indicate that the newly expressed 
substance is possible toxin(s) and/or allergen(s), further assessment should be 
conducted according to Annex III and/or IV . 
 
C) Evaluation of metabolites   
Some recombinant-DNA microorganisms may have been modified in a manner that 
could result in new or altered levels of various metabolites in the food.  a. Where altered residue or metabolite levels are identified in foods, consideration should be given to the potential impacts on human health using conventional procedures for establishing the safety of such metabolites. 
b. New or altered levels of metaboli tes produced by a recombinant-DNA 
microorganism may change the population of microorganisms in mixed culture, potentially increasing the risk for growth  of harmful organisms or accumulation of 
harmful substances. Possible effects of ge netic modification of a microorganism 
on other microorganisms should be assessed when a mixed culture of microorganisms is used for food processing. 
 
D) Nutritional modification
 
Foods derived from recombinant-DNA microorganisms that have undergone modification to intentionally alter nutritional quality or functionality should be subjected to additional nutritional assessm ent to assess the consequences of the 
changes and whether the nutrient intakes are likely to be altered by the introduction 
of such foods into the food supply.
 
a. The likely intake of the food derived from the recombinant-DNA microorganism should be estimated. The expected intake of the food should be used to assess the nutritional implications of the altered nutrient profile both at customary and maximal levels of consumption.
  
b. Although the recombinant-DNA microorga nism components may be individually 
assessed as safe, the impact of the change on the overall nutrient profile should be determined.
 
c. When the modification results in a food product with a composition that is 
significantly different from its conventional counterpart, it may be appropriate to use additional conventional foods or f ood components (i.e. foods or food 
components whose nutritional composition is  closer to that of the food derived 
from recombinant-DNA microorganism) as appropriate comparators to assess the nutritional impact of the food. 
d. It is also important to ascertain to what extent the modified nutrient is bioavailable and remains stable with time, processing and storage.
 
e. Animal feeding studies may be warranted for foods derived from recombinant-DNA microorganisms if changes in the bioa vailability of nutrients are expected or 
if the composition is not comparable to conventional foods. 
 
 21f. Foods designed for health benefits, may require an assessment beyond the scope 
of these guidelines such as specific nutrit ional, toxicological or other appropriate 
studies. 
g. In consideration of geographical and cultural variation in food consumption patterns, the nutrient and the populations affected should be identified for nutritional changes to a specific food.
 
h. Attention should be paid to the partic ular physiological characteristics and 
metabolic requirements of specific populati on groups such as infants, children, 
pregnant and lactating women, the elderl y and those with chronic diseases or 
compromised immune systems.  
i. If the characterization of th e food indicates that the available data are insufficient 
for a thorough safety assessment, properly designed animal studies could be requested on the whole foods. 
 
E) Antibiotic resistance and gene transfer
21 
a. Antibiotic resistance a) When the microorganisms contain antibiot ic resistance genes, describe the 
location of the genes in the cell;  
b) any indication of the presence of pl asmids, transposons, and integrons 
containing such resistance genes should be specifically addressed.  
b. Gene transfer a) Note the location of the genetic material to be inserted into the genome. Chromosomal integration of the inserted genetic material may be preferable to 
localization on a plasmid; 
b) where the recombinant-DNA microorganism will remain viable in the gastrointestinal tract, genes should be avoided in the genetic construct that 
could provide a selective advantage to r ecipient organisms to which the genetic 
material is unintentionally transferred; and 
c) sequences that mediate integration into other genomes should be avoided in constructing the introduced genetic material.  
 
F) Viability and residence of microorganisms in the human gastrointestinal tract For applications in which recombinant-DNA microorganisms used in production remain viable in the final food product, the following information must be provided: a. Demonstration of the viability or residence time of the microorganism alone and within the respective food matrix in the digestive tract. 
b. Assessments of the impact on the intestinal microflora in appropriate system.  
 
G) Food processing a. The potential effects of food processi ng, including home preparation, on foods 
derived from recombinant-DNA microorganisms should be considered. 
 G. OTHER CONSIDERATIONS  
 The safety assessment should be reviewed in the light of new scientific information that calls into question the conclusions of the original safety assessment.  
                                                
 
21 Please refer to the “Guidelines on the Assessment of the Health Foods for I mproving the Gastrointestinal 
Tract Functions” issued by DOH  for more details.  
 22 
II. LITERATURES AND REFERENCES 
 
A. Documentation regarding the approval and co nsumption of the GMO in other countries. 
B. Any other information to demonstrate safety of the GMO as food. 
C. For safety assessment of unintended effects, refer to Annex V . 
 23ANNEX III - SAFETY ASSESSMENT OF POSSIBLE 
TOXICITY  
 
 
I. Current dietary exposure and possible e ffects on population sub-groups should be 
considered.  
 
II. Appropriate oral toxicity studies22 may need to be carried out in cases where the protein 
present in the food is not similar to proteins  that have previously been consumed safely 
in food, and taking into account its biological function in the organism  where known.  
 III. Potential toxicity of non-protein substances that  have not been safely consumed in food 
should be assessed on a case-by-case basis depending on the identity and biological 
function in the organism  of the substance and dietary expo sure. The type of studies to be 
performed may include studies on metabolism, toxicokinetics, sub-chronic toxicity, chronic toxicity/carcinogenicity, reproducti on and development toxicity according to the 
traditional toxicological approach.  
 IV. Assessment of possible toxicity of the newl y expressed substances in recombinant-DNA 
organisms may require the isolation of the new substance from the recombinant-DNA 
organism , or the synthesis or production of the su bstance from an alternative source, in 
which case the material should be show n to be biochemically, structurally, and 
functionally equivalent to that produced in the recombinant-DNA organism .  
 
 
                                                 
22 Please refer to the “Guidelines on the Safety Assessment of the H ealth F oods” issued by DOH  or the OECD 
Guidelines for the Testing of Chemicals.  
 24ANNEX IV - SAFETY ASSESSMENT OF POSSIBLE 
ALLERGENICITY  
 
 
I. Serum screening  
A. For those proteins that originate from a source known to be allergenic, or have 
sequence homology with a known allergen , specific serum screenings should be 
performed where sera are available.  
A) A minimum of 8 relevant sera is required to achieve a 99% certainty that the new protein is not an allergen in the case of a major allergen
23; and  
B) a minimum of 24 relevant sera is required to achieve the same level of certainty in the case of a minor allergen
24.  
B. In the case of a newly expressed protein derived from a known allergenic source, a negative result in in vitro immunoassays
25 may not be considered sufficient, but should 
prompt additional testing, such as the possible use of skin test and ex vivo protocols. A 
positive result in such tests would indicate a potential allergen.  
C. Assessment of possible allergenicity of the newly expressed proteins in recombinant-
DNA organisms  may require the isolation of the new proteins from the recombinant-
DNA organism , or the synthesis or production of the proteins from an alternative 
source, in which case the material should be shown to be biochemically, structurally, 
and functionally equivalent to that produced in the recombinant-DNA organism .  
 
II. Other considerations  
A. The types of food processing would be a pplied and its effects on the presence of the 
protein in the final food product; and  
 
B. Other scientifically so und methods and tools26 may be considered in assessing the 
allergenicity potential of the newly expressed proteins.  
 
 
  
                                                
 
23 A major allergen is defined as one to  which more than 50 % of individuals sensitive to that substance react in 
IgE-specific immunoassays.  
24 A minor allergen is defined as one to  which less than 50 % of individuals sensitive to that substance react in 
IgE-specific immunoassays.  
25 A negative result in in vitro immunoassays may not be considered suffi cient; a positive resu lt in other tests 
would indicate a potential allergen.  
26 These may include targeted serum screening (i.e. the assess ment of binding to IgE in sera of individuals with 
clinically validated allergic responses to broadly-re lated categories of foods); the development of international 
serum banks; use of animal models; and examination of newly expressed proteins  for T-cell epitopes and 
structural motifs associated with allergens.  
 25ANNEX V - SAFETY ASSESSMENT OF UNINTENDED 
EFFECTS  
 
 
Safety assessment of unintended effects should include data and information on the 
insertion of DNA sequences, subsequent conventional breeding of the recombinant-DNA 
organism , and/or the random insertion of DNA sequences into the organism  genome which 
may cause disruption or silencing of existing genes, activation of silent genes, or 
modifications in the expression of existing genes.  
 
Comparative safety assessment process on GM  foods derived from recombinant-DNA 
organisms can be applied to identify, if any, unintended effects.  
 
I. Provide the following information:  
A. the phenotypic characteristics of the organism ; and  
B. the formation of new or changed patterns of metabolites. For example, the expression 
of enzymes at high levels may give rise to  secondary biochemical effects or changes in 
the regulation of metabolic pathways and/or altered levels of metabolites.  
 II. The biological relevance and potential eff ects on food safety of unintended effects 
should be evaluated.  
 26