Food chemicals codex ( PDFDrive )

EIGHTH EDITION FOOD CHEMICALS CODEX FCC 8 By authority of the United States Pharmacopeial Convention Prepared by the Council of Experts and published by the Board of Trustees THE UNITED STATES PHARMAC.

Prepared by the Council of Experts and published by the Board of Trustees

THE UNITED STATES PHARMACOPEIAL CONVENTION

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FCC 8 Preface / v

Preface

contains monographs for ingredients that may not be

FCC 8

currently marketed in the United States

This section provides general information about the Eighth

USP acquired FCC from the NAS in 2006 and assumed Edition of the Food Chemicals Codex (FCC) and background

responsibility for its ongoing development and publication.information on the United States Pharmacopeial Convention

To continue the work of the Food and Nutrition Board of(USP) Additional information about the specific uses of this

IOM, USP formed a Food Ingredients Expert Committee

compendium is provided in the General Provisions and

within its Council of Experts This Expert Committee is

Requirements section (page 1).

responsible for approving all new and revised standards in

FCC.

MISSION

FCC is published in continuing pursuit of the mission of USP: FCC 8

To improve the health of people around the world through

The Eighth Edition of FCC (FCC 8) includes more than

public standards and related programs that help ensure the

1,100 monographs It also contains more than 150 General

quality, safety, and benefit of medicines and foods

Tests and Assays, providing procedures frequently cited in

monographs, sometimes with acceptance criteria, in order

FCC began after the passage of the 1958 Food Additives chapter with up-to-date relevant informational materials onAmendment to the United States (U.S.) Federal Food, Drug, method validation and various analytical techniques,

and Cosmetic Act Although the U.S Food and Drug reference tables and information on current Good

Administration (FDA) had, by regulations and informal Manufacturing Practices Additions, deletions, and other

statements, defined in general terms the quality revisions of text from the FCC Seventh Edition are indicated

requirements for food additives, food colors, substances on page xix in the Admissions section The FCC and its

generally recognized as safe for use in foods (GRAS) and Supplements become effective 90 days from the official date

other food chemicals in the US market prior to 1958 (prior- of publication, unless otherwise noted

sanctioned articles), these requirements were not sufficiently

Monograph Elements

specific to serve as release, procurement, and acceptance

Each FCC 8 monograph represents the documentary

stan-specifications for manufacturers and users of food chemicals

dard for an article, manifested by specifications that speakTherefore, regulators, industry and other interested parties

to the quality and safety of the food ingredient Each recognized the need for a compendium of standards

mono-graph includes, when available, the following: empiricaldesigned especially for food chemicals, comparable to the

formula, structural formula, and formula weight; description

United States Pharmacopeia for drugs and the National

of the substance, including physical form, odor (flavoring

Formulary for excipients, which would define the quality of

agents only), and solubility (see the descriptive terms forfood-grade chemicals in terms of identity, strength, and

solubility in the General Provisions and Requirements section);

purity The National Academy of Sciences (NAS) was

function; packaging and storage; labeling; identification; requested to develop this compendium and published the

as-say (or a quantitative test to serve as an asas-say); impurities

first edition of the FCC in 1966 Subsequent editions were

(inorganic and organic); specific tests; and other published by the NAS in 1972, 1981, 1996, and 2003,

require-ments The specifications provided, taken together, through the Food and Nutrition Board of the Institute of

repre-sent a compositional understanding of the substance

Medicine (IOM), which formed a Committee on Food

Chemicals Codex to elaborate the FCC.

Substances included in the first edition were limited to FCC revisions are published biennially in new editions, in

chemicals added directly to foods to achieve a desired Supplements published in intervening years and, when

function Subsequent editions added: (a) processing aids circumstances warrant, as Expedited Standards or Immediatesuch as enzymes, extraction solvents, filter media, and boiler Standards.

water additives; (b) foods, such as fructose and dextrose;

and (c) functional ingredients that affect not the foods to Supplements

which they are added, but the human body when the food The First Supplement to FCC 8 will be published in

comprehensive compendium of standards for these articles, date of publication, unless otherwise noted The Index in

collectively termed food ingredients The introduction of each Supplement is cumulative and includes citations to thenew food ingredients as well as constant changes in biennial revision The contents of the Supplement are inte-manufacturing processes and advances in analytical and grated into the following edition of FCC, along with new

metrological sciences lead to a need for continuous revision revisions that have been adopted since the Supplement to

of the FCC Because of its regulatory status in countries the previous compendium.

other than the United States, and its worldwide use, the FCC

Revision Type Symbol Subscript

Expedited Standards

Expedited Standards are revisions that the Food Ingredients New Text Adopted in a new text 1S, 2S, 3S (FCC biennial



Expert Committee determines, for public health or other

reasons, should become effective prior to publication of the New Text Adopted in FCC  new text  FCC biennial edition

next edition of the FCC or Supplement Proposed expedited

The following table shows symbols and effective dates for

standards are posted on the FCC Forum website for a

com-FCC 8 and its Supplements:

ment period of 90 days If there are no significant

com-ments, they become effective on the date posted on the

USP website, unless otherwise noted These revisions will be

2 June 1, 2013  and 2S(FCC8)

Immediate Standards

3 December 1, 2013  and 3S(FCC8)

Immediate Standards are revisions that the Food Ingredients

Expert Committee determines should be made available

im-mediately because of an urgent public health need These

FCC REVISION PROCESS

standards are posted as final on the USP website without

prior public notice and comment and are effective upon The FCC is revised on an ongoing basis in accordance with

website publication unless a delayed effective date is speci- USP Policies and Rules and Procedures Users of the FCC arefied These standards will be incorporated into the next requested and encouraged to submit suggestions for

analytical methods, and to review and comment upon

Errata are text published in the FCC or its Supplements that

do not accurately reflect the intended standards as ap-proved by the Food Ingredients Expert Committee A list of Food Ingredients Expert Committee

The Food Ingredient Expert Committee (FIEC) is part oferrata and corresponding corrections to an edition of the

USP‘s Council of Experts and is the scientific

decision-FCC or to a Supplement are published on USP‘s website, and

making body for the FCC Its principal functions include the incorporated into the next published edition of the FCC or

following:

Supplement Errata shall not be subject to public notice and

• To propose means by which FCC standards may be kept

comment

current in reflecting food-grade quality on the basis ofingredient safety, good manufacturing practices, and

Print and Electronic Presentations

advances in analytical capabilities

The FCC and its Supplements are available in print form and

• To provide information on issues relating to standards for

in an Internet version that allows individual registered users

particular substances and analytical test procedures

to access the FCC online The Internet format provides

ac-• To recommend the establishment of Expert Panels

cess to FCC content, along with extensive search options It

consisting of a committee member and other experts or

is continuously and cumulatively updated to integrate the

specialists to address specific issues relevant to

content of Supplements For users of the print edition, the

monograph development and to report their findings and

Supplements are included with the purchase of the FCC.

advisory recommendations to the full committee

Users of the FCC print edition must retain the Supplements

• To evaluate comments submitted by interested parties on

and review the FCC portion of the USP website in order to

any aspect of proposed FCC standards.

have up-to-date information

• To approve final standards before their publication in the

existing food-grade ingredients

marizes the types of symbols and the associated subscripts

The FIEC meets regularly to discuss food ingredients topics,used in FCC publications:

including technical and policy issues relevant to the FCC

Public Participation in FCC Revisions

Text Deletion Adopted as Effective Date

Although the FIEC is the ultimate decision-making body for

an Expedited or Immedi- • •

ate Standard FCC standards, these standards are developed by an

excep-tional process of public involvement and substantial

interac-Text Deletion Adopted in  1S, 2S, 3S (FCC biennial



a Supplement edition) tion between USP and its stakeholders, both domestically

and internationally Participation in the revision process

re-Text Deletion Adopted in FCC biennial edition

 

also from scientific, technical, and trade organizations

New Text Adopted as an Effective Date

Expedited or Immediate •new text•

Standard

FCC 8 Preface / vii

Figure 1 Public Review Process

monographs or those needing updating, contain informa- of FCC standards All proposals will have a 90-day comment

tion submitted voluntarily by manufacturers and other inter- period Figure 1 shows the public review and comment

pro-ested parties At times, USP staff may develop information cess and its relationship to standards development

to support a monograph through a Request for Revision USP

has developed a document titled Guideline for Submitting Re- Working with Government Agencies

quests for Revision to FCC, which is available at www.usp.org. USP works in many ways with government agencies in the

To facilitate the continuous revision of FCC and ensure an United States and abroad, including the FDA, to promote

open, transparent, and participatory revision process, USP good communications and optimal interactions The USP

representa-monographs, General Tests and Assays, and other draft doc- tives to participate in FIEC meetings, enabling continuing

uments via the FCC Forum The Forum is available free of interactions between the regulators‘ scientific staff and

Ex-charge For more information, visit www.usp.org/fcc pert Committee activities Staff in the FDA Centers, who areComments received are considered by the FIEC, who de- responsible for review of USP compendial activities, providetermine whether changes should be made to the proposed specific links and opportunities for exchange of comments.revisions based on those comments Proposed standards are The Center for Food Safety and Applied Nutrition is the

finalized when the FIEC votes to make them effective text in center that links FDA and USP in the areas of food

ingredi-FCC Thus, the USP standards-setting process gives those ents and FCC.

who manufacture, regulate, and use food ingredients the

and Advisory Groups, which act in an advisory capacity to

manufacturers, vendors, and users of food chemicals FCC

The composition of the USP Convention membership isstandards serve as the basis for many buyer and seller

designed to ensure a global representation from all contractual agreements

sec-tors of health care, with an emphasis on practitioners,

In the United States, the first edition of FCC was given

given USP’s practitioner heritage (see the History section).

quasi-legal recognition in July 1966 by means of a letter of

Voting Delegates of Convention member organizationsendorsement from FDA Commissioner James L Goddard,

elect USP’s President, Treasurer, other members of thewhich was reprinted in the book The letter stated that “the

Board of Trustees, and the Council of Experts They also

FDA will regard the specifications in the Food Chemicals

adopt resolutions to guide USP’s strategic direction and

Codex as defining an ‘appropriate food grade’ within the

amend USP’s Bylaws The 2010 meeting of the USP meaning of Sec 121.101(b)(3) and Sec 121.1000(a)(2) of

Con-vention occurred in April 2010 in Washington, DC A the food additive regulations, subject to the following

list-ing of all current Votlist-ing Delegates of the USP qualification: this endorsement is not construed to exempt

Conven-tion is included in the People secConven-tion.

any food chemical appearing in the Food Chemicals Codex

Board of Trustees

from compliance with requirements of Acts of Congress or

USP’s Board of Trustees is responsible for the with regulations and rulings issued by the Food and Drug

manage-ment of the business affairs, finances, and property ofAdministration under authority of such Acts.”

USP During its 5-year term, the Board defines USP’s Subsequently, various additional specifications from

stra-tegic direction through its key policy and operational

de-previous FCC editions were also incorporated by reference in

cisions A listing of the members of the 2010–2015 Board

the U.S Code of Federal Regulations to define specific safe

of Trustees is included in the People section.

ingredients under Title 21, in various parts of Sections 172,

173, and 184 It is anticipated that FDA will from time to Council of Experts

time continue to update its regulatory references to the FCC. The Council of Experts is the standards-setting body ofUSP will work diligently to assure that the FCC contains USP For the 2010–2015 cycle it is composed of 21

monographs for all substances added to foods in the United members, elected to 5-year terms by USP’s Convention,States, including all ingredients that are marketed as food each of whom chairs an Expert Committee These Chairs,additives and color additives under an FDA regulation in turn, elect the members of their Expert Committees.following a successful petition of FDA, ingredients that are The Expert Committees are responsible for the content ofaffirmed to be GRAS, and ingredients that are marketed USP’s official and authorized publications (see Figure 2).under approvals issued prior to the 1958 Food Additive The Executive Committee of the Council of Experts in-

In Canada, in the absence of national specifications, the direction, is an appeals body, and performs other

func-Fourth edition of the FCC, as amended from time to time, is tions that support the Council of Experts’ operations.officially recognized in the Canadian Food and Drug

Regulations under Section B.01.045(b) as the reference for Expert Panels to the Council of Experts

For Australia and New Zealand, the Food Standards Panels to assist the Council of Experts by providing Australia New Zealand recognizes the Seventh Edition of the sory recommendations to particular Expert Committees

advi-FCC as a primary source of identity and purity specifications in response to a specific charge consistent with the for substances added to food in Standard 1.3.4 Identity and pert Committee’s Work Plan Expert Panels are continu-

In Israel, the Public Health Regulations state that those People section.

who produce, import, market, or store a food additive must Stakeholder Forums and Project Teams

comply with the requirements established in the latest USP may form several domestic and international

Stake-edition of FCC or in the latest Stake-edition of the Compendium of holder Forums and Project Teams during the 2010–2015

Food Additive Specifications published by the Joint FAO/WHO cycle, including the Food Ingredients and Dietary

receive comments on USP’s standards-setting activities

GENERAL INFORMATION REGARDING Depending on the topic, a Stakeholder Forum may create

Project Teams to work on selected topics USP also holds

USP

Standards and Science Symposia in various regionsthroughout the world to promote scientific exchanges on

USP GOVERNANCE, STANDARDS-SETTING, AND

ADVI-topics relating to USP compendia

SORY BODIES

International Standards and Science Symposia

USP’s governing, standards-setting, and advisory bodies

in-• North Americaclude the USP Convention, the Board of Trustees, the Coun-

• India/West Asiacil of Experts and its Expert Committees, Expert Panels (for-

• China/East Asiamerly known as Advisory Panels), and staff Additional

• Latin Americavolunteer bodies include Stakeholder Forums, Project Teams,

FCC 8 Preface / ix

Figure 2 2010–2015 USP Council of Experts

partici-pate and vote, so long as any conflicts have been

headquarters and throughout the world, including an

ac-count management office in Basel, Switzerland, and

labo-Confidentiality and Document Disclosure

ratory facilities in Hyderabad, India; Shanghai, China; and

Members of the Council of Experts, Expert Committees, andS˜ao Paulo, Brazil

Expert Panels sign confidentiality agreements, in keepingwith USP’s Confidentiality Policy and the confidentiality pro-

USP POLICIES, RULES, AND PROCEDURES visions of the Rules and Procedures of the Council of Experts.

The USP Document Disclosure Policy, available on USP’s

standards-USP’s Articles of Incorporation, its Constitution and Bylaws, setting process by making information available to the and the Rules and Procedures of the 2010–2015 Council of lic, yet provides protection to manufacturers and others

pub-Experts are available on USP’s website (www.usp.org) Col- who submit confidential information to USP.

lectively, these documents serve USP volunteers and staff as

activities

United States Pharmacopeia and the National Conflicts of Interest

For-mulary— The United States Pharmacopeia (USP) and

Na-USP’s Conflict of Interest provisions require all members of

tional Formulary (NF) are compendia of science-based

stan-the Council of Experts, its Expert Committees, Expert Panels,

dards for drug and biologic dosage forms, drug substances,Board of Trustees, and key staff to disclose financial or other

excipients, medical devices, and dietary supplements Theseinterests that may interfere with their duties as USP volun-

standards are set by Expert Committees following public teers Members of the Board of Trustees, Council of Experts,

no-tice and opportunity for comment through publication inand its Expert Committees are not allowed to take part in

the final discussion or vote on any matter in which they

the free Pharmacopeial Forum The USP and NF are recog- industry documents, and other tools and resources It isnized as official compendia of the United States in the Fed- published every two years, as a hardcover print edition.eral Food, Drug, and Cosmetic Act, and also are recognized

Com-in the laws of many countries around the world The USP

pendium (MC) includes monographs, general chapters, and

and the NF are separate compendia although they are

pub-reference materials for suitable chemical and biologicallished in the same volume

medicines and their ingredients approved by national

regu-Chromatographic Columns— This comprehensive refer- latory authorities The purpose of the MC is to help ensure ence, previously titled Chromatographic Reagents, provides that these medicines are of good quality by providing up-detailed information needed to conduct chromatographic to-date, relevant public standards and reference materials

procedures found in USP–NF Chromatographic Columns lists MC standards are available to manufacturers, purchasers,

na-the brand names of na-the column reagents cited in every pro- tional regulatory authorities, and others to ensure posal for new or revised gas- or liquid-chromatographic ana- ity of a medicine to MC standards through testing The MC lytical procedures that have been published in PF since does not include standards for foods or for traditional

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column reagents were used to validate analytical procedures

USP Catalog— Use of official USP Reference Standards that have become official The branded column reagents list

pro-motes uniform quality of drugs, food ingredients, and dietary

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supplements and supports first-, second-, and third-party

test-USP Dictionary— The USP Dictionary of USAN and Inter- ing of all manufactured and compounded articles The

publi-national Drug Names provides, in a single volume, the most cation listing the collection of official USP Reference Standardsup-to-date United States Adopted Names of drugs; official can be accessed on the USP website at www.usp.org

USP–NF names; nonproprietary, brand, and chemical names; and is available in print form by contacting USP Sales andgraphic formulas; molecular formulas and weights; CAS reg- Marketing staff at 301-816-8237 The listing identifies newistry numbers and code designations; drug manufacturers; items, replacement lots, lots of a single item that are simul-

and pharmacologic and therapeutic categories The Diction- taneously official, lots deleted from official status, and a

pre-ary helps to ensure the accuracy of the following: product view of items eventually to be adopted Purchase order labeling; reports, articles, and correspondence; FDA regula- formation is included, and the names of distributors whotory filings; and pharmaceutical package inserts It is pub- can facilitate international availability of these items are sug-lished annually and is recognized by FDA as the official gested The USP Reference Standards program benefits from

in-source for established drug names (See Nomenclature.) the widespread voluntary contribution of suitable materials

and test data from manufacturers USP advances this

unoffi-USP Dietary Supplements Compendium— The Dietary cial material to official status via careful characterization

Supplements Compendium combines, in a single volume, studies and collaborative testing, followed by review and

ap-USP–NF standards for dietary supplements, standards and in- proval by the appropriate Expert Committee.

formation from the Food Chemicals Codex, regulatory and

FCC 8 Contents / iii

Contents

PREFACE v

PEOPLE xi

ADMISSIONS xviii

ANNOTATED xix

GENERAL PROVISIONS AND REQUIREMENTS APPLYING TO SPECIFICATIONS, TESTS, AND ASSAYS OF THE FOOD CHEMICALS CODEX 1

MONOGRAPH SPECIFICATIONS 9

PROVISIONAL MONOGRAPH SPECIFICATIONS 1209

GENERAL TESTS AND ASSAYS 1213

Appendix I: Apparatus for T est and Assays 1217

Appendix II: Physical Tests and Determinations 1221

A Chromatograhy 1221

B Physicochemical Properties 1230

C Others 1242

Appendix III: Chemical Tests and Determinations 1262

A Identification Tests 1262

B Limit Tests 1264

C Others 1279

Appendix IV: Chewing Gum Base 1298

Appendix V: Enzyme Assays 1303

Appendix VI: Essential Oils and Flavors 1336

Appendix VII: Fats and Related Substances 1341

Appendix VIII: Oleoresins 1357

Appendix IX: Rosins and Related Substances 1360

Appendix X: Carbohydrates (Star ches, Sugars, and Related Substances) 1364

Appendix XI: Flavor Chemicals (Other Than Essential Oils) 1375

Appendix XII: Microbiological Tests 1381

Appendix XIII: Adulterants and Contaminants in Food Ingredients 1384

Appendix XIV: Markers for Authenticity T esting 1388

SOLUTIONS AND INDICATORS 1393

GENERAL INFORMATION 1409

INDEX 1613

• F LUORIDE, Fluoride Limit Test, Method III, Appendix IIIB

6-Methyl-1,2,3-oxathiazine-4(3H)-one-2,2 Dioxide Potassium Acceptance criteria: NMT 3 mg/kg

Sample solution: 2 g in 20 mL of water

Control: 2 µg Pb (2 mL of Diluted Standard Lead

Solution)

Acceptance criteria: NMT 1 mg/kg Organic Impurities

Standard: 4-hydroxybenzoic acid ethyl ester

crystalline powder It is freely soluble in water and very Chromatographic system, Appendix IIA

Packaging and Storage: Store in well-closed containers Column: 25-cm × 4.6-mm (id) stainless steel, or

C18 silica gel, or equivalent

• A Sample solution: 0.3 g in 1 mL of glacial acetic acid . P ROCEDURE Injection volume: 20 µL

Elution: Isocratic

Analysis: Add a few drops of sodium cobaltinitrite TS to Suitability requirements: The resolution, R, between

Acceptance criteria: The Sample solution shows an other than that caused by acesulfame potassium

Reference standard: USP Acesulfame Potassium RS Acceptance criteria: The sum of the areas of all peaks

Acceptance criteria: The spectrum of the sample three times the elution time of acesulfame potassium,

exhibits maxima at the same wavelengths as those in except for the acesulfame potassium peak, does not

analysis of the Dilute sample solution (NMT 20 µg/g of

ASSAY

UV-active compounds)

Sample: 200–300 mg, previously dried at 105° for 2 h SPECIFIC TESTS

Analysis: Dissolve the Sample in 50 mL of glacial aceticacid in a 250-mL flask [N • L OSS ON D RYING, Appendix IIC: 105° for 2 h

slow.] Add 2 or 3 drops of crystal violet TS, and titrate • PH, pH Determination, Appendix IIB

that persists for at least 30 s [C AUTION —Handle Acceptance criteria: Between 5.5 and 7.5

perchloric acid in an appropriate fume hood.] Perform a

blank determination (see General Provisions), and make

any necessary correction Each mL of 0.1 N perchloric

acid is equivalent to 20.12 mg of CHKNOS

10 / Acetaldehyde Diethyl Acetal / Monographs FCC 8

Function: Flavoring agent First Published: Prior to FCC 6

IDENTIFICATION

Appendix IIIC

Acceptance criteria: The spectrum of the sample

exhibits relative maxima at the same wavelengths asthose of the spectrum below

Acetaldehyde Diethyl Acetal occurs as a colorless to pale

• R EFRACTIVE I NDEX , Appendix II: At 20°yellow liquid

Acceptance criteria: Between 1.379 and 1.384 Odor: Ethereal, fruity

Solubility: Soluble in propylene glycol, vegetable oils;

method (see General Provisions).

slightly soluble in water

Acceptance criteria: Between 0.821 and 0.827 Boiling Point: ∼102°

Acetaldehyde Diethyl Acetal

UNII: GO1N1ZPR3B [acetaldehyde]

First Published: Prior to FCC 6

Acetaldehyde occurs as a flammable, colorless liquid It may

Solubility: Miscible in alcohol, organic solvents, water Boiling Point: ∼21°

Function: Flavoring agent

IDENTIFICATION SPECIFIC TESTS

exhibits relative maxima at the same wavelengths as • S PECIFIC G RAVITY : Determine at 0° ± 0.05° by means of a

gravity at 0°/20° (see General Provisions).

Acceptance criteria: NLT 99.0% of C2H4O • R ESIDUE ON E VAPORATION, M-16, Appendix XI

First Published: Prior to FCC 6

Function: Flavoring agent

p-Methoxyacetophenone • I NFRARED S PECTRA, Spectrophotometric Identification Tests,

Appendix IIIC

Acceptance criteria: The spectrum of the sample

exhibits relative maxima at the same wavelengths asthose of the spectrum below

Acetanisole occurs as a colorless to pale yellow fused solid

12 / Acetanisole / Monographs FCC 8

• L EAD, M-9, Appendix XI

Acceptance criteria: 10 mg/kg

Acetanisole

phenolphthalein TS and titrate with 1 N sodium

First Published: Prior to FCC 6

hydroxide Each mL of 1 N sodium hydroxide isequivalent to 60.05 mg of C2H4O2

Acceptance criteria: NLT 99.5% and NMT 100.5%

C2H4O2 by weight

IMPURITIES

UNII: Q40Q9N063P [acetic acid]

SPECIFIC TESTS

Acetic Acid, Glacial, occurs as a clear, colorless liquid It boils Sample: 19 mL (20 g)

at about 118° When well-diluted with water (e.g., 1:100), Analysis: Evaporate the Sample in a tared dish on a

it has a vinegar odor and taste It is miscible with water, steam bath and dry at 105° for 1 h

glass-stoppered container and add 0.1 mL of 0.1 N

.

Last Revision: Second Supplement, FCC 7

Acetyl Methyl CarbinolDimethylketol

3-Hydroxy-2-butanone

FEMA: 2008FEMA: 2008

UNII: BG4D34CO2H [acetoin]

UNII: BG4D34CO2H [acetoin]

DESCRIPTION DESCRIPTION

Acetoin Monomer occurs as a colorless to pale yellow liquid.Acetoin Dimer occurs as a white to pale yellow powder

It can contain some variable amount of its dimer

Odor: Odorless

Odor: Buttery Solubility: Soluble in hot propylene glycol; slightly soluble

Solubility: Miscible in alcohol, propylene glycol, water;

in weak alkali; insoluble or practically insoluble in most

insoluble or practically insoluble in vegetable oilssolvents

Boiling Point: ∼148°

Function: Flavoring agent

Function: Flavoring agent

ASSAY

IDENTIFICATION

• I NFRARED S PECTRA, Spectrophotometric Identification Tests,

XI

Appendix IIIC

Acceptance criteria: NLT 96.0% of C4H8O2

Acceptance criteria: The spectrum of the sample

exhibits relative maxima at the same wavelengths asthose of the spectrum below

14 / Acetoin Monomer / Monographs FCC 8

Acetoin Monomer

Sample solution: 1 mg/mL First Published: Prior to FCC 6

Analysis: Place 10 mL of the Sample solution into a

glass-stoppered flask, add 25 mL of sodium hydroxide2-Propanone

TS, and allow the mixture to stand for 5 min Add 25Dimethyl Ketone

mL of 0.1 N iodine, stopper the flask, allow thecontents to stand in a cold, dark place for 10 min, andadd 30 mL of 1 N sulfuric acid Titrate the excess iodinewith 0.1 N sodium thiosulfate, using starch TS as the

indicator Perform a blank determination (see General

CAS: [67-64-1]

of 0.1 N iodine is equivalent to 0.9675 mg of C3H6O.UNII: 1364PS73AF [acetone]

Acceptance criteria: NLT 99.5% and NMT 100.5%

chloroform, and with most volatile oils

• L EAD, Lead Limit Test, Atomic Absorption Spectrophotometric

Function: Extraction solvent

Graphite Furnace Method, Method I, Appendix IIIB

Packaging and Storage: Store in tight containers remote

Analysis: Mix the Sample with 10 mL of water, add 5

Analysis: To both the Sample solution and 10 mL of the

mL of 1 N sodium hydroxide, warm, and add 5 mL of

Standard solution, add 0.15 mL of a 5% solution of

5,5-iodine TS

dimethyl-1,3-cyclohexanedione in alcohol, and

Acceptance criteria: A yellow precipitate of iodoform

evaporate on a steam bath until the Acetone isforms

volatilized Dilute both to 10 mL with water and coolquickly in an ice bath while stirring vigorously

Acceptance criteria: Any turbidity produced by the Acceptance criteria: The solution remains clear for at

Sample solution does not exceed that produced by the least 30 min

Standard solution (NMT 0.002%). • S PECIFIC G RAVITY : Determine by any reliable method (see

Analysis: Add 0.2 mL of 10% phosphoric acid and 0.25 • S UBSTANCES R EDUCING P ERMANGANATE

1 mL of each Control solution and Sample solution Analysis: Transfer the Sample into a glass-stoppered

Allow the mixtures to stand for 15 min, then add 0.3 cylinder, add 0.05 mL of 0.1 N potassium

mL of 100 mg/mL sodium bisulfite solution to each, permanganate, mix, and allow to stand for 15 min

and shake until colorless Slowly add 5 mL of ice-cold Acceptance criteria: The pink color does not entirely

the addition Add 0.1 mL of 10 mg/mL chromotropic • W ATER, Water Determination, Appendix IIB

acid solution, mix, and digest on a steam bath for 20 Analysis: Use freshly distilled pyridine instead of

Acceptance criteria: Any violet color produced by the Acceptance criteria: NMT 0.5%

Sample solution does not exceed that produced by the

Control solution (NMT 0.05%).

Analysis: Evaporate the Sample to dryness at 60° Add 3 First Published: Prior to FCC 6

drops of a solution of 100 mg of sodium nitrite in 5

mL of sulfuric acid to the residue, allow the mixture to

stand for about 3 min, and then carefully add 3 mL of UNII: 3O959710YK [acetone peroxide]

2 N sodium hydroxide

Acetone Peroxides, usually mixed with an edible carrier such

SPECIFIC TESTS as cornstarch, occur as a fine, white, free-flowing powder

• A CIDITY (AS ACETIC ACID)

They are a mixture of monomeric and linear dimeric

Sample: 38 mL

acetone peroxides (mainly 2,2-hydroperoxypropane), with

Analysis: Mix the Sample with an equal volume of

minor proportions of higher polymers

carbon dioxide-free water, add 0.1 mL of

Function: Bleaching agent; maturing agent; dough

phenolphthalein TS, and titrate with 0.1 N sodium

conditionerhydroxide

Packaging and Storage: Store in tightly closed Acceptance criteria: NMT 0.1 mL is required to

containers in a cool, dry place, preferably below 24°

produce a pink color (NMT 0.002%)

Avoid exposure to the skin and eyes.]

Sample: 23 mL

water, add 0.1 N sulfuric acid until a red color just • P ROCEDURE

Acceptance criteria: NMT 0.1 mL of 0.1 N sulfuric acid sulfuric acid, allow to stand for a few minutes, and add

Acceptance criteria: Within a range of 1°, including

Analysis: Evaporate the Sample to dryness in a tared Analysis: Transfer the Sample into a 250-mL beaker, add

dish on a steam bath, dry the residue at 105° for 30 50 mL of 10% sulfuric acid, allow to stand for at least 3

[NOTE—Use an Abb´e or other refractometer of equal or P, as g of hydrogen peroxide equivalents per 100 g of

Acceptance criteria: Between 1.358 and 1.360 at 20°

P = V × N × 0.017 × 100/W

Sample: 38 mL

16 / Acetone Peroxides / Monographs FCC 8

0.017 = milliequivalent weight of hydrogen peroxide Solubility: Very soluble in most fixed oils, propylene glycol;

Multiply the value P so obtained by 1.6 to convert to water; insoluble or practically insoluble in glycerin

Acceptance criteria: A sample yields an amount of Solubility in Alcohol, Appendix VI: One mL dissolves in 5

hydrogen peroxide equivalent to NLT 16.0% of acetone mL of 50% alcohol

Sample solution: Prepare as directed for organic Acceptance criteria: The spectrum of the sample

Acceptance criteria: Between 1.533 and 1.535

Acetylbenzene

method (see General Provisions).

Methyl Phenyl Ketone

Acceptance criteria: Between 1.025 and 1.028

OTHER REQUIREMENTS

Acceptance criteria: Passes test

• S OLIDIFICATION P OINT , Appendix IIB

3-Acetyl-2,5-dimethyl Furan

IDENTIFICATION

First Published: Prior to FCC 6

• I NFRARED S PECTRA, Spectrophotometric Identification Tests,

Appendix IIIC

exhibits relative maxima at the same wavelengths asthose of the spectrum below

• R EFRACTIVE I NDEX , Appendix II: At 20°

3-Acetyl-2,5-dimethyl Furan occurs as a yellow liquid

Solubility: Soluble in alcohol, most fixed oils, propylene Acceptance criteria: Between 1.027 and 1.048

glycol; slightly soluble in water

Boiling Point: ∼83° (11 mm Hg)

18 / 3-Acetyl-2,5-dimethyl Furan / Monographs FCC 8

3-Acetyl-2,5-dimethyl Furan

N-Acetyl-L-Methionine and add 100 mL of water, 5 g of dibasic potassium

phosphate, 2 g of monobasic potassium phosphate, and

First Published: Prior to FCC 6

2 g of potassium iodide Mix well to dissolve, add 50.0

mL of 0.1 N iodine, stopper the flask, and mix Allow to

N-Acetyl-L-2-amino-4-(methylthio)butyric Acid

stand for 30 min, add starch TS indicator, and thentitrate the excess iodine with 0.1 N sodium thiosulfate.Perform a residual blank titration Each mL of 0.1 Niodine is equivalent to 9.563 mg C7H13NO3S

Acceptance criteria: NLT 98.5% and NMT 101.5%

C7H13NO3S, calculated on the dried basis

UNII: 9J12WX5B6A [n-acetylmethionine]

• L EAD, Lead Limit Test, Appendix IIIB

Sample Solution: Prepare as directed for organic

DESCRIPTION

compounds

N-Acetyl-L-Methionine occurs as a colorless or lustrous,

Control: 5 µg Pb (5 mL of Diluted Standard Lead

white, crystalline solid or a white powder It is soluble in

Solution)

water, in alcohol, in alkali solutions, and in dilute mineral

Acceptance criteria: NMT 5 mg/kg

acids, but practically insoluble in ether

Packaging and Storage: Store in tightly closed, light- • L OSS ON D RYING , Appendix IIC: 105° for 2 h

• O PTICAL ( S PECIFIC) R OTATION , Appendix IIB

IDENTIFICATION

Sample: 20 mg/mL (sample previously dried), made to

100 mL

Tests, Appendix IIIC

Acceptance criteria: [α]D20 between −18.0° and

Sample preparation: Mineral oil mull

−22.0°, on the dried basis

Acceptance criteria: The spectrum of the sample

• R ESIDUE ON I GNITION ( S ULFATED A SH) , Appendix IIC

exhibits relative maxima at the same wavelengths as

N-Acetyl-L-Methionine (Mineral Oil Mull)

2-Acetyl Thiazole

IDENTIFICATION

Appendix IIIC

Acceptance criteria: The spectrum of the sample

exhibits relative maxima at the same wavelengths asthose of the spectrum below

ASSAY

2-Acetyl Thiazole occurs as a colorless to pale yellow liquid • R EFRACTIVE I NDEX , Appendix II: At 20°

Solubility: Soluble in propylene glycol, vegetable oils; • S PECIFIC G RAVITY : Determine at 25° by any reliable

Solubility in Alcohol, Appendix VI: One mL dissolves in 1

mL of 95% ethanol

20 / 2-Acetyl Thiazole / Monographs FCC 8

2-Acetyl Thiazole

Packaging and Storage: Store in well-closed containers First Published: Prior to FCC 6

IMPURITIES

Sample: 10 g Acceptance criteria: NMT 2 mg/kg

SPECIFIC TESTS

• A CID V ALUE, Method II, Appendix VII

INS: 472a

Acceptance criteria: NMT 6

UNII: 5Z17386USF [diacetylated monoglycerides]

Acetylated Monoglycerides occur as clear, thin liquids or Acceptance criteria: The result should conform to the

solids, ranging in color from white to pale yellow They representations of the vendor

consist of partial or complete esters of glycerin with a • I ODINE V ALUE , Appendix VII

mixture of acetic acid and edible fat-forming fatty acids Acceptance criteria: The result should conform to the

They may be manufactured by the interesterification of representations of the vendor

edible fats with triacetin and glycerin in the presence of • R EICHERT -M EISSL V ALUE , Appendix VII

catalytic agents, followed by molecular distillation, or by Acceptance criteria: Between 75 and 200

the direct acetylation of edible monoglycerides with acetic • S APONIFICATION V ALUE , Appendix VII

anhydride and without the use of a catalyst or molecular Acceptance criteria: The result should conform to the

distillation They are insoluble in water, but are soluble in representations of the vendor

alcohol, in acetone, and in other organic solvents, the

extent of solubility depending on the degree of

esterification and the melting range

. IDENTIFICATION

3-Acetylpyridine • I NFRARED S PECTRA, Spectrophotometric Identification Tests,

Appendix IIIC

First Published: Prior to FCC 6

Acceptance criteria: The spectrum of the sample

exhibits relative maxima at the same wavelengths asMethyl Pyridyl Ketone

those of the spectrum below

Acceptance criteria: Between 1.530 and 1.540

DESCRIPTION • S PECIFIC G RAVITY : Determine at 25° by any reliable

3-Acetylpyridine occurs as a colorless to yellow liquid method (see General Provisions).

Solubility: Soluble in acids, alcohol, ether, water

OTHER REQUIREMENTS

Boiling Point: ∼230°

• W ATER, Water Determination, Method I, Appendix IIB

Function: Flavoring agent

2-Acetylpyrrole occurs as a white to pale brown fine

Odor: Bready Solubility: Insoluble or practically insoluble in propylene

glycol, vegetable oils, water

Boiling Point: ∼220°

22 / 2-Acetylpyrrole / Monographs FCC 8

Solubility in Alcohol, Appendix VI: One g dissolves in 6 FEMA: 3126

UNII: GR391IBU5C [2-acetylpyrazine]

mL of ethanol

Function: Flavoring agent

DESCRIPTION ASSAY 2-Acetylpyrazine occurs as colorless to pale yellow crystals

Function: Flavoring agent

OTHER REQUIREMENTS

exhibits relative maxima at the same wavelengths asthose of the spectrum below

ASSAY

2-Acetylpyrazine

Acceptance criteria: NLT 99.0% of C6H6N2OMethyl Pyrazinyl Ketone

. Sample stock solution: Dissolve sample, as needed

Acid Hydrolysates of Proteins with 20% aqueous sodium chloride, to obtain a

solution with a solids content of 36%

First Published: Prior to FCC 6

Sample preparation: Transfer a 20-g aliquot of the Last Revision: Second Supplement, FCC 7

Sample stock solution into a 20-mL Extrelut NT column

(EM Science, Gibbstown, NJ), or equivalent, and allowAcid-Hydrolyzed Proteins

it to equilibrate for 15 min Elute the column with 150Hydrolyzed Vegetable Protein (HVP)

mL of ethyl acetate, collecting the eluent in a 250-mLHydrolyzed Plant Protein (HPP)

short-neck, round-bottom flask with a 24/40 joint

Hydrolyzed (Source) Protein Extract

Using a rotary evaporator at 50°, concentrate theAcid-Hydrolyzed Milk Protein

eluent to a volume of approximately 3 mL Add 0.5 mL

of Internal standard solution to the eluent, transfer this

DESCRIPTION

mixture to a 4-dram screw-cap vial, and dilute to aAcid Hydrolysates of Proteins occur as liquids, pastes,

volume of 5.0 mL

powders, or granules They are composed primarily of

Chromatographic system, Appendix IIA

amino acids, small peptides (peptide chains of five or fewer

Mode: Gas chromatography

amino acids), and salts resulting from the essentially

Detector: Electrolytic conductivity detector [NOTE—complete hydrolysis of peptide bonds in edible

Operate the detector in the halogen mode.]

proteinaceous materials, catalyzed by food-grade acids and

Column: 30-m × 0.53-mm (id), fused-silica column, or

/or heat Cleavage of peptide bonds typically ranges from

equivalent, coated with 1-µm Supelcowax 10 or an

a low of 85% to essentially 100% In processing, the

equivalent bonded carbowax column fitted with aprotein hydrolysates may be treated with safe and suitable

50-cm retention gap of 0.53-mm, deactivated, fusedalkaline materials The edible proteinaceous materials used

silica, or equivalent

as raw materials are derived from corn, soy, wheat, yeast,

Temperature

peanuts, rice, or other safe and suitable vegetable or plant

Column: Hold at 170° for 5 min, then increase at

sources, or from milk

5°/min to 250°, hold at 250° for 10 min

Function: Flavoring agent; flavor enhancer

liquid and paste samples to dryness in a suitable tared

Carrier gas: Helium

container; then, as for the powdered and granular forms,

Reactant gas: Hydrogen

dry to constant weight at 105° (See General Provisions.)]

Solvent: 1-Propanol

1-Propanol: 0.5 mL/min through the cell or at the

• L EAD Sample solution: Prepare as directed for organic, Lead Limit Test, Appendix IIIB Injection volume: 1.0 µL

Injection type: Use a capillary injector operated in the

Acceptance criteria: NMT 3 mg/kg, on the dried basis by venting flow from the column at all times, except

Standard stock solution: 125 µg/mL of reagent-grade Analysis: Separately inject Standard solution A,

3-chloropropane-1,2-diol (3-MCPD) in ethyl acetate Standard solution B, Standard solution C, and the

Diluted standard solution: 6.25 µg/mL of 3-MCPD in Sample preparation into the chromatograph and

Internal standard solution: 10 µg/mL of 1-chlorotet- area ratios of 3-MCPD to the Internal standard

Standard solution A: 2 mL of Diluted standard solution and 2.5 mL of Internal standard solution diluted to 25 versus the µg of 3-MCPD in each Standard solution to

obtain the standard curve From the chromatogram

mL with ethyl acetate (contains 0.5 µg/mL 3-MCPD) of the Sample preparation, measure the area ratio of

Standard solution B: 8 mL of Diluted standard solution 3-MCPD to the Internal standard solution and, using

and 2.5 mL of Internal standard solution diluted to 25 the standard curve, determine the amount of

3-mL with ethyl acetate (contains 2.0 µg/mL 3-MCPD) MCPD, in µg, in the 20-g aliquot of Sample stock

Standard solution C: 16 mL of Diluted standard solution solution taken.

and 2.5 mL of Internal standard solution diluted to 25 Acceptance criteria: NMT 1 mg/kg, on the dried

24 / Acid Hydrolysates of Proteins / Monographs FCC 8

Diluted standard solution: 1 µg/mL of DCP in Diluent TN = percentage of Total Nitrogen, determined

Standard solutions: Pipet 1, 2, 3, and 4 mL portions of • A MMONIA N ITROGEN , Appendix IIIC

Diluted standard solution, into separate 50-mL Acceptance criteria: NMT 1.5%, on the dried basis

volumetric flasks Add 1.0 mL of Internal standard • G LUTAMIC A CID , Appendix IIIC

solution to each and dilute with Diluent to volume. Acceptance criteria: NMT 20.0% as glutamic acid Sample solution: Dissolve 5.0 g of the sample in a (C5H9NO4) and NMT 35.0% of the total protein, both

solution Quantitatively transfer this solution to an • I NSOLUBLE M ATTER

equivalent After 15 min, elute the column with three Analysis: Transfer the Sample into a 250-mL Erlenmeyer

20-mL portions of Diluent, and collect all of the eluate. flask, add 75 mL of water, cover the flask with a watchCarefully evaporate the eluate to less than 4 mL Add glass, and boil gently for 2 min Filter the solution

1.0 mL of Internal standard solution, and dilute with through a tared filtering crucible, dry at 105° for 1 h,

Diluent, as necessary, to bring the final volume to 5.0 cool, and weigh

Mode: Gas chromatography with a split injector Standard solution: 1.91 µg/mL of potassium chloride Detector: Electrolytic conductivity detector (corresponds to 1.0 µg/mL of potassium ion)

Column: 50-m × 0.2-mm (id), fused-silica column Sample stock solution: Transfer 1.00 ± 0.05 g of

Column: Hold at 115° for 10 min, then increase at warming the solution if necessary to complete solution

[NOTE—Precondition the column by heating it at Sample solution: 1:300 (v/v) dilution of the Sample

Analysis: Separately inject each of the Standard Acceptance criteria: The absorbance of the Sample

solutions and the Sample solution into the solution does not exceed that of the Standard solution.

chromatograms Calculate the area ratios of DCP to • S ODIUM

Internal standard solution for each Standard solution. Standard stock solution: 254.2 µg/mL of sodium

Standard solution to obtain the standard curve From Standard solution: 12.71 ng/mL of sodium chloride

the chromatograph of the Sample solution, measure made from the Standard stock solution (corresponds to the area ratio of DCP to the Internal standard solution 5 ng/mL of sodium ion)

and, using the standard curve, determine the amount Sample stock solution: Transfer 1.00 ± 0.05 g of

Acceptance criteria: NMT 0.05 mg/kg, on the dried Ash in a muffle furnace at 550° for 2–4 h Allow the ash

warming the solution if necessary to complete solution

SPECIFIC TESTS of the residue Filter the solution through acid-washed

Acceptance criteria: NLT 3.0%, on the dried basis filter paper with hot water, dilute to volume, and mix.

• α-A MINO N ITROGEN/ T OTAL N ITROGEN P ERCENT R ATIO Sample solution: 1:4000 (v/v) dilution of the Sample

Analysis: Using a suitable atomic absorption

Result = 100[(AN – P)/(TN – P)]

spectrophotometer, determine the absorbance of the

Standard solution and the Sample solution at 589.0.

Acceptance criteria: The absorbance of the Sample Analysis: Transfer the appropriate Sample into a tared

solution does not exceed that of the Standard solution. 250-mL Erlenmeyer flask, and record the weight to the

approximately 2 g of potassium iodide, place the flaskover a magnetic stirrer, and stir until the potassiumiodide crystals dissolve (about 1 min) Add 1 mL of 6 N

hydrochloric acid, and stir for 30 s While continuously

Acidified Sodium Chlorite Solutions

stirring, titrate the liberated iodine with standardized

First Published: Prior to FCC 6

0.025 N sodium thiosulfate (Na2S2O3) When most ofthe brown iodine color has faded, add 2 mL of starchindicator solution, and titrate to a clear endpoint,

chlorous acid (HClO2) ASC Solutions are produced by

and suitable acid to achieve a pH within the range 2.3 to

3.9 depending on the intended use

Function: Antimicrobial agent in processing water used to V = volume of titrant (mL)

spray, dip, rinse, or store food before processing, to be N = normality of the sodium thiosulfate titrant

followed by rinsing in potable water or by blanching, Mr = molecular weight of sodium chlorite, 90.44

cooking, or canning; sanitizer for hard surfaces; broad- F = conversion factor for mg/g to ppm, 1000

spectrum bactericide, virucide, fungicide, and sporicide W = weight of the sample taken (g)

because chlorine dioxide gas will generate in the solution [NOTE—The concentration of sodium chlorite also can

alternatively be determined using ion chromatography

2.2 ]

Control: 10 µg of Pb (10 mL of Diluted Standard Lead depending on the application

50-mL beaker; add 10 mL of water, 1 mL of 20%

First Published: Prior to FCC 6

sulfuric acid, and 1 mL of a 40 mg/mL potassium

permanganate solution Cover the beaker with a watch

Achilleic Acid

• PH, pH Determination, Appendix IIB

chlorine dioxide gas, do not adjust the pH below 2.3.]

Acceptance criteria: Between 2.3 and 3.9

[NOTE—The pH is chosen depending on the application

It controls the concentration of metastable chlorous

acid, which rapidly breaks down into chlorine dioxide, C

[NOTE—See 21 CFR 173.325; “Determination of Sodium UNII: 93371T1BXP [aconitic acid]

Chlorite: 50 ppm to 1500 ppm,” Alcide Corporation.]

1 Hautman, Daniel P and Munch, David J “Method 300.1: Determination of

Sample: For solutions containing 40 to 250 ppm, use a

inorganic anions in drinking water by ion chromatography, Revision 1.0.” U.S.

100-g sample; for those containing 250 to 500 ppm,

Environmental Protection Agency, Office of Ground Water and Drinking

use a 50-g sample; for those containing 500 to 1100 Water 1997 Online Available: http://www.epa.gov/OGWDW/methods/

ppm, use a 20-g sample; for those containing 1100 to sourcalt.html [accessed October 19, 2007].

method II In: Standard Methods for the Examination of Water and Wastewater,

20th Ed Baltimore, MD: APHA/AWWA/WEF Pp 4-73 and 4-79.

26 / Aconitic Acid / Monographs FCC 8

tube in a stream of water and transfer the acid solution

DESCRIPTION

into a color comparison tube View the tube vertically

Aconitic Acid occurs in the leaves and tubers of Aconitum

against a white background and compare to the same

napellus L (Fam Ranunculaceae) and various species of

volume of the Control in a similar matching tube.

Achillea and Equisetum, in beet root, and in sugar cane It

Acceptance criteria: The color of the Sample solution is

may be synthesized by the dehydration of citric acid by

not darker than that of the Control.

sulfuric or methanesulfonic acid Aconitic Acid from the

• R ESIDUE ON I GNITION ( S ULFATED A SH), Method I, Appendix

above sources has the “trans” configuration It has a

IICmelting point of 195° to 200° with decomposition It is

Sample: 4 g

practically odorless and has a winy taste It is soluble in

Acceptance criteria: NMT 0.1%

water and in alcohol and is slightly soluble in ether

• W ATER, Water Determination, Appendix IIB

Function: Flavoring substance; adjuvant

• I NFRARED Sample preparation: Neat as a potassium bromide A BSORPTION S PECTRUM 5 ′-Adenylic Acid

First Published: First Supplement, FCC 7

dispersion

Acceptance criteria: The Sample preparation exhibits

Adenosine 5′-monophosphateinfrared absorption bands at 3030, 2630, and

Adenylic acid

1720 cm−1

AMP

• V ISIBLE A BSORPTION S PECTRUM

Adenosine 5′-phosphoric acid

Sample solution: Aqueous solution

Acceptance criteria: The Sample solution exhibits major

absorption peaks at 411 and 432 nm, with little or no

absorption at 389 nm

ASSAY

Analysis: Dissolve the Sample in 40 mL of water, add UNII: 415SHH325A [adenosine phosphate]

phenolphthalein TS, and titrate with 1 N sodium

DESCRIPTION

hydroxide Each mL of 1 N sodium hydroxide is

5′-Adenylic Acid occurs as colorless or white crystals, or as aequivalent to 58.04 mg of C6H6O6

white, crystalline powder It is very slightly soluble in

Acceptance criteria: NLT 98.0% and NMT 100.5% of

water, and practically insoluble in alcohol It is produced

C6H6O6, calculated on the anhydrous basis

by enzymatic cleavage of yeast ribonucleic acid (RNA) with

IMPURITIES a 5′-phosphodiesterase followed by heat treatment, further

• L EAD, Lead Limit Test, Atomic Absorption Spectrophotometric Function: Source of 5′-Adenylic Acid

Graphite Furnace, Method I, Appendix IIIB Packaging and Storage: Store in tight containers

Acceptance criteria: NMT 0.5 mg/kg

IDENTIFICATION SPECIFIC TESTS • A . I NFRARED A BSORPTION, Spectrophotometric Identification

Analysis: Neutralize 10 mL of Sample solution with 6 N Sample and standard preparation: A

Analysis: Transfer the Sample into a 22- × 175-mm test Standard solution in the Assay.

tube previously rinsed with 10 mL of 95% sulfuric acid

ASSAY

and allowed to drain for 10 min Add 10 mL of 95%

sulfuric acid, agitate the tube until solution is complete,

Mobile phase: 0.1 M potassium dihydrogen phosphate

and immerse the tube in a water bath at 90° ± 1° for

(KH2PO4) in degassed water, adjusted with 0.1 M

60± 0.5 min, keeping the level of the acid below the

level of the water during the heating period Cool the

dipotassium hydrogen phosphate (K2HP04) to a pH of Standard solutions: 0.05, 0.1, 0.2, 0.5, 1, and 2 µg/mL

Standard solution: 0.02 mg/mL of USP 5’-Adenylic Acid Diluent

RS in Mobile phase [NOTE—Ultra-sonication for 15 min Sample: 5 g

at 30° may be necessary to aid in complete dissolution.] Sample solution: Dissolve the Sample in 40 mL of 10% Sample solution: 0.02 mg/mL in Mobile phase [NOTE— nitric acid in a 100-mL volumetric flask, and dilute withUltra-sonication for 15 min at 30° may be necessary to water to volume

Column: 25 cm × 4.6-mm; packed with 5-µm reversed Setup: Use a suitable ICP–OES configured in a radial

Suitability requirement 1: The relative standard auxiliary gas set to flow at 2.25 L/min The sample isdeviation of the 5’-adenylic acid area responses from delivered to the spray chamber by a multi-channel

between the 5′-adenylic acid peak and all other for 20 s prior to analysis A 40-s read delay is also

Analysis: Separately inject equal volumes of the Standard fluid flow equilibration after the high-speed flush,

solution and Sample solution into the chromatograph, prior to the first analytical read of the sample

retention time for 5′-adenylic acid is 27.5 min.] Analysis: Generate a standard curve using Diluent as a

less than 0.999.]

Result = (rU/rS) × (CS/CU) × 100 Similarly, analyze the Sample solution on the ICP.

Calculate the concentration (mg/kg) of arsenic in the

Sample solution

Standard solution

Acceptance criteria: 98.0%–103.0%, calculated on the F = final volume of the Sample solution, 100 mL

IMPURITIES [NOTE—When water is specified as a diluent, use

specified, use nitric acid of a grade suitable for trace Diluent: 4% nitric acid in water

element analysis with as low a content of arsenic as Standard stock solution: 100 µg/mL of cadmium

Standard stock solution: 100 µg/mL of arsenic Standard solutions: 0.005, 0.05, 0.1, 0.2, 0.5, 1, and 2

prepared by diluting a commercially available 1000 mg µg/mL of cadmium, from the Standard stock solution

Sample: 5 g

Sample solution: Dissolve the Sample in 40 mL of 10%

nitric acid in a 100-mL volumetric flask, and dilute with

1 YMC-Pack ODS-AQ (YMC Europe GmbH, Dinslaken, Germany), or

28 / 5 ′-Adenylic Acid / Monographs FCC 8

Spectrophotometric system, Plasma Spectrochemistry, specified, use nitric acid of a grade suitable for trace

Setup: Same as that described in the test for Arsenic, Diluent: 4% nitric acid in water

Analysis: Generate a standard curve using Diluent as a prepared by diluting a commercially available 1000 mg

correlation coefficient for the best-fit line should not be Standard solutions: 0.025, 0.05, 0.1, 0.2, 0.5, 1, and 2

Calculate the concentration (mg/kg) of cadmium in the Sample: 5 g

nitric acid in a 100-mL volumetric flask, and dilute with

Spectrophotometric system, Plasma Spectrochemistry,

C = concentration of cadmium in the Sample

Appendix IIC

solution determined from the standard

Mode: ICP–OES

W = weight of the Sample taken (g)

but set to scan for mercury at 194.164 nm

F =final volume of the Sample solution, 100 mL

Analysis: Generate a standard curve using Diluent as a

deionized ultra-filtered water When nitric acid is

Similarly, analyze the Sample solution on the ICP.

specified, use nitric acid of a grade suitable for trace

Calculate the concentration (mg/kg) of mercury in theelement analysis with as low a content of lead as

Sample taken:

practical.]

Standard stock solution: 100 µg/mL of lead prepared

of lead, from the Standard stock solution diluted with W = weight of the Sample taken (g)

Sample solution: Dissolve the Sample in 40 mL of 10% Organic Impurities

nitric acid in a 100-mL volumetric flask, and dilute with • E THANOL

Spectrophotometric system, Plasma Spectrochemistry, sodium hydroxide Add 10 mL of this solution to a

Setup: Same as that described in the test for Arsenic, Add 10 mL of this solution to a 20-mL headspace vial,

Analysis: Generate a standard curve using Diluent as a Chromatographic system, Appendix IIA

pressure-correlation coefficient for the best-fit line should not be loop headspace autosampler

Column temperature: 20 min at 40°; increase to

Injection port temperature: 140°

C = concentration of lead in the Sample solution

Detector temperature: 250°

mL)

Flow rate: 2.5 mL/min

W = weight of the Sample taken (g)

Headspace unit: 2.5 mL/min

F = final volume of the Sample solution, 100 mL

[NOTE—When water is specified as a diluent, use

deionized ultra-filtered water When nitric acid is 2 CP-Select 624 CB (Varian-Chrompack, Palo Alto, CA), or equivalent.

Transfer temperature: 90° SPECIFIC TESTS

Suitability requirement: The relative standard Sample preparation: Proceed as directed using a 10-g

deviation of the ethanol peak area responses from sample and incubating at 30–35° for 18–24 h

Analysis: Separately inject equal volumes of the • E NTEROBACTER SAKAZAKII (Cronobacter Spp.), Appendix XIIC Standard solution and Sample solution into the Sample preparation: Proceed as directed using a 10-g

measure the peak responses [NOTE—The approximate Acceptance criteria: Negative in 10 g

Acceptance criteria: The peak area from the Sample Sample preparation: Dissolve 25 g of the sample at a

solution does not exceed that from the Standard sample/broth ratio of 1/8, and proceed as directed

solution (NMT 100 mg/kg). Acceptance criteria: Negative in 25 g

Mobile phase and Chromatographic system: Prepare Method), Appendix XIIB

Sample solution: 1.0 mg/mL [NOTE—Ultra-sonication • T OTAL Y EASTS AND M OLDS C OUNT, Method I (Plate Count

for 15 min at 30° may be necessary to aid in complete Method), Appendix XIIB

Standard solution: Mixture of USP Disodium

5′-Uridylate RS, USP 5′-Adenylic Acid RS, USP 5′-Cytidylic

Acid RS, USP Disodium Guanylate RS, and USP

Sample: Standard solution

Suitability requirement 1: The relative standard

Hexanedioic Aciddeviation of the 5’-adenylic acid peak area responses

1,4-Butanedicarboxylic Acidfrom replicate injections is NMT 2.0%

Suitability requirement 2: The resolution, R, between

the 5’-adenylic acid peak and all other nucleotide

peaks is NLT 2.0

Analysis: Separately inject equal volumes of the

Standard solution and Sample solution into the

chromatograph, and measure the responses for all

nucleotide peaks on the resulting chromatograms, UNII: 76A0JE0FKJ [adipic acid]

except the peak from 5’-adenylic acid [NOTE—The

approximate retention times are 4.6 min (5′-cytidylic DESCRIPTION

acid), 6.2 min (5′-uridylic acid), 10.3 min (5′-guanylic Adipic Acid occurs as white crystals or a crystalline powder.acid), 11.5 min (5′-inosinic acid), and 27.5 min (5′- It is not hygroscopic It is freely soluble in alcohol, solubleadenylic acid).] Separately calculate the percentage of in acetone, and slightly soluble in water.

each analyte (5′-cytidylic acid, 5′-guanylic acid, 5′- Function: Buffer; neutralizing agent

inosinic acid, and 5′-uridylic acid) in the sample taken: Packaging and Storage: Store in well-closed containers.

rU = peak area of the analyte from the Sample

Tests, Appendix IIIC solution

Reference standard: USP Adipic Acid RS

rS = peak area of the analyte from the Standard

Sample and standard preparation: K

solution

Acceptance criteria: The spectrum of the sample

CS = concentration of analyte in the Standard

exhibits maxima at the same wavelengths as those in

solution (mg/mL)

the spectrum of the Reference standard.

CU = concentration of analyte in the Sample

Acceptance criteria: The sum of the percentages for all • P ROCEDURE

anhydrous basis

30 / Adipic Acid / Monographs FCC 8

Analysis: Mix the Sample with 75 mL of recently boiled Function: Stabilizer; emulsifier; thickener

stoppered Erlenmeyer flask, add phenolphthalein TS,

IDENTIFICATION

and titrate with 0.5 N sodium hydroxide to the first

• A . P ROCEDURE

appearance of a faint pink endpoint that persists for at

Analysis: Place a few fragments of unground sample or

least 30 s, shaking the flask as the endpoint is

a small amount of the powder on a slide, add a fewapproached Each mL of 0.5 N sodium hydroxide is

drops of water, and examine microscopically

• L EAD, Lead Limit Test, Flame Atomic Absorption Sample: 1 g

Spectrophotometric Method, Appendix IIIB Analysis: While stirring continuously, boil the Sample

Acceptance criteria: A clear liquid results that congeals

SPECIFIC TESTS between 32° and 39° to form a firm, resilient gel that

IIB

Analysis: Transfer the Sample into a tared 125-mL Sample solution: Prepare as directed for organic

fusing with 5 g of potassium pyrosulfate or bisulfate, Acceptance criteria: NMT 3 mg/kg

followed by boiling in 2 N sulfuric acid and rinsing with • L EAD, Lead Limit Test, Appendix IIIB

water Melt the sample completely over a gas burner, Sample solution: Prepare as directed for organic

starts, lower or remove the flame to prevent the sample Control: 5 µg Pb (5 mL of Diluted Standard Lead

from boiling and to keep it burning slowly until it is Solution)

furnace for 30 min or until the carbon is completely

SPECIFIC TESTS

removed, then cool and weigh

• A SH ( A CID -I NSOLUBLE) , Appendix IIC Acceptance criteria: NMT 0.002%

Acceptance criteria: NMT 0.5%, calculated on the dried

• W ATER, Water Determination, Appendix IIB

basis

Acceptance criteria: NMT 0.2%

Acceptance criteria: NMT 6.5%, calculated on the dried

basis

to 5 mL of the solution

Sample: 7.5 g

DESCRIPTION Analysis: Add sufficient water to the Sample to make

Agar is commercially available as white to pale yellow 500 g, boil for 15 min, and readjust to the original

strips, or in cut, flaked, granulated, or powdered forms 200 mL, heat almost to boiling, filter while hot throughAgar is a generic name given to a group of related a tared filtering crucible, rinse the container with severalmolecules with a repeating unit of agarobiose formed portions of hot water, and pass the rinsings through thebasically by D-and L-galactose units interlinked with α-1,3 crucible Dry the crucible and its contents at 105° to

D-galactopyranose unit contains a sulfate ester group It is Acceptance criteria: The weight of the residue does not

considering as such the red seaweed from phylum • L OSS ON D RYING , Appendix IIC: 105° for 5 h

Rodophyta, which belong to the Gelidiceae, Gracilariaceae, Sample preparation: Cut unground sample into 2- toand Ahnpheltiaceae families It is insoluble in cold water, 5-mm pieces before drying.

• S TARCH Sample preparation: Mineral oil mull

Analysis: Boil 100 mg of sample in 100 mL of water, Acceptance criteria: The spectrum of the sample

cylinder, fill to volume with water, mix, and allow to

Analysis: Dissolve the Sample in 3 mL of formic acid and

stand at about 25° for 24 h Pour the contents of the

50 mL of glacial acetic acid Add 2 drops of crystalcylinder through moistened glass wool, allowing the

violet TS and titrate with 0.1 N perchloric acid to awater to drain into another 100-mL graduated cylinder

blue-green endpoint Perform a blank determination

Acceptance criteria: NMT 75 mL of water is obtained.

(see General Provisions), and make any necessary

correction Each mL of 0.1 N perchloric acid isequivalent to 8.909 mg of C3H7NO2

C3H7NO2, calculated on the dried basis

DL-2-Aminopropanoic Acid

IMPURITIES

Inorganic Impurities

• L EAD, Lead Limit Test, Appendix IIIB

Sample preparation: Prepare as directed for organic

compounds

Control: 5 µg Pb (5 mL of Diluted Standard Lead

DL-Alanine occurs as a white crystalline powder It is freely • L OSS ON D RYING , Appendix IIC: 105° for 3 h

soluble in water, but sparingly soluble in alcohol The pH Acceptance criteria: NMT 0.3%

of a 1:20 aqueous solution is between 5.5 and 7.0 It melts • R ESIDUE ON I GNITION ( S ULFATED A SH) , Appendix IIC

with decomposition at about 198° It is optically inactive Sample: 1 g

Packaging and Storage: Store in well-closed,

light-resistant containers

IDENTIFICATION

Tests, Appendix IIIC

Analysis: Dissolve the Sample in 3 mL of formic acid and

Last Revision: FCC 6

50 mL of glacial acetic acid Add 2 drops of crystalviolet TS and titrate with 0.1 N perchloric acid to a

L-2-Aminopropanoic Acid

blue-green endpoint Perform a blank determination

(see General Provisions), and make any necessary

correction Each mL of 0.1 N perchloric acid isequivalent to 8.909 mg of C3H7NO2 [C AUTION —Handle

perchloric acid in an appropriate fume hood.]

Acceptance criteria: NLT 98.5% and NMT 101.5% of

• L EAD, Lead Limit Test, Appendix IIIB

L-Alanine occurs as a white crystalline powder It is freely

Sample solution: Prepare as directed for organic

soluble in water, sparingly soluble in alcohol, and insoluble

compounds

in ether The pH of a 1:20 aqueous solution is between 5.5

Control: 5 µg Pb (5 mL of Diluted Standard Lead

and 7.0

Solution)

Function: Nutrient

Acceptance criteria: NMT 5 mg/kg Packaging and Storage: Store in well-closed, light-

• O PTICAL ( S PECIFIC) R OTATION , Appendix IIB

Tests, Appendix IIIC

Sample: 10 g, previously dried Reference standard: USP L-Alanine RS

Analysis: Dissolve the Sample in sufficient 6 N Sample and Standard preparation: K

hydrochloric acid to make 100 mL

Acceptance criteria: The spectrum of the sample

Acceptance criteria

exhibits maxima at the same wavelengths as those in

[α]D20 between +13.5° and +15.5°, on the dried basis; or

the spectrum of the Reference standard.

[α]D25 between +13.2° and +15.2°, on the dried basis Acceptance criteria: NLT 20% and NMT 23% of carbon

IMPURITIES

Inorganic Impurities

• A RSENIC, Arsenic Limit Test, Appendix IIIB

Acceptance criteria: NMT 3 mg/kg

(C6H8O6)n Formula wt, calculated 176.13 • L EAD, Lead Limit Test, Appendix IIIB

Formula wt, actual (avg.) 200.00 Sample solution: Prepare as directed for organic

Solution)

Alginic Acid occurs as a white to yellow-white, fibrous

powder It is a hydrophilic colloidal carbohydrate extracted SPECIFIC TESTS

from various species of brown seaweeds (phaeophyceae) • L OSS ON D RYING , Appendix IIC: 105° for 4 h

with dilute alkali It may be described chemically as a linear Acceptance criteria: NMT 15.0%

glycuronoglycan consisting mainly of β-1,4 linked D- • R ESIDUE ON I GNITION ( S ULFATED A SH) , Appendix IIC

mannuronic and L-guluronic acid units in the pyranose ring Sample: 3 g

form Alginic Acid is insoluble in water, readily soluble in Acceptance criteria: NMT 8.0%, calculated on the driedalkaline solutions, and insoluble in organic solvents The pH basis

of a 3:100 suspension in water is between 2.0 and 3.4

Function: Stabilizer; thickener; emulsifier

Packaging and Storage: Store in well-closed containers.

Analysis: Add 1 mL of calcium chloride TS to 5 mL of

Sample solution. L-α-Aspartyl-N-(2,2,4,4-tetramethyl-3-thietanyl)-D

-Acceptance criteria: A voluminous gelatinous precipitate alaninamide, hydrated

forms

• B . P ROCEDURE

Sample solution: 1:150 in 0.1 N sodium hydroxide

Analysis: Add 1 mL of 2 N sulfuric acid to 5 mL of

UNII: 6KI9M51JOG [alitame]

Analysis: Place the Sample in a test tube Add 5 mL of

water, 1 mL of a freshly prepared 1:100 solution of

DESCRIPTION

naphtholresorcinol:ethanol, and 5 mL of hydrochloric

Alitame occurs as a white, odorless, crystalline powderacid Heat the mixture to boiling, boil gently for about

having an intensely sweet taste One method of production

3 min, and then cool to about 15° Transfer the is through a multi-step synthesis involving the reaction

contents of the test tube into a 30-mL separatory

between two intermediates, (S)-[2,5-dioxo-(4-thiazolidine)]

funnel with the aid of 5 mL of water, and extract with

acetic acid and

(R)-2-amino-N-(2,2,4,4-tetramethyl-3-15 mL of isopropyl ether Perform a blank

thietanyl)propanamide The final product is isolated and

determination (see General Provisions).

purified through crystallization of an

alitame/4-Acceptance criteria: The isopropyl ether extract from

methylbenzenesulfonic acid adduct followed by additional

the Sample exhibits a deeper purple hue than that from

purification steps, and finally recrystallization from water asthe blank

the 2.5 hydrate It is freely soluble in water and alcohol,and the pH of a 5% solution is between 5.0 and 6.0

ASSAY

Function: Sweetener; flavor enhancer

Packaging and Storage: Store in tight containers in a Analysis: Each mL of 0.25 N sodium hydroxide

cool place

consumed in the assay is equivalent to 25 mg of

(CHO) (equiv wt 200.00)

34 / Alitame / Monographs FCC 8

Sample solution: 5 mg/mL

IDENTIFICATION

Dilute sample solution: 0.5 mg/mL from the Sample

• A . I NFRARED A BSORPTION, Spectrophotometric Identification

solution Tests, Appendix IIIC

Chromatographic system, Appendix IIA

Reference standard: USP Alitame RS

Mode: High-performance liquid chromatography

Sample and standard preparation: K

Detector: UV 217 nm Acceptance criteria: The spectrum of the sample

Column: 15-cm × 0.39-cm NovaPak C18 reverse phase

exhibits maxima at the same wavelengths as those in

ion-pair (Waters, or equivalent)

the spectrum of the Reference standard.

Flow rate: 1.0 mL/min [NOTE—Maintain the Mobile

• B . P ROCEDURE

phase at a pressure and flow rate capable of giving the

Sample: 10 mg

elution times listed under Analysis.]

Analysis: To 5 mL of a solution containing 300 mg of

Injection size: 100 µL

ninhydrin in 100 mL of n-butanol and 2 mL of glacial

System suitability

acetic acid, add the Sample, and heat to gentle reflux.

Sample: Dilute standard solution B (three replicates)

Acceptance criteria: An intense blue-violet color is

Suitability requirement: The relative standard

triplicate The retention times for the beta-isomer,

Analysis: To 5 mL of a freshly prepared 0.001 M

alitame, and alanine amide should be approximately 6,

potassium permanganate solution, add the Sample, and

10, and 15 min, respectively If a column of a differentmix thoroughly

make or length is used, the retention times may vary

Acceptance criteria: The purple solution changes to

proportionally to the times listed.]

brown

Equilibrate the column by pumping Mobile phase

ASSAY through it until a drift-free baseline is obtained Inject

[NOTE—In this procedure, alitame and its impurities, into the chromatograph, and record the

alanine amide (N-(2,2,4,4-tetramethyl-3-thietanyl)-D- chromatograms Calculate the average peak areas foralaninamide) and beta-isomer (L-β-aspartyl-N-(2,2,4,4- alitame from both chromatograms

tetramethyl-3-thietanyl)-D-alaninamide hydrate) [2:5]), Calculate the weight percentage for alitame in the

performance liquid chromatography.]

Result = (rDU/rDS) × (CDS/CDU) × 100

Solution A: Dissolve 0.69 g of sodium phosphate

monobasic monohydrate and 4.32 g of

1-rDU = peak response for alitame from the Dilute

octanesulfonate, sodium in 200 mL of water Adjust

sample solution

with 85% phosphoric acid to a pH of 2.5, then dilute

rDS = peak response for alitame from Dilute

with water to 1000 mL Pass through a 0.22-µm

standard solution B

Millipore filter, or equivalent

CDS = concentration of alitame in Dilute standard

Mobile phase: Acetonitrile and Solution A (1:4) [NOTE—

solution B, corrected for water content and

Degas by sonication under aspirator vacuum for 2 min.]

purity (mg/mL)

Standard solution A: Transfer 25 mg each of a suitable

CDU = concentration of the Dilute sample solution,

alanine amide reference standard and a suitable

beta-corrected for water (mg/mL)isomer reference standard to a 500-mL volumetric flask,

Acceptance criteria: 98.0%–101.0% of alitame, on the

using 50 mL of methanol to aid in dissolution Dilute

anhydrous basiswith water to volume [NOTE—Store in a refrigerator.]

Standard solution A to a 50-mL volumetric flask, and Inorganic Impurities

Standard solution B: Transfer 50 mg of USP Alitame RS Graphite Furnace Method, Appendix IIIB

dissolve, then add 5 mL of Dilute standard solution A, Acceptance criteria: NMT 1 mg/kg

and dilute with water to volume

Dilute standard solution B: Transfer 5 mL of Standard

solution B to a 50-mL volumetric flask, and dilute with

water to volume

Organic Impurities

• A LANINE A MIDE AND B ETA -I SOMER

Solution A, Mobile phase, Standard solution A, Dilute

standard solution A, Standard solution B, Dilute

standard solution B, Sample solution, Dilute sample

solution, and Chromatographic system: Proceed as

directed in the Assay.

Analysis: Proceed as directed in the Assay Inject the

UNII: WZB9127XOA [fd&c red no 40]

Sample solution and Standard solution B into the

Calculate the average peak areas for the beta-isomer Allura Red occurs as a red-brown powder or granule It is

6-hydroxy-5-[(2-methoxy-5-Calculate the weight percentage of alanine amide and methyl-4-sulfophenyl)azo]-2-naphthalenesulfonic acid It

acid and dark red in base It is insoluble in ethanol

CS = concentration of the analyte in Standard

Analysis: Adjust the pH of three aliquots of the Sample

solution B, corrected for water content and

solution to pH 1, pH 7, and pH 13 Measure the

purity (mg/mL)

absorbance intensities (A) and wavelength maxima of

CU = concentration of the Sample solution,

these solutions with a suitable UV-visiblecorrected for water (mg/mL)

spectrophotometer

Acceptance criteria

Acceptance criteria Alanine amide: NMT 0.2% on the anhydrous basis

pH 1: A = 0.83 at 490 nm (Both neutral and acid

Beta-isomer: NMT 0.3% on the anhydrous basis

solutions exhibit a shoulder at about 410 nm.)

• T OTAL C OLOR, Color Determination, Methods I and II,

• O PTICAL ( S PECIFIC) R OTATION, Appendix IIB

Appendix IIIC: Both methods must be used.

Sample solution: 10 mg/mL, on the as-is (undried)

Method I Spectrophotometric

basis

Sample: 175 to 225 mg Acceptance criteria: [α]D25 between +40° and +50°

Analysis: Transfer the Sample into a 1-L volumetric

• W ATER, Water Determination, Appendix IIB

flask; dissolve in and dilute to volume with water

Acceptance criteria: 11%–13%

Determine as directed at 502 nm using 0.052 L/

(mg·cm) for the absorptivity (a) for Allura Red.

Method II TiCl3 Titration

Sample: 0.2 g

Allura Red1

Analysis: Determine as directed using 8.06 as the

Acceptance criteria: The average of results obtained

Class: Monoazo

IMPURITIES

Inorganic Impurities

Sample solution: Prepare as directed for organic

compounds

Acceptance criteria: NMT 3 mg/kg

1 To be used or sold for use to color food that is marketed in the United

• L EAD, Lead Limit Test, Appendix IIIB

States, this color additive must be from a batch that has been certified by the

U.S Food and Drug Administration (FDA) If it is not from an FDA-certified Sample solution: Prepare as directed for organic

batch, it is not a permitted color additive for food use in the United States, compounds

even if it is compositionally equivalent The name FD&C Red No 40 can be

Control: 10 µg Pb (10 mL of Diluted Standard Lead

applied only to FDA-certified batches of this color additive Allura Red is a

Solution)

common name given to the uncertified colorant See the monograph entitled

FD&C Red No 40 for directions for producing an FDA-certified batch. Acceptance criteria: NMT 10 mg/kg

36 / Allura Red / Monographs FCC 8

Sample solution: 250 mg/mL in 0.1 M disodium borate AS = area of the densitometer curve for the

Analysis: Use an injection volume of 20 µL for the Acceptance criteria

6,6’-Oxybis(2-naphthalenesulfonic acid), Disodium Higher and Lower Sulfonated Subsidiary Colors (as

6-Hydroxy-2-naphthalenesulfonic acid, Sodium • W ATER -I NSOLUBLE M ATTER, Color Determination, Appendix

• LAppendix IIICOSS ON DRYING (VOLATILE MATTER), Color Determination, Allyl α-Ionone

First Published: Prior to FCC 6

• CHLORIDE, Sodium Chloride, Color Determination,

Appendix IIIC

Allyl Ionone

• SULFATES (AS SODIUM SALTS), Sodium Sulfate, Color

Determination, Appendix IIIC

Acceptance criteria: NMT 15.0% in combination

Standard solution: 20 mg/mL of purified Allura Red Allyl α-Ionone occurs as a colorless to yellow liquid

(free of subsidiary colors) and 1 mg/mL each of lower Odor: Fruity, woody

and higher sulfonated subsidiary colors [NOTE—Store in Solubility: Soluble in alcohol; insoluble or practically

Analysis: Prepare a 20- × 20-cm glass plate coated with mL of 90% alcohol to give a clear solution.

a 0.25-mm layer of Adsorbent Spot aliquots of the Function: Flavoring agent

Sample solution and the Standard solution side-by-side 3

standards may be run simultaneously.] When the plate • I NFRARED S PECTRA, Spectrophotometric Identification Tests,

has air dried for 15 min, develop it in an unlined tank Appendix IIIC

equilibrated with the Developing solvent system for at Acceptance criteria: The spectrum of the sample

least 20 min Allow the solvent front to reach to within exhibits relative maxima at the same wavelengths as

3 cm from the top of the plate Allow the plate to dry those of the spectrum below

in a fume hood, and by visual inspection, compare the

ASSAY

intensities of the lower and higher sulfonated subsidiary

colors with those in the Standard solution If the

XI

subsidiary colors in the Sample solution appear more

Acceptance criteria: NLT 88.0% of C16H24O

concentrated than those in the Standard solution,

set to monitor the absorbance maximum of each

• R EFRACTIVE I NDEX , Appendix II: At 20°Calculate the percentage of each of the subsidiary Acceptance criteria: Between 1.502 and 1.507

colors, if present above 0.1%, by the formula:

method (see General Provisions).

Result = (A × p)/AS

Acceptance criteria: Between 0.926 and 0.932

OTHER REQUIREMENTS

Acceptance criteria: NMT 0.1%

Allyl α-Ionone

Allyl Cyclohexanepropionate exhibits relative maxima at the same wavelengths as

those of the spectrum below

First Published: Prior to FCC 6

Acceptance criteria: Between 1.457 and 1.462

DESCRIPTION • S PECIFIC G RAVITY : Determine at 25° by any reliable

Allyl Cyclohexanepropionate occurs as a colorless liquid method (see General Provisions).

Solubility: Miscible in alcohol, chloroform, ether; insoluble

OTHER REQUIREMENTS

or practically insoluble in glycerin, water

Solubility in Alcohol, Appendix VI: One mL dissolves in 4

38 / Allyl Cyclohexanepropionate / Monographs FCC 8

Allyl Cyclohexanepropionate

those of the spectrum below

First Published: Prior to FCC 6

• R EFRACTIVE I NDEX , Appendix II: At 20°

Allyl Heptanoate occurs as a colorless to pale yellow liquid • S PECIFIC G RAVITY : Determine at 25° by any reliable

Solubility in Alcohol, Appendix VI: One mL dissolves in 1

OTHER REQUIREMENTS

mL of 95% alcohol

Function: Flavoring agent

Allyl Heptanoate

those of the spectrum below

First Published: Prior to FCC 6

UNII: 3VH84A363D [allyl hexanoate]

• R EFRACTIVE I NDEX , Appendix II: At 20°

Allyl Hexanoate occurs as a colorless to light yellow liquid • S PECIFIC G RAVITY : Determine at 25° by any reliable

Solubility: Miscible in alcohol, most fixed oils; insoluble or Acceptance criteria: Between 0.884 and 0.890

practically insoluble in propylene glycol, water

OTHER REQUIREMENTS

Boiling Point: ∼185°

Solubility in Alcohol, Appendix VI: One mL dissolves in 6