TQF Specifications and Standards for Product Test Items TQF-PCS-126 Scope of Type of Test Cat

TQF Specifications and Standards for Product Test Items TQF-PCS-126 Scope of Type of Test Cat. Test Item Criterion Remarks Certification Test Necessity 1)

Total plate count (CFU/g) Refer to food  The following foods with functional factory self-  claims shall individually comply regulation  with the requirements in Annex 1:  1. Functional foods containing 3 Coliform group (MPN/g) Below 10  CoQ10 Microbiology E. coli (MPN/g) Negative  2. Mushroom functional foods containing β-glucan

Business 3. Functional foods containing γ- guide to Aminobutyric acid Plasticizers reducing  4. Fish oil functional foods plasticizers in

Analysis Analysis Chemical Chemical containing ω-3 fatty acids foods 5. Foods containing soybean Content of functional Refer to the  lecithin composition specifications 6. Foods containing soybean of individual isoflavones functional 7. Mushroom functional foods foods containing water soluble crude

polysaccharides Self-claimed composition Refer to food  8. Functional foods containing

factory self- phytosterols regulation 9. Functional foods containing dietary fibers 10. Functional foods containing resistant maltodextrins 11. Ganoderma tsugae functional

Functional Foods Functional foods containing triterpenoid

26. 12. Functional foods containing lactic acid bacteria (LAB)

Analysis 13. Functional foods containing catechins 14. Functional foods containing lycopene

Composition 15. Functional foods containing spirulina 16. Functional foods containing anthocyanidins 17. Functional foods containing proanthocyanidins 18. Functional foods containing glucosamine 19. Functional foods containing fructans 20. Functional foods containing cereal β-glucans 21. Functional foods containing green algae

Note: (1) Test items are classified into:  for “sanitation standard” and  for “quality specifications”. If it is necessary to add “concerned item”, the Certification Body will make a professional decision based on the practical requirements. (2) Sanitation standards and quality specifications are the reference for on site audit, addition of new products, TQF Specifications and Standards for 1 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PCS-126 and surveillance audit. (3) After an update of sanitation regulations or national standards, the updated version shall prevail. (4) When a combination of food additives is used in the same scope of application, the sum of the usage dose of each food additive divided by the standard dose for usage (that is, the dosage use / dosage standard) shall not be greater than 1. (5) This table outlines only the certified category and the scope of certification for TQF Specifications and Stnadards for Product Test Items, but it does not include all products in the scope of certification. (6) Where there is product claim in the labeling, the food factory shall submit relevant data or test reports. (7) Food factory self-regulation: When a particular item is not mentioned in the sanitation regulations and national standards, product specification and plant control standard shall be taken by the food factory self- regulation procedure.

TQF Specifications and Standards for 2 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

“Annex 1 Specifications and Criteria for Functional Food” 1. Specification for functional food containing CoQ10

I. Scope This specification applies to foods containing solid CoQ10 ingredients.

II. Definitions CoQ10: Coenzyme Q10, CoQ10 or Ubiquinone, exists in every animal cell and is a natural substance that can be independently synthesized by animals. As a liposoluble substance mostly found in the heart, the liver, and the kidney, CoQ10 mainly produces energy ATP through reduction-oxidation reactions to quickly revitalize the energy supply system of cells and neutralize the free radicals produced from the energy production process so as to protect cells against free radical damage (Nelson & Cox, 2000). The amount of CoQ10 in the human body reaches a peak at age 20 and begins to decline from then on. The amount of CoQ10 in the human body also drops when a person often exercises vigorously, works excessively, lives in high tension, lives in a highly-polluted environment, or has an illness or psychological stress. Due to its anti-oxidation function, many studies found that CoQ10 can help strengthen the human heart (Folkers et al., 1985), reduce the risk of cardiovascular diseases (Rundek et al., 2004), lower hypertension risk (Burke et al., 2001), enhance energy and mental strength (Lee, 2000), fight cancer (Portakal et al., 2000), and prevent skin aging (Blatt et al., 1998).

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The content of functional compostion of CoQ10 shall be labeled.

V. Supplementation (I) This test method is designed for testing the total CoQ10 content only and cannot test the content of cis-CoQ10 or trans- CoQ10 or verify the content of oxidized or reduced CoQ10. (II) This Specification applies only to products containing solid CoQ10 and cannot quantify CoQ10 in liquid products. Therefore, this specification is only applicable to solid CoQ10 products.

TQF Specifications and Standards for 3 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

VI. Reference 1. 李寧遠，2000，補充 CoQ10 對耐力運動員及健康人之抗氧化效力及運動表現的影響，行政院國科會科學技術資料中心， PC8910-1298。 2. Blatt, T., Whittle, M., Maksiuk, T., Wolber, R., Keyhani, R., Untiedt, S., Kielholz, J., Gohla, S., Hoppe, U., Schachtschabel, D., Schreiner, V. and Stab, F. 1998. Modulation of the oxidative stress response in aging skin by pro- and anti-oxidants. J. Investig. Dermatol. 110：532. 3. Burke, BE., Neuenschwander, R. and Olson, RD. 2001. Randomized, double-blind, placebo-controlled trial of coenzyme Q10 in isolated systolic hypertension. South Med. J. 94：1112-1117. 4. Folkers, K., Vadhanavikit, S. and Mortensen, S. A. 1985. Biochemical rationale and myocardial tissue data on the effective therapy of cardiomyopathy with coenzyme Q10. Proc. Natl. Acad. Sci., U.S.A., 82：901-904. 5. Nelson, D. L. and Cox, M. M. 2000. Lehninger Principles of Biochemistry, Worth Publ., 3rd ed., pp. 659-672. 6. Portakal, O., Ö zkaya, Ö ., Inal, M. E., Bozan, B., Koşan, M. and Sayek, I. 2000. Coenzyme Q10 concentrations and antioxidant status in tissues of Breast cancer patients. Clin. Biochem. 33：279-284. 7. Rundek, T., Naini, A., Sacco, R., Coates, K. and Dimauro, S. 2004. Atorvastatin decreases the coenzyme Q10 level in the blood of patients at risk for cardiovascular disease and stroke. Arch. Neurol. 61： 889-892.

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TQF Specifications and Standards for Product Test Items TQF-PST-001

2. Specification for Mushroom Functional Food Containing β-glucan

I. Scope This specification applies to foods made with mushroom ingredients containing β-glucan.

II. Definitions (I) Mushrooms Mushrooms refers to edible fungi such as glossy ganoderma, brown mushroom, mushroom, Volvariella volvacea, enokitake, Antrodia cinnamomea, Wolfiporia extensa, tremella fuciformis, and Hericium erinaceus. (II) Mushroom β-glucan Polysaccharides in mushrooms are stored in the fungus’ cell wall which is composed of two types of polysaccharides: (a) cellulose in the form of compact physical fibers and (b) α-glucan, β-glucan, and glycoprotein in the form of a matrix. The physiological activity of polysaccharides is correlated with the characteristics of their structure, composition, mass, and degree. Functional polysaccharides that can fight tumors and enhance immunity found in related mushroom studies fall into the following five categories: (1) homoglucans are linked by β-(1-3), α-(1-3), β-(1- 6), and α-(1-6) bonds with β-(1-6), β-(1-2) or α-(1-4) branches; (2) heteroglucans are linked by glucan bonds with xylose, arabinose, galactose, mannose and xylose branches; (3) heterogalactans are linked by galactan bonds with glucose, trehalose, xylose, and arabinose branches; (4) other heteroglycans are linked by xylan, fucosan, and mannan bonds with glucose, mannose, and galactose branches; and (5) polysaccharide-protein or peptide complexes. The β- glucan in this specification refers to the polysaccharides of mushrooms containing physiological activity that can fight tumors and enhance immunity with β-glycosidic bond-linked glucan as the main chain. (III) Mushroom functional food containing β-glucan Mushroom functional food refer to the products with physiological activity containing the sporocarps or mycelia obtained from fermentation with edible mushroom fungi or liquids. Mushroom functional food in this specification specifically refer to functional food containing at least 3% β-glucan (dry weight) from glossy ganoderma, Agaricus blazei Murill, and Hericium erinaceus.

Item Max. Level (dry weight) Lead Below 3 ppm Cadmium Below 2 ppm

TQF Specifications and Standards for 5 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

International Common Name Common Name Max. Level (ppm) Remarks Allethrin Allethrin 3.0 Insecticide Carbendazim Bavistin 0.5 Germicide Diazinon Diazinon 0.2 Insecticide Dicloran Dicloran 0.5 Germicide Malathion Malathion 2.0 Insecticide Prochloraz Prochloraz 0.5 Germicide Pyrethrins Pyrethroid 1.0 Insecticide Thiabendazole Mertect 5.0 Germicide

(2) The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” by the FDA and the latest version of the relevant laws and regulations. (II) The content of functional composition of β-glucan shall be labeled. .

V. Supplementation (I) Currently there is no publicly recognized method for analyzing β-glucan across the world, and the guidelines for β- glucan analysis are as follows: 1. Amongst currently found polysaccharides from mushroom, those with verified structure, available for trade, and clinically used to fight tumors or enhance immunity as an auxiliary cancer therapy include letinan, schizophyllan, PSK Krestin, PSP polysaccharides peptide, and Grifola D. Letinan and schizophyllan are glucans linked by a β- (1-3) bond with β-(1-6) branch; PSK Krestin is linked by β-(1-3) and β-(1-4) bonds with a β-(1-6) branch, with glucose as the major monosaccharide. PSK contains trehalose and PSP contains arabinose and rhamnose. 2. As defined in the specification, mushrooms contain a wide variety of physiologically active polysaccharides with complex structures. This structure diversity makes quantitative analysis difficult. Therefore, no method for the routine analysis is currently available to quantify functional polysaccharides. While β-(1-3)- D-glucan is the common structure of commercialized polysaccharide preparations, methods for quantitative analysis are mostly developed based on this structure, including: (1) Immune response method: First, target polysaccharides for quantification are purified for animals to produce antibodies through immune response. After purification, antibodies are chemically modified for quantifying target polysaccharides. Although this method can better quantify the content of functional polysaccharides, the antibodies used in the test are effective only on the corresponding functional polysaccharides and cannot be applied to the quantification of functional polysaccharides with other structures. In addition, as the process for purifying polysaccharides and preparing antibodies is time- consuming and the quantification cost is expensive, the method is not suitable for routine analysis. However, by developing commercialized antibody reagents for functional polysaccharides of different structures can turn it into a quick and accurate method for testing functional polysaccharides. (2) Aniline blue fluorometric method: The assay quantifies β-(1-3)- D-glucan by forming a fluorescent polymer with aniline blue and β-(1-3)- D-glucan. Although it is a quick and simple method, it cannot quantify the real content of functional polysaccharides and the results are prone to interference. Therefore, it is suitable for the preliminary test at the development phase and the quality control of a single product. (3) Enzymatic hydrolysis with high performance liquid chromatography (HPLC): After breaking down (1-3)- β-D-glucan into small molecule saccharides with (1-3)-β-D-glucanases through hydrolysis, the content of small molecule saccharides is quantified to calculate the total content. This method has greater interference resistance than the fluorometric method. Although the measured polysaccharide content is lesser than the content of functional polysaccharides, with appropriate hydrolysis conditions to fully hydrolyze (1→3)-β- D-glucan, the method can test the content of (1→3)-β-D-glucan more accurately. However, it is not easy to fully hydrolyze (1→3)-β-D-glucan with enzymes. TQF Specifications and Standards for 6 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

3. Based on the above descriptions, there is currently no method that can accurately quantify the content of polysaccharides. After concluding different methods, this specification recommends Megazyme’s “mushroom and yeast beta-glucan assay” kit which first hydrolyzes total polydextrose with acid and enzymes into glucose to quantify glucose content before deducting α-glucan to calculate β-glucan content. (II) The methods for analyzing the relevant items are as follows:

Item Method Heavy Metals Test Methods for Heavy Metals in Foods - Cadmium Test (2) (Shu-Shou-Shi-Zi-No. 0929206232 dated 4 April 2003) Test Methods for Heavy Metals in Foods - Lead Test (2) (Shu-Shou-Shi-Zi-No. 0929227157 dated 23 December 2003) AOAC999.10, AOAC999.11 Pesticide Residue Test Methods for Pesticide Residues in Foods (Shu-Shou-Shi-Zi-No. 0949424750 dated 24 August 2005) General Plate Count Microbiological Test Methods for Foods - Plate Count Test CNS10890 Coliform Group Microbiological Test Methods for Foods - Coliform Group Test CNS10984 E. coli Microbiological Test Methods for Foods-E. coli Test (Wei- Shu-Shi-Zi-No. 0900025538 dated 20 April 2001) Pathogen Refer to MOHW announcements and CNS. Pathogen Refer to MOHW announcements and CNS.

VI. Reference 1. Megazyme Mushroom and Yeast Beta-glucan AssayManual (2007) 2. Zhang, M., Cui, S.W., Cheung, P.C.K. and Wang, Q. Anti-tumor polysaccharides from mushrooms: a review on their isolation process, structural characteristics and anti-tumor activity. Trends in food science and technology. 18:4-19. (2007) 3. AOAC International. 2003. Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.10. William Horwitz ed. Gaithersburg, MD U.S.A 4. AOAC International. 2003. Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.11. William Horwitz ed. Gaithersburg, MD U.S.A 5. 經濟部標準檢驗局：食品微生物之檢驗法-生菌數之檢驗 CNS10890(1991) 6. 經濟部標準檢驗局：食品微生物之檢驗法-大腸桿菌群之檢驗 CNS10984 (1998)

TQF Specifications and Standards for 7 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

3. Specification for Functional Food Containing γ-Aminobutyric Acid

I. Scope This specification applies to natural foods claimed to contain rich γ-aminobutyric acid (GABA) or foods made of ingredients from natural foods produced by GABA enrichment technology.

II. Definitions (I) γ-aminobutyric acid (GABA): Gamma-aminobutyric acid (γ-aminobutyric acid, GABA), chemical formula C4H9NO2, is a non-protein-structured amino acid containing an amine (-NH2) at the gamma position of the carbon chain of butyric acid. It is extremely water soluble but insoluble in alcohol, ether, and benzene. It is heat-resistant and can resist head damage to facilitate processing. As a neurotransmitter, ionized GABA is extensively distributed in animals and plants that can lower blood pressure and inhibit spasms and depression. (II) GABA enrichment technology: Ordinary plants contain very low GABA content. Many studies found that with only some stimulation, such as germination, cold or thermal shock, calcium fortification, and pH adjustment, GABA will quickly and massively accumulate in plants due to the defense effect to achieve GABA enrichment. GABA can also be enriched by means of microbiological fermentation in appropriate culturing conditions. (III) The functional food containing γ-aminobutyric acid in this specification include fermented or germinated foods and cereals, such as natural foods including germinated brown rice, GABA tea and fermented soybeans, and foods made with their derivatives.

III. Product specifications (I) Appearance: Products shall retain their original flavors and colors and contain no decay, color change, odor, contamination, mold, or foreign matter. (II) Content of specific composition: 1. Comply with the labeling. 2. The minimum GABA content in germinated brown rice and fermented soybeans shall be 15 mg/100 g; and the minimum GABA content in GABA tea shall be 150 mg/100 g. (III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the Food and Drug Administration (hereafter referred to as FDA) and the latest version of regulations relating to food sanitation standardsFDA. (IV) Heavy metal ： 1. Germinated brown rice products shall comply with the latest version of the “Standard for the Tolerance of Heavy Metals in Rice” promulgated by the FDA.

Item Max. Level Mercury 0.05 ppm Cadmium 0.4 ppm Lead 0.2 ppm

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TQF Specifications and Standards for Product Test Items TQF-PST-001

(VII) Other: Comply with the latest version of the “Sanitation Standard for the Tolerance of Mycotoxins in Foods” promulgated by the FDA.

V. Supplementation (I) The methods for analyzing the relevant items are as follows:

Item Method Heavy metal 1. Test Methods for Heavy Metals in Foods - Cadmium Test (2) (Shu-Shou-Shi-Zi-No. 0929206232 dated 4 April 2003) 2. Test Methods for Heavy Metals in Foods - Lead Test (2) (Shu-Shou-Shi-Zi-No. 0929227157 dated 23 December 2003) 3. Test Methods for Heavy Metals in Foods - Mercury Test (2) (Shu-Shou-Shi-Zi-No. 0939300138 dated 8 January 2004) 4. AOAC999.10 5. AOAC999.11 Pesticide Test Methods for Pesticide Residues in Foods-Methods for Testing Multiple Residues (3) residue (Shu-Shou-Shi-Zi-No. 0949424750 dated 23 December 2003) Aflatoxin Test Methods for Pesticide Residues in Foods CNS 4090 N6097(25 August 1997) Plate Count Microbiological Test Methods for Foods - Plate Count Test CNS 10890 N6186 (26 February 1991) Coliform Microbiological Test Methods for Foods - Coliform Group Test CNS 10984 N6194 (29 September 1998) Group E. coli Microbiological Test Methods for Foods - E. coli Test (Wei-Shu-Shi-Zi-No.0900025538 dated 20April 2001) Pathogen Refer to MOHW announcements and CNS.

(II) Introduction to amino acid test methods 1. Amino acid is the basic unit of protein. Its components carboxyl (-COOH) and amine (-NH2) can be reacted with acids and alkalines respectively. Therefore, amino acids are an amphoteric substance. From literature over the years, we can conclude methods for analyzing amino acids into five categories: chemical analysis, enzymatic analysis, isotopic analysis, microbiological analysis, and chromatography. Nitrite and ninhydrin are the common reagents used in chemical analysis. The former analyzes the content of nitrogen production, while the latter tests its colorimetric level. There are many other reagents that can cause sedimentation and color change with a specific amino acid to measure its content. In enzymatic analysis, enzymes are used to promote decarboxylation in amino acids to produce carbon dioxide. The isotope dilution technique is applied to test the amino acid in hydrolyzed protein by adding marked amino acid in hydrolyzed substances to calculate the degree of dilution of isotopes to quantify amino acids in original hydrolyzed substances. Microbiological analysis quantifies amino acid as the growth-limiting factor in microorganisms. In earlier chromatography practice, silica gels or filter papers were used as the stationary phase to separate amino acids. Later on, some scholars separated amino acids with starch columns and resin columns. Moore and others (1958) developed automatic amino acid analysis by quantifying the color change of amino acids separated with resin columns after reaction with ninhydrin, which is the basic model of the amino acid analyzer nowadays. The automatic amino acid analysis proposed by Moore et al. (1958) used a two- column system, i.e. separating acidic and neutral amino acids with “long” resin columns and alkaline amino acids with “short” resin columns. Today, the two-column system has been simplified into the single-column system. In 1966, Japan announced a new method for analyzing amino acids called “ligand analysis”. This method forms amino acid compounds with positive ions such as Ni2+, Co2+, Cu2+, Cd2+, or Zn2+ before exchanging resin absorption with concentrated acid ions to separate and quantify different types of amino acids. Hitachi has developed an automatic amino acid analyzer with this theory. TQF Specifications and Standards for 9 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

2. Some scholars have introduced a method which produces fluorescent derivatives through the derivatization of o- phthalaldehyde (OPA) and 2-mercaptoethanol (2-ME) with GABA before analyzing with capillary electrophoresis or fluorescence spectrometry. As OPA has quick derivatization, in-capillary derivatization is used to enable direct derivatization and separation of samples in the capillary. 3. Scholars have compared three common pre-column derivatization methods for testing GABA with HPLC, using reagents including HN, PITC and o-phthalaldehyde (OPA) to derivatize samples before analyzing the GABA content in samples with HPLC for comparison. The results show that the effect of HN derivatization is not good on samples with complex composition, such as tea, germinated brown rice, and germinated seeds, as the GABA peak is unidentified in HPLC spectrum; PITC derivatization is prone to interference and thus overestimates samples containing high protein amino acids such as germinated brown rice and germinated seeds; and OPA derivatization has a good separation effect of the GABA peak of all samples in the experiment, and the derivatization process is simple and fast, thus recommended for use in relevant tests. 4. Amino acid automatic analysis spectrum (1) Amino acid automatic analysis spectrum (germinated brown rice)

0.12

Ala

g-ABA 0.10

0.08

0.06

0.04 Val

Intensity (AU) Intensity

NH3

Glu Gly

0.02 Arg Lys

Leu

Asp

1Mehis

Cys

P-Ser

Ser

Thr

b-Ala

Ile Hylys

Met

Phe Orn

Tau Trp

EOHNH2

Car

Tyr

Asn PEA 0.00

0 10 20 30 40 50 60 70 80 90 100 110 Retention Time (min)

(2) Amino acid automatic analysis spectrum (control item)

Val

Cys 0.15

Met

Tau Asp

0.10 Orn

g-ABA

Ile

Ala

Gly

Thr

Cit

Ser

Cysthi

Phe

Leu

Lys

b-Ala

Hylys

Urea

b-AiBA

3Mehis

Tyr

Intensity (AU) Intensity

Arg

1Mehis

His 0.05 Glu

P-Ser

Trp

Asn

Sar a-ABA EOHNH2

a-AAA

PEA

Car

NH3

Ans

0.00

0 10 20 30 40 50 60 70 80 90 100 110 Retention Time (min)

VI. Reference 1. AOAC International. 2003. Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.11. William Horwitz ed. Gaithersburg, MD U.S.A 2. AOAC International. 2003. Lead, Cadmium, Zinc, Copper, and Iron in Foods by Atomic Absorption Spectrophotometry after Microwave Digestion. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.10. William Horwitz ed. Gaithersburg, MD U.S.A 3. Chen-Ying; Shen-Yan and Yao-Huiyuan, 2006. Preparation of gamma-amino butyric acid by means of endogenous protease in rice bran and glutamate decarboxylase. Cereal-&-Feed-Industry. (2): 23-24. TQF Specifications and Standards for 10 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

4. C. Simo; A. Rizzi; C. Barbas and A. Cifuentes, 2005. Chiral capillary electrophoresis mass spectrometry of amino acids in foods. Electrophoresis. 26(7-8): 1432-1441. 5. Fang-Kemin; Li-Zaigui; Yuan-Hancheng and Meng-Ying, 2006. Determination of amma-aminobutyric acid in germinated brown rice by high pressure liquid chromatography. Food-Science,-China., 27(4): 208-211. 6. Gonzalez Paramas; Gomez Barez; Cordon Marcos; GarciaVillanova and Sanchez, 2006. HPLC-fluorimetric method for analysis of amino acids in products of the hive (honey and bee-pollen). Food-Chemistry. 95(1): 148- 156. 7. Handoyo,-T and Morita,-N, 2006. Structural and functional properties of fermented soybean (tempeh) by using Rhizopus oligosporus. International-Journal-of-Food-Properties. 9(2): 347-355. 8. H. F. Wang; Y. S. Tsai; M. L. Lin and S. M. Ou, 2006. Comparison of bioactive components in GABA tea and green tea produced in Taiwan. Food-Chemistry. 96(4): 648-653. 9. Hua-Zhang; Jing-Wang; De-Chang-Xu and Chang-Hua-Sun, 2005. Determination of gamma-amino butyric acid as functional factor in germinated brown rice. Journal of Henan University of Technology Natural Science Edition. 26(5): 25-27. 10. K. B. Park and S. H. Oh, 2005. Production and characterization of GABA rice yogurt. Food Science and Biotechnology. 14(4): 518-522 . 11. Komatsuzaki,-N; Tsukahara,-K; Toyoshima,-H; Suzuki,-T; Shimizu,- N and Kimura,-T, 2007. Effect of soaking and gaseous treatment on GABA content in germinated brown rice. Journal-of-Food-Engineering. 78(2): 556-560. 12. N. Komatsuzaki; J. Shima; S. Kawamoto; H. Momose and T. Kimura, 2005. Production of gamma-aminobutyric acid (GABA) by Lactobacillus paracasei isolated from traditional fermented foods. Food Microbiology. 22(6): 497- 504. 13. 經濟部標準檢驗局：食品微生物之檢驗法－生菌數之檢驗 CNS10890(1991) 14. 經濟部標準檢驗局：食品微生物之檢驗法－大腸桿菌群之檢驗 CNS10984(1998)

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TQF Specifications and Standards for Product Test Items TQF-PST-001

4. Specification for Fish oil Functional Food Containing ω-3 Fatty Acid

I. Scope This specification applies to fish oil foods containing ω-3 fatty acids.

II. Definitions (I) Fatty acid It is a hydrocarbon formed by 3-25 single-bond or double-bond carbon atoms and comprising three elements: carbon (C), hydrogen (H), and oxygen (O) with methyl (CH3-) at the carbon chain side and carboxyl (-COOH) on the other side, mostly without branches and only a small number have branches or rings. Most natural fatty acids are even carbon fatty acids. By chain length, fatty acids include short-chain fatty acids (SCFAs) (C4-C6), medium-chain fatty acids (MCFAs) (C8-C10), and long-chain fatty acids (LCFAs) (C12 and above). By saturation or double-bond count, fatty acids include saturated fatty acids, monounsaturated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs). (II) Omega-3 fatty acids (ω-3 fatty acids) It is the first double bond of fatty acids located on the third carbon from the methyl side and with two or more bonds, including α-linolenic acid (ALA, C18:3), eicosapentaenoic acid (EPA, 20:5), and docosahexaenic acid (DHA, 22:6). ω-3 fatty acids can help regulate blood lipids, resist inflammation, increase phospholipid-derived fatty acids (PLFAs) in cell walls, resist arrhythmia, stabilize atheromatous plaque, reduce the mortality of cardiovascular diseases, and prevent sudden cardiac death (SCD) (Kris-Etherton et al., 2004). (III) Fish oil health foods containing ω-3 fatty acids Fish oil foods using fish oils extracted from edible fish with ordinary extraction methods. (IV) The minimal level of ω-3 fatty acids in products shall more than 30 g/100 g. (V) Omega-3 fatty acids specified in the foregoing items shall be in the form of triglycerides, with purity calculated by the grand total of α-ALA, EPA, and DHA. (MOHW Specification for Fish Oil Health Foods 0960403055: Fish Oil Specification Draft)

III. Product specifications (I) Appearance Products shall retain their original flavors and colors and contain no decay, color change, odor, contamination, mold, or foreign matter. (II) Specific composition 1. The content shall comply with the “Specification for Fish Oil Health Foods” promulgated by the Food and Drug Administration (Hereafter referred to as FDA). 2. The minimal level of ω-3 fatty acids in products shall be 30 g/100 g. (III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the FDA and the latest version of regulations relating to food sanitation standards. (IV) Heavy metal The content shall comply with the relevant food sanitation standards promulgated by the FDA: the maximum level of heavy metals shall be 20 ppm (by lead) and the maximum level of arsenic shall be 2 ppm. (V) Pesticide residue The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA. (VII) Other 1. Polychlorinated biphenyls (PCBs): Below 0.5ppm 2. Dioxin: 0.75 pg/g fat 3. Peroxide value (POV): Below 10 meq/kg

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TQF Specifications and Standards for Product Test Items TQF-PST-001

V. Supplementation (I) Manufacturers complying with this specification and holding a health food permit shall indicate the following items and text in a noticeable location on the package: 1. Warning: Patients taking anticoagulants, such as aspirin, antihyperlipidemic agents; hemophilia patients with coagulopathy; pregnant women; and diabetic patients are advised to consult their health professionals before using this product. 2. Precautions A. Please consult a physician, pharmacist or dietician before using this product. Balanced diet and appropriate exercise are the foundation of physical health. B. This product does not cure or correct diseases of any kind. Patients should seek medical attention. C. Do not overdose.

VI. Reference 1. 中華民國國家標準：食用油脂之檢驗方法 CNS 14529 2. 中華民國國家標準：食用油脂之檢驗方法 CNS 14769 3. 中華民國國家標準：食品中殘留農藥檢驗方法 CNS 13570 4. 中華民國國家標準：食品中戴奧辛及多氯聯苯殘留量檢驗方法 CNS 14758 5. 中華民國國家標準：食品微生物之檢驗方法─大腸桿菌之檢驗 CNS 10951 6. 中華民國國家標準：食品微生物之檢驗方法─大腸桿菌群之檢驗 CNS 10984 7. 日本保健食品規格標準 8. Kris-Etherton, P.M., Hecker, K.D. and Binkoski, A.E. 2004. Polyunsaturated fatty acids and cardiovascular health. Nutr Rev. 62(11):414-426. 9. Official Methods of Analysis AOCS Ce-1b-89 10. Official Methods of Analysis：AOAC 965.35 11. Official Methods of Analysis：AOAC 971.21 12. Official Methods of Analysis：AOAC 986.15 13. Official Methods of Analysis：AOAC 990.05

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TQF Specifications and Standards for Product Test Items TQF-PST-001

5. Specification for Foods Containing Soybean Lecithin

I. Scope This specification applies to foods containing soybean lecithin.

II. Definitions Soybean lecithin: Soybean lecithin refers to the phosphatides mixed with phosphatidyl choline (PC), phosphatidyl ethanol amine (PE), phosphatidyl inositol (PI), phosphatidyl serine (PS), phosphatidic acid (PA), and sphingolipids contained in soybeans. Phosphatides refer to a compound formed by phospholipids from a phosphate group and the third carbon group in triglycerides and other composition, such as forming PC with choline and PE with ethanolamine. In general, soybean lecithin is the by-product from the degumming process in soybean oil refinement or is directly extracted from soybeans. Foods containing soybean lecithin refers to foods made with soybean lecithin and other ingredients through processing, with a minimum level of lecithin at 10%. It is generally believed that lecithin can maintain cell membrane integrity; normal liver and kidney functions; prevent arteriosclerosis, heart disease, and liver disease; reduce blood lipids and liver fat; prevent choleliths and senile dementia.

III. Product specifications (I) Appearance Products shall retain their original flavors and colors and contain no decay, color change, odor, contamination, mold, or foreign matter. (II) Content of specific composition 1. Comply with the labeling. 2. Minimum content of soybean lecithin shall be 10%. (III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the Food and Drugs Administration (hereafter referred to as FDA) and the latest version of regulations relating to food sanitation standards. (IV) Heavy metal 1. Total heavy metal (by lead): Below 20 ppm. 2. Arsenic (As): Below 3 ppm (by As). 3. Lead (Pb): Below 10 ppm. (V) Pesticide residue Complies with the “Standards for Pesticide Residue Limits in Foods” for soybeans and dry soybeans in the food sanitation standards. See Supplementation (1) for details. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA. (VII) Other 1. Aflatoxins: The maximum level of aflatoxins including B1, B2, G1, and G2 shall be below 10 ppb as specified in the “Edible oils and fats” from the “Sanitation Standard for the Tolerance of Mycotoxins in Foods” of “Sanitation Standard for General Foods”. 2. Polychlorinated biphenyls (PCBs): Not detected as specified in the “Standard for the Tolerance of Polychlorinated Biphenyl in Foods”. 3. Peroxide value (POV): Below 30 meq/kg for ingredients and fat/oil processed products and not applicable for non-fat/non-oil processed foods. 4. Acid Value (AV): Below 40 mgKOH/g for ingredients and fat/oil processed products acid value and not applicable for non-fat/non-oil processed foods.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” by the FDA and the latest version of relevant laws and regulations. (II) The content of the functional composition in foods containing soybean lecithin shall be labeled.

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TQF Specifications and Standards for Product Test Items TQF-PST-001

V. Supplementation None.

VI. Reference 1. 林虔宏, 2001. 蛋黃卵磷脂微膠囊之製作及其物理性質之探討. 國立中興大學畜產學系碩士論文。 2. Spios, E.F., and Szuhaj, B.F., 1996. Lecithins. Bailey's industrial oil and fat products. John Wiley & Sons Inc, New York.：311-395. 3. 日本健康食品產品規格標準，第 5 章健康食品，序號 13 大豆卵磷脂含有食品規格基準：595-598。 4. 李敏雄，1997。卵磷脂之製備及應用。食品科學 24(6)：728-735。 5. A.O.C.S. Official Method 7b-91。 6. Official Methods of Analysis：AOAC 972.25。 7. Official Methods of Analysis：AOAC 952.13。 8. 中華民國國家標準：食用油脂之檢測方法---銅含量之測定 CNS 14769。 9. 中華民國國家標準：食用油脂之檢測方法---鉛含量之測定 CNS 4529。 10. 中華民國國家標準：食品中殘留農藥檢驗方法 CNS 13570。 11. 中華民國國家標準：食用油脂之檢測方法---酸價之測定 CNS 3647。 12. 中華民國國家標準：食品中黃麴毒素之檢測方法 CNS 4090。 13. 中華民國國家標準：食品中多氯聯苯及戴奧辛殘留量之檢測方法 NS 14758。 14. 中華民國國家標準：食品微生物之檢測方法---大腸桿菌群之檢驗方法 CNS 10984。 15. 中華民國國家標準：食品微生物之檢測方法---大腸桿菌之檢驗方法 CNS 10951。 16. 中華民國國家標準：食品微生物之檢測方法---生菌數之檢驗方法 CNS 10890。 17. Juneja, L.R., Koketsu, M., Kawanami, H., Sasaki, K., Kim, K., and Yamamoto, T. 1994. Large-scale preparation of sialic acid and its 101 derivatives from chalaza and delipidated egg yolk. Egg uses and processing technologies: new developments. chapter 16. CAB international. p192-206. 18. 美國藥物食品管理局(FDA)聯邦法規(Code of Federal Regulation) CFR 582.1400，http://www.fda.gov/。 19. 美國藥物食品管理局(FDA) ，http://www.fda.gov/。 20. 美國黃豆出口協會台灣辦事處網站，http://www.asaim.org.tw/。

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TQF Specifications and Standards for Product Test Items TQF-PST-001

6. Specification for Foods Containing Soybean Isoflavones

I. Scope This specification applies to foods made with ingredients containing soybean isoflavones.

II. Definitions Soybean isoflavones: These refer to the compound of 12 glycosyl group and non-glycosyl group isoflavones, including daidzin, genistin, glycitin, daidzein, genistein, glycitein, 6”-O-acetyldaidzin, 6”-O-acetylgenistin, 6”-O-acetylglycitin, 6”-O-malonyldaidzin, 6”-O-malonylgenistin, and 6”-O-malonylglycitin. Foods containing soybean isoflavones refer to foods made with soybean isoflavones and other ingredients through processing, with the minimum level of soybean isoflavones at 1%. Soybean isoflavones are phytoestrogens with functions similar to that of human estrogen. Therefore, they can help relieve menopause symptoms, reduce the incidence rate of cardiovascular disease in women after menopause, and prevent osteoporosis. Some isoflavones, such as genistein, have good antioxidation effect. In addition, soybean isoflavones can reduce the functionality of total cholesterol and LDL in blood, and can prevent cancer as reported in some literature.

III. Product specifications (I) Appearance Products shall retain their original flavors and colors and contain no decay, color change, odor, contamination, mold, or foreign matter. (II) Content of specific composition 1. Comply with the labelling. 2. Minimum content of soybean isoflavones shall be 1%. (III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the Food and Drug Administration (hereafter referred to as FDA) and the latest version of regulations relating to food sanitation standards. (IV) Heavy metal The content shall comply with the “Act Governing Food Safety and Sanitation” by the FDA and the latest version of regulations: 1. Total heavy metal (by lead): Below 20 ppm. 2. Arsenic (As): Below 2 ppm (by As). (V) Pesticide residue Complies with the “Standards for Pesticide Residue Limits in Foods” for soybeans and dry soybeans in the “food sanitation standards”. See Supplementation (1) for details. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA. (VII) Other 1. Aflatoxins: The maximum level of aflatoxins including B1, B2, G1, and G2 shall be below 10 ppb as specified in the “Edible oils and fats” from the “Sanitation Standard for the Tolerance of Mycotoxins in Foods” of “Sanitation Standard for General Foods”. 2. Polychlorinated biphenyls (PCBs): Not detected as specified in the “Standard for the Tolerance of Polychlorinated Biphenyl in Foods”.

IV. Labeling 1. Labeling shall comply with the “Act Governing Food Safety and Sanitation” by the FDA and the latest version of the relevant laws and regulations. 2. The content of the health functional composition in foods containing soybean isoflavones shall be labeled.

V. Supplementation None.

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VI. Reference 1. 中華民國國家標準：食品中殘留農藥檢驗方法 CNS 13570。 2. 中華民國國家標準：食用油脂之檢測方法---酸價之測定CNS 3647。 3. 中華民國國家標準：食品中黃麴毒素之檢測方法 CNS 4090。 4. 中華民國國家標準：食品中多氯聯苯及戴奧辛殘留量之檢測方法 CNS 14758。 5. 中華民國國家標準：食品微生物之檢測方法---大腸桿菌群之檢驗方法 CNS 10984。 6. 中華民國國家標準：食品微生物之檢測方法---大腸桿菌之檢驗方法 CNS 10951。 7. 中華民國國家標準：食品微生物之檢測方法---生菌數之檢驗方法 CNS 10890。 8. 陳武介, 黃豆異黃酮與其生理機能。美國黃豆出口協會http://www.asaim.org.tw/。 9. 黃玉玲，1999。脫脂黃豆對大白鼠體內異黃酮素含量及抗脂質氧化之影響。輔仁大學食品營養學系碩士論文。 10. Brandi, M.L., 1992. Flavonoids：Biochemical effects and therapeutic application. Bone Miner. 19：3S-14S. 11. Barnes, S., Grubbs, C., Setchell, K. and Carlson, J., 1990. Soybean inhibit mammary tumors in models of breast cancer. Prog. Clin. Biol. Res. 347：239-253. 12. Brouns, F., 2002. Soya isoflavones: a new and promising ingredient for the health foods sector. Food Research International 35：187–193. 13. Cao, G., Sofic, E. and Prior, R.L., 1997. Antioxidant and prooxidant behavior of flavonoids: structure-activity relationships. Free Radic. Biol. Med. 22：749-760. 14. Eldridge, A.C. and Kwolek, W.F., 1983. Soybean isoflavones: effect of environment and variety on composition. J Agri Food Chem 31：394-396. 15. Farmakalidis, E. and Murphy, P.A., 1985. Isolation of 6"-O-acetyldaidzein and 6"-O-acetylgenistein from toasted defatted soy flakes. J Agric. Food Chem. 33: 385-389. 16. Knight, D.C. and Eden, J. A., 1996. A review of the clinical effect of phytoestrogens. Obstet. Gynecol. 87：897- 904. 17. Official Methods of Analysis：AOAC 972.25。 18. Official Methods of Analysis：AOAC 952.13。 19. Official Methods of Analysis：AOAC 985.01。 20. Official Methods of Analysis：AOAC 972.25。 21. Wang, H.J. and Murphy, P.A., 1996. Mass balance study of isoflavones during soybean processing. J Agri Food Chem 44：2377-2383.

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TQF Specifications and Standards for Product Test Items TQF-PST-001

7. Specification for Mushroom Functional Food Containing Water Soluble Crude Polysaccharides I. Scope This specification applies to foods containing water soluble crude polysaccharides with mushroom ingredients.

II. Definitions (I) Mushrooms Mushrooms refers to edible fungi such as glossy ganoderma, brown mushroom, mushroom, Volvariella volvacea, enokitake, Antrodia cinnamomea, Wolfiporia extensa, tremella fuciformis, and Hericium erinaceus. (II) Mushroom Water Soluble Crude Polysaccharides Polysaccharides in mushrooms are stored in the fungus’ cell wall which is composed of two types of polysaccharides: (a) cellulose in the form of compact physical fibers and (b) α-glucan, β-glucan, and glycoprotein in the form of a matrix. The physiological activity of polysaccharides is correlated with the characteristics of their structure, composition, mass, and degree. Functional polysaccharides that can fight tumors and enhance immunity found in related mushroom studies fall into the following five categories: (1) homoglucans are linked by β-(1-3), α-(1-3), β- (1-6), and α-(1-6) bonds with β-(1-6), β-(1-2) or α-(1-4) branches; (2) heteroglucans are linked by glucan bonds with xylose, arabinose, galactose, mannose and xylose branches; (3) heterogalactans are linked by galactan bonds with glucose, trehalose, xylose, and arabinose branches; (4) other heteroglycans are linked by xylan, fucosan, and mannan bonds with glucose, mannose, and galactose branches; and (5) polysaccharide-protein or peptide complexes. The mushroom water soluble crude polysaccharides in this specification refer to physiologically active water soluble crude polysaccharides from mushrooms that can help resist tumors and enhance immunity. (III) Mushroom functional food containing water soluble crude polysaccharides Mushroom functional food refer to the products with physiological activity containing the sporocarps or mycelia obtained from fermentation with edible mushroom fungi or liquids. Mushroom FHCs in this specification specifically refer to FHCs containing at least 3% water soluble crude polysaccharides (dry weight) from glossy ganoderma and Agaricus blazei Murill.

Item Max. Level (dry weight) Lead Below 3 ppm Cadmium Below 2 ppm

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TQF Specifications and Standards for Product Test Items TQF-PST-001

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” by the FDA and the latest version of the relevant laws and regulations. (II) The content of water soluble crude polysaccharides shall be labeled.

V. Supplementation (I) Currently, there are no publicly-recognized methods for analyzing water soluble crude polysaccharides across the world. This method is based on “A Study on the Analytical Method for Water-soluble Crude Polysaccharides in Ling-Zhi” by Lu, Gong, and Fu (2001) (153-161, 39(2), 2001, Taiwan Journal of Agricultural Chemistry and Food Science). After separating and collecting over 10,000 molecules of crude polysaccharides with capillary columns in molecular sieve chromatography, they quantified the total content of crude polysaccharides. The method can clearly distinguish and completely retain over 10,000 molecules of crude polysaccharides. (II) Literature review found that the quantity of molecules contained in functional polysaccharides with anti- cancer or immunity-enhanced characteristics found in related studies on 28 types of mushrooms in the past 30 years is over 10,000. This provides a discriminatory point to eliminate the majority of polysaccharides that do not contain functional molecules added to or contained in products. (III) Methods of analysis of related test items are as follows: Item Method Heavy metal Test Methods for Heavy Metals in Foods - Cadmium Test (2) (Shu-Shou-Shi-Zi-No. 0929206232 dated 4 April 2003) Test Methods for Heavy Metals in Foods - Lead Test (2) (Shu-Shou-Shi-Zi-No. 0929227157 dated 23 December 2003) AOAC999.10, AOAC999.11 Pesticide residue Test Methods for Pesticide Residues in Foods (Shu-Shou-Shi-Zi-No. 0949424750 dated 24 August 2005) General Plate Count Microbiological Test Methods for Foods-General Plate Count Test CNS10890 Coliform Group Microbiological Test Methods for Foods - Coliform Group Test CNS10984 E. coli Microbiological Test Methods for Foods - E. coli Test (Wei-Shu- Shi-Zi-No.0900025538 dated 20 April 2001) Pathogen Refer to MOHW announcements and CNS.

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TQF Specifications and Standards for Product Test Items TQF-PST-001

VI. Reference 1. 水可溶性粗多醣分析方法依據期刋文獻:呂淑芳、宮昭雲、傅偉光。靈芝中水溶性粗多醣分析方法之研究。台灣農業化學與食品科學，2001，39(2)，153-161。 2. Zhang, M., Cui, S.W., Cheung, P.C.K. and Wang, Q. 2007. Anti-tumor polysaccharides from mushrooms: a review on their isolation process, structural characteristics and anti-tumor activity. Trends in food science and technology. 18:4-19. 3. AOAC International. 2003. Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.10. William Horwitz ed. Gaithersburg, MD U.S.A 4. AOAC International. 2003. Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.11. William Horwitz ed. Gaithersburg, MD U.S.A

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TQF Specifications and Standards for Product Test Items TQF-PST-001

8. Specification for Functional Food Containing Phytosterols

I. Scope This specification applies to foods containing phytosterols.

II. Definitions (I) Phytosterols Phytosterols, or plant sterols, are formed by three six-membered rings and one five-membered ring and are aliphatic tetracyclic alcohols like cholesterols, with very similar chemical structures. However, phytosterols have 28-29 carbon elements, while cholesterols have 27 carbon elements. Free sterols (FSes) are found naturally in plant foods such as vegetable oils, nuts, seeds, alyce clover, vegetables, and fruit. Common phytosterols include stigmasterol, β-sitosterol, and campesterol. (II) Steryl esters In addition to FSes, plants contain other forms of phytosterols. By combining hydroxyl in C-3 with LCFAs or phenolic acid to form steryl esters (SEs) or with C-1 of monosaccharides to form steryl glycosides (SG). (III) Physiological functions of phytosterols Phytosterols can reduce cholesterol absorption and promote the synthesis of LDL receptor protein to reduce low density lipoprotein cholesterol (LDL-C). (IV) The minimum level of FSes in products shall be 1.4 g/100 g (based on the phytosterol content in Taishan Phytosterol Sunflower Oil).

III. Product specifications (I) Appearance Products shall retain their original flavors and colors and contain no decay, color change, odor, contamination, mold, or foreign matter. (II) Content of specific composition 1. Comply with the labeling. 2. The minimum content of FSes in products shall be more than 1.4 g/100 g. (III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the Food and Drug Administration (hereafter referred to as FDA) and the latest version of regulations relating to food sanitation standards. (IV) Heavy metal The content shall comply with the relevant food sanitation standards promulgated by the FDA: the maximum level of heavy metals shall be 20 ppm (by lead) and the maximum level of arsenic shall be 2 ppm. (V) Pesticide residue The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA. (VII) Other 1. Polychlorinated biphenyls (PCBs): Below 0.5 ppm 2. Dioxin: 0.75 pg/g fat 3. Peroxide value (POV): Below 10 meq/kg

V. Supplementation (I) Manufacturers complying with this specification and holding a health food permit shall indicate the following items TQF Specifications and Standards for 21 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

and text in a noticeable location on the package: 1. Warning: Patients taking anticoagulants, such as aspirin, antihyperlipidemic agents; hemophilia patients with coagulopathy; pregnant women; and diabetic patients are advised to consult their health professionals before using this product. 2. Precautions: A. Please consult a physician, pharmacist or dietician before using this product. Balanced diet and appropriate exercise are the foundation of physical health. B. This product does not cure or correct diseases of any kind. Patients should seek medical attention. A. Do not overdose.

VI. Reference 1. 中華民國國家標準：食用油脂之檢驗方法 CNS 14529 2. 中華民國國家標準：食用油脂之檢驗方法 CNS 14769 3. 中華民國國家標準：食用調合植物油 CNS 總號 7525 類號 N5181 4. 中華民國國家標準：食品中殘留農藥檢驗方法 CNS 13570 5. 中華民國國家標準：食品中黃麴毒素檢驗方法 CNS 4090 (20) 6. 中華民國國家標準：食品中戴奧辛及多氯聯苯殘留量檢驗方法 CNS 14758 7. 中華民國國家標準：食品微生物之檢驗方法─大腸桿菌之檢驗 CNS 10951 8. 中華民國國家標準：食品微生物之檢驗方法─大腸桿菌群之檢驗 CNS 10984 9. 日本保健食品規格標準 10. 中華民國國家標準： CNS 3192 包裝食品標示 11. Cargill Method：QC_SE008_Rev01 12. de Jone, A. Plat, J. and Mensink, R.P. 2003. Metabolic effects of plant sterols and stanols (review). J. Nutr. Biochem. 14:362-369. 13. Denke M. A. 2002. Dietary prescriptions to control dyslipidemias. CircμLation. 105(2):132-135. 14. Official Methods of Analysis：AOAC 971.21 15. Official Methods of Analysis：AOAC 976.26 16. Official Methods of Analysis：AOAC 986.15 17. Official Methods of Analysis：AOAC 990.05 18. Simons L. A. 2002. Additive effect of plant sterol-ester margarine and cerivastatin in lowering low-density lipoprotein cholesterol in primary hypercholesterolemia. American Journal of Cardiology. 90(7):737-740.

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9. Functional Food Containing Dietary Fibers

I. Scope This specification applies to foods claimed to contain total dietary fibers or added total dietary fibers.

II. Definitions (I) Dietary fibers Total dietary fibers are polysaccharides and lignin that cannot be digested by human enzymes. They mainly come from the non-starch carbohydrates and lignin in plant tissues. Most polysaccharides are cell wall polysaccharides including cellulose, hemicellulose, and pectin. Total dietary fibers are the sum of water-soluble dietary fibers and non-water-soluble dietary fibers.

Composition of dietary fibers 1. A cellulose is a glucose-unit long-chain linear polymer linked by β-1,4-glycosidic bond without side chains. The polymer molecule is formed by 100-200 glucose units. Polymers in parallel are combined into micro cellulose by hydrogen bonds with sturdy molecules. It is an extremely stable insoluble substance that cannot be digested in the human gastrointestinal tract. 2. A hemicellulose is a complex substance containing various complex polysaccharides. Its main chain is often formed by xylose, mannose, and galactose, with arabinose, uronic acids, and galactose in the branched chain. It exists in the cell wall like the cellulose and lignin. 3. Alpha-1,4-galacturonan is the main structure of pectin, others include neutral sugars like rhamnose, fucose, xylose, and galactose. Pectin is a matrix polysaccharide in the cell wall. 4. Lignin is a non-carbohydrate substance, a high-molecular aromatic compound bonded by about 40 phenols with strong molecules. 5. Non-cell-wall polysaccharides include hydrophilic colloids, such as seaweed polysaccharides and plant colloids. (II) The foods containing dietary fibers in this specification refers to foods with a minimum level of 3 g of dietary fibers in every 100 g of solid (semi-solid) foods or 1.5 g of dietary fibers in every 100 ml of liquid foods.

III. Product specifications (I) Appearance Products shall retain their original flavors and colors and contain no decay, color change, odor, contamination, mold, or foreign matter. (II) Content of specific composition 1. Comply with the labeling. 2. The foods containing dietary fibers in this specification refers to foods with a minimum level of 3 g of dietary fibers in every 100 g of solid (semi-solid) foods or 1.5 g of dietary fibers in every 100 ml of liquid foods. (III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the Food and Drug Administration (hereafter referred to as FDA) and the latest version of regulations relating to food sanitation standards. (IV) Limits of heavy metal and minerals 1. Beverages (excluding natural fruit and vegetable juices and concentrated fruit and vegetable juices) (1) Arsenic: Below 0.2 ppm. (2) Lead: Below 0.3 ppm. (3) Zine: Below 5 ppm. (4) Copper: Below 5 ppm. (5) Tin: Canned, below 250 ppm. (6) Stibium: Below 0.15 ppm. 2. Other forms of foods shall comply with the food sanitation standards promulgated by the FDA. The content shall comply with the relevant food sanitation standards: the maximum level of heavy metals shall be 20 ppm (by lead) and the maximum level of arsenic shall be 2 ppm. (V) Pesticide residue

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TQF Specifications and Standards for Product Test Items TQF-PST-001

The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA.

V. Supplementation (I) AOAC 985.29 “Total Dietary Fiber in Foods, Enzymatic-Gravimetric Method” applies to dietary fiber analysis. The principle of this method is to remove fats (>10%) in foods. After drying, -amylase, protease, and amyloglucosidase are used remove starch and protein in the samples. Then, quardruple of hot alcohol are added to the filtered solution obtained from the previous enzymatic treatment to settle soluble dietary fibers. After filtering and drying, the amount of dietary fibers is obtained after removing the residual ash and protein in the residue. (II) The methods for analyzing the relevant items are as follows: Item Method Heavy Metals and Test Methods for Heavy Metals in Foods - Cadmium Test (2) (Shu-Shou-Shi-Zi-No. Minerals 0929206232 dated 4 April 2003) Test Methods for Heavy Metals in Foods - Lead Test (2) (Shu-Shou-Shi-Zi-No. 0929227157 dated 23 December 2003) Heavy method analysis methods in the food additive specifications and test methods CNS 3971 N6093 Heavy Metal Test Method for Canned Foods - Test of Tin Content CNS 12869 N6231 Mineral Test Methods in Infant Formulas - Test of Copper, Iron, Magnesium, Manganese, Potassium, Sodium, and Zinc. AOAC999.10--Lead, Cadmium, Zinc, Copper, and Iron in Foods by Atomic Absorption Spectrophotometry after Microwave Digestion. AOAC999.11--Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. AOAC946.16--Antimony in Foods. Pesticide residue Test Methods for Pesticide Residues in Foods CNS13570 General Plate Microbiological Test Methods for Foods CNS10890 Count Coliform Group Microbiological Test Methods for Foods CNS10984 E. coli Microbiological Test Methods for Foods CNS10951

VI. Reference 1. 中華民國國家標準：食品微生物之檢驗法-生菌數之檢驗 CNS10890(1991) 2. 中華民國國家標準：食品微生物之檢驗法-大腸桿菌群之檢驗 CNS10984 (1998) 3. AOAC International. 2003. Total Dietary Fiber in Foods, Enzymatic- Gravimetric Method. In. “Official Methods of Analysis of AOAC International” 17th ed. 985.29. William Horwitz ed., Gaithersburg, MD U.S.A.. 4. AOAC International. 2003. Total, Soluble and Insoluble Dietary Fiber in Foods and Food Products, Enzymatic Gravimetric Method. In. “Official Methods of Analysis of AOAC International” 17th ed. 991.43. William Horwitz ed. Gaithersburg, MD U.S.A 5. AOAC International. 2003. Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.11. William Horwitz ed. Gaithersburg, MD U.S.A 6. AOAC International. 2003. Lead, Cadmium, Zinc, Copper, and Iron in Foods by Atomic Absorption Spectrophotometry after Microwave Digestion. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.10. William Horwitz ed. Gaithersburg, MD U.S.A TQF Specifications and Standards for 24 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

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7. AOAC International. 2003. Antimony in Food, Spectrophotometric Method. In. “Official Methods of Analysis of AOAC International” 17th ed. 46.16 William Horwitz ed. Gaithersburg, MD U.S.A

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10. Specification for Functional Foods Containing Resistant Maltodextrins

I. Scope This specification applies to foods clamming to contain resistant maltodextrins or foods added with resistant maltodextrins.

II. Definitions (I) Resistant maltodextrins Formed by glucose and combined by glycosidic bonds including α-1,4, α-1,6, α/β-1,2, α/β-1,3, and β-1,6, resistant maltodextrins have highly branched structures. In general, resistant maltodextrins are made with maize (corn), potatoes, and other starch. After heating, ingredients are hydrolyzed with amylase to remove parts that can be digested, and resistant maltodextrins are obtained after desalination, bleaching, and tempering. Resistant maltodextrins do not have a specific composition. Resistant maltodextrins in foods can be obtained by adding, processing (extrusion), or tradition maltodextrins. Resistant maltodextrins are considered as dietary fibers in Japan, New Zealand, and Australia. Traditionally, dietary fibers are defined as polysaccharides and lignin that cannot be digested by human enzymes. They mainly come from the non-starch carbohydrates and lignin in plant tissues. Most polysaccharides are cell wall polysaccharides including cellulose, hemicellulose, and pectin. Currently, dietary fibers are defined based on their physiological functions. According to American Association of Cereal Chemists (AACC, 2001), dietary fibers are the edible part or similar carbohydrates in plants that can resist the decomposition and absorption by the human intestinum tenue to partial or total decomposition by the human intestinum crissum. Dietary fibers include polysaccharides, oligosaccharides, lignin, and other related plant substances. Physiologically, they help regulate intestinal track health, promote intestinal peristalsis, shorten transport time, soften stools, and increase stool volume to facilitate defecation. Partially fermented by enteric bacteria at the end of the gastrointestinal track, they can produce SCFAs and lower the pH value of intestinal track to promote probiotic growth in the intestinal track. Resistant maltodextrins can also slow saccharide absorption in the intestinal track, moderate blood glucose rise after eating, and absorb sodium taurocholate to reduce cholesterols and fats in plasma and the liver. (II) Functional foods containing resistant maltodextrins Functional foods containing resistant maltodextrins refer to products containing physiological functions by adding or producing resistant maltodextrins in foods during processing. The content of dietary fibers quantified by AOAC 2001.03 analysis is the indicative composition of resistant maltodextrins. Foods containing resistant maltodextrins in this specification refer to foods with a minimum level of 3 g of dietary fibers in every 100 g of solid (semi-solid) foods or 1.5 g of dietary fibers in every 100 ml of liquid foods.

III. Product specifications (I) Appearance Products shall retain their original flavors and colors and contain no decay, color change, odor, contamination, mold, or foreign matter. (II) Specific composition 1. Comply with the labeling. 2. The foods containing dietary fibers in this specification refers to foods with a minimum level of 3 g of dietary fibers in every 100 g of solid (semi-solid) foods or 1.5 g of dietary fibers in every 100 ml of liquid foods. (III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the Food and Drug Administration (hereafter referred to as FDA) and the latest version of regulations relating to food sanitation standards. (IV) Limits of heavy metal and minerals 1. Beverages (excluding natural fruit and vegetable juices and concentrated fruit and vegetable juices) (1) Arsenic: Below 0.2 ppm. (2) Lead: Below 0.3 ppm. (3) Zine: Below 5 ppm. (4) Copper: Below 5 ppm. (5) Tin: Canned, below 250 ppm. (6) Stibium: Below 0.15 ppm. 2. Other forms of foods shall comply with the food sanitation standards promulgated by the FDA: the maximum TQF Specifications and Standards for 26 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

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level of heavy metals shall be 20 ppm (by lead) and the maximum level of arsenic shall be 2 ppm. (V) Pesticide residue The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA.

V. Supplementation (I) Currently there is no method that can detect resistant maltodextrins alone from the food substrate. As a variety of dietary fibers, resistant maltodextrins can be indicated by the dietary fibers quantified by AOAC2001.03 analysis contained in foods with resistant maltodextrins. The principle of this analysis method is to remove fats (>10%) in foods. After drying, starch and protein in samples are removed by -amylase, protease, and amyloglucosidase. Then, four additions of hot alcohol are added to the filtered solution obtained from the said enzymatic treatment to settle soluble dietary fibers (resistant maltodextrins with higher molecular mass will be settle in this procedure). After filtering and drying, insoluble dietary fibers+high molecule soluble dietary fibers are obtained after removing the residual ash and protein in the residue. The content of resistant maltodextrins with lower molecular mass will be obtained by HPLC analysis. The sum of both will be the content of total dietary fibers in foods. (II) The methods for analyzing the relevant items are as follows: Item Method

Heavy Metals and Test Methods for Heavy Metals in Foods - Cadmium Test (2) (Shu-Shou-Shi-Zi-No. Minerals 0929206232 dated 4 April 2003) Test Methods for Heavy Metals in Foods - Lead Test (2) (Shu-Shou-Shi-Zi-No. 0929227157 dated 23 December 2003) Heavy method analysis methods in the food additive specifications and test methods CNS 3971 N6093 Heavy Metal Test Method for Canned Foods - Test of Tin Content CNS 12869 N6231 Mineral Test Methods in Infant Formulas - Test of Copper, Iron, Magnesium, Manganese, Potassium, Sodium, and Zinc. AOAC999.10--Lead, Cadmium, Zinc, Copper, and Iron in Foods by Atomic Absorption Spectrophotometry after Microwave Digestion. AOAC999.11--Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. AOAC946.16--Antimony in Foods. Pesticide residue Test Methods for Pesticide Residues in Foods CNS13570 General Plate Microbiological Test Methods for Foods CNS10890 Count Coliform Group Microbiological Test Methods for Foods CNS10984 E. coli Microbiological Test Methods for Foods CNS10951

TQF Specifications and Standards for Product Test Items TQF-PST-001

Gravimetric Method. In. “Official Methods of Analysis of AOAC International” 17th ed. 991.43. William Horwitz ed. Gaithersburg, MD U.S.A 5. AOAC International. 2003. Total dietary fiber in foods containing Resistant Maltodextrin.Enzymatic- gravimetric method and liquid chromatography determination. In. “Official Methods of Analysis of AOAC International” 17th ed. 2001.03. William Horwitz ed. Gaithersburg, MD U.S.A 6. AOAC International. 2003. Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.11. William Horwitz ed. Gaithersburg, MD U.S.A 7. AOAC International. 2003. Lead, Cadmium, Zinc, Copper, and Iron in Foods by Atomic Absorption Spectrophotometry after Microwave Digestion. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.10. William Horwitz ed. Gaithersburg, MD U.S.A 8. AOAC International. 2003. Antimony in Food, Spectrophotometric Method. In. “Official Methods of Analysis of AOAC International” 17th ed. 46.16 William Horwitz ed. Gaithersburg, MD U.S.A 9. Food Standards Australia New Zealand (2004), Final assessment report. Application A491 ： Resistant Maltodextrin as Dietary Fibre.

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11. Specification for Ganoderma Tsugae Functional Foods Containing Ganoderic Acid

I. Scope This specification applies to ganoderic acid foods made with ingredients containing the sporocarp of ganoderma tsugae.

II. Definitions (I) Ganoderma tsugae functional foods containing ganoderic acid Ganoderma tsugae functional foods refer to products containing biologically active composition made with the sporocarp of Ganoderma tsugae. (II) Ganoderic acid Triterpenoids with a lanostane backbone structure are the bittering constituent of glossy ganoderma. Based on different backbone carbon numbers, triterpenoids include C30 ganoderic acid, C27 lucidenic acid, and C24 lucidone. Ganoderic acid is the main triterpenoid in Ganoderma tsugae. Studies found that ganoderic acid can effectively inhibit the activity of angiotensin converting enzyme (ACE) to lower blood pressure.

Item Max. Level (dry weight) Lead Below 3 ppm Cadmium Below 2 ppm

2. The content shall comply with the relevant food sanitation standards promulgated by the FDA: the maximum level of heavy metals shall be 20 ppm (by lead) and the maximum level of arsenic shall be 2 ppm. (V) Pesticide residue 1. The pesticide residue shall comply with the mushroom standard in “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. International Common Name Common Name Max. Level (ppm) Remarks Allethrin Allethrin 3.0 Insecticide Carbendazim Bavistin 0.5 Germicide Diazinon Diazinon 0.2 Insecticide Dicloran Dicloran 0.5 Germicide Malathion Malathion 2.0 Insecticide Prochloraz Prochloraz 0.5 Germicide Pyrethrins Pyrethroid 1.0 Insecticide Thiabendazole Mertect 5.0 Germicide TQF Specifications and Standards for 29 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

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2. The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA.

IV. Labeling 1. Labeling shall comply with the “Act Governing Food Safety and Sanitation” by the FDA and the latest version of the relevant laws and regulations. 2. The content of ganoderic acid shall be labeled.

V. Supplementation (I) Item Method

Heavy metal 1. Test Methods for Heavy Metals in Foods - Cadmium Test (2) (Shu-Shou-Shi-Zi-No. 0929206232 dated 4 April 2003) 2. Test Methods for Heavy Metals in Foods - Lead Test (2) (Shu-Shou-Shi-Zi-No. 0929227157 dated 23 December 2003) 3. AOAC999.10 4. AOAC999.11 Pesticide Test Methods for Pesticide Residues in Foods residue (Shu-Shou-Shi-Zi-No. 0949424750 dated 24 August 2005) General Plate Microbiological Test Methods for Foods-General Plate Count Test CNS10890 (1991) Count Coliform Microbiological Test Methods for Foods - Coliform Group Test CNS10984 (1998) Group E. coli Microbiological Test Methods for Foods - E. coli Test (Wei-Shu-Shi-Zi-No.0900025538 dated 20 April 2001) Pathogen Refer to MOHW announcements and CNS.

(II) Methods for analyzing ganoderic acid: (1) The method introduced by Lu, Yang, Gong, and Fu (2005) in “A Quantitative Method for Triterpenoids in Ganoderma Tsugae” is applied to analyze ganoderic acid compounds. (2) Method description: The ganoderic acid compound is one of the important functional composition in glossy ganoderma. Currently, 119 types of ganoderic acid compounds have been separated and identified from the sporocarp and mycelium of ganoderma lucidum, including ganoderic acids, lucidenic acids, ganodermic acids, ganoderenic acids, lucidones, ganoderal, and ganoderiols(1). While there is a wide variety of ganoderic acid compounds, no standard product is currently available for commercial trade. As they must be purified from samples, the process is time-consuming, the yield is low, and quantification is difficult. This method calculates the relative response factors of the peak area of a ganoderic acid compound. Then, from the relationship of the relative response factors, the content and total content of other ganoderic acid compounds can be obtained from a single standard product, simplifying procedure and reducing used amount of single product. The difference between the calculated results and the quantification outcome of individual standard products is small, suggesting that this method can reduce experimental costs and shorten analysis time to efficiently quantify the content of ganoderic acid compounds in Ganoderma tsugae. (3) Structure verification of ganoderic acid compounds: The results of HPLC on the sporocarp of Ganoderma tsugae show that the spectrum of Ganoderma tsugae from different sources is extremely similar. After extracting ganoderic acid compounds from the sporocarp of Ganoderma tsugae using ethanol ultrasonic agitation, the HPLC spectrum is shown in Figure 1. The figure shows that over 20 ganoderic acid compounds can be detected from Ganoderma tsugae. After comparing with related literature(1), the signal of peak 2 should be ganoderic acid C, peak 3 should be ganoderic acid C6, peak 4 should be ganoderic acid G, peak 6 should be ganoderic acid B, peak 9 should be ganoderic acid A, peak 10 should be ganodeneric

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acid D, and peak 11 should be ganoderic acid D.

VI. Reference 1. 食品微生物之檢驗法－生菌數之檢驗 CNS10890(1991) 2. 中華民國國家標準：食品微生物之檢驗法－大腸桿菌群之檢驗 CNS10984(1998) 3. 水野卓、川合正允，菇類的化學．生化學。pp.237-248。國立編譯館，台北，台灣(1996) 4. 黃惠君，食藥用菇的營養與藥用價值。食品工業，36(5):25-32(2004) 5. 呂淑芳、楊筱冬、宮昭雲、傅偉光，松杉靈芝中類三萜化合物定量方法。台灣農業化學與食品科學， 43(3):200-205 (2005) 6. AOAC International. 2003. Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.10. William Horwitz ed. Gaithersburg, MD U.S.A 7. AOAC International. 2003. Determination of Lead, Cadmium, Copper, Iron, and Zinc in Foods. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.11. William Horwitz ed. Gaithersburg, MD U.S.A 8. D. H. Chen and K. D. Chen: Determination of Ganoderic acids in triterpenoid constituents of Ganoderma tsugae. J.Food Drug Anal., 11(3):195-201(2003)。

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12. Specification for Functional Foods Containing Lactic Acid Bacteria (LAB)

I. Scope This specification applies to foods containing active lactic acid bacteria (LAB).

II. Definitions Lactic acid bacteria (LAB) is a group of numerous and jumbled bacteria series and not a term used in microbial taxonomy. In general, LAB is a collective term referring to bacteria that can produce a large amount of lactic acid through fermentation with carbohydrates. They usually carry the common characteristics described below: (1) gram- positive bacteria (without mobility), (2) complex nutritional needs for growth: carbohydrates, amino acid, nucleic acid derivatives, vitamins, and a wide variety of auxins; (3) usually lack catalase activity and cytochrome; (4) anaerobic, microaerophilic, aerotolerant anaerobic or facultative anaerobic bacteria. In general, they can live in an aerobic environment, but better in an anoxic state. There are also obligate anaerobes. After long-term human consumption and science research, some LAB are known as normal bacteria series in the human intestinal track (Edwards, 1993; Kleessen et al., 1995) with physiological functions (Sarrela et al., 2002). Suitable LAB supplementation, such as Bifidobacterium and Lactobacillus, can promote intestinal health which is the potential of probiotics. However, different strains of the same bacterium functionally vary. The table below shows the LAB for use in foods approved by the Department of Health (DoH) in September 2008. The actual types of LAB for use in foods shall be refer to the latest announcement by the Food and Drug Administration (hereafter referred to as FDA).

Table 1 DoH List of LAB Allowed for Use in Foods Bacillus coagulans Lactobacillus kefir Bifidobacterium bifidum Lactobacillus lactis Bifidobacterium breve Lactobacillus lactis subsp. lactis Bifidobacterium infantis Lactobacillus paracasei Bifidobacterium lactis Lactobacillus plantarum Bifidobacterium longum Lactobacillus reuteri Bifidobacterium adolescentis Lactobacillus rhamnosus Enterococcus faecalis Lactobacillus salivarius Enterococcus faecium Lactobacillus sporogenes Lactobacillus acidophilus Lactobacillus pentosus Lactobacillus bifidus Lactobacillus johnsonii Lactobacillus brevis Lactococcus lactis subsp. cremoris Lactobacillus bulgaricus Lactococcus lactis subsp. lactis Lactobacillus casei Lactococcus lactis subsp. lactis biovar diacetylactis Lactobacillus casei subsp. rhamnosus Leuconostoc mesenteroides subsp. cremoris Lactobacillus cremoris Sporolactobacillus inulinus Lactobacillus delbrueckii Streptococcus lactis Lactobacillus delbrueckii subsp. bulgaricus Streptococcus salivarius subsp. thermophilus Lactobacillus fermentum Streptococcus thermophilus Lactobacillus gasseri Streptococcus faecalis Lactobacillus helveticus

III. Product specifications (I) Appearance Products shall retain their original flavors and colors and contain no decay, color change, odor, contamination, mold, or foreign matter. (II) Content of specific composition 1. Comply with indications. 2. There are three tpyes of specifications: Product Form LAB Content (1) Powder, capsule Above 109 CFU/g TQF Specifications and Standards for 32 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

(2) Tablet/pill Above 108 CFU/g (3) Liquid or gel Above 107 CFU/g (or mL)

3. LAB contained in products shall be one of those listed in the “DoH List of LAB Allowed for Use in Foods” (III) Microbial limits Shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (IV) Heavy metals The content shall comply with the relevant food sanitation standards. If no standards applicable, the maximum level of heavy metals shall be 20 ppm (by lead) and the maximum level of arsenic shall be 2 ppm. (V) Pesticide residue The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The scientific name and total count of LAB shall be labeled. (III) Warning 1. For dairy products containing LAB, the following warning shall be labeled: “Those with lactose intolerance should seek advice from their physicians or dieticians before use.”

V. Test Methods Refer to CNS 14760 Methods of Test for Food Microorganisms-Test of Lactic Acid Bacteria

VI. Supplementation (I) This LAB analysis method aims to test the total LAB count. (II) The species of LAB shall be verified. The accuracy of strains or strains sub-cultured more than five times shall be verified every year.

VII. Reference 1. 中華民國國家標準：發酵乳 CNS 3058。 2. 中華民國國家標準：食品微生物之檢驗法-生菌數之檢驗 CNS 10890。 3. 中華民國國家標準：乳品檢驗法-乳酸菌之檢驗 CNS 14760。 4. Edwards, C. 1993. Interactions between nutrition and intestinal microflora. Proc. Nitr. Soc. 52: 375-382. 5. Kleessen, B., Bunke, H., Tovar, K., Noack, J. and Sawatzki, G. 1995. Influence of two infant formulas and human milk on the development of the faecal flora in newborn infants. Acta Paediatr. 84: 1347-1356. 6. Sarrela, M., Lahteenmaki, L., Crittenden, R., Salminen, S. and Mattila-Sandholm, T. 2002. Gut bacteria and health foods-the European perspective. Int. J. Food Microbiol. 78: 99-117.

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13. Specification for Functional Foods Containing Catechins

I. Scope This specification applies to solid, powdered or liquid foods containing catechins.

II. Definitions Catechin is a type of polyphenol and the source of the bitter and puckering tastes in tea and a factor determining tea quality. There are eight types of catechin, including epigallocatechin (EGC), catechin (CT), epicatechin (EC), epigallocatechin gallate (EGCG), gallocatechin gallate (GCG), epicatechin gallate (ECG), gallocatechin (GC), and catechin gallate (CG). Catechin content labeling refers to the total content of the above eight compounds.

Product Form Content of Catechin Compounds (1) Liquid Above 25 mg/100 mL (2) Capsule or tablet/pill Above 150 mg/g (3) Solid Above 5 mg/g

(III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the Food and Drug Admnistration (hereafter referred to as FDA) and the latest version of regulations relating to food sanitation standards. (IV) Heavy metals The content shall comply with the relevant food sanitation standards. If no standards applicable, the maximum level of heavy metals shall be 20 ppm (by lead) and the maximum level of arsenic shall be 2 ppm. (V) Pesticide residue The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The content of catechins in products shall be labeled.

V. Supplementation Refer to CNS 15022, N 6384 Method for Test of “Catechins”.

VI. Reference (1) 中華民國國家標準 CNS 15022, N 6384 兒茶素之檢驗方法。 (2) Ikeda, I. 2008. Multifunctional effects of green tea catechins on prevention of the metabolic syndrome. Asia Pac. J. Clin. Nutr. 17 (S1): 273-274. (3) Inan, S., Takanom, M., Yanamoto, M., Murakami, D., Tajika, K., Yodogawa, K., Yokoyama, S., Ohno, N., Ohba, T., Sano, J ., Ibuki, C., Seino, Y. and Mizuno, K. 2007. Tea catechin consumption reduces circulating oxidized low- density lipoprotein. Int. Heart J. 48(6): 725-732.

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(4) Kao to Launch Healthya® Water: First Sports Drink Approved as a ‘Food for Specified Health Use’ in the Body Fat Category. http://www.kao.co.jp/en/news/2006/n20060516-01hc.htmL. (5) Kimura, M., Umegaki, K., Kasuya, Y., Sugisawa, A. and Higuchi, M. 2002. The relation between single=double or repeated tea catechin ingestions and plasma antioxidant activity in humans. European Journal of Clinical Nutrition. 56: 1186–1193. (6) Green tea and weight loss. 2005. Nutrition Research Newsletter. http://findarticles.com/p/articles/mi_m0887/is_/ai_n13648421. (7) Murase, T., Nagasawa, A., Suzuki, J., Hase, T. and Tokimitsu, I. 2002. Beneficial effects of tea catechins on diet- induced obesity: stimulation of lipid catabolism in the liver. International Journal of Obesity. 26: 1459–1464. (8) Nagao, T., Komine, Y., Soga, S., Meguro, S., Hase, T., Tanaka, Y. and Tokimitsu, I. 2005. Ingestion of a tea rich in catechins leads to a reduction in body fat and malondialdehyde-modified LDL in men. Am J Clin Nutr. 81: 122–129. (9) Nagao, T., Hase, T. and Tokimitsu, I. 2007. A green tea extract high in catechins reduces body fat and cardiovascular risks in humans. OBESITY. 15(6): 1473-1483. (10) Spanos, G. A., Wrolstad, R. E. and Heatherbell, D. A. 1990. Influence of processing and storage on the phenolic composition of apple juice. J Agric. Food Chem. 38: 1572.

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14. Specification for Functional Foods Containing Lycopene

I. Scope This specification applies to foods containing natural lycopene.

II. Definitions Lycopene, molecular formula C40H56 and molecular mass 536.9, is a pigment and a strong antioxidant at the same time. As a type of carotenoid, it can eliminate free radicals or oxidants two times of β-carotene, four times of vitamin C, and 100 times of vitamin E. Therefore, it has caught the attention of the field of medical, health food, and beverage.

Prduct Form Lycopene Content (1) Liquid Above 7 mg/100 mL (2) Capsule or tablet/pill Above 20 mg/100 g (3) Solid Above 30 mg/100 g

(III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the Food and Drug Administration (hereafter referred to as FDA) and the latest version of regulations relating to food sanitation standards. (IV) Heavy metals The content shall comply with the relevant food sanitation standards. If no standards applicable, the maximum level of heavy metals shall be 20 ppm (by lead) and the maximum level of arsenic shall be 2 ppm. (V) Pesticide residue The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The content of lycopene shall be labeled.

V. Test Methods Refer to CNS 14836, N 6374 Method for Test of “Lycopene Content in Foods”.

VI. Supplementation (I) The content of trans-lycopene is the outcome of this test method.

VII. Reference (1) 中華民國國家標準 CNS 14836, N 6374「食品中茄紅素含量」之檢驗方法。 (2) Agarwal, S. and Rao, A. V. 1998. Tomato lycopene and low density lipoprotein oxidation: a human dietary intervention study. Lipids, 33, 981–984. TQF Specifications and Standards for 36 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

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(3) Carlo, L. V. 2002. Tomatoes, lycopene intake, and digestive tract and female hormone-related neoplasms. Exp. Biol. Med. 227: 860-863. (4) Cohen, L. A., Zhao, Z., Pittman, B. and Khachik, F. 1999. Effect of dietary lycopene on Nmethylnitrosourea-induced mammary tumorigenesis. Nutr. Cancer. 34: 153–159. (5) Howard, D., Sesso, J. E., Buring, S. M., Zhang, E. P., Norkus and Gaziano, J. M. 2005. Dietary and Plasma Lycopene and the Risk of Breast Cancer ,Cancer Epidemiology Biomarkers & Prevention. 14: 1074-1081. (6) Livny, O., Kaplan, I., Reifen, R., Polak-Charcon, S., Madar, Z. and Schwartz, B. 2002. Lycopene inhibits proliferation and enhances gap-junction communication of KB-1 human oral tumor cells. J. Nutr. 132(12): 3754-9. (7) Nahum, A., Hirsch, K., Danilenko, M., Watts, C. K., Prall, O. W., Levy, J. and Sharoni, Y. 2001. Lycopene inhibition of cell cycle progression in breast and endometrial cancer cells is associated with reduction in cyclin D levels and retention of p27(Kip1) in the cyclin E-cdk2 complexes. Oncogene 20:3428-3436. (8) Narisawa, T., Fukaura, Y., Hasebe, M., Nomura, S., Oshima, S., Sakamoto, H., Inakuma, T., Ishiguro, Y., Takayasu, J. & Nishino, H. 1998. Prevention of N-methylnitrosourea- induced colon carcinogenesis in F344 rats by lycopene and tomato juice rich in lycopene. Jpn. J. Cancer Res. 89: 1003–1008. (9) Okajima, E., Ozono, S., Endo, T., Majima, T., Tsutsumi, M., Fukuda, T., Akai, H., Denda, A., Hirao, Y., Okajima, E., Nishino, H., Nir, Z. and Konishi, Y. 1997. Chemopreventive efficacy of piroxicam administered alone or in combination with lycopene and - carotene on the development of rat urinary bladder carcinoma after N-butyl-N-(4- hydroxybutyl)nitrosamine treatment. Jpn. J. Cancer Res. 88: 543–552. (10) Palan, P., et al. 1996. "Plasma levels of beta-carotene, lycopene, canthaxanthin, retinol, and alpha- and gamma- tocopherol in cervical intraepithelial neoplasia and cancer," Clinical Cancer Research. 2: 181-1851. (11) Rao, A.V. and Agarwal, S. 1998. Bioavailability and in vivo antioxidant properties of lycopene from tomato products and their possible role in the prevention of cancer. Nutr. Cancer. 31: 199–203. (12) Rao, A.V. & Agarwal, S. 1999. Role of lycopene as antioxidant carotenoid in the prevention of chronic diseases: a review. Nutr. Res. 19: 305–323. (13) Winn, D. M,, Ziegler, R. G., Pickle, L. W., Gridley, G., Blot, W. J. and Hoover, R. N. 1984. Diet in the etiology of oral and pharyngeal cancer among women from the southern United States. Cancer Research. 44(3): 1216-1222. (14) Yeh, S. L., Huang, C. S. and Hu, M. L. 2005. Lycopene enhances UVA-induced DNA damage and expression of heme oxygenase-1 in cultured mouse embryo embryo fibroblasts. Eur J Nutr. 44: 365-70. (15) Young, I. S. and Woodside, J. V. 2001. Antioxidants in health and disease. J Clin Pathol. 54: 176-86.

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15. Specification for Functional Foods Containing Spirulina

I. Scope This specification applies to foods containing edible spirulina, including spirulina powder, spirulina granules, and spirulina tablets.

II. Definitions (I) Spirulina: Oscillatoriaceae, Oscillatoriales, Cyanobacteria, Cyanophyta; 300-500 μm spiral multicellular organism in green or blue color. Spirulina contains rich protein (53-62%), carbohydrates (17-25%), fat (4-6%), minerals (8- 13%), and moisture (3-6%). Other composition of spirulina include fatty acids, multivitamins, β-carotene, zeaxanthin, chlorophyll a, and phycocyanins. (II) Prior particle: Processed by manually cultured and sprayed dry. (III) Spirulina foods: Spirulina and prior particle spirulina processed foods for human consumption and without other composition than spirulina. (IV) Spirulina compound food: Prior particle spirulina processed foods with other additives for human consumption, containing at least 50% of spirulina prior particle. (V) Spirulina compound processed food: Prior particle spirulina processed foods with other additives for human consumption, containing at least 30% but below 50% of spirulina prior particle.

Item Spirulina Foods Spirulina Compound Foods Spirulina Compound Processed Foods Phycocyanins Above 2000 mg/100 g Above 1000 mg/100 g Above 600 mg/100 g Chlorophyll a Above 500 mg/100 g Above 250 mg/100 g Above 150 mg/100 g Total carotenoids Above 100 mg/100 g Above 50 mg/100 g Above 30 mg/100 g

Item Max. Level (ppm) Lead 1.0 Cadmium 1.0 Mercury 0.5 Arsen 1.0 Note: This is calculated based on the fresh Spirulina containing moisture of 85%.

(V) Pesticide residue The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA. (VII) Other

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Maximum level of pheophorbide: Existing pheophorbide below 50 mg/100 g; total pheophorbide below 100 mg/100 g. 1. This specification applies only to Spirulina containing moisture below 70% (dry weight). 2. “mg/100 g” refers to the amount (mg) of Existing pheophorbide or total pheophorbide in every 100 g of product.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The content of spirulina shall be labeled.

V. Supplementation (I) Edible green algae refers to green algae and spirulina containing moisture below 7%. (II) Total pheophorbides refer to the sum of existing pheophorbides and chlorophyll amylase activity. (III) Heavy metals, arsenic, and pheophorbides are calculated by dry weight.

VI. Reference (1) 中華民國國家標準：「食用藍藻」之檢驗方法 CNS 14721 (2) 中華民國國家標準：「食品中水分」之檢驗方法 CNS 5033 (3) 中華民國國家標準：「食品中粗蛋白質」之檢驗方法 CNS 5035 (4) 中華民國國家標準：﹝食用綠藻﹞第 4.4 節,葉綠素 a 之測定 CNS 4204 (5) U.S. Food and Drug Administration Center for Food Safe and Applied Nutrition Office of Additive Safe ,October 6,2003;GRAS Notice No.GRN 000127. (6) U.S. Food and Drug Administration Center for Food Safe and Applied Nutrition Office of Additive Safe ,June 12,2002;GRAS Notice No.GRN 000127.

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16. Specification for Functional Foods Containing Anthocyanidins

I. Scope This specification applies to (1) fruit juice, beverages, and alcoholic products; and (2) capsule and tablet/pill foods containing anthocyanins.

II. Definitions Anthocyanins are the largest group of water-soluble pigments in flora and the major component of red, blue, and purple colors in fruit, vegetables, flowers and plant tissues or products. As a flavonoid, anthocyanin is a very important pigment in nature. It is different from other flavonoids, as it can form flavylium (2-phenylbenzopyrylium) cation. There are six common types of anthocyanins in plants: pelargonidin, cyanidin, delphinidin, peonidin, petunidin, and malvidin, where cyanidin is the most common anthocyanin ligand.

Product Form Anthocyanin Content (1) Fruit juice, beverages, and Above 70 ppm (cyanidin-3-glucoside equivalent) alcoholic products (2) Capsule and tablet/pill Above 3 % products

(III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the Food and Drug Administration (hereafter referred to as the FDA) and the latest version of regulations relating to food sanitation standards. (IV) Heavy metals The content shall comply with the relevant food sanitation standards. If no standards applicable, the maximum level of heavy metals shall be 20 ppm (by lead) and the maximum level of arsenic shall be 2 ppm. (V) Pesticide residue The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA. (VII) Other Alcoholic products shall also comply with the sanitation standards for alcohol.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The content of anthocyanin shall be labeled.

V. Supplementation 1. Fruit juice, beverages, and alcoholic products: Refer to AOAC Official Method 2005.02 Total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines. 2. Capsule and tablet/pill products: Manufacturers shall propose the test methods for experts to approve.

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VI. Reference (1) AOAC International. 2005. Total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines. In. “Official Methods of Analysis of AOAC International” 18th ed. 2005.02. Horwitz, H. ed. Washington DC. U.S.A. (2) Christine, A. W. and Renee, J. G. 2004. Anthocyanins and other flavonoids. Nat. Prod. Rep., 21: 539-573. (3) Dangles, O., Saito, N. and Brouillard, R. 1993. Anthocyanin intramolecular copigment effect. Phytochemistry. 34: 119-124. (4) Harborne, J. B. and Williams, C. A. 2000. Advances in flavonoid research since 1992. Phytochemistry. 55(6): 481- 504. (5) Mazza, G. and R. Brouillard. 1987. Colour stability and structural transformations of cyanidin 3,5-diglucoside and four 3-deoxyanthocyanins in aqueous solutions. J. Agric. and Food Chem. 35: 422-426. (6) Mazza, G. 1995. Anthocyanins in grapes and grape products. Crit. Rev. Food sci. nutr. 35(4): 341-371. (7) Mazza, G., Kay, C. D., Cottrell, T. and Holub, B. J. 2002. Absorption of anthocyanins from blueberries and serum antioxidant status in human subjects. J. Agri. Food. Chem. 50: 7731-7737. (8) Mazza, G., Juan, E. C. and Colin, D. K. 2004. Methods of analysis for anthocyanins in plants and biological fluids. JAOAC Int. 87(1): 129-145.

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17. Specification for Functional Foods Containing Proanthocyanidins

I. Scope This specification applies to beverages containing proanthocyanidins.

II. Definitions Proanthocyanidin is a polyphenol compound generally found in fruits such as grapes, cranberries, and blueberries. It is also called condensed tannins. It is formed by a flavan-3-ols unit and divided by the degree of polymerization into: monomeric, oligomeric, and polymeric proanthocyanidins. Proanthocyanidin with polymerization below 10 is often called oligomeric proanthocyanidin (OPC).

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The content of proanthocyanidin shall be labeled.

V. Supplementation Currently, there is no publicly recognized method for analyzing proanthocyanidins. The following method is recommended: Namiko, N., Tsuji, S. and Tosuhide T. 2003. Analysis of proanthocyanidins in grape seed extracts, health foods and grape seed oils. J. health sci. 49(1):45-54.

VI. Reference (1) 張雲評，2008，原花青素之心血管保健功效及萃取技術，食品工業︰40(12): 26-36. (2) Gu, L. Klem, M. A., Hammerstone, J. F., Beecher, G., Holden, J., Haytowitz, D., Gebhardt, S. and Prior, R. L. 2003. Concentrations of proanthocyanidins in common foods and estimations of normal consumption. J Nutr. 134(3): 613- 617. (3) Namiko, N., Tsuji, S. and Tosuhide, T. 2003. Analysis of proanthocyanidins in grape seed extracts, health foods and grape seed oils. J. Health Sci. 49(1): 45-54. (4) USDA，ARS. USDA database for the proanthocyanidin content of selected foods. http://www.nal.usda.gov/fnic/foodcomp(最後瀏覽日期︰2009 年 7 月) (5) Weber, H. A., Hodges, A. E., Guthrie, J. R., O’Brien, M. O., Robaugh, D. Clark, A. P., Harris, R. K., Algaier, J. W. and Smith, C. S. 2007. Comparison of proanthocyanidins in commercial antioxidants:grape seed and pine bark

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extracts. J. Agric. Food Chem. 55: 148-156.

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18. Specification for Functional Foods Containing Glucosamine

I. Scope This specification applies to tablet/pilll and liquid foods containing glucosamine hydrochloride.

II. Definitions Glucosamine, chemical structure 2-Amino-2-deoxy-D-glucose and molecular mass 179.17 Da, is an aminosugar. It is often distributed in nature in the form of mucopolysaccharides. Although rich glucosamine is found in crustaceans, chitin is difficult for human absorption. Industrially, natural glucosamine can be produced by acid hydrolysis or enzymatic hydrolysis of chitin or chitosan. It is commercially sold as a health food or additive. Based on the effect of human absorption, glucosamine falls into three categories: (a) glucosamine sulfate is the easiest to absorb and only used for medicinal purposes and not used as raw materials or finished products in food factory; (b) glucosamine free base without sodium; and (c) glucosamine hydrochloride. Without claiming treatment effect, both (b) and (c) are managed as foods.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The content of glucosamine hydrochloride shall be labeled. (III) Warning message for chitin extract products: “Not suitable for vegetarians/vegans and may cause allergy.”

V. Supplementation The results of this experience are the results of glucosamine hydrochloride.

VI. Reference (1) 蘇南雄、李敏雄(1998)，Listonella damsela NTU-FC-6 幾丁質酵素之生產與基本性質之探討，中國農業化學會誌，3(1):65-76。 (2) 美國家庭醫師學會(American Academy of Family Physicians, AAFP)，Alternative Therapies for Traditional Disease States: Osteoarthritis,2003. http://www.aafp.org/afp/20030115/339.html (最後瀏覽日期 98 年 8 月) (3) AOAC Official Method 2005.01 Glucosamine in Raw Materials and Dietary Supplements Containing Glucosamine TQF Specifications and Standards for 44 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

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Sulfate and/or Glucosamine Hydrochloride High-Performance Liquid Chromatography with FMOC-Su Derivatization. (4) Jong, C. Y. Studies on Toxicity of Glucosamine in Rats, National Taiwan Ocean University Department of Food Science NTOU, Taipei, 2001. (5) Leu, S. 1998. The prevention and cure of deformability osteoarthritis, NewFood Industry, 40: 11-15. (6) Morelli, V., Naquin, C. and Weaver, V. 2003. Alternative Therapies for Traditional Disease States: Osteoarthritis. Am Fam Physician. 67: 339-44. (7) The United States Pharmacopeia Convention, 2009. The Pharmacopeia of United States of America 32 United States Pharmacopeia Convention,Inc., Rockville,U.S.A.

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19. Specification for Functional Foods Containing Fructans

I. Scope This specification applies to foods containing fructans.

II. Definitions Fructan is a polymer often found in Asterales, Liliales, Poales, Campanulales, Ericale, and Dipsacales for energy storage. Scholars found that fructan is often formed by one glucose linked with multiple fructose, with a degree of polymerization up to 200 in plants and 100,000 in bacteria. However, there is also fructan containing only fructose. Fructans by bonding are divided into three types: inulin type, levan type, and graminan type. Currently, fructans in food products are mainly extracted from chicory, and some from jerusalem artichoke. This is because both plants contain rich fructans (approx. 15-20%), thus becoming the main source of fructan products. Specific foods, such as onions, burdocks, asparagus, chives, bananas, and wheats also contain rich fructans.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The content of fructans shall be labeled.

V. Supplementation Refer to AOAC Official Method 999.03 Measurement of Total Fructan in Foods - Enzymatic /Spectrophotometric Method

VI. Reference (1) AACC. 2001. AACC Report. The definition of dietary fiber. Cereal Food World. 46(3): 112-126. (2) AOAC International. 2003. Measurement of Total Fructan in Foods - Enzymatic/ Spectrophotometric Method. In. “Official Methods of Analysis of AOAC International” 17th ed. 999.03. William Horwitz ed. Gaithersburg, MD. U.S.A. (3) Flamm, G., Glinsmann, W., Kritchevsky, D., Prosky, L. and Roberfroid, M. 2001. Inulin and oligosaccharide as dietary fiber: a review of the evidence. Crit. Rev. Food. Sci. Nutri. 41(5): 353-362. (4) Frank, A. 2002. Technological functionality of inulin and oligofructose. Brit. J. Nutr. 87: 287-291. (5) Hipsley, E. H. 1953. Dietary ”fibre” and pregnancy toxaemia. Br Med J. 2: 420-422. (6) Letexier, D., Diraison, F. and Beylot, M. 2003. Addition of inulin to moderately high-carbohydrate diet reduces TQF Specifications and Standards for 46 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

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hepatic lipogenesis and plasma triacylglycerol concentrations in humans. Am. J. Clin. Nutr. 77: 559-564. (7) Pool-Zobel, B. L. 2005. Inulin-type fructans and reduction in colon cancer risk: review of experimental and human data. Brit. J. Nutr. 93: S73-S90. (8) Roberfroid, M. 1993. Dietary fiber, inulin and oligofructose: a review comparing their physiological effects. Crit. Rev. Food. Sci. Nutr. 33(2): 103-148. (9) Roberfroid, M. B. and Delzenne, N. M. 1998. Dietary fructans. Annu. Rev. Nutr. 18: 117-143. (10) Tungland, B. C. and Meyer, D. 2002. Nondigestible oligo-polysaccharides dietary fiber: Their physiology and role in human health and food. Compr. Rev. Food. Sci. Food Safety. 3: 73-77.

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20. Specification for Functional Foods Containing Cereal β-Glucans

I. Scope This specification applies to foods and beverages containing cereal β-glucans.

II. Definitions (I) β-glucans in barley or oat is formed by β-1→3 and β-1→4 linked with glucose. (II) Foods processed with cereals, including oat, barley, rye, and other cereals.

Product Form Content of β-Glucan (1) Solid Above 2 g/100 g (2) Liquid Above 200 mg/100 mL

(III) Microbial limits The content of microorganisms shall comply with the “Act Governing Food Safety and Sanitation” by the Food and Drug Administration (hereafter referred to as the FDA) and the latest version of regulations relating to food sanitation standards. (IV) Heavy metals The content shall comply with the relevant food sanitation standards. If no standards applicable, the maximum level of heavy metals shall be 20 ppm (by lead) and the maximum level of arsenic shall be 2 ppm. (V) Pesticide residue The pesticide residue of all ingredients shall comply with the latest “Standards for Pesticide Residue Limits in Foods” promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The content of β-glucan shall be labeled.

V. Supplementation (I) Solid products: Refer to AOAC Official Method 995.16 (II) Liquid products: Refer to EBC (European brewery convention) Methods 8.13.1

VI. Reference (1) AOAC Official Method 995.16. 1995. β-D-glucan in barely and oats. (2) EBC 8.13.1. High molecular weight beta-glucan content of wort: enzymatic method.

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21. Specification for Functional Foods Containing Green Algae

I. Scope This specification applies to foods containing green algae.

II. Definitions (I) Green algae Genera Chlorella include Chlorella vulgaris and Chlorella sorokiniana, with the size about 2-10 µm. They are unicellular in a spherical or elliptical shape with a cell wall. (II) Green algae powder Green alga powder produced by hot-air drying and spray drying of Genera Chlorella cultured by artificial photosynthesis or non-photosynthesis. (III) Green algae foods Foods processed with a minimum of 95% of green algae original powder. (IV) Green algae compound foods Foods processed with a minimum of 50% of green algae original powder. (V) Green algae compound processed foods Foods processed with at least 30% but below 50% of green algae original powder.

Green Algae Green Algae Compound Item Green Algae Foods Compound Foods Processed Foods Protein Above 50 g/100 g Above 25 g/100 g Above 15 g/100 g Chlorophyll Above 1500 Above 750 mg/100 g Above 450 mg/100 g mg/100 g Vitamin B2 Above 2 mg/100 g Above 1 mg/100 g Above 0.6 mg/100 g Iron Above 10 mg/100 g Above 5 mg/100 g Above 3 mg/100 g Green Algae Hot Above 1.5 Above 1.5 Above 1.5 Water Extract Indicative value: OD260 nm

Item Max. Level (ppm) Lead 1.0 Cadmium 1.0 Mercury 0.5 Arsen 1.0 Note: This is calculated based on the fresh green algae containing moisture of 85%.

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promulgated by the FDA. (VI) Packaging Packaging shall comply with the latest “Sanitation Standard for Food Utensils, Containers and Packages” promulgated by the FDA. (VII) Other Maximum level of pheophorbides (reduced in proportion with green alga foods, green algae compound foods, and green alga compound processed foods: 1. Existing pheophorbides: Below 60 mg/100 g, 30 mg/100 g, and 18 mg/100 g 2. Total pheophorbides: Below 80 mg/100 g, 40 mg/100 g, and 24 mg/100 g 3. This specification applies only to green algae containing moisture below 70% (dry weight). 4. “mg/100 g” refers to the amount (mg) of Existing pheophorbide or total pheophorbide in every 100 g of product. 5. Where there are inconsistencies between this specification and current laws and regulations, the latter shall prevail.

IV. Labeling (I) Labeling shall comply with the “Act Governing Food Safety and Sanitation” and the latest version of the relevant laws and regulations. (II) The content of green algae shall be labeled.

V. Test Methods Refer to CNS 4202, N5134 Method for Test of “Edible Green Algae”.

VI. Supplementation (I) Edible green algae refer to green algae and spirulina containing moisture below 7% and complying with the “Edible Alga Sanitation Standards” and other regulations for food sanitation management. (II) Total pheophorbides refer to the sum of existing pheophorbides and chlorophyll amylase activity. (III) Heavy metals, arsenic, and pheophorbides are calculated by dry weight. (IV) Test methods of related items

Item Method Moisture CNS 5033 Methods for Test of Moisture in Food Crude protein CNS 5035 Methods for Test of Crude Protein in Food Crude ash CNS 5034 Methods for Test of Crude Ash in Food

VII. Reference (1) 日本健康營養食品協會：健康補助食品規格基準集。 (2) 經濟部標準檢驗局：包裝食品標示 CNS 3192。 (3) 中華民國國家標準 CNS 4202, N5134「食用綠藻」之檢驗方法。 (4) 中華民國國家標準 CNS 5033「食品中水分」之檢驗方法。 (5) 中華民國國家標準 CNS 5035「食品中粗蛋白質」之檢驗方法。 (6) 中華民國國家標準 CNS 5034「食品中粗灰分」之檢驗方法。

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Functional Food (Mushroom and Fermented) Ingredients Review Form (for the special requirements of microorganisms only) -Basic Data- Company name: Personel in charge: Title: Address: Contact person: Title: Phone: Fax: Email: -Background Data- I. Sources of ingredients (hereafter referred to as “ingredient”) for mushroom or fermented foods: Domestic (Self-produced Purchased) Import II. Role of ingredient in the final product: Main ingredient Secondary ingredient Food additive Others III. Variety and manufacture of ingredient: Mushroom Foods (go to (1)) Fermented Foods (go to (2)) (1) Mushroom Foods:

Scientific Name: Common Name:

Name

Bacteria Bacteria

Ecological collection of sporocarps Artificial culture of sporocarps (Natural Artificial cultured) (Wood block Plastic bag Soil Others)

Manufacture Wood-based (Tree: ) Soil-based (Soil: ) Insect-based (Insect: ) Symbiosis (Host: )

Substrate Please specify: Place, time and environment of collection or the record of culture (including culture

composition and pesticide, if used), and so on.

Others

(2) Fermented Foods: (Note: Fermentation of mushroom mycelia is also an ingredient of fermented function foods)

Scientific Name: Common Name:

Name

Bacteria Bacteria Pure bacterial fermentation Compound bacteria fermentation

Solid culture Liquid culture

Manufacture Use of ordinary substrates in fermentation Use of other functional elements in (The fermentation substrate is not functional) fermentation

Substrate (Functional elements such as medicinal herbs) Ingredients contain viable Ingredients contain no viable

Viable

Please specify:

Others

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TQF Specifications and Standards for Product Test Items TQF-PST-001

IV. Form of ingredient during warehousing: (not final product form) Liquid (Extract Fermented solution Solid (Original Slice Powder) Concentrated solution V. Final processing of ingredients: (Please specify the manufacturing process of ingredients, such as powdering sporocarps or spray drying after extraction, purely fermented mycelia or filtered purely fermented solution or direct freeze drying of culture solution, etc.)

VI. Status of application for health food certification in Taiwan of the ingredient’s derivatives Approved (Certification items: ) Pending (Certification items: ) Preparation (Certification items: ) Not applied for VII. References (Please attach supporting data for assessment, such as analysis data, bacterial source or verification data, and safety and function effectiveness data issued by a credited unit, and published literature, etc.)

TQF Specifications and Standards for 52 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

Item Specifications of Functional Ingredients of Mushroom Sporocarps I. Background Data of Mushroom Sporocarps of Ingredients 1.1. Name of microorganism Common name: Scientific name: (Yes No Certificate No.: ) 1.2. Strain source (please specify):

1.3. Strain safety: GRAS strain Traditional edible strain Newly developed strain (Yes No Safety Certificate No.: ) Others II. Background Data of Functional Composition (or special nutrients) in Ingredients 2.1. Indicative functional constituent: No Yes (name of constituent ) 2.2. References of function effectiveness: No Yes (Ancient books Related literature Self-developed) III. Sources of Sporocarp Ingredients 3.1 Self-produced (Purchased starter Self-prepared starter) (Answer items 4 and 5) 3.2 Purchased (Domestic Import) (Answer item 5) IV. Culturing Process of Self-Produced Ingredients(skip for purchased ingredients) 4.1. Starter control items 4.1.1. Purchased starter acceptance criteria (skip for self-prepared starters) 4.1.1a. Strain name confirmation: No Yes (Document of proof: ) Test method: (Acceptance criteria: ) 4.1.1b. Strain activity assessment: No Yes (Document of proof: ) Test method: (Acceptance criteria: ) 4.1.1c. Other test items: (Document of proof: ) Test method: (Acceptance criteria: ) 4.1.1d. Sampling plan: No Yes (If yes, outline the plan)

4.1.2. Quality control for self-prepared starters (skip for purchased starters) 4.1.2a. Strain storage: Liquid nitrogen -80℃  Sterile water Subculture Others ( ) 4.1.2b. Strain verification: No Yes (Document of proof: ) Test method: (Qualification criteria: ) 4.1.2c. Activity assessment: No Yes Test method: (Qualification criteria: ) 4.1.2d. Other test items Test method: (Qualification criteria: ) 4.1.2e. Supplementary description:

4.2. Control items in culturing process 4.2.1. Culture Process SOP No Yes (Document No.: ) 4.2.2. Test items and methods in the culture process Mycelium culture quantity: Test method: pH value: Test method: Sporocarp appearance Test method: Indicative constituent 1: Test method: TQF Specifications and Standards for 53 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

Indicative constituent 2: Test method: Other: Test method: 4.2.3. Test items at different stages 4.2.3a. Seed culture stage Mycelium culture quantity: pH value Others (please specify):

4.2.3b. Culture stage Substrate verification (e.g. heavy metal, drug, and pesticide residues in plastic bags) Sporocarp appearance: Indicative constituent:

Other (Please specify):

4.2.4. Indicators at yield end-point and qualification criteria Sporocarp appearance: Qualification criteria: Indicative constituent: Qualification criteria: Other (Please specify): Qualification criteria:

4.3. Subsequent harvesting and processing processes 4.3.1. Processing SOP No Yes (Document No.: ) 4.3.2. Test items and methods of processing process Indicative constituent: Other: Test method:

4.3.3. Indicators at processing end-point and qualification criteria Indicative constituent: Qualification criteria: Other: Qualification criteria:

4.3.4. Ingredient form at processing end-point Liquid (Extract Concentrate) Solid (Original Slice Powder Nanoscale) V. Warehousing Criteria and Temporary Storage of Ingredients 5.1. Temporary Storage SOP No Yes (Document No.: ) Temperature ( ℃) Humidity ( ％～％) Storage Time ( ) 5.2. Warehousing (or Acceptance) Criteria 5.2.1. General Food Tests Plate count Heavy Metals Pesticide Residue 5.2.2. Strain confirmation Yes (Test method: Qualification criteria: ) No 5.2.3. Indicative constituent Yes (Test method: Qualification criteria: ) No 5.2.4. Other (Test method: Qualification criteria: )

TQF Specifications and Standards for 54 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

5.3. Post-processing Process of Ingredients 5.3.1. Processing SOP No Yes (Document No.: ) 5.3.2. Test items and methods of processing process Indicative constituent: Other: Test method:

5.3.3. Indicators at processing end-point and qualification criteria Indicative constituent: Qualification criteria: Other: Qualification criteria:

5.3.4. Ingredient form at processing end-point: Liquid (Extract Concentrate) Solid (Original Slice Powder Nanoscale) VI. Other

TQF Specifications and Standards for 55 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

Item Specifications of Fermented Functional Ingredients (Note: Fermentation of mushroom mycelia is also an ingredient of fermented functional foods) I. Background Data of Mushroom Sporocarps of Ingredients 1.1. Name of microorganism Common name: Scientific name: (Yes No Certificate No.: ) 1.2. Strain source (please specify):

1.3. Strain safety: GRAS strain Traditional edible strain Newly developed strain (Yes No Safety Certificate No.: ) Others II. Background Data of Functional Composition (or special nutrients) in Ingredients 2.1. Indicative functional constituent: No Yes (name of constituent ) 2.2. References of function effectiveness: No Yes (Ancient books Related literature Self-developed) III. Sources of Sporocarp Ingredients 3.1 Self-produced (Purchased starter Self-prepared starter) (Answer items 4 and 5) 3.2 Purchased (Domestic Import) (Answer item 5) IV. Culturing Process of Self-Produced Ingredients (skip for purchased ingredients) 4.1. Starter control items 4.1.1. Purchased starter acceptance criteria (skip for self-prepared starters) 4.1.1a. Strain name confirmation: No Yes (Document of proof: ) Test method: (Acceptance criteria: ) 4.1.1b. Strain viable count: No Yes (Document of proof: ) Test method: (Acceptance criteria: ) 4.1.1c. Strain activity assessment: No Yes (Document of proof: ) Test method: (Acceptance criteria: ) 4.1.1d. Other test items: (Document of proof: ) Test method: (Acceptance criteria: ) 4.1.1e. Sampling plan: No Yes (If yes, outline the plan)

4.1.3. Quality control for self-prepared starters (skip for purchased starters) 4.1.2a. Strain storage: Liquid nitrogen -80℃  Sterile water      Subculture Others ( ) 4.1.2b. Strain verification: No Yes (Document of proof: ) Test method: (Qualification criteria: ) 4.1.2c. Viable count: No Yes Test method: (Qualification criteria: ) 4.1.2d. Activity assessment: No Yes Test method: (Qualification criteria: ) 4.1.2e. Other test items Test method: (Qualification criteria: ) 4.1.2f. Supplementary description:

4.2. Fermentation control items 4.2.1. Fermentation SOP □No □Yes (Document No.: ) TQF Specifications and Standards for 56 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

4.2.2. Test items and methods of fermentation Bacteria quantity: Test method: pH value: Test method: Residual sugar: Test method: Dissolved oxygen value: Test method: Indicative constituent 1: Test method: Indicative constituent 2: Test method: Other: Test method: 4.2.3. Test items at different stages 4.2.3a. Seed culture stage Bacteria quantity: pH value: Other (Please specify): 4.2.3b. Production stage Bacteria quantity: pH value: Residual sugar: Dissolved oxygen value: Indicative constituent: Other (Please specify):

4.2.4. Indicators at fermentation end-point and qualification criteria Bacteria quantity: Qualification criteria: pH value: Qualification criteria: Indicative constituent: Qualification criteria: Other (Please specify): Qualification criteria:

4.3. Subsequent Recovery Processing Process 4.3.1. Processing SOP No Yes (Document No.: ) 4.3.2. Test items and methods of processing process Bacteria quantity pH value Indicative constituent 1 Indicative constituent 2 (same as 4.2.2.) Other: Test method:

4.3.3. Test Items of Different Stages Bacteria quantity: Indicative constituent: Other:

4.3.4. Indicators at processing end-point and qualification criteria Bacteria quantity: Qualification criteria: Indicative constituent: Qualification criteria: Other: Qualification criteria:

4.3.5. Ingredient form at processing end-point: With viable Without viable Liquid (Extract Concentrate) Solid (Block Powder Nanoscale) V. Warehousing Criteria and Temporary Storage of Ingredients 5.1. Temporary Storage SOP No Yes (Document No.: ) Temperature ( ℃) Humidity ( ％～％) Storage Time ( )

5.2. Warehousing (or Acceptance) Criteria

TQF Specifications and Standards for 57 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]

TQF Specifications and Standards for Product Test Items TQF-PST-001

5.2.1. General Food Tests Plate count Heavy Metals Others:

5.2.2. Strain confirmation Yes (Test method: Qualification criteria: ) No

5.2.3. Indicative constituent Yes (Test method: Qualification criteria: ) No

5.2.4. Other (Test method: Qualification criteria: )

5.3. Post-processing Process of Ingredients 5.3.1. Processing SOP No Yes (Document No.: ) 5.3.2. Test items and methods of processing process Bacteria quantity pH value Indicative constituent 1 Indicative constituent 2 (same as 4.2.2.) Other: Test method:

5.3.3. Test items at different stages Bacteria quantity: Indicative constituent: Other:

5.3.4. Indicators at processing end-point and qualification criteria Bacteria quantity: Qualification criteria: Indicative constituent: Qualification criteria: Other: Qualification criteria:

5.3.5. Ingredient form at processing end-point: With viable Without viable Liquid (Extract Concentrate) Solid (Block Powder Nanoscale) VI. Other

TQF Specifications and Standards for 58 / 58 © Taiwan Quality Food Association Product Test Items Ver. 2.0 [Functional Foods]