1FINAL DRAFT  
STANDARDS FOR ETHNIC FLOUR–BASED CONFECTIONERIES ( POLVORON, 
PIAYA, AND BARQUILLOS ) 
 
1. SCOPE 
 
This standard shall apply to ethnic flour-based confecti oneries, specifically polvoron, 
piaya, and barquillos , in suitable packaging materials or containers. 
 
2. DEFINITION OF TERMS 
 
For the purpose of this standard, the following terms sh all mean: 
 
Confectionery - a group of food items primarily made of sugar and ot her 
sweeteners, and includes candies, caramels, toffees, and choc olate bars.  It is a 
generic term for sweetened food products.  Sugar confecti onery refers to products 
such as sweets, candy and chocolates.  These products are sh elf-stable and usually 
have water activity below 0.85. (Dictionary of Food S cience and Technology, 
International Food Information Service, Blackwell Pub lishing, UK, 2005.)  
 
Container - any form of packaging material, which completely or par tially encloses 
the food (including wrappers). A container may enclose  the food as a single item or 
several units or types of prepackaged food when such is pre sented for sale to the 
consumer. 
 
Current Good Manufacturing Practices (cGMP) - a quality assurance system 
aimed at ensuring that products are consistently manufactu red, packed or repacked 
or held to a quality appropriate for the intended u se. It is thus concerned with both 
manufacturing and quality control procedures.  
 
Flavor and flavoring substances - substances which are added to impart flavor 
which are either natural, nature identical or artifici al flavoring substances (A.O. No.  288-B s. 1984; Rules and Regulations governing the Lab eling of Prepackaged Food 
Products distributed in the Philippines). 
(a) natural flavor  
flavoring substances derived through appropriate physica l processes from spices, 
herbs, fruit or fruit juices, vegetable or vegetable j uices, edible yeast, bark, bud, root, 
leaf or plant materials, meat, fish, poultry, eggs, d airy products or fermentation 
products thereof. 
(b) nature-identical flavoring substances  
substances chemically derived from aromatic materials or o btained synthetically, 
which are chemically identical to substances present in' natural products intended for 
human consumption. 
(c) artificial flavoring substances  
substances that impart flavor but which have not been i dentified in natural products 
or natural sources of flavorings. 
 
Flour - a powdery substance that is the product of grinding or mi lling of cereal grains 
(such as but not limited to wheat, rice, barley, and ma ize), root crops and legumes. 
 
Food - any processed substance which is intended for human consump tion and 
includes drink for man, beverages, chewing gum and any su bstances which have 
been used as an ingredient in the manufacture, prepar ation or treatment of food. 
(RA 9711 Food and Drug Administration (FDA) Act of 20 09). 
 
Food additives - any substance the intended use of which results or may 
reasonably be expected to result, directly or indirectl y, in its becoming a component 
or otherwise affecting the characteristics of any food (in cluding any substance 
intended for use in producing, manufacturing, packing, p rocessing, preparing, 
treating, packaging, transporting, or holding food; an d including any source of 
radiation intended for any such use), if such substance is not generally recognized, 
among experts qualified by scientific training and exp erience to evaluate its safety, 
as having been adequately shown through scientific proce dures to be safe under the 
conditions of the intended use (R.A. 3720. Food, Drug and Cosmetic Act).    3 
Food and Drug Administration or FDA  - formerly known as Bureau of Food and 
Drug (BFAD) of the Department of Health (DOH); which  was renamed in accordance 
to RA 9711 (Food and Drug Administration (FDA) Act of  2009). 
 
Food standard - a regulatory guideline that defines the identity of a given food 
product (i.e. its name and the ingredients used for i ts preparation) and specifies the 
minimum quality factors and, when necessary, the require d fill of the container. It 
may also include specific labeling requirements other tha n or in addition to the 
labeling requirements generally applicable to all pre packaged foods. 
 
Ingredient  - any substance including food additive, used as a compon ent in the 
manufacture or preparation of a food and present in the final product in its original or 
modified form. 
 
Label - includes any tag, brand, mark, pictorial, or other descri ptive script, written, 
printed, marked, embossed or impressed on, or attached to the container. 
 
Labeling - any written, printed or graphic matter (1) upon any article or any of its 
container or wrappers and/or (2) accompanying the package d food. 
 
Lot - food produced during a period of time and under more  or less the same 
manufacturing condition indicated by a specific code. 
 
Milk - the normal mammary secretion of milking animals obtaine d from one or more 
milkings without either addition to it or extraction f rom it, intended for consumption as 
liquid milk or for further processing (CODEX STAN 206-1 999). 
 
Milk product - a product obtained by any processing of milk, which may co ntain 
food additives, and other ingredients functionally ne cessary for the processing 
(CODEX STAN 206-1999). 
  4Muscovado sugar 
a non-centrifugal sugar which is primarily composed of su crose and molasses. It 
includes three forms, namely: amorphous; lump or molded  locally known as 
“panocha”  or “ sangkaka ”; and thick, concentrated syrup locally known as “ inuyat”. 
(BAFPS Draft Standards for Muscovado Sugar)   
 
Packaging - the process of packing that is part of the production cycle  applied to a 
bulk product to obtain the finished product. It also ref ers to any material, including 
painted material, employed in the packaging of a prod uct including any outer 
packaging used for transportation of shipment. Packaging  materials are referred to 
as primary or secondary according to whether or not they are intended to be in direct 
contact with the product. 
 
Rancidity - formation of off-flavors in food due to lipid oxidat ion (oxidative rancidity) 
and/or release of free fatty acids by lipolysis (hydrol ytic rancidity). 
 
Water activity - the ratio of vapor pressure of water in the food substr ate to the 
vapor pressure of pure water at the same temperature ( Jay et. al. , 2005). It is also a 
measure of water available for chemical reactions and m icrobial growth (Fennema, 
1996). 
 
3. DESCRIPTION OF PRODUCTS 
 
3.1 Product Definition 
Ethnic flour-based confectioneries are confectionery prod ucts with flour and sugar as 
basic ingredients. The ethnic flour-based confectionerie s specifically covered by this 
standard are the following: 
 
3.1.1 Polvoron –The product is usually made from a mixture of toasted flour, sugar, 
butter and milk or milk products. Other ingredients such as pinipig, chocolate and 
nuts could be added. The product mixture may be formed  into oval or round shapes 
or by using appropriate polvoron molders.   5 
3.1.2 Piaya  – Piaya  is a traditional flat, disc- shaped unleavened pastry  with 
muscovado sugar filling. Other ingredients such as but not  limited to purple yam and 
pandan could also be added. 
 
3.1.3 Barquillos  – Barquillos  may also be called as wafer rolls. The product is a 
wafer with thin and delicate texture, traditionally made using a special wafer iron 
(barquillera ) or baked in an oven, and then immediately rolled to form cylinders. 
Barquiron  is a variant of barquillos  with polvoron filling. 
 
3.2 Process Description 
The product shall have undergone a process sufficient to ensure quality and shelf 
life stability at ambient conditions and shall be packed  in any suitable packaging 
material and/or container.  
 
4 ESSENTIAL COMPOSITION AND QUALITY FACTORS 
 
4.1  Raw Materials 
  
4.1.1 Basic Ingredients 
(a) Flour – May come from cereal grains, root crops, or  legumes, and must conform 
to RA 8976 (Philippine Food Fortification Act of 2000 ) (for wheat flour), and other 
applicable food standards. 
 
(b) Sugar and other sweeteners –  May include table sugar, muscovado sugar, corn 
syrup, and other similar food items, and must conform to  RA 8976 (Philippine Food 
Fortification Act of 2000) (for refined sugar), and other applicable food standards. In 
some cases, sweetened condensed milk and similar milk produ cts also act as the 
source of sugar and/or sweetener.  
 
(c) Milk and milk products – Applicable as basic ingredient only for polvoron. Must 
conform to requirements prescribed by PNS/BAFPS 36:2008  (Philippine National  6Standard for Fresh Milk), FDA A.O. No. 132 s. 1970 (Re gulation Prescribing the 
Standard of Identity and Quality of Milk and Milk Pro ducts, B-4.12-01), and other 
applicable food standards.  
 
(d) Butter, margarine, lard, oil, and other similar food items  – Applicable as basic 
ingredient for polvoron and piaya. Must comply with RA 8976 (Philippine Food 
Fortification Act of 2000) (for cooking oil), FDA A.O. No. 230 s. 1974 (Regulation: B-
4 Definition and Standards for Food; B-4.15 Fats and Oils, B-4.15-01 Shortening), 
FDA A.O. No. 243 s. 1975 (Regulation: B-4 Definition  and Standards of Food; B-
4.18 Margarine), and other applicable food standards . 
 
4.1.2 Optional Ingredients 
(a) Eggs  –Must be fresh eggs, and must comply with the requirem ents prescribed by 
PNS/BAFPS 35: 2005 (Philippine National Standards f or Table Egg), and other 
applicable food standards. Other forms of processed eggs can be used. 
 
(b) Fruit, vegetables, nuts, and root crops – May include  fresh items or preserves 
and must conform to all applicable food standards 
 
(c) Flavor and Flavoring Substances - All flavor/flavor ing substances as defined in 
subsection 2.4 shall be food grade, and must comply with  FDA’s B.C. No. 016 s.2006 
(Updated List of Food Additives). 
 
(d) Potable Water – Must be water fit for human consum ption and potability 
determined by health authorities as cited in Philippin e National Standards for 
drinking water (Department of Health A. O. No. 2007- 0012. Philippine National 
Standards for Drinking Water 2007). 
 
(e) Sesame Seeds – Must conform to applicable food stand ards. 
 
4.2 Quality Criteria 
        7     4.2.1 General Requirements 
 
(a) Water Activity – The water activity (a W) for polvoron, piaya  and barquillos should 
not be greater than 0.60 at 25 oC. 
     
  (b) Microbiological limits  
The microbiological limits of ethnic flour-based confec tioneries shall conform to the 
standards set for Processed Foods, under the food descriptio n Breakfast Cereals 
and Snack Foods (Bureau Circular No. 1-A, 2004; Guidel ines for the Assessment of 
Microbiological Quality of Processed Foods). The microbiol ogical limits of ethnic 
flour-based confectioneries shall be as follows*: 
      Table 1. Microbiological limits of ethnic flour-based confec tioneries 
Test Microorganism n C m M 
Yeast and Molds, cfu/g 5 2  10  10 3 
Coliforms, MPN/g 5 2 10 10 2 
  n – number of samples tested 
  c – maximum number of samples > m but  not more t han M 
m – guide level 
  M – Maximum acceptance level 
 
4.2.2 Types of Defects 
(a) Odor/flavor/color 
A sample unit affected by objectionable odors, flavors a nd colors which are 
indicative of rancidity and yeast and mold growth. 
(b) Presence of yeasts and molds 
The presence of yeasts and molds may signify spoilage an d may be due to improper 
handling of raw materials and finished products during processing, storage, and 
distribution. 
(c) Foreign matter 
The presence in the sample unit of any matter, which h as not been derived from 
ingredients or processing aids used, does not pose a threa t to human health and is 
readily recognized without magnification or is present at a level determined by 
magnification method or any equivalent methods that i ndicates non-compliance with 
good manufacturing practices and sanitation practices. 
  84.2.3 Classification of “Defectives” 
A container that has any of the type of defects set in 4 .2.2 shall be considered as 
“defective”. 
 
4.2.4 Lot Acceptance 
A lot shall be considered as meeting the applicable qua lity requirements when the  
number of “defectives”, as defined in sub-section 4.2.3,  does not exceed the  
acceptance number of the appropriate sampling plan. 
 
5 FOOD ADDITIVES 
 
 
5.1 Food additives when used shall be in accordance with the regulations 
established by the Food and Drugs Administration (FDA)  (Bureau Circular No. 016 
s.2006. Updated List of Food Additives) and/or the Cod ex Alimentarius Commission.  
 
The following food additives listed in, but not limit ed to, Table 2, may be used for the 
manufacture of ethnic flour-based confectioneries. 
 
5.2 All others that have not been included in the abo ve list shall be allowed as carry-
over provided they are approved by FDA regulation (B .C. No. 016 s. 2006; Updated 
List of Food Additives) and shall be in accordance to th e Section 4 of the Preamble 
of the General Standard for Food Additives (GFSA) (C odex Stan 192-1995, Rev. 5 
(2004)).  These additives include those that are used for the raw materials and other 
ingredients.  
 
 Table 2. Food Additives for Ethnic flour-based confectioneri es ( polvoron, piaya, 
 and barquillos ) (B.C. No.016 s. 2006. Updated List of Food Additive s)  
Function  Food Additive  Max. level of usage  
Anti-caking agent Polydimethylsiloxane 10 mg/kg b 
Antioxidant Ascorbyl Esters 200 mg/kg (As ascorbyl stearate) d 
BHA 100 mg/kg (Fat or oil basis) b 
200 mg/kg (Fat or oil basis)  b 
50 mg/kg c 
BHT 200 mg/kg (Fat or oil basis)  b 
50 mg/kg c 
25 mg/kg d  9Gallate, propyl 200 mg/kg (Fat or oil basis)  b 
Tertiary Butylhyroquinone 200 mg/kg (Fat or oil bas is)  b 
200 mg/kg c 
Tocopherols 500 mg/kg (Fat or oil basis) b 
Color Allura Red AC 348 mg/kg b 
Amaranth 100 mg/kg  b 
Annatto Extracts 25 mg/kg (As total bixin or norbix in) b 
Brilliant Blue FCF 300 mg/kg b 
Caramel Colour, Class III GMP  b 
GMP  c 
GMP  d 
Caramel Colour, Class IV GMP  b 
GMP  c 
Carotenoids GMP  c 
200 mg/kg  d 
Fast Green FCF 100 mg/kg  b 
100 mg/kg  d 
Sunset Yellow FCF 400 mg/kg b 
Preservative Benzoates 1500 mg/kg (As benzoic acid)   b 
Hydroxybenzoates, p- 2000 mg/kg (As p-hydroxybenzoi c acid)  
b 
300 mg/kg (As p-hydroxybenzoic acid)  
d 
Stabilizer Polysorbates 10000 mg/kg  b 
5000 mg/kg (Flour basis) c 
5000 mg/kg  d 
Sorbates 2000 mg/kg (As sorbic acid) b 
Sorbitan Esters of Fatty Acids 20000 mg/kg a 
5000 mg/kg (Flour basis) c 
10000 mg/kg d 
Sweetener Alitame 300 mg/kg a 
Acesulfame Potassium 3500 mg/kg b 
Aspartame 10000 mg/kg b 
5000 mg/kg d 
Saccharin 3000 mg/kg  b 
2000 mg/kg d 
Sucralose 1500 mg/kg  b 
a Based on the Food Category System: 5.0 Confectioner y;  
b Based on the Food Category System:5.2 Sugar-based confectionery including hard and soft candy, 
nougats, etc. other than food categories  05.1, 05.3, and 05.4;  
c Based on the Food Category System: 7.1.2. Crackers,  excluding sweet crackers  
d Based on the Food Category system: 7.2 Fine bakery wares 
 
6  CONTAMINANTS  
  10  The products covered by this standard shall comply with the maximum limits for 
contaminants and the maximum residue limits for pesticid es and veterinary drugs 
established by the Codex Alimentarius Commission. 
 
7 HYGIENE 
 
7.1    It is recommended that the product covered by the pr ovisions of this standard 
be prepared and handled in accordance with the approp riate sections of the 
Recommended International Code of Practice – General Principles of Food Hygiene 
(CAC/RCP 1 – 1969, Rev. 4-2003), Code of Hygienic Pr actice for Milk and Milk 
Products (CAC/RCP 57-2004), Recommended Internationa l Code Of Hygienic 
Practice for Egg Products CAC/RCP 15-1976 (amended 1978 , 1985) , and/or the 
FDA A.O. No. 153 s. 2004 - Guidelines, Current Good M anufacturing Practices in 
Manufacturing, Packing, Repacking or Holding Food 
 
7.2    When tested by appropriate methods of sampling and  examination, the product: 
-    shall be free from filth that may pose a hazard  to health; 
-     shall be free from parasites which may represent a hazard to health; 
-  shall not contain any substance originating from micro organisms in amounts  
  which may represent a hazard to health; 
-  shall be free from microorganisms capable of develop ment under normal  
  conditions of storage; and 
-     shall be free from container integrity defects wh ich may compromise the hermetic  
     seal. 
 
8 PACKAGING AND LABELING 
 
 
8.1    The packaging used for ethnic flour-based confectioneri es should be made of 
suitable food-grade materials, and should be clean, an d hygienic. The packaging 
materials used should not adversely affect product qualit y and safety.  
  11  The product must also be packed and sealed in suitable co ntainers that will provide 
additional protection against contamination. The packagi ng material used should be 
able to withstand mechanical, chemical and thermal stresse s encountered during 
normal product handling and distribution. 
  
8.2  The net weight of any sample unit shall be in accordan ce with FDA’s B.C. No. 6-
A s. 1998 (Permissible Net Content Variation in Prepacka ged Food). 
 
8.3 Labeling 
Each container shall be labeled and marked with the fol lowing information in 
accordance with FDA’s Labeling Regulation (A.O. 88-B s. 1984; Rules and 
Regulations governing the Labeling of Prepackaged Food  Products distributed in the 
Philippines): 
(a) The name of the product shall be known as Polvoron (Molded  Toasted Flour with 
Butter), Piaya (Native Pastry with Melted Sugar Fillings) , or Barquillos  (Wafer Rolls). 
Additional descriptors pertaining to the ingredients use d or the product form may 
also be included (e.g. Barquiron, Purple yam piaya; Chocolate  polvoron, Pandan-
flavored barquillos ). Other local or regional names referring to products  similar to 
those defined under subsection 3.1 may also be included, provided that these 
names are acceptable in the area of distribution, and i n accordance with the 
customs or practices of the area or country in which the pr oduct is distributed. 
(b) The complete list of ingredients and food additive s used in the preparation of the 
product in descending order of proportion. 
(c)  The net quantity of content by weight in the met ric system. Other systems of 
measurement required by importing countries shall appe ar in parenthesis after the 
metric system unit. 
(d) The name and address of the manufacturer, packer an d/or distributor of the food. 
(e) Open date marking  
The words “Best/Consume Before” or “Expiry Date” or “ Use by” indicating end of 
period at which the product shall retain its optimum quality attributes at defined 
storage conditions. 
(f) Lot or code number identifying product lot.  12  (g) The words “Product of the Philippines”, or the coun try of origin if imported. 
(h)  Additional requirements 
A pictorial representation of raw material or end-pro duct on the label should not 
mislead the consumer with respect to the raw material or  end-product so illustrated. 
(i) Optional information 
Any special conditions for the storage of the food may be indicated in the label. 
Where practicable, storage instructions shall be in close proximity to the open date 
marking . 
 
8.4 Nutrition Labeling  
Nutrition labeling shall conform to the established re gulations of FDA and/or 
authority.  
 
 
9 METHODS OF ANALYSIS AND SAMPLING 
 
 
9.1 Method of Sampling 
Sampling shall be in accordance with the FAO/WHO Codex  Alimentarius Sampling 
Plans for Prepackaged Foods - CAC/RM 42-1969, Codex Ali mentarius Volume 13, 
1994. 
 
9.2 Determination of Water Activity  
According to the AOAC Official Methods of Analysis, 16th ed., 1995. Method No. 
978.18. 
  
 9.3 Enumeration of Yeast and Mold Count 
 According to the USFDA Bacteriological Analytical Manua l (2001) 
 
 9.4 Enumeration of Coliform and E.coli 
 According to the USFDA Bacteriological Analytical Manua l (2001) 
 
 10 REFERENCES  13   
Abiva, C. C. 2001. A Quick Guide to Filipino Food and Cooking . Anvil Publishing, 
Inc. Pasig City, Philippines. 
 
A.O. No. 88-A s. 1984. Regulatory Guidelines Concerning Food Additives . 
Department of Health. San Lazaro Compound, Sta. Cru z, Manila, Philippines. 
 
A.O. No. 88-B s. 1984. Rules and Regulations governing the Labeling of 
Prepackaged Food Products distributed in the Philip pines . Bureau of Food and 
Drugs. Department of Health. Alabang, Muntinlupa Cit y, Philippines. 
 
A.O. No. 132 s. 1970. Regulation Prescribing the Standard of Identity and   
Quality of Milk and Milk Products (B-4.12-01). Department of Health. San Lazaro 
Compound, Sta. Cruz, Manila, Philippines. 
 
A.O. No. 230 s. 1974. B-4 Definition and Standards of Food; B-4.15 Fats a nd 
Oils; B-4.15-01 Shortening. Department of Health. San Lazaro Compound, Sta. 
Cruz, Manila, Philippines. 
 
A.O. No. 243 s. 1975. B-4 Definition and Standards of Food; B-4.18 Margar ine. 
Department of Health. San Lazaro Compound, Sta. Cru z, Manila, Philippines. 
 
A.O. No. 153 s. 2004. Guidelines, Current Good Manufacturing Practice in 
Manufacturing, Packing, Repacking or Holding Food. Department of Health. San 
Lazaro Compound, Sta. Cruz, Manila, Philippines. 
 
A. O. No. 2007-0012. Philippine National Standards for Drinking Water 20 07. 
Department of Health, Manila, Philippines 
 
Association of Analytical Chemists. Official Methods of Analysis Manual.  16 th  ed., 
1995. AOAC International. 481 North Frederick Ave., S uite 500, Gaithersburg, MD 
20877-2417. U.S.A. 
 
BAFPS. Draft Standards for Muscovado Sugar . Bureau of Agriculture and 
Fisheries Product Standards. Department of Agriculture. Visayas Ave., Quezon City. 
 
B.C. No. 6-A s. 1998. Permissible Net Content Variation in Prepackaged Fo od . 
Bureau of Food and Drugs. Department of Health. Alab ang, Muntinlupa City, 
Philippines. 
 
B.C. No. 01-A s. 2004. Guidelines for the Assessment of Microbiological Qu ality 
of Processed Foods . Bureau of Food and Drugs. Department of Health. Al abang, 
Muntinlupa City, Philippines. 
 
B.C. No.016 s. 2006. Updated  List of Food Additives. Bureau of Food and Drugs, 
Department of Health. Alabang, Muntinlupa City, Phi lippines. 
  14  Belitz, H.-D., W. Grosch, and P. Schieberle. 2009. Food Chemistry . 4 th  revised and 
extended ed. Springer-Verlag, Heidelberg, Germany.   
 
Codex Alimentarius Commission. 1995. FAO/WHO Codex Alimentarius 
Commission Manual . Food and Agriculture Organization. Viale delle Ter me di 
Caracalla, 00100 Rome, Italy. 
 
DOH. 2000. Implementing Rules and Regulations for the  Food Fortification Act of 
2000 (RA 8976 s. 2006). Department of Health. San La zaro Compound, Sta. Cruz, 
Manila, Philippines. 
 
Dictionary of Food Science and Technology , International Food Information 
Service, Blackwell Publishing, UK, 2005  
 
FNRI. 1997. The Philippine Food Composition Tables . Food and Nutrition 
Research Institute, Department of Science and Technology , Bicutan, Taguig, 
Philippines. 
 
Food, definition . ALINORM 04/27/41, para. 88 and Appendix VI. 2005.  Codex 
Alimentarius Commission. Food and Agriculture Organiza tion. Viale delle Terme di 
Caracalla, 00100 Rome, Italy. 
 
Konkel, P. J. 2001. Confectionery Products . Chapter 56. In Compendium of 
Methods for the Microbiological Examination of Food s . 4 th  ed. edited by F.P. 
Downes and K. Ito. American Public Health Association, Wa shington, DC, USA. 
 
PNS/BAFPS 35:2005. Philippine National Standard: Table Egg.  Bureau of 
Product Standards, Department of Trade and Industry, Makati City, Philippines. 
 
PNS/BAFPS 36:2008. Philippine National Standard: Fresh Milk.  Bureau of 
Product Standards, Department of Trade and Industry, Makati City, Philippines 
 
R.A. 3720. Food, Drug and Cosmetic Act. Bureau of Food and Drugs. Department 
of Health. Alabang, Muntinlupa City, Philippines. 
 
R.A. 9711. Food and Drug Administration (FDA) Act of 2009 . Bureau of Food and 
Drugs. Department of Health. Alabang, Muntinlupa Cit y, Philippines.      
 
Vaclavik, V. and E. Christian. 2008. Essentials of Food Science . 3 rd  edition. 
Springer Science+Business Media, LLC., New York, USA. 
 
 
 
 
 
 
 
  15  ANNEX 1 
 
FAO/WHO Alimentarius Sampling Plan for Prepackaged Foods 
(AQL=6.5) CAC/RM 42-1969  
 
1. Sampling Plan 1 (Inspection Level I, AQL = 6.5) 
1.1 Net Weight is equal to or Less Than 1 kg (2.2 lb) 
 
 
1.2 Net Weight is Greater than 1 kg (2.2 lb) but not more than 4.5 kg (10 lb) 
 
Lot size (N)  Sample size (n)  Acceptance No. (c)  
2,400 or less 6 1 
2,841 –15,000 13 2 
15,001 – 24,000 21 3 
24,001 – 42,000 29 4 
44,001 – 72,000 48 6 
72,001 – 120,000 84 9 
More than 120,000 126 13 
 
 
1.3 Net Weight Greater Than 4.5 kg (10 lb) 
 
 
 
 
 
 
 
 
 Lot size (N)  Sampl e size (n)  Acceptance No. (c)  
4,800 or less 6 1 
4,801 –24,000 13 2 
24,001 – 48,000 21 3 
48,001 – 84,000 29 4 
84,001 – 144,000 48 6 
144,001 – 240,000 84 9 
More than 240,000 126 13 
Lot size (N)  Sample size (n)  Acceptance No. (c)  
600 or less 6 1 
601 –2,000 13 2 
2,001 – 7,200 21 3 
7,201 – 15,000 29 4 
15,001 – 24,000 48 6 
24,001 – 42,000 84 9 
More than 42,000 126 13  16  2.Sampling Plan 2 (Inspection Level II, AQL = 6.5) 
 
2.1 Net Weight is Equal To or Less Than 1 kg (2.2 lb) 
 
Lot size (N)  Sample size (n)  Acceptance No. (c)  
4,800 or less 13 2 
4,801 –24,000 21 3 
24,001 – 48,000 29 4 
48,001 – 84,000 48 6 
84,001 – 144,000 84 9 
144,001 – 240,000 126 13 
More than 240,000 200 19 
 
2.2 Net Weight is Greater Than 1 kg (2.2 lb) but not more than 4.5 kg  (10 lb) 
 
Lot size (N)  Sample size (n)  Acceptance No. (c)  
2,400 or less 13 2 
2,841 –15,000 21 3 
15,001 – 24,000 29 4 
24,001 – 42,000 48 6 
44,001 – 72,000 84 9 
72,001 – 120,000 126 13 
More than 120,000 200 19 
 
2.3  Net Weight Greater Than 4.5 kg (10 lb) 
     
 
 
 
 
 
 
 
  
 
 
 
 
 Lot size (N)  Sample size (n)  Acceptance No. (c)  
600 or less 13 2 
601 –2,000 21 3 
2,001 – 7,200 29 4 
7,201 – 15,000 48 6 
15,001 – 24,000 84 9 
24,001 – 42,000 126 13 
More than 42,000 200 19  17  ANNEX 2 
Determination of Water Activity (AOAC 978.18) 
  
A. Principle 
Water activity, a w, is ratio of vapor pressure of H 2O in product to vapor pressure of pure H 2O at 
same temperature. It is numerically equal to 1/100 of relative humidity (RH) generated by 
product in closed system. RH can be calculated from direct measurement of  
partial vapor pressure or dew point or measured indire ctly by sensors whose physical or electric 
characteristics are altered by RH to which they are expo sed. Instruments are checked or 
calibrated on basis of RH generated by standard salt slush es.  
 
B. Instruments and Systems 
(Select 1 of following instruments or systems to perform  test. Each has different application 
limitations because of interferences from other volatile  components of products being 
measured. check with instrument manufacturer for more specific limitations.)  
 
(a) Change in electrical conductivity of immobilized sa lt solution. – Instrument available from 
Beckman Industrial, Rosemount Analytical Div., 89 Comm erce Rd, Cedar Grove, NJ 07009; 
Nova Sina AG, Andreastrasse 7-11, CH 8050, Zurich,Switz erland; Rotronic Instrument 
Corp., 160 E. Main St, Huntington, NY 11743. Immobi lized salt sensors are affected by 
polyols such as glycerol and glycol and by volatile amin es 
(b) Change in electrical capacitance of polymer thin fi lms. – Instrument available from General 
Eastern Instruments, 50 Hunt St, Watertown, MA 02172.  Polymer thin film sensors are 
affected by CH 3COOH.  
(c) Dew point by chilled mirror technique. – Instrument  available from EG&G, Environmental 
Equipment Division, 217 Middlesex Turnpike, Burlingto n, MA 01803 or General Eastern 
Instruments. Dew point measurements can be affected by co ndensables with lower critical 
temperature than H 2O.  
(d) Longitudinal change in dimensions of water-sorbing fiber. – Instrument available from G Lufft 
Metallbarometerfabrik, D-7, Postfach 692, Neue Weinste ige 22, Stuttgart, Germany.  
(e) Partial water vapor pressure by manometric system. – Partial H 2O vapor pressure 
measurements can be made useless by living products that re spire, such as grains or nuts; 
by active fermentation; or by products that expand e xcessively when subjected to high 
vacuum.  
(f) Relative weight of moisture sorbed by anhydrous hy drophilic solid, e.g., microcrystalline 
cellulose.-see J. Agr. Food chem. 22, 326(1974).  
 
C. Apparatus and Reagents 
(As needed for instrument or system selected.)  
(a) Dew point instrument. – Equipped to measure temp erature to ±0.1°. See 978.18B(c).  
(b) Forced-draft cabinet. – Constant temperature, set t o maintain 25 ± 1°; capacity ≥0.06 m 3 (2 cu 
ft); with access port to accommodate instrument sensor lea ds. Use in conjunction with (c).  
(c) Insulated box with cover. – Large enough to hold t est container, (e), and small enough to fit 
in forced-draft cabinet, (b); with access port to accommod ate instrument sensor leads. 
Protect test container from short-term temperature flu ctuations.  
(d) Manometric system. – Sensitive to pressure differenti al of ± 0.01 mm Hg (1.33 Pa). See 
978.18B(e).  
(e) Test containers. – 120 or 240 mL (4 or 8 oz) wide- mouth or Mason glass jars with Al- or 
Teflon-lined screw caps and gaskets. Check integrity of  cap seals and sensor leads by any 
means available, e.g., ability of system to hold vacuum , using Tesla coil.   18  (f) Water bath. – Capable of maintaining temperatur e constant within 0.1° at 25±1°; capacity 
sufficient to hold measuring chamber of selected apparatu s.  
(g) Hydrophilic solid. – Microcrystalline cellulose, Type PH-101 (FMC Corp.,Pharmaceutical and 
Bioscience Division, 1735 Market St, Philadelphia, PA 19 103, or equivalent).  
(h) Reference salts. – ACS reagent grade, fine crystal.  see Table 978.18.  
 
D. Preparation of Reference Salt Slushes 
 
Place selected reference salt in test container to depth of ca 4 cm for more soluble salts (lower 
aw), to depth of ca 1.5 cm for less soluble salts (higher a w), and to intermediate depth for 
intermediate salts. Add H 2O in ca 2 mL increments, stirring well with spatula afte r each addition, 
until salt can absorb no more H 2O as evidenced by free liquid. Keep free liquid to m inimum 
needed to establish saturation of salt with H 2O. Slushes are ready for use upon completion of 
mixing, and are usable indefinitely (except for some h igh a w salts susceptible to bacterial 
attack), if contained in manner to prevent substantial  evaporation losses. Some slushes, e.g., 
NaBr, may solidify gradually by crystal coalescence, with no effect on a w.  
 
E. Calibration 
 
Select ≥ 5 salts to cover a w range of interest or range of sensor being used. Measu re humidity 
generated by each salt slush in terms of instrument read out, as in 978.18F. Plot readout against 
aw values given in Table 978.18 for selected salts, using cross-section paper scaled for reading 
to 0.001 a w unit. Draw best average smooth line through plotted points. Use this calibration line 
to translate sensor instrument readout of samples to a w or to check vapor pressure or dew point 
instruments for proper functioning.  
 
F. Determination 
 
Place calibration slush or sample in forced-draft cabinet , (b), or H 2O bath, (f), until temperature 
is stabilized at 25±1°. Transfer salt slush or sample to test container, (e), seal container with 
sensing device attached, and place in temperature control  device. Use volume of sample or 
slush >1/20 total volume sample container plus any associa ted void volume of sensing system, 
but not so much as to interfere with operation of syst em. Record instrument response at 15, 30, 
60, and 120 min after test container is placed in tem perature control device, or record response 
on strip chart. Two consecutive readings, at indicated i ntervals, which vary by <0.01 a w unit are 
evidence of adequately close approach to equilibrium. Continue readings at 60-min intervals, if 
necessary. Convert last reading to a w by calculation from physical measurements or by 
reference to calibration line. Make all measurements wi thin range of calibration points; do not 
extrapolate calibration line. Make all measurements in  same direction of change, and, if required 
by properties of sensor, expose sensor to controlled RH b elow ambient before starting each 
measurement.   19  ANNEX 3 
Enumeration of Yeast and Mold Count 
Enumeration of Yeasts and Molds in Food--Dilution P lating Technique (USFDA, 2001)  
A. Equipment and materials   
1. Basic equipment (and appropriate techniques) for p reparation of sample homogenate 
2. Equipment for plating samples 
3. Incubator, 25°C 
4. Arnold steam chest 
5. pH meter 
6. Water bath, 45 ± 1° C 
 
B. Media and reagents  
 
1. Dichloran rose bengal chloramphenicol (DRBC) agar 
2. Dichloran 18% glycerol (DG18) agar 
3. Plate count agar (PCA), standard methods; add 100 mg chloramphenicol/liter when this 
medium is used for yeast and mold enumeration. This me dium is not efficient when 
"spreader" molds are present. 
4. Malt agar (MA) 
5. Malt extract agar (Yeasts and Molds) (MEAYM) 
6. Potato dextrose agar (PDA), dehydrated; commerciall y available 
 
C. Procedures  
Sample preparation  
Analyze 25-50 g from each subsample; generally, larger sample sizes increase 
reproducibility and lower variance compared with small samples. Test individual subsamples 
or composite according to respective Compliance Program f or the food under analysis. Add 
appropriate amount of 0.1% peptone water to the wei ghed sample to achieve 10 -1 dilution, 
then homogenize in a stomacher for 2 min. Alternativ ely, blending for 30-60 sec can be used 
but is less effective. Make appropriate 1:10 (1+9) di lutions in 0.1% peptone water. Dilutions 
of 10 -6 should suffice. 
Plating and incubation of sample  
Spread-plate method . Aseptically pipet 0.1 ml of each dilution on pre- po ured, solidified 
DRBC agar plates and spread inoculum with a sterile, bent glass rod. DG18 is preferred 
when the water activity of the analyzed sample is less than 0.95. Plate each dilution in 
triplicate. 
Pour-plate method . Use sterile cotton-plugged pipet to place 1.0 ml por tions of sample 
dilution into prelabeled 15 x 100 mm Petri plates ( plastic or glass), and immediately add 20-
25 ml tempered DG18 agar. Mix contents by gently swir ling plates clockwise, then 
counterclockwise, taking care to avoid spillage on dish l id. After adding sample dilution, add 
agar within 1-2 min; otherwise, dilution may begin t o adhere to dish bottom (especially if 
sample is high in starch content and dishes are plastic) an d may not mix uniformly. Plate 
each dilution in triplicate. 
From preparation of first sample dilution to pouring  or surface-plating of final plate, no more 
than 20 min (preferably 10 min) should elapse. Note: Spread plating of diluted sample is  20  considered better than the pour plate method. When th e pour plate technique is used, fungal 
colonies on the surface grow faster and often obscure tho se underneath the surface, 
resulting in less accurate enumeration. Surface plating gives a more uniform growth and 
makes colony isolation easier. DRBC agar should be used for spread plates only. 
Incubate plates in the dark at 25°C. Do not stack pl ates higher than 3 and do not 
invert. Note:  Let plates remain undisturbed until counting. 
Counting of plates  
Count plates after 5 days of incubation. If there is no growth at 5 days, re-incubate for 
another 48 h. Do not count colonies before the end of the incubation period because 
handling of plates could result in secondary growth fro m dislodged spores, making final 
counts invalid. Count plates containing 10-150 colonies. If mainly yeasts are present, plates 
with 150 colonies are usually countable. However, if su bstantial amounts of mold are 
present, depending on the type of mold, the upper cou ntable limit may have to be lowered at 
the discretion of the analyst. Report results in colony  forming units (CFU)/g or CFU/ml based 
on average count of triplicate set. Round off counts to two significant figures. If third digit is 6 
or above, round off to digit above (e.g., 456 = 460 ); if 4 or below, round off to digit below 
(e.g., 454 = 450). If third digit is 5, round off t o digit below if first 2 digits are an even number 
(e.g., 445 = 440); round off to digit above if firs t 2 digits are an odd number (e.g., 455 = 460). 
When plates from all dilutions have no colonies, repor t mold and yeast counts (MYC) as less 
than 1 times the lowest dilution used. 
Isolate individual colonies on PDA or MA, if further a nalysis and species identification is 
necessary. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  21  ANNEX 4 
Enumeration of E.coli and Coliform Count 
(Enumeration of Escherichia coli  and the Coliform Bacteria (USFDA, 2001)) 
Conventional Method for coliforms, fecal coliforms and E. coli  
A. Equipment and materials 
1. Covered water bath, with circulating system to maint ain temperature of 45.5 ± 0.2°C. Water 
level should be above the medium in immersed tubes. 
2. Immersion-type thermometer, 1-55°C, about 55 cm l ong, with 0.1°C subdivisions, certified 
by National Institute of Standards and Technology (NI ST), or equivalent 
3. Incubator, 35 ± 1.0°C 
4. Balance with capacity of > 2 kg and sensitivity of 0.1 g 
5. Blender and blender jar 
6. Sterile graduated pipets, 1.0 and 10.0 mL 
7. Sterile utensils for sample handling 
8. Dilution bottles made of borosilicate glass, with pol yethylene screw caps equipped with 
Teflon liners. Commercially prepared dilution bottles containing sterile Butterfield's 
phosphate buffer can also be used. 
9. Quebec colony counter, or equivalent, with magnifyi ng lens 
10. Longwave UV light [~365 nm], not to exceed 6 W. 
11. pH meter 
B. Media and Reagents  
Brilliant green lactose bile (BGLB) broth, 2% (M25 ) 
Lauryl tryptose (LST) broth (M76 ) 
EC broth (M49 ) 
Levine's eosin-methylene blue (L-EMB) agar (M80 ) 
Butterfield's phosphate-buffered water (R11 ) or equivalent diluent (except for shellfish) 
Lauryl tryptose MUG (LST-MUG) broth (M77 ) 
Peptone Diluent, 0.1% (R56 ) 
MPN - Presumptive test for coliforms, fecal colifor ms and E. coli  
Weigh 50 g food into sterile high-speed blender jar. (see Chapter 1 and current FDA compliance 
programs for instructions on sample size and compositing) Frozen samples can be softened by 
storing it for < 18 h at 2-5°C, but do not thaw. Add 450 mL of Butte rfield's phosphate-buffered 
water and blend for 2 min. If <50 g of sample are av ailable, weigh portion that is equivalent to 
half of the sample and add sufficient volume of sterile  diluent to make a 1:10 dilution. The total 
volume in the blender jar should completely cover the blades. 
Prepare decimal dilutions with sterile Butterfield's p hosphate diluent. Number of dilutions to be 
prepared depends on anticipated coliform density. Shake  all suspensions 25 times in 30 cm arc 
or vortex mix for 7 s. Do not use pipets to deliver <10% of their total volume. Transfer 1 mL 
portions to 3 LST tubes for each dilution for at least  3 consecutive dilutions. Hold pipet at angle 
so that its lower edge rests against the tube. Let pip et drain 2-3 s. Not more than 15 min should 
elapse from time the sample is blended until all dilut ions are inoculated in appropriate media.  22  NOTE:  Use 5-tube MPN for analysis of shellfish and shellfish ha rvest waters. 
Incubate LST tubes at 35°C. Examine tubes and record r eactions at 24 ± 2 h for gas, i.e., 
displacement of medium in fermentation vial or efferv escence when tubes are gently agitated. 
Re-incubate gas-negative tubes for an additional 24 h and examine and record reactions again 
at 48 ± 2 h. Perform confirmed test on all presumptiv e positive (gas) tubes. 
MPN - Confirmed test for coliforms  
From each gassing LST tube, transfer a loopful of suspen sion to a tube of BGLB broth, avoiding 
pellicle if present. Incubate BGLB tubes at 35°C and ex amine for gas production at 48 ± 2 h. 
Calculate most probable number (MPN) of coliforms based  on proportion of confirmed  gassing 
LST tubes for 3 consecutive dilutions. 
MPN - Confirmed test for fecal coliforms and E. coli  
From each gassing LST tube from the Presumptive test, tr ansfer a loopful of each suspension to 
a tube of EC broth (a sterile wooden applicator stick ma y also be used for these transfers). 
Incubate EC tubes 24 ± 2 h at 45.5 °C and examine for  gas production. If negative, reincubate 
and examine again at 48 ± 2 h. Use results of this tes t to calculate fecal coliform MPN. To 
continue with E. coli  analysis, proceed to Section F under Enumeration of Esch eria coli and the 
Coliform Bacteria of the USFDA Bacteriological Analyt ical Manual (2001). The EC broth MPN 
method may be used for seawater and shellfish since it con forms to recommended procedures 
(1). (Caution: see Note below). 
NOTE:  Fecal coliform analyses are done at 45.5± 0.2°C for al l foods, except for water testing 
and in shellfish and shellfish harvest water analysis, whi ch uses an incubation temperature of 
44.5± 0.2°C.