Relação de metodologias para análises de alimentos

1. By Authority Of
THE UNITED STATES OF AMERICA
Legally Binding Document
By the Authority Vested By Part 5 of the United States Code § 552(a) and
Part 1 of the Code of Regulations § 51 the attached document has been duly
INCORPORATED BY REFERENCE and shall be considered legally
binding upon all citizens and residents of the United States of America.
HEED THIS NOTICE: Criminal penalties may apply for noncompliance.
Official Incorporator:
THE EXECUTIVE DIRECTOR
OFFICE OF THE FEDERAL REGISTER
WASHINGTON, D.C.
Document Name:
CFR Section(s):
Standards Body:
e
AOAC: Official Methods of Analysis, 1980
21 CFR 131.150(c)
AOAC International

3. OFFICIAL
METHODS OF ANALYSIS
OF THE
ASSOCIATION OF OFFICIAL
ANALYTICAL CHEMISTS
WILLIAM HORWITZ, Editor
THIRTEENTH EDITION, 1980
PUBLISHED BY THE
ASSOCIATION OF OFFICIAL ANALYTICAL CHEMISTS
PO Box 540, BENJAMIN FRANKLIN STATION
WASHINGTON, DC 20044

4. Direct inquiries related to the scientific content of Official Methods of Analysis to:
Editor, Official Methods of Analysis
Association of Official Analytical Chemists
Box 540, Benjamin Franklin Station
Washington, DC 20044 USA
Direct inquiries related to the procurement of Official Methods of Analysis, supplements (Changes in Methods). Journal of the
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COPYRIGHT 1920,1925, 1931, 1936, 1940, 1945, 1950, 1955, 1960, 1965, BYTHE
ASSOCIATION OF OFFICIAL AGRICULTURAL CHEMISTS
AND 1970, 1975, 1980 BY THE
ASSOCIATION OF OFFICIAL ANALYTICAL CHEMISTS
The methods of the Association were also copyrighted in 1916, when they
were published in the Journal of the Association of
Official Agricultural Chemists
Library of Congress Catalog Card Number: 20-21343
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ii

5. Preface to Thirteenth Edition
The most noticeable physical change in this thirteenth edition
of Official Methods of Analysis of the Association of Official
Analytical Chemists ("The Book of Methods") is its size. A survey
of users of the Book of Methods revealed an overwhelming
desire to maintain the compendium as a single volume. The
easiest way to do this was to increase the size of the page.
Users also expressed a desire for a system that will keep the
same reference number for a given method from edition to
edition. A practical system that will maintain the continuity of
the numbering system and the organizational structure of the
methods within a chapter has not yet been devised.
Approximately 175 new methods have been added during the
current five year period (1974-1978); 83 methods have been
deleted, replaced, or surplused. The approval of an average of
only 35 new methods per year represents a marked decline from
the 70 per year of the four year period of the previous edition
(1970-1973) and the 50 per year of the five year period of the
tenth edition (1965-1969). The decline is undoubtedly the result
of a number of factors. Chief among them are the greater
complexity of modern methods, requiring a large investment in
resources that is not readily mobilized to fit an Associate
Referee's schedule; and the fact that government agencies are
attempting to obtain compliance, especially of the newer stat-
utes, by promulgation of regulations and by auditing rather than
by laboratory examinations.
The greatest activity, as measured by approval of new meth-
ods, is in the field of pesticide formulations, partly as a result
of the active implementation of the cooperative agreement
with the Collaborative International Pesticide Analytical Council
(CIPAC). Other active areas include extraneous materials, vita-
mins and other nutrients, dairy products, and microbiological
methods. In fact, examination for and by biological constituents
(filth, microbiological assays, and examination for food-borne
pathogens) comprises approximately 17% of the new methods
adopted.
A comparison of the types of methods adopted between this
and the previous edition reveals that gas-liquid chromatography
has overtaken spectrophotometry in its various forms (visible,
ultraviolet, and fluorescent) as the most predominant quanti-
tative technique in Official Methods. High pressure (or perform-
ance) liquid chromatography has spurted from none to 6% of
the adopted methods within the relatively short period of five
years. Electrometric methods (potentiometric, polarographic,
and ion-selective) are now sufficiently numerous to deserve a
separate category. Infrared methods are no longer a major
factor in quantitation; they are now mainly used for identification
and confirmation. A comparison of the present and previous
editions is presented in Table 1. The figures given are only
rough approximations because of the arbitrariness often re-
quired in classifying a method and in deciding when a new
method or revision is sufficiently independent to warrant con-
tributing to the statistics.
iii
Table 1. Classification of new methods approved by the AOAC
in the thirteenth and twelfth editions
Method classification
Gas-liquid chromatography
Spectrophotometry
Titrimetric
Filth, isolation
Automated
Biological and microbiological
High pressure liquid chromatography
Atomic absorption
Electrometric
Chromatography (thin layer, etc.)
Gravimetric
Miscellaneous (physical, qualitative,
etc.)
13th edition
(1980)
18%
16
10
8
7
9
6
4
5
3
2
11
12th edition
(1975)
14%
20
7
14
5
8
9
3
6
13
The most interesting new collaboratively studied method is
the mass spectrometric method for the detection of adulteration
of honey with high fructose corn sirup. Corn sirups (from a
monocotyledonous plant) have a distinctly different 13C/12
C ratio
than sirups from most dicotyledonous plants, which are the
source of most honeys. The isotope ratio mass spectrometer
required forthis determination is a highly specialized instrument,
even in the field of mass spectrometry. Despite the rarity of the
instrumentation, sufficient laboratories participated in the col-
laborative study to establish the reproducibility of the method.
A problem that has arisen is how to handle the numerous
individual instruments of diverse design and manufacture that
have been developed to automate a particular determination.
Even if this problem is solved by incorporating all the available
instruments into the initial collaborative study, the problem
returns with the first "new and improved" modification. Different
instrument designs and their subsequent modifications have
been handled in the infrared determination of milk constituents
by providing performance specifications which must be met by
the basic instrument in general, when compared to a reference
method or reference sample. In addition, the user must satisfy
himself that his particular instrument also meets the perform-
ance specifications by frequent comparisons with the reference
method or sample. This requirement eliminates the need for
repeated collaborative studies every time a manufacturer rede-
signs or modifies his basic equipment, and in addition provides
a continuous quality control technique on the performance of
the instrument, method, and laboratory.
It should not be overlooked that automation is not confined
to physical and chemical determinations. The microbiological
chapter contains three collaboratively studied instrumental
methods for somatic cell count and an instrumental method for
distributing a liquid sample for plate counting. Biochemical
diagnostic kits have also been evaluated collaboratively for their

6. effectiveness in identifying Salmonella and other enteric isolates.
A criticism that is often leveled at the AOAC validation
mechanism is that it is too slow to keep up with the pace of
requirements for methods by regulatory agencies and the
regulated industry. This objection overlooks the point that the
speed with which a method is validated is almost completely in
the hands of those needing the validated method, rather than
in the hands of the AOAC. The method must be tested for
ruggedness, and the directions tested for clarity; samples must
be prepared and distributed; laboratories must analyze the
samples and report the results. Dubious results must be inves-
tigated and occasionally samples must be reanalyzed. The data
must be tabulated and analyzed, and a report written. There is
no way to short-cut the process of obtaining intra- and interlab-
oratory variability except to conduct the necessary experimental
work and perform the necessary statistical calculations. Only
then is the recommendation subject to the time restrictions of
the AOAC mechanism. To ensure that the AOAC mechanism of
annual approval is not holding up use of needed methods, a
new temporary class of methods has been introduced entitled
"Interim First Action." These are methods which, together with
their supporting studies, have been received between annual
meetings and have been sent through the customary reviewing
procedure. To be designated Interim First Action, a method
must have been approved by the appropriate Referee and
Subcommittee, and by the Chairman of the Committee on
Official Methods. The method only lacks the formal vote by
those AOAC members having regulatory authority over the
commodity involved. Such formal approval is usually provided
at the next annual meeting, at which time the method becomes
"First Action." Approximately five interim methods are approved
annually. Several such methods appear in this volume, depend-
ing upon the time of their submission and review in relation to
the editorial status of the chapter to which they are assigned.
The current status of these Interim First Action methods, as well
as the status of any method, can be found by consulting the
latest cumulative index to Changes in Methods, which appears
as the final pages of each March issue of the Journal.
For those users of AOAC methods who may not be familiar
with the procedures by which a method is included in this
volume, we are reprinting a paper prepared for the Joint
International Symposium, "The Harmonization of Collaborative
Studies," held in London, England 9-10 March 1978. The
document was drafted by the AOAC Committee on Collaborative
Studies: Elwyn D. Schall, Chairman; Charles W. Gehrke, William
Horwitz, Anthony J. Malonski, James P. Minyard, Jr., Forrest W.
Quackenbush, and Ernest S. Windham. This paper appeared in
Analytical Chemistry (March 1978), 50, 337A-340A.
The publication of this "Book of Methods" is possible only
because of the extensive cooperation of thousands of analysts
who have volunteered to direct, participate in, and review the
numerous collaborative studies that form the basis for inclusion
of the several thousand methods that appear herein. Special
recognition is due to Mrs. Betty Johnson who has prepared an
entirely new index for this edition.
28 September, 1979
William Horwitz, Ph.D.
Editor
Abstract from
Preface to First Edition
"In presenting this revision of the official and tentative methods of analysis of the Association of Official Agricultural Chemists, it
is appropriate to give a brief statement of the organization of the Association, its purpose, and the procedure by which the methods
are adopted.
"Membership in the Association is institutional and includes the State Departments of Agriculture, the State Agricultural Colleges
and Experiment Stations, the Federal Department of Agriculture, and the Federal, State, and City offices charged with the enforcement
of food, feed, drug, fertilizer, insecticide and fungicide control laws.
"The Association was founded at Philadelphia, Pa., September 9, 1884, by the following representative agricultural chemists of that
time, the organization being the result of a series of informal meetings held the immediately preceding years:
"Prof. H. W. Wiley, Chemist of the Department of Agriculture, Washington, D.C.
Mr. Clifford Richardson, Assistant Chemist of the Department of Agriculture, Washington, D.C.
Mr. Philip E. Chazal, State Chemist of South Carolina.
Dr. Chas. W. Dabney, Jr., State Chemist of North Carolina.
Dr. W. J. Gascoyne, State Chemist of Virginia.
Dr. E. H. Jenkins, Connecticut Experiment Station.
Prof. John A. Meyers, State Chemist of Mississippi.
Prof. H. C. White, State Chemist of Georgia.
Mr. C. DeGhequier, Secretary National Fertilizer Association.
Dr. Schumann, Dr. Lehmann, Mr. Gaines and others."
iv

7. Contents
Preface to Thirteenth Edition ......................... .
Preface to First Edition .............................. .
List of Illustrations .................................. .
Collaborative Study Procedures of the Association of
Official Analytical Chemists ........................ .
Definitions of Terms and Explanatory Notes .......... .
CHAPTER
1. Agricultural Liming Materials .................... .
Calcium Silicate Slags ........................ .
Elemental Analysis ........................... .
2. Fertilizers ....................................... .
Free Water ................................... .
Phosphorus .................................. .
Nitrogen ..................................... .
Potassium ................................... .
Other Elements ............................... .
Peat ......................................... .
3. Plants .......................................... .
Metals ....................................... .
Nonmetals ................................... .
Other Constituents ........................... .
Pigments .................................... .
Tobacco ..................................... .
4. Disinfectants .................................... .
Phenol Coefficient ............................ .
Use-Dilution Method .......................... .
Available Chlorine Germicidal Equivalent
Concentration .............................. .
Sporicidal Test ............................... .
Fungicidal Test ............................... .
Germicidal and Detergent Sanitizers ........... .
Germicidal Spray Products .................... .
Water Disinfectants for Swimming Pools ....... .
Tuberculocidal Activity ........................ .
Bacteriostatic Activity of Laundry Additives .... .
5. Hazardous Substances .......................... .
Paint ......................................... .
Preparations Containing Fluorides ............. .
Preparations Containing Methanol ............. .
Preparations Containing Phenol ............... .
Soda Lye .................................... .
Earthenware ................................. .
6. Pesticide Formulations .......................... .
General Methods ............................. .
Arsenic .................................... .
Lead ....................................... .
Copper .................................... .
Zinc ....................................... .
Fluorine .................................... .
PAGE
iii
iv
ix
xii
xv
2
3
7
9
9
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18
20
27
31
31
38
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49
51
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56
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59
60
61
61
63
64 '
65
67
69
69
69
70
71
71
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72
72
72
74
74
75
75
v
Contamination by Organochlorine Pesticides ..
Volatility of Ester Forms of Hormone-Type
Herbicides ............................... .
Inorganic and Organometallic Pesticides and
Adjuvants ................................ .
Paris Green ................................ .
Lead Arsenate .............................. .
PAGE
77
77
78
78
78
Calcium Arsenate. . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
Zinc Arsenite. . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . 79
Copper Carbonate .. . . . . . . . . . . . . . . . . . . . . . . . . . 79
Copper Naphthenate . . . . . . . . . . . . . . . . . . . . . . . . . 79
Bordeaux Mixtures .......................... 80
Potassium Cyanate. . . . . . . . . . . . . . . . . . . . . . . . . . 81
Cyanides. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
Lime Sulfur Preparations. . . . . . . . . . . . . . . . . . . . . 81
Hypochlorites ............................... 82
Chloramine T ............................... 83
Mineral Oils. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
Soap....................................... 83
Mineral Oil-Soap Emulsions. . . . . . . . . . . . . . .. . . 83
Mercurial Seed Disinfectants. . . . . . . . . . . . . .. . . 84
Ant Poisons and Rodenticides . . . . . . . . . . . . . . . . 85
Fumigants .................................. 85
Pesticides Related to Natural Products and Their
Synergists ................................ 86
Allethrin .................................... 86
Derris and Cube Powder. . . . . . . . . . . . . . . . . .. . . 87
Pyrethrins . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . 89
Tobacco and Tobacco Products .............. 90
Halogenated Pesticides ........................ 91
Aldrin, Dieldrin, and Endrin .................. 91
Amiben ..................................... 93
BHC ........................................ 93
Balan or Trifluralin .......................... 95
Captan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96
Chlordane. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97
Chlorotoluron, Chloroxuron, or Metoxuron . . . . 99
Dichlobenil .. . . .. . ... . . . .. .. . .. . . ... . . . .. ... . 100
DDT.. . . . ... . . ..... . .. . . . .. . ... . . . . .. . .... . . 100
Dacthal ..................................... 102
Dicamba .................................... 102
2,4-D ....................................... 103
Dalapon .................................... 104
Dicofol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 104
Fluometuron ................................ 105
Folpet ...................................... 105
Heptachlor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105
Picloram-2,4-D .............................. 106
Sodium Trichloroacetate ......... " .. . . .... .. 107
Nonhalogenated Pesticides .................... 107
Aldicarb .................................... 107
Amitrole .................................... 107
Carbaryl .................................... 108
DDVP ................... '" ..... " . . ... . .. . . 108
Diquat and Paraquat ............. " . . . . . . . . . . 109

8. PAGE
Dithiocarbamates . . . . . . . . . . . . . . . . . . . . . .. . . . . . 110
Thiram ............................... '" . ... 111
Dodine...................................... 111
Formaldehyde. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
Ethion ...................................... 112
Formothion ................................. 113
Malathion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
Parathion ................................... 116
Phorate ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
Sulfoxide ................................... 120
Tetraethylpyrophosphate .................... 120
Organic Thiocyanates . . . . . . . . . . . . . . . . . . . . . . . . 120
Thiocarbamates ............................. 121
Triazines, Diazinon, and Dichlorobenzilates . . . . 121
Triphenyltin Compounds. . . . . . .. . . . . . . .. . . . . . 122
Quaternary Ammonium Compounds ........... 123
7. Animal Feed..................................... 125
Nitrogen ...................................... 125
Other Constituents ............................ 132
Preservatives. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141
8. Baking Powders and Baking Chemicals ........... .
9. Beverages: Distilled Liquors ..................... .
Spirits ....................................... .
Cordials and Liqueurs ........................ .
10. Beverages: Malt Beverages and Brewing Materials
Beer ......................................... .
Malt ......................................... .
Cereal Adjuncts .............................. .
Hops ........................................ .
Brewing Sugars and Sirups ................... .
Wort ......................................... .
Yeast ........................................ .
Brewers' Grains
11. Beverages: Wines ............................... .
Preservatives ................................. .
Flavors ...................................... .
12. Beverages: Nonalcoholic and Concentrates ....... .
13. Cacao Bean and Its Products .............. " ..... .
Shell ......................................... .
Cacao Products Processed with Alkalies ....... .
Chocolate Liquor ............................. .
Fat .......................................... .
Dairy Ingredient Constituents ................. .
Saccharine Ingredients Other Than Lactose .... .
Starch ....................................... .
143
147
147
159
162
162
171
177
178
180
181
182
183
185
192
193
194
199
200
205
205
205
206
207
208
Chocolate Products. . . . . . . . . . . . . . . . . . . . . . . . . . . . 209
Theobromine. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 209
14. Cereal Foods. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . 211
Wheat Flour. . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . 211
Wheat, Rye, Oats, Corn, Buckwheat, Rice, and
Barley and Their Products Except Cereal
Adjuncts .................................... 220
Soybean Flour ................................ 223
Bread....... . . . .... . ..... ..... ... . . . . . .... . ... 223
Baked Products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226
PAGE
Baked Products Other Than Bread (Not
Containing Fruit) ............................ 227
Fig Bars and Raisin-Filled Crackers ............. 227
Macaroni, Egg Noodles, and Similar Products. . . 228
15. Coffee and Tea ....................... " . . . . . .. . . . 233
Green Coffee. . . . . . . . . . . . . . .. . . . . . . . . . . . . . .. . . . 233
Roasted Coffee . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233
Tea........................................... 235
16. Dairy Products. . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . 238
Sampling ..................................... 238
Milk.......................................... 240
Automated Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . 246
Cryoscopic Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . 249
Penicillins in Milk. . . . . . . . . . .. . . . . . . . . . . . . . . . . . . 255
Cream........................................ 256
Evaporated Milk. . . . . . . . . . . . . . . . . . . .. . . . . . .. . . . 258
Sweetened Condensed Milk ........ " . . . . . .. . . . 259
Dried Milk, Nonfat Dry Milk, and Malted Milk ... 259
Butter. .... ... .... . .. . . . . .... . ... . ....... . ..... 260
Cheese....................................... 264
Ice Cream and Frozen Desserts. . . . . . . . . . . . . . . . . 272
17. Eggs and Egg Products. . . . . . . .. . . . . . . . . . . . . . . . . . . 275
18. Fish and Other Marine Products ....... " . . . .. ..... 285
19. Flavors .......................................... 306
Alcohol ....................................... 306
Vanilla Extract and Its Substitutes .............. 306
Lemon, Orange, and Lime Extracts and Flavors. 313
Lemon and Orange Oils. . . . .... . ..... ... . ... . . . 316
Almond Extract. . .... . ... . . .. ... . ....... . ..... . 318
Cassia, Cinnamon, and Clove Extracts. ... . .... . 319
Ginger Extract. . ..... ..... . .... . . .... . . . . .... .. 319
Peppermint, Spearmint, and Wintergreen
Extracts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 319
Anise and Nutmeg Extracts .................... 319
Other Extracts and Toilet Preparations. . . . . . . . . . 319
20. Food Additives: Direct. . . . . . . . . . . . . . . . . . . . . . . . . . . . 323
Acidulants .................................... 323
Antioxidants .................................. 323
Chemical Preservatives ........................ 325
Emulsifying Agents. . . . . . . . . . . . . . . .. . . . . . . .. . . . 343
Enzymes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 344
Miscellaneous. . . . . . . . . . . . . . . . . . . . .. . . . . . . .. . . . 345
Nonnutritive Sweeteners. . . . . . . . . . . . . . . . . . . . . . . 347
Nutrients. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 352
Solvents ...................................... 352
21. Food Additives: Indirect. . .. .... . .... .. . . . ... ..... 354
22. Fruits and Fruit Products ......................... 359
Moisture. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361
Solids........................................ 362
Potassium .................................... 363
Sodium....................................... 364
Phosphorus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 364
Betaine ....................................... 365
Fruit Acids .................................... 366
Sugars........................................ 369
Anthocyanins ................................. 370
vi

9. PAGE PAGE
Lemon Juice .................................. 372 Carbaryl .................................... 483
Sulphenone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 485
23. Gelatin, Dessert Preparations, and Mixes . . . . . . . . . . 374 Perthane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 485
DDT........................................ 485
24. Meat and Meat Products ......................... 376 Dichlone ................. . . . . . . . . . . . . . . . . . . . 487
Meat.......................................... 376 Guthion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 488
Meat Extracts and Similar Products. . . . . . . . . . . . . 383 Dodine...................................... 488
Ethylenethiourea ............................ 488
25. Metals and Other Elements as Residues in Foods .. 385 Glyodin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 489
Antimony ..................................... 385 Hexachlorobenzene and Mirex ............... 490
Arsenic ....................................... 385 Malathion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 490
Cadmium ..................................... 388 Maleic Hydrazide .................. . . . . . . . . . . 490
Copper....................................... 391 Methoxychlor ............................... 491
Fluorine. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 392 Mirex . . . . . . . .. . .. . . ... . . . . . . .. . . . . ... . .. . . . . 492
Lead.......................................... 397 Monofluoroacetic Acid. . . . . . . . . . . . . . . . . . . . . . . 492
Manganese ................................... 405 Naphthaleneacetic Acid. . . . . . . . . . . . . .. . . . . . . . 493
Mercury ...................................... 405 Nicotine .................................... 494
Nickel. . .. . ... ... . .... . .. . . .. . ... . ... . . . .. . ... . 409 Parathion ................................... 495
Selenium ..................................... 409 Piperonyl Butoxide .......................... 495
Tin........................................... 410 Thiram...................................... 495
Titanium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 411
Zinc.......................................... 411 30. Spices and Other Condiments .................... 497
Spices........................................ 497
26. Natural Poisons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 414
Mycotoxins ................................... 414
Aflatoxins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 415
Prepared Mustard ............................. 500
Dressings for Foods ........................... 501
Vinegars. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . 503
Minicolumn Detection Methods .............. 417
Quantitative Methods. . . . . . . . . . . . . . . . . . . . .. . . 418
Aflatoxin M, . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 427
Ochratoxins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 428
Patulin. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 430
Sterigmatocystin ............................ 431
Zearalenone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 432
Marine Toxins. . . . . . . . . . . . . . . . . . . . . .. . . . . . . .. . . 433
Phytotoxins ................................... 433
31. Sugars and Sugar Products. . . . . . . . . . . . . . . . . . . . . . . 506
Sugars and Sirups. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 506
Molasses and Molasses Products. .. . . . .. . .. . .. . 516
Confectionery. ... .. . . . .. . . .. . .. . .. . . . .. . . . .. . . 519
Honey........................................ 520
Maple Products ............................... 528
Sugar Beets. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 532
Starch Conversion Products. . . . . . . . . . . . . . . . . . . . 532
27. Nuts and Nut Products ........................... 435
32. Vegetable Products, Processed. . . . .. . . . .. .. . . .. .. . 537
Canned Products .............................. 537
Peanut Butter ................................. 435
Shredded Coconut. . .. . . . . . . . . . . . . . . . . . . . . . . . . . 436
Dried Vegetables .............................. 542
Frozen Vegetables. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 543
28. Oils and Fats. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 437 33. Waters; and Salt . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 547
.. ?hysical Constants ............................ 437 Water......................................... 547
Chemical Constants ........................... 438 Salt........................................... 565
Acid Constituents. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 441
Sterols. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 452 34. Color Additives .................................. 568
Tests for Specific Oils. . . . . . . . . . . . . . . . . . . . . . . . . . 456 Separation and Identification of Color Additives
Chick Edema Factor ........................... 459 in Foods, Druq!'; and Cosmetics. . . . . . . . . .. . . . 568
Mono- and Diglycerides . . . . . . . . . . . . . . . . . . . . . . . . 461 Analysis Of r brcial Synthetic Organic Color
t:'laxseed ...................................... 464 Additives .....:.............................. 570
29. Pesticide Residues ............................... 466 35. Cosmetics ....................................... 582
Multiresidue Methods ......................... 466 General Methods. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 582
General Method for Organochlorine and Deodorants and Anti-Perspirants ............... 583
Organophosphorus Pesticides. . . . . . . . . . . . . . 466 Dipilatories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 587
Multiple Residue Methods for Face Powders ................................. 587
Organophosphorus Pesticides. . . . . . . . . . . . . . 476 Hair Preparations. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . 587
Multiple Residue Methods for Fumigants ..... 480 Suntan Preparations. . . . . . . . . . . . . . . . . . . . . . . . . . . 589
Multiple Residue Method for Aryl N- Vanishing Cream. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . 589
Methylcarbamate Insecticides .............. 480 Bioassay. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 590
Methods for Individual Residues ............... 481
Benzene Hexachloride ....................... 481 36. Drugs: General. . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . 591
Biphenyl .................................... 481 General Directions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 591
Aramite . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 483 Solvents ...................................... 591
Captan ...................................... 483 Halogenated Compounds. . . . . . . . . . . . . . . . . . . . . . 592
vii

10. PAGE
Inorganic Drugs ............................... 594
Mydriatics and Myotics ........................ 604
Microscopic Tests ............................. 604
37. Drugs: Acidic .................... '" ........ '" .. 614
Acids......................................... 614
Antipyretic Drugs. .... . ... . ... ... . . ..... ..... .. 615
Hypnotic Drugs ................. " . .. . ... ..... . 619
Phenolic Drugs. . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . 625
Sulfonamide Drugs. .. . . . . . . . . . . . . . . . . . . . . . .. . . 629
Other Imide Drugs. . . . . . . . . . . . . . . . . .. . . . . . . .. . . 631
38. Drugs: Alkaloid and Related Bases ................ 636
Alkaloids, Opium. . . .. . . . . . . . . . . . . . . . . . . . . . . . . . 636
Alkaloids Other Than Opium ................... 639
Other Bases. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 649
39. Drugs: Neutral ................................... 667
Steroids and Related Hormones. . . . . . . . . . . . . . . . 667
Nonalkaloidal Vegetable Drugs and Their
Derivatives. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 674
Miscellaneous. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 678
40. Drugs: Illicit. .. .. ... . . ..... ... .... . . .... . . . ... .. . . 685
41. Drugs and Feed Additives in Animal Tissues. . . . . . . 688
ANOT......................................... 688
Arsenic ....................................... 688
Clopidol ...................................... 689
Decoquinate .................................. 690
Ethoxyquin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 691
Melengestrol Acetate .......................... 691
Nalidixic Acid ................................. 693
Zoalene . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 694
42. Drugs in Feeds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 695
Qualitative Tests .............................. 695
Arsenic ....................................... 695
Cyzine ........................................ 696
Aklomide ..................................... 697
p-Aminobenzoic Acid. . . . . . .. . . . . . . . . . . . . . . . . . . 697
Enheptin ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 698
Amprolium ............................... " . . . 698
Arsanilic Acid ................................. 699
Bithionol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 699
Buquinolate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 699
Cadmium Anthranilate. . . . . . . . . . . . . . . . . . . . . . 700
Carbadox ..................................... 700
Decoquinate .................................. 700
Dibutyltin Dilaurate ., ..................... " . . . 701
Diethylstilbestrol .............................. 702
Nitrophenide ....... .
Nitarsone .......... .
Phenothiazine ...... .
Piperazine .......... .
Pyrantel Tartrate ... .
Racephenicol ....... .
Reserpine ....... " ..
Ronnel .............. ,
Roxarsone .......... ,
Sulfonamide Drugs ..
Thiabendazole ....... ,
Zoalene ............. ,
Antibiotics .......... ,
43. Vitamins and Other Nutri
Chemical Methods ...
Microbiological Metho
Bioassay Methods ...
Nutritionally Related C
44. Extraneous Materials: IsCl
General ............ .
Beverages and Bevera
Dairy Products ..... .
Nuts and Nut Product~
Grains and Their Prodl
Baked Goods ....... .
Breakfast Cereals ... .
Eggs and Egg Product
Poultry, Meat, and Fisl
Products ......... .
Fruits and Fruit Produ(
Sugars and Sugar Prol
Vegetables and Vegetc
Spices and Other ConI
Animal Excretions .. .
Miscellaneous ...... .
45. Forensic Sciences ..... .
Latent Fingerprints ..
Glass Fragments ....
Mineral Wool Insulatio
Voice Print Identificati(
46. Microbiological Methods
Eggs and Egg Product
Frozen, Chilled, Precoe
Commercial Sterility 0
Thermophilic Bacterial
Clostridium perfringen
Salmonella . ........ .
Optical Somatic Cell C

11. FIGURE
1:01.
2:01.
2:02.
2:03.
2:04.
2:05.
2:06.
2:07.
2:08.
3:01.
3:02.
3:03.
3:04.
3:05.
3:06.
3:07.
4:01.
4:02.
6:01.
6:02.
6:03.
6:04.
6:05.
6:06.
6:07.
6:08.
6:09.
7:01.
7:02.
7:03.
Illustrations
Apparatus for Automatic Filtration and Measurement of Lime Solutions ........................................... .
Sampling Pattern ................................................ " ............................................. .
Sampling Cup ..... " ................................... , ........ " ............................................. .
Missouri and Indiana Weighted Restricted-fill Fluid Fertilizer Sampling Bottles Designed to Fill While Being Lowered
(and Raised) in Storage Tanks ................................................................................... .
Sampling Apparatus for Ammoniacal Solutions, Including "Quick Coupler" for Attaching to Storage Tanks .......... .
Apparatus for Control of Water Pressure ......................................................................... .
Flow Diagram for Automated Analysis for Phosphorus ........................................................... .
Flow Schematic for K20 in Fertilizers ............................................................................ .
Apparatus for Elution of Resin Column .......................................................................... .
Suction Device Used in Micro Method for Determining Calcium ................................................... .
Schematic Drawing of Air Flow System Used in Semiautomated Analysis for Fluoride ............................. .
Flow Diagram for Semiautomated Analysis for Fluoride .......................................................... .
Schematic Drawing of Microdistillation Apparatus ................................................ , ............... .
Microdistillation Column ........................................................................................ .
Apparatus for Determining Lignin ............................................................................... .
Distillation Apparatus ........................................................................................... .
Transfer Loop and Manner of Using in Phenol Coefficient Technic ................................................ .
Stainless Steel Spindle for Winding Test Fabric .................................................................. .
Delivery Assembly for Sampling Pressurized Containers .......................................................... .
Apparatus for Distilling Arsenious Chloride ...................................................................... .
Apparatus for Determining Fluorine ............................................................................. .
Partition Column and Solvent Evaporator ........................................................................ .
Melting Point Apparatus ........................................................................................ .
Distillation Apparatus ........................................................................................... .
Carbon Disulfide Evolution Apparatus .................................................................... ..... .
Absorption System for Thiram ............................................................................. : .... .
Filtration Apparatus ............................................................................................ .
Flow Diagram for Semiautomated Analysis for Crude Protein ..................................................... .
Agitator ........................................................................................................ .
Oklahoma State Filter Screen ................................................................................... .
PAGE
2
7
7
7
8
9
12
20
26
32
41
42
43
43
48
54
57
67
72
73
76
94
94
99
110
111
122
128
131
133
7:04. Modified California State Buchner Funnel... .... . ... . ... . .. . . . ... .... . .... . ... .... . . . .... ... . . ... ... .. . .... ... ... . 133
7:05. Continuous Heater for Distilled Water, 1.25% Alkali, and 1.25% Acid ............................................... 133
8:01. Chittick Apparatus for Gasometric Determination of Carbon Dioxide ............................... :-................ 143
9:01. 100 mL and 50 mL Pycnometers ................................................................................. 148
9:02. Williams Tube. . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149
9:03. Steam Distillation Flask. . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156
9:04. Apparatus for Determining Hydrogen Cyanide in Distilled Spirits................................................... 158
10:01. Absorption Buret................................................................................................ 166
11 :01. Distillation Apparatus for Chemical Determination of Alcohol in Wine .............................................. 186
11 :02. Volatile Acid Still (Cash Still) .......................... " ............ '" ..................................... '" . . 188
11 :03. Carbon Dioxide Apparatus; Manometric Method ...................................... '" ............... '" . ...... 190
11 :04. Carbon Dioxide Apparatus; Volumetric Method ........ '" ................ '" . '" . . . . ...... . . ...... ... .... ... . .. . . 192
12:01. Continuous Extraction Apparatus .......... " . '" ............................ ,. . . ..... ... . . ... . .. . ... ... . . ... ... .. 195
13:01. Spiral Vessel Sections. . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . .. . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203
13:02. Stone Cells ..................................................................................................... 204
13:03. Graph Used in Correcting Cuprous Oxide for Effect of Sucrose ........................................ '" . . .. . ... .. 207
16:01. Apparatus for Drying Pentabromacetone by Aspiration ............................................................ 241
16:02. Liquid Extractor ...................................... '" . " ....................................... , ... . . .. .... .. 242
16:03. Flow Diagram for Determination of Fat in Milk ......... '" ......... , ..... '" . .... . . .... ... . .... ... ... . .. . . ... . .. . . 247
16:04. Modified Jones Reductor ........................................................................................ 266
16:05. Modified Plastic Desiccator ...................................................................................... 273
18:01. Extraction Apparatus............................................................................................ 291
18:02. Steam Distillation Assembly ..................................................................................... 292
ix

12. FIGURE PAGE
18:03. Muscle Bath .................................................................................................... 294
18:04. Electrophoresis Cabinet. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . 300
18:05. Disc Electrophoresis Apparatus .................................................................................. 300
18:06. Key to Identification of Canned Salmon Species by Scale Characteristics ........................................... 302
18:07. Sockeye (Red) Salmon Scale (Oncorhynchus nerka) ............................................................... 303
18:08. Chinook (King) Salmon Scale (Oncorhynchus tshawytscha) ........................................................ 304
18:09. Coho (Silver) Salmon Scale (Oncorhynchus kisutch) . . . . .. . . . . . .. .. . . . ... . . .. . . ... . . . . ... . . .. . .. ... . .. . . . .. . . ... . . . 304
18:10. Chum Salmon Scale (Oncorhynchus ketal ........................................................................ 304
18:11. Pink Salmon Scale (Oncorhynchus gorbuscha) ........ ............................... .... ................... ...... 304
19:01. Wilson Flask.................................................................................................... 315
19:02. Oil Separator Trap. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . 320
20:01. Steam Distillation Apparatus. . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 326
20:02. Placement of Slides on Template for Applying Silica Gel Coating .................................................. 327
20:03. Steam Distillation Apparatus. . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . 338
20:04. Apparatus for Modified Monier-Williams Method for Sulfur Dioxide. . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . 339
20:05. Alternative S02 Absorber. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . .. . . . . 339
20:06. Gas Chromatograms of TBS, DBS, and TBS + DBS . . . . . .. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 346
20:07. Microsampling Die .............................................................................................. 348
20:08. GLC Effluent Collection Trap for Determination of Cyclohexylamine ................................................ 350
21 :01. Test Cell. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . 356
21:02. Oven Rack...................................................................................................... 357
22:01. Apparatus for Determining Volume of Frozen Fruits by Displacement............................................... 359
22:02. Capillary Viscometer ............................................................................................ 361
22:03. Electrical Circuit Diagram for Dried Fruit Moisture Tester.. . . . . ... . . .. . . ... . . .... . . .... .. .. . . ... . ... . ... . . . .. . . ... . 361
22:04. Connecting Tube Adapter for Direct Distillation ................................................................... 370
23:01. 0.5 Inch Bloom Gelometer Plunger ............................................................................... 374
23:02. 1.0 Inch Bloom Gelometer Plunger. . . . . .. .. . ... .. .. . . ... ... . . . ... . ... . . . .. . . . .. . . . . . ... ... . . . ... . .. . ... . . . .... ... 375
24:01. Helix Inlet Manifold. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . 377
24:02. Phosphorus Analytical Manifold. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . 378
24:03. Nitrogen Analytical Manifold. . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . 379
25:01. Arsenic Apparatus. . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . . . . . . 386
25:02. Apparatus for Flameless Atomic Absorption Analysis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 406
25:03. Digestion Vessel ................................................................................................ 407
25:04. Special Digestion Apparatus for Mercury Residues. . . . . . . . . . . . . .. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . 408
26:01. Spotting and Scoring Patterns for 2-Dimensional TLC Plates. . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . 424
26:02. Plate Scraper for Removing Adsorbent From TLC Plates .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 424
26:03. Schematic Showing Parts of Egg and Modes of Test Material Introduction. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . 426
28:01. Titer Stirring Assembly. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . .. . . . 439
28:02. Liquid-Liquid Extractor. . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . 442
28:03. Apparatus for Determining Reichert-Meissl and Polenske Values ................................................... 443
28:04. Constant Temperature Bath and Accessories. . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . 444
28:05. Distribution Heads; Manifold. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445
28:06. Glass Micro Filter for Sterol Acetate Precipitates .................................................................. 452
28:07. Crystalline Forms of Free Sterols. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . 453
28:08. Apparatus for Determining Melting Point ......................................................................... 458
29:01. KCI Thermionic Detector Coil for In-series Dual Detection System .................................................. 468
29:02. KCI Thermionic Detector Coil for Parallel and In-series Split Dual Detection Systems ................................ 469
29:03. In-series Split Dual Detection System. . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . .. . . . . . . .. . . . . . . . . . . . . . . . . . . 469
29:04. Parallel Dual Detection System. . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . 469
29:05. In-series Dual Detection System. .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . . . . . . 469
29:06. Sweep Co-distillation Apparatus. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 477
29:07. Evaporative Concentrator. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . .. . . . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . 484
29:08. Distillation Apparatus for Maleic Hydrazide Determination ......................................................... 491
30:01. Apparatus for Volatile Oil in Spices. . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . 498
31 :01. Fractionator..................................................................................................... 506
31 :02. Carbon Combustion and Purification System. . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . 527
31 :03. Distillation Apparatus for Determining Formaldehyde in Maple Sirup.. . . .... . ... .. . . . ... . ... . . ... . .. . . ... . ... . ... . . 530
33:01. Apparatus for Determination of Mercury by Flameless Atomic Absorption. . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . 560
33:02. Phosphorus Manifold. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 563
34:01. Titanous Chloride Titration Apparatus ............................................................................ 574
34:02. Sulfiding Apparatus ............................................................................................. 579
36:01. Apparatus for Determining Arsenic in Iron-Arsenic Tablets. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . 594
36:02. Flow Diagrams for Semiautomated Analysis for Ferrous Sulfate. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . 601
37:01. Bromine Apparatus.............................................................................................. 619
x

13. FIGURE PAGE
37:02. Flow Diagram for Automated Analysis for Acenocoumarol, Dicumarol, Phenprocoumon, Potassium Warfarin, and
Sodium Warfarin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 629
38:01. Flow Diagram for Semiautomated Fluorometric Analysis for Reserpine............................................. 646
38:02. Flow Diagram for Phenylephrine Hydrochloride ................................................................... 661
38:03. Assembly of Debubbler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 662
39:01. Flow Diagram for Semiautomated Analysis for Prednisolone or Prednisone. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 674
39:02. Flow Diagram for Automated Analysis for Digoxin ................................................................ 675
39:03. Scrubber Trap for Ammonia Distillation .......................................................................... 679
39:04. Flow Diagram for Automated Analysis of Methenamine and Methenamine Mandelate. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 683
43:01. Flow Diagram for Automated Analysis for Niacin and Niacinamide.. .... . .... ..... . . . ........ . .... .... ... . . ... . ... . 745
43:02. RRR-a-Tocopherol . . . . . . . .. . . . . . . .. . . . . . .. . . . . . . .. . . . . .. . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . 752
43:03. Line Test Chart. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 771
43:04. Radii Sections Scored According to Line Test Chart ............................................................... 772
44:01. Mechanical Butter Stirrer ........................................................................................ 781
44:02. Wildman Trap Flask .......................................................................... , .... ........ . ... .. 781
44:03. Rot Fragment Counting Slide .................................................................................... 782
44:04. Sediment Filtering Apparatus, Unassembled ...................................................................... 786
44:05. Sediment Filtering Apparatus, Assembled ........................................................................ 786
44:06. Rot Fragments from Tomato Puree ...................... , . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . 800
44:07. Rot Fragments from Tomato Puree. . . . . . .. . . . . . . .. . . . . ... . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . . . . . 801
44:08. Mold Filaments in Tomato Products. . . . . .. . . . . . . .. . . . . . .. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . 802
44:09. Insect Penetration of Packaging .................................................................................. 817
46:01. Plastic Template Schematic for Microslide Assembly.............................................................. 842
46:02. Arrangement of Antisera and Homologous Reference Enterotoxins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 843
46:03. Examples of 4 Possible Reactions in Bivalent Detection System .................................................... 844
46:04. Effect of Amount of Enterotoxin in Test Preparation on Development of Reference Line of Precipitation .............. 845
46:05. Appearance of Microslide Gel Diffusion Test as Monovalent System ............................................... 845
46:06. Precipitate Patterns in Microslide Gel Diffusion Test Demonstrating Nonspecific (Atypical) Lines of Precipitation. . . . . . 845
46:07. Optical Somatic Cell Counter Flow Diagram ....................................................... , . . . .. . . . . . . . . . . 846
46:08. Optical Somatic Cell Counter Flow Diagram. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 848
46:09. Optical Somatic Cell Counter Flow Diagram. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 849
47:01. Combustion Tube............................................................................................... 853
47:02. Carbon and Hydrogen Apparatus.... . . . .......... ... ......... ........... .... ..... . . . ... . .... .... ....... ....... . .. 855
47:03. Upper Section of Distilling Apparatus. . . . .. . . . . . . . . . . . . ... . . . . . . .. . . . . . .. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . 857
47:04. Gravimetric Setup for Oxygen Determination ..................................................................... 859
47:05. Quartz Reaction Tube and Filling for Oxygen Determination ....................................................... 859
47:06. Details of Modified Clark Apparatus .............................................................................. 862
47:07. Modified Clark Apparatus. . . . . . . .. . . . . . . .. . . . . . . . . . . . . ... . . . . . . .. . . . . . .. . . . . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . 863
48:01. Cross-section of Marinelli Beaker. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 869
xi

14. Collaborative Study Procedures of the Association of
Official Analytical Chemists
The Association of Official Analytical Chemists (AOAC) is a
unique, nonprofit scientific organization whose primary purpose
is to serve the needs of government regulatory and research
agencies for analytical methods. The goal of the Association is
to provide methods which will perform with the necessary
accuracy and precision under usual laboratory conditions (1).
Since its formation in 1884 the AOAC has provided a mechanism
to select methods of analysis from published literature or
develop new methods, collaboratively test them through inter-
laboratory studies, approve them, and publish the approved
methods for a wide variety of materials relating to foods, drugs,
cosmetics, agriculture, forensic science, and products affecting
the public health and welfare. Its membership is composed of
scientists from Federal, State, Provincial, and other regulatory
bodies who work within the AOAC's established procedures as
researchers, methods collaborators, and committee members.
Although most of the members are from North America, many
nations throughout the world are represented.
The AOAC has almost a century of experience in utilizing the
collaborative study as a means of determining the reliability of
analytical methods for general purposes and, especially, for
regulatory purposes. In fact, the AOAC's major contribution to
analytical science has been to bring the collaborative study
technique for the validation of analytical methods to a high
degree of perfection. In such a study, laboratories analyze
identical sample sets which cover the range of applicability of
a method previously selected as being useful and practical. The
purpose of the study is to establish the characteristics of the
methods with respect to accuracy, precision, sensitivity, range,
specificity, limit of detection, limit of reliable measurement,
selectivity, practicality, and similar attributes, as required.
ORGANIZATION AND PROCEDURES FOR AOAC
COLLABORATIVE STUDIES
The collaborative study is organized and directed by an analyst
designated as the Associate Referee for the specific subject
under investigation. Currently, some 600 Associate Referees
appointed by the Association are responsible for as many topics.
An Associate Referee is selected for his knowledge, interest,
and experience in the subject matter field. He operates under
the scientific guidance, support, and administrative supervision
of a General Referee, who is in turn responsible for a product
area. The Associate Referee reviews the literature and selects
one or two of the better analytical methods available, modifying
them as needed. Alternatively, he may develop or adapt a
method used in his laboratory for the analyte and matrix under
study, testing it thoroughly in his laboratory before designing
a collaborative study. The General Referee is kept informed of
such preliminary studies.
The samples analyzed in a collaborative study are normally
prepared and distributed to the participants by the Associate
Referee. The Association follows the recommendations of You-
den (2) that not fewer than five laboratories participate and that
a minimum of six sample materials be sent to each. These are
xii
minima and, in practice, both are usually exceeded. In addition,
a reference or practice sample is included, where possible.
Laboratories with at least some experience in the general
subject matter are selected as collaborators. Because the objec-
tive of the study is to standardize the method, as contrasted to
standardizing the analyst (3). all analysts are instructed to follow
the method exactly as written even though they may not concur
with the Associate Referee's selection among possible alterna-
tives. The level ofthe analyte in the samples is usually unknown
to the participants.
All individual results obtained by the collaborators are re-
ported to the Associate Referee, who compiles and evaluates
them. Since statistical treatment of the data is considered
essential in a rigorous evaluation of the method for accuracy,
precision, sensitivity, and specificity, it is now required for all
studies. The Association considers this of such importance that
it provides statistical assistance in all cases where it is otherwise
unavailable to the Associate Referee. A statistical manual (4) is
also provided.
The Associate Referee makes the initial judgment on the
performance of the method. If he recommends approval, it
passes to the General Referee and then to a committee of
experts. If both recommend approval, the method is presented
at the Association's annual business meeting for vote by the
membership.
Approved methods and supporting data are published in the
Journal of the Association of Official Analytical Chemists. They
are subject to scrutiny and general testing by other analysts for
at least a year before final adoption. They may be modified and
restudied collaboratively as needed, should feedback from gen-
eral use reveal flaws in the method or in its written set of
directions. Approved methods are included in the Association's
"Official Methods of Analysis", a book of some 1000 pages
which is updated every 4-5 years.
The preceding summary of AOAC's modus operandi recog-
nizes the need for healthy skepticism toward results obtained
by analytical methods which have not undergone such rigorous
scrutiny and interlaboratory testing of their accuracy, precision,
dependability, specificity, and practicality.
SELECTION OF METHODS FOR STUDY
A certain degree of variability is associated with all measure-
ments. Much of the research on analytical chemistry is an
attempt to minimize that variability. But there are many different
types of variability in analytical work. We often find that when
we attempt to minimize one kind, we must necessarily permit
expansion in another kind. In practical analytical chemistry, the
problem often comes down to which variability is to be mini-
mized.
Some examples ofthis point may be helpful. In atomic weight
determination, everything-especially practicality-is sacrificed
for accuracy. A high degree of accuracy and practicality is
required in the assay of precious metals, but the fire assay used
is generally applicable to little else besides metals and minerals.

15. In clinical chemistry, within-laboratory precision (repeatability)
is critical, and often is of greater interest to clinical laboratories
than absolute accuracy or agreement with the values of other
laboratories (reproducibility). In drug analysis, a high degree of
accuracy is required in the therapeutic range because the
analytical values determining the identity, strength, quality, and
purity of pharmaceutical preparations, as laid down in phar-
macopoeial specifications, are directly related to clinical value.
With polynuclear hydrocarbons, specificity is important, since
some of these compounds are carcinogenic while others are
not. In applying the famous Delaney clause of the United States
Federal Food, Drug, and Cosmetic Act, all attributes of the
analytical methods are secondary to the detection of extremely
small concentrations (detectability), or to exhibiting a high
degree of response for small changes in concentration (sensi-
tivity).
There is a very special case involving accuracy, where the
"true value" is determined by the method of analysis. Many
legal specifications and standards for food and agricultural
products define ill-defined components such as moisture, fat,
protein, and crude fiber in terms of reference methods. There-
fore, the precision of these methods becomes the limiting factor
for their performance. In fact, most analyses involved in com-
mercial transactions require primarily that the buyer and seller
agree on the same value (analytically and economically), re-
gardless of where it stands on an absolute scale.
The point of these examples is that although methods of
analysis are characterized by a number of attributes-accuracy,
precision, specificity, sensitivity, detectability, dependability,
and practicality-no method is so flawless that all these qualities
can be maximized simultaneously. For any particular analysis,
the analyst must determine, on the basis of the purpose of the
analysis, which attributes are essential and which may be
compromised.
Unfortunately, the literature is replete with examples indicat-
ing that an individual analyst, and especially the originator of
a method of analysis, is not an unbiased judge of the relative
merits of the methods of analysis which he develops and uses.
In our experience, the collaborative study provides impartial
data on the suitability of the method. The data, in many cases,
speak for themselves.
The collaborative study, or ring test or round robin test, as it
is called in other organizations, provides the basic information
on the performance of analytical methods. The extent of the
information will depend on the number of samples provided,
the number of analyses performed, and the number of labora-
tories participating. The data should be unbiased because the
composition of the samples is known only to the administrator
of the study. Some of the requirements of the study and their
relationship to the characteristics and attributes of the method
are as follows:
(1) Accuracy. Samples must be of defined composition (by
spiking, by formulation, or by analytical consensus).
(2) Specificity. Samples should contain related analytes.
(3) Sensitivity. Samples should differ from each other or from
negative samples by a known amount.
(4) Applicability. Samples should include the concentration
range and matrix components of interest.
(5) Blanks. Samples should include different matrices with
"none" of the component of interest.
(6) Precision. Instructions should request replicate analyses
by the same or different analysts in the same laboratory,
preferably on different days. By far a better procedure is to
include "blind" (unknown to the analyst) replicate samples in
the series.
(7) Practicality. Instructions should request information as to
xiii
the actual and elapsed time required for the analyses; the
availability of reagents, equipment, and standards; and any
necessary substitutions. When practice samples are included,
the number of analyses required to achieve the stated recovery
and repeatability, should be reported.
PROCEDURAL DETAILS OF COLLABORATIVE STUDY
As numerous beginners in this field have discovered, much
preliminary work must be done before sending out samples:
(1) The method must be chosen and demonstrated to apply
to the matrices and concentrations of interest.
(2) The critical variables in the method should have been
determined and the need for their control emphasized [a rug-
gedness test (5) is useful for this purpose].
(3) The method should be written in detail by the Associate
Referee and tested by an analyst not previously connected with
its development.
(4) Unusual standards, reagents, and equipment must be
available from usual commercial sources of supply, or sufficient
quantities must be prepared or obtained to furnish to the
participants.
(5) The samples must be identical and homogeneous so that
the analytical sample error is only a negligible fraction of the
expected analytical error.
(6) A sufficient number of samples must be prepared to cover
typical matrices and the concentration range of interest (toler-
ance, maximum or minimum specifications, likely levels of
occurrence, etc.).
(7) Samples must be stable and capable of surviving the
rigors of commercial transportation.
(8) Reserve samples should be prepared and preserved to
replace lost samples and to permit reanalysis of samples con-
sidered as outliers to attempt to discover the cause of abnormal
results.
(9) The instructions must be clear. They should be reviewed
by someone not connected with the study to uncover potential
misunderstandings and ambiguities.
(10) If the analyte is subject to change (e.g., bacterial levels,
nitroglycerin tablets), provision must be made for all participants
to begin the analysis at the same time.
(11) Practice samples of a known and declared composition
should be furnished with instructions not to analyze the un-
knowns until a specified degree of recovery and repeatability
(or other attribute) has been achieved.
(12) Provision should be made when necessary for submis-
sion of standard curves, tracings of recorder charts, or photo-
graphs of thin-layer plates in order to assist in determining
possible causes of error.
OTHER TYPES OF INTERLABORATORY STUDIES
This type of collaborative study, which is designed to deter-
mine the characteristics of a method, must be carefully distin-
guished from other types of interlaboratory studies which by
design or through ignorance provide other kinds of information.
The most important types of other studies are:
(1) Those studies which require the collaborators to investi-
gate the variability of parts of methods or applicability to
different types of samples. (An interlaboratory study is usually
an inefficient way of obtaining this type of information.)
(2) Those studies which permit an analyst to use any method
he desires. Such studies invariably produce such a wide scatter
of results that the data are of little value for evaluation of
methods. They may be useful in selecting a method from a
number of apparently equivalent methods, provided the purpose

16. is emphasized beforehand and the participants provide a de-
scription of the method used in order to permit a correlation of
the details of the methods with apparent biases and variabilities.
(3) Those studies which are used for quality control purposes,
whose participants are not permitted sufficient time to gain
familiarity with the method, or who permit deviations to enter
into the performance of the analyses on the grounds that the
deviation is obviously an improvement which could not possibly
affect the results of the analysis, or who claim to have a superior
method.
With this background information, it is now appropriate to
introduce the following definitions which were agreed upon as
part of the guidelines for collaboration between the AOAC and
the Collaborative International Pesticide Analytical Council L~d.
(CIPAC) (6).
Collaborative study. An analytical study involving a number
of laboratories analyzing the same sample(s) by the same
method(s) for the purpose of validating the performance of the
method(s).
Preliminary interlaboratory study. An analytical study in which
two or more laboratories evaluate a method to determine if it
is ready for a collaborative study.
Laboratory performance check. The analysis of very carefully
prepared and homogeneous samples, normally of known active
ingredient content, to establish or verify the performance of a
laboratory or analyst.
SUMMARY
The collaborative study i's an experiment designed to evaluate
the performance of a method of analysis through the analysis
of a number of identical samples by a number of different
laboratories. With proper design, it provides an unbiased eval-
uation of the performance of a method in the hands of those
analysts who will use it. A collaborative study must be distin-
guished from those studies designed to choose a method or to
determine laboratory or analyst performance.
REFERENCES
(1) AOAC, "Handbook of the AOAC", 4th ed., AOAC, Box 540,
Benjamin Franklin Station, Washington, D.C. 20044, 1977.
(2) W. J. Youden, "Accuracy of Analytical Procedures", J. Assoc.
Off. Anal. Chern., 45, 169-73 (1962).
(3) Harold Egan, "Methods of Analysis; An Analysis of Meth-
ods", ibid., 60,260-7 (1977).
(4) W. J. Youden and E. H. Steiner, "Statistical Manual of the
AOAC: Statistical Techniques for Collaborative Tests. Plan-
ning and Analysis of Results of Collaborative Tests", AOAC,
Box 540, Benjamin Franklin Station, Washington, D.C. 20044,
1975.
(5) W. J. Youden, "The Collaborative Test", J. Assoc. Off. Anal.
Chern., 46, 55-62 (1963).
(6) "Guidelines for Collaboration Between the Association of
Official Analytical Chemists (AOAC) and the Collaborative
International Pesticide Analytical Council Ltd. (CIPAC)", ibid.,
57,447-9 (1974).
BIBLIOGRAPHY
Daniel Banes, "The Collaborative Study as a Scientific Concept",
J. Assoc. Off. Anal. Chern., 52,203-06 (1969).
William Horwitz, "Problems of Sampling and Analytical Meth-
ods", ibid., 59, 1197-203 (1976).
Reprinted with permission from: Analytical Chemistry (March 1978) 50, 337A-340A.
Published 1978 American Chemical Society
xiv

17. Definitions of Terms and Explanatory Notes
Reagents
(1) Term "H20" means distilled water, except where otherwise
specified, and except where the water does not mix with the
detn, as in "H20 bath."
(2) Term "alcohol" means 95% ethanol by vol. Alcohol of
strength x% may be prepd by dilg x mL 95% alcohol to 95 mL
with H20. Absolute alcohol is 99.5% by vol. Formulae of specially
denatured alcohols (SDA) used as reagents are as follows:
SDA No. 100 parts alcohol plus
1 5 wood alcohol
2-B
3-A
12-A
13-A
23-A
30
0.5
5
5
10
10
10
benzene or rubber
hydrocarbon solv.
MeOH
benzene
ether
acetone
MeOH
"Reagent" alcohol is 95 parts SDA 3-A plus 5 parts isopropanol.
(3) Term "ether" means ethyl ether, peroxide-free by follow-
ing test: To 420 mL ether in separator add 9.0 mL 1% NH4V03
in H2S04 (1+16). Shake 3 min and let sep. Drain lower layer into
25 mL g-s graduate, dil. to 10 mL with H2S04 (1 +16), and mix.
Any orange color should not exceed that produced by 0.30 mg
H20 2(1 mL of soln prepd by dilg 1 mL 30% H20 2to 100 mL with
H20) and 9.0 mL 1% NH4V03in H2S04 (1 + 16). Peroxides may be
eliminated by passing .;;700 mL ether thru 10 cm column of
Woelm basic alumina in 22 mm id tube.
(4) Reagents listed below, unless otherwise specified, have
approx. strength stated and conform in purity with Recom-
mended Specifications for Analytical Reagent Chemicals of
American Chemical Society:
Assay
Sulfuric Acid . . . . . . . . . . . . . . . . . . . . . . . .. 95.0-98.0% H2S04
Hydrochloric acid .................... 36.5-38.0% HCI
Nitric acid ........................... 69.0-71.0% HN03
Fuming nitric acid. . . . . . . . . . . .. . . . . . .. ;,,90% HN03
Acetic acid . . . . . . . . . . . . . . . . . . . . . . . . . .. ;,,99.7% HC2H30 2
Hydrobromic acid ......... " ..... . ... 47.0-49.0% HBr
Ammonium hydroxide. . . . . . . . . . . . . . .. 28-30% NH3
Phosphoric acid. . . . . . . . . . . . . . . . . . . . .. ;,,85% H3P04
Where no indication of diln is given, reagent is of concn given
above.
(5) All other reagents and test solns, unless otherwise de-
scribed in text, conform to requirements of American Chemical
Society. Where such specifications have not been prepd, use
highest grade reagent. When anhyd. salt is intended, it is so
stated; otherwise the crystd product is meant.
(6) Unless otherwise specified, phenolphthalein (phthln) used
as indicator is 1% alc. soln; Me orange is 0.1 % aq. soln; Me red
is 0.1 % alc. soln.
(7) Directions for stdzg reagents are given in Chapter 50.
(8) Unusual reagents not mentioned in reagent sections or
cross referenced, other than common reagents normally found
in laboratory, are italicized first time they occur in method.
(9) Com. prepd reagentsolns must be checked for applicability
to specific method. They may contain undeclared buffers, pre-
servatives, chelating agents, etc.
xv
(10) In expressions (1 +2), (5+4), etc., used in connection with
name of reagent, first numeral indicates vol. reagent used, and
second numeral indicates vol. H20. For example, HCI (1 +2)
means reagent prepd by mixing 1 vol. HCI with 2 vols H20.
When one of reagents is solid, expression means parts by wt,
first numeral representing solid reagent and second numeral
H20. Solns for which the solv. is not specified are aq. solns.
(11) In making up solns of definite percentage, it is understood
thatx g substance is dissolved in H20 and dild to 100 mL. Altho
not theoretically correct, this convention will not result in any
appreciable error in any of methods given in this book.
(12) Chromic acid cleaning soln is prepd by (1) adding 1 L
H2SO. to ca 35 mL satd aq. Na2Cr207 soln; or (2) adding 2220 mL
(9 Ib) H2S04 to ca 25 mL satd aq. Cr03 soln (170 g/100 mL).
Reagents may be tech. grade. Use only after first cleaning by
other means (e.g., detergent) and draining. Mixt. is expensive
and hazardous. Use repeatedly until it is dild Of has a greenish
tinge. Discard carefully with copious amts of H20.
(13) All calcns are based on table of international atomic
weights, 52.001.
Apparatus
(14) Burets, vol. flasks, and pipets conform to following
Federal specifications (available from General Services Admin.,
Specification Activity 3F1, Washington Navy Yard, Bldg. 197,
Washington, DC 20407):
Buret NNN-B-00789a May 19, 1965
Flask, vol. NNN-F-00289d Feb 7, 1977
Pipet, vol. NNN-P-395c March 13, 1970
Pipet, measuring NNN-P-350c July 16,1973
See also NBS Circular 602, "Testing of Glass Volumetric Ap-
paratus" (available as Com 73-10504 from NTIS, Springfield, VA
22151).
(15) Standard taper ($) glass joints may be used instead of
stoppers where the latter are specified or implied for connecting
glass app.
(16) Sieve designations, unless otherwise specified, are those
described in Federal Specification RR-S-366e, Nov 9,1973 (avail-
able from General Services Admin.). Designation" '100-mesh'
(or other number) powder (material, etc.)" means material
ground to pass thru std sieve No. 100 (or other number).
Corresponding international std and US std sieves are given in
Table 1.
(17) Term "paper" means filter paper, unless otherwise spec-
ified.
(18) Term "high-speed blender" designates mixer with 4
canted, sharp-edge, stainless steel blades rotating at the bottom
of 4-lobe jar at 10,000-12,000 rpm, or with equiv. shearing
action. Suspended solids are reduced to fine pulp by action of
blades and by lobular container, which swirls suspended solids
into blades. Waring Blendor, or equiv., meets these require-
ments.
(19) "Flat-end rod" is glass rod with one end flattened by
heating to softening in flame and pressing vertically on flat
surface to form circular disk with flat bottom at end.
(20) Designation and pore diam. range of fritted glassware
are: extra coarse, 170-220 /-Lm; coarse, 40-60; medium, 10-15;
fine, 4-5.5; Jena designations and pore diam. are: 1, 110 /-Lm;
2, 45; 3, 25; 4, 8.

18. (21) Unless otherwise indicated, temps are expressed as
degrees Centigrade.
Table 1. Nominal Dimensions of Standard Test Sieves (U.S.A.
Standard Series)
Sieve Designation
Nominal Nominal
International Sieve Wire
Standard' U.S.A. Opening, Diameter,
(ISO) Standard inches mm
12.5 mmb '/2 in.b 0.500 2.67
11.2 mm 7/'6 in. 0.438 2.45
9.5 mm 3/8 in. 0.375 2.27
8.0 mm 5/,6 in. 0.312 2.07
6.7 mm 0.265 in. 0.265 1.87
6.3 mmb
'/. in." 0.250 1.82
5.6 mm No. 3'/2 0.223 1.68
4.75 mm No.4 0.187 1.54
4.00 mm No.5 0.157 1.37
3.35 mm No.6 0.132 1.23
2.80 mm No.7 0.111 1.10
2.38 mm No.8 0.0937 1.00
2.00 mm No. 10 0.0787 0.900
1.70 mm No. 12 0.0661 0.810
1.40 mm No. 14 0.0555 0.725
1.18 mm No. 16 0.0469 0.650
1.00 mm No. 18 0.0394 0.580
850 "me No. 20 0.0331 0.510
710 "m No. 25 0.0278 0.450
600 "m No. 30 0.0234 0.390
500 "m No. 35 0.0197 0.340
425 "m No. 40 0.0165 0.290
355 "m No. 45 0.0139 0.247
300 "m No. 50 0.Q117 0.215
250 "m No. 60 0.0098 0.180
212 "m No. 70 0.0083 0.152
180 "m No. 80 0.0070 0.131
150 "m No. 100 0.0059 0.110
125 "m No. 120 0.0049 0.091
106 "m No. 140 0.0041 0.076
90 "m No. 170 0.0035 0.064
75 "m No. 200 0.0029 0.053
63 "m No. 230 0.0025 0.044
53 "m No. 270 0.0021 0.037
a These standard designations correspond to the values for test sieve
apertures recommended by the International Organization for Standard-
ization, Geneva, Switzerland.
b These sieves are not in the standard series but they have been
included because they are in common usage.
c 1000 I-'m = 1 mm.
Standard Operations
(22) Operations specified as "wash (rinse, ext. etc.) with two
(three, four, etc.) 10 mL (or other voL) portions H20 (or other
solv.)" mean that the operation is to be performed with indicated
vol. of solv. and repeated with same vol. of solv. until number
of portions required have been used.
(23) Definitions of terms used in methods involving spectro-
photometry are those given in JAOAC 37, 54(1954). Most
important principles and definitions are:
(a) More accurate instrument may be substituted for less
accurate instrument (e.g., spectrophtr may replace colorimeter)
where latter is specified in method. Wavelength specified in
method is understood to be that of max. absorbance (A). unless
no peak is present.
(b) Absorbance(s) (A).-Neg. logarithm to base 10 of ratio of
transmittance (T) of sample to that of ref. or std material. Other
xvi
names that have been used for quantity represented by this
term are optical density, extinction, and absorbancy.
(e) Absorptivity(ies) (a).-Absorbance per unit concn and cell
length. a = A/be, where b is in cm and c in gil, or a = (A/be)
x 1000, if c is in mg/L. Other names that have been used for
this or related quantities are extinction coefficient, specific
absorption, absorbance index, and £16';'".
(d) Transmittance(s) (T).-Ratio of radiant power transmitted
by sample to radiant power incident on sample, when both are
measured at same spectral position and with same slit width.
Beam is understood to be parallel radiation and incident at right
angles to plane parallel surface of sample. If sample is soln,
solute transmittance is quantity usually desired and is detd
directly as ratio of transmittance of soln in cell to transmittance
of solv. in an equal cell. Other names that have been used for
this quantity are transmittancy and transmission.
(e) Standardization.-Spectrophtr may be checked for accu-
racy of wavelength scale by ref. to Hg lines: 239.95, 248.3,
253.65, 265.3, 280.4, 302.25, 313.16, 334.15, 365.43, 404.66,
435.83, 546.07, 578.0, and 1014.0 nm. To check consistency of
absorbance scale, prep. soln of 0.0400 g K2CrO./L 0.05N KOH
and det. absorbance at following wavelengths in 1 cm cell: 230
nm, 0.171; 275,0.757; 313.2,0.043; 375, 0.991; 400, 0.396. See
"Standards for Checking the Calibration of Spectrophotome-
ters," Letter Circular LC-l017, reissued Jan 1967, NBS.
(24) Least square treatment of data and calculation of regres-
sion lines.-This technic finds the best fitting straight line for
set of data such as std curve. It calcs that straight line whose
sum of squares of vertical deviations (usually A) of observations
from the line is smaller than corresponding sum of squares of
deviations from any other line. Equation of straight line is:
Y = a + bX,
where a is intercept at Y axis (X = 0). and b is slope of line.
Least square estimates of constants are:
b = I (X, Y,) - [(IX,I Y,)/n]
IXf - (IX,)2/n
a=Y-bX,
where I = "sum of" the n individual values of indicated
operation, and X and Yare the averages of the X and Y points.
Example: To find "best" straight line relating A (Y) to concn
(X):
Observation Concn Absorbance
No. CI X, Y, X~ X,V,
1 80 1.270 6400 101.6
2 60 1.000 3600 60.0
3 40 0.700 1600 28.0
4 30 0.550 900 16.5
5 20 0.250 400 5.0
6 10 0.100 100 1.0
7 0 0.050 0 0.0
Totals:
n=7 IX, = 240 IY, = 3.92 IXl = 13000 I(X,Y,) = 212.1
x= IX,!n = 240/7 = 34.29
Y = IY,!n = 3.92/7 = 0.56
b = 212.1 - (240)(3.92)/7 77.7 = 0.0163
13000 - (240)2/7 4771
a = 0.56 - 0.0163(34.29) = 0.001
Best equation is then:
Y = 0.00 + 0.0163X
If for sample, A = 0.82, corresponding concn (X) would be:
X = (Y - 0.00)/0.0163 =0.82/0.0163 = 50.3.

19. Many scientific and statistical calculators are preprogrammed
to perform this calcn.
(25) Common safety precautions are given in Chapter 51.
Editorial Conventions
(26) For sake of simplicity, abbreviations CI and I instead of
CI2 and 12 are used for chlorine and iodine. Similar abbreviations
have been used in other cases (0, N, H). The same abbreviation
may also be used for the ion where no ambiguity will result.
(27) Reagents and app. referenced with only a letter, e.g., (c).
will be found in the reagent or apparatus section of that method.
(28) To conserve space, most of the articles and some prep-
ositions have been eliminated.
(29) Names and addresses of manufacturers and suppliers,
and trade names of frequently mentioned materials, are fur-
nished below solely as a matter of identification and conven-
ience, without implication of approval, endorsement, or certifi-
cation. The same products available from other suppliers or
other brands from other sources may serve equally well if
proper tests indicate their use is satisfactory. These firms when
mentioned in a method are given by name only (without
addresses).
Manufacturers and Suppliers
Ace Glass, Inc., PO Box 688, 1430 N West Blvd, Vineland, NJ
08360
Aldrich Chemical Co., Inc., 940 W St. Paul Ave, Milwaukee, WI
53233
Allied Chemical Corp., Specialty Chemicals Div., PO Box 1087R,
Morristown, NJ 07960
Aluminum Company of America, 1501 Alcoa Bldg, Pittsburgh,
PA 15219
American Cyanamid Co., Agricultural Div., PO Box 400,
Princeton, NJ 08540
American Instrument Co., Div. of Travenol Laboratories, Inc.,
8030 Georgia Ave, Silver Spring, MD 20910
(ASBC) American Society of Brewing Chemists, 3340 Pilot Knob
Rd, St. Paul, MN 55121
(ATCC) American Type Culture Collection, 12301 Parklawn Dr,
Rockville, MD 20852
Analabs Inc., 80 Republic Dr, North Haven, CT 06473
Applied Science Laboratories, Inc. (Applied Science Division,
Milton Roy Co.). PO Box 440, State College, PA 16801
Baird-Atomic, Inc., 125 Middlesex Tnpk, Bedford, MA 01730
J. T. Baker Chemical Co., 222 Red School Ln, Phillipsburg, NJ
08865
Barber-Colman Co., see Searle Analytic, Inc.
Bausch & Lomb, Inc., Analytical Systems Div., 820 Linden Ave,
Rochester, NY 14625
BBL, Div. of Bioquest, PO Box 243, Cockeysville, MD 21030
Beckman Instruments, Inc., 2500 Harbor Blvd, Fullerton, CA
92634
Becton, Dickinson, & Co., Rutherford, NJ 07070
Bio-Rad Laboratories, 32nd and Griffin Ave, Richmond, CA 94804
Brinkmann Instruments, Inc., Cantiague Rd, Westbury, NY 11590
Burdick & Jackson Laboratories, Inc., 1953 S Harvey St, Muske-
gon, MI 49442
Burrell Corp., 2223 Fifth Ave, Pittsburgh, PA 15219
Calbiochem, 10933 N Torrey Pines Rd, La Jolla, CA 92037
Carborundum Co., PO Box 423, Niagara Falls, NY 14302
Cenco Inc., 2600 5 Kostner Ave, Chicago, IL 60623
Coleman Instruments Division, Perkin-Elmer Corp., 2000 York
Rd, Oak Brook, IL 60521
Corning Glass Works, Laboratory Products Dept., Corning, NY
14830
xvii
Curtin Matheson Scientific, Inc., PO Box 1546, Houston, TX
77001
Ddco Laboratories, PO Box 1058A, Detroit, MI 48232
Dohrmann Div. of Envirotech Corp., 3240 Scott Blvd, Santa
Clara, CA 95050
Dow Chemical Co., Ag-Organics Dept., PO Box 1706, Midland,
MI48640
Dow Corning Corp., Midland, MI 48640
E. I. du Pont de Nemours & Co., Wilmington, DE 19898
Eastman Kodak Co., Eastman Organic Chemicals, 343 State St,
Rochester, NY 14650
Eaton-Dikeman Co., Mt. Holly Springs, PA 17065
Elanco Products Co., Div. of Eli Lilly Co., Elanco Analytical
Laboratory, Dept. MC757, Indianapolis, IN 46206
Fisher & Porter Co., Lab Crest Scientific Div., County Line Rd,
Warminster, PA 18974
Fisher Scientific Co., 711 Forbes Ave, Pittsburgh, PA 15219
Floridin Co., Berkeley Springs, WV 25411
Foss America Inc., PO Box 504, Route 82, Fishkill, NY 12524
GAF Corp., 140 W 51st St, New York, NY 10020
G.B. Fermentation Industries, Inc., 1 N Broadway, Des Plaines,
IL 60016
Geigy Chemical Corp., Saw Mill River Rd, Ardsley, NY 10502
Hamilton Co., PO Box 17500, Reno NV 89510
Hess & Clark Laboratories, Div. of Rhodia, Inc., 7th and Orange
Sts, Ashland, OH 44805
Hewlett-Packard Co., 1501 Page Mill Rd, Palo Alto, CA 94304
Hoffman-La Roche, Inc., Nutley, NJ 07110
ICI-America, Inc., Chemical Research Dept., Wilmington, DE
19899
ICN-K&K Laboratories, Inc., 121 Express St, Plainview, NY
11803
ICN Pharmaceuticals, Inc., Life Sciences Group, 26201 Miles Rd,
Cleveland, OH 44128
johns-Manville Products Corp., Greenwood Plaza, Denver, CO
80217
Kimble Products, Owens-Illinois, PO Box 1035, Toledo, OH 43666
Kontes Glass Co., Spruce St, Vineland, NJ 08360
Labconco Corp., 8811 Prospect Ave, Kansas City, MO 64132
Eli Lilly & Co., 740 5 Alabama St, Indianapolis, IN 46206
Mallinckrodt Chemicals Works, Science Products Div., 2nd &
Mallinckrodt Sts, St. Louis, MO 63147
MC/B Manufacturing Chemists, 2909 Highland Ave, Norwood,
OH 45212
Matheson Scientific, Inc., see Curtin Matheson Scientific, Inc.
Merck & Co., Inc., 126 E Lincoln Ave, Rahway, NJ 07065
Miles Laboratories, Inc., Elkhart, IN 46514
Monsanto Chemical Co., 800 N Lindberg Blvd, St. Louis, MO
63166
(NBS) National Bureau of Standards, Washington, DC 20234
(NF) National Formulary, see USP
New York Laboratory Supply Co., 510 Hempstead Tnpk, West
Hempstead, NY 11552
Orion Research Inc., 380 Putnam Ave, Cambridge, MA 02139
Perkin-Elmer Corp., 702-G Main Ave, Norwalk, CT 06856
Phillips Chemical Co., Division of Phillips Petroleum Co., Spe-
cialty Chemicals, Drawer '0', Borger, TX 79007.
Pierce Chemical Co., PO Box 117, Rockford, IL 61105
H. Reeve Angel & Co., Inc., 9 Bridewell PI, Clifton, NJ 07014
Rohm & Haas Co., Independence Mall West, Philadelphia, PA
19105
Salsbury Laboratories, Charles City, IA 50616
Sargent-Welch Scientific Co., 7300 N Linder Ave, Skokie, IL
60076
(S&S) Schleicher & Schuell, Inc., 543 Washington St, Keene, NH
03431

20. Schoeffellnstrument Corp., 24 Booker St, Westwood, NJ 07675
SGA Scientific, Inc., 735 Broad St, Bloomfield, NJ 07003
Scientific Products, Div. of American Hospital Supply Corp.,
1430 Waukegan Rd, McGaw Park, IL 60085
Searle Analytic, Inc., 2000 Nuclear Dr, Des Plaines, IL 60018
Shell Oil Co., PO Box 2463, Houston, TX 77001
Sigma Chemical Co., PO Box 14508, Sl. Louis, MO 63178
G. Frederick Smith Chemical Co., PO Box 23344, Columbus, OH
43223
Sterwin Chemicals, Inc., 90 Park Ave, New York, NY 10016
Supelco, Bellefonte, PA 16823
Technicon Instruments Corp., 511 Benedict Ave, Tarrytown, NY
10591
Arthur H. Thomas Co., Vine St at 3rd, PO Box 779, Philadelphia,
PA 19105
Ultra-Violet Products, Inc., 5100 Walnut Grove Ave, San Gabriel,
CA 91778
Union Carbide Corp., Chemicals and Plastics, 270 Park Ave, New
York, NY 10017
Union Carbide Corp., Agricultural Products and Services, PO
Box 1906, Salinas, CA 93901
Uniroyal Chemical, Elm St, Naugatuck, CT 06770
The Upjohn Co., Kalamazoo, MI 49001
(USDA) U.S. Department of Agriculture, Office of Information,
Washington, DC 20250
(USP) United States PharmacopeiaI Convention, Inc., 12601
Twinbrook Pkwy, Rockville, MD 20852
Varian Aerograph, 2700 Mitchell Dr, Walnut Creek, CA 94598
Varian Instrument Div., 611 Hansen Way, Palo Alto. CA 94303
Velsicol Chemical Corp., 341 E Ohio St, Chicago, IL 60611
VWR Scientific, PO Box 3200, San Francisco, CA 94119
Wallerstein Co., see G. B. Fermentation Industries, Inc.
Waters Associates, Inc., Maple St, Milford, MA 01757
Winthrop Laboratories, Special Chemicals Dept., 90 Park Ave,
New York, NY 10016
Trade Names
Amberlite. Ion exchange resins. Rohm and Haas Co.
Anakrom. Gas chromatography supports. Analabs, Inc.
Celite. Diatomaceous products. Johns-Manville Products Corp.
Chromosorb. Chromatographic supports and packings. Johns-
Manville Products Corp.
Dowex. Ion exchange resins. Dow Chemical Co.
Florisil. Chromatographic adsorbents. Floridin Co.
Gas-Chrom. Gas chromatography solid supports. Applied Sci-
ence Laboratories, Inc.
Hyf/o Super-Cel. Diatomaceous products. Johns-Manville Prod-
ucts Corp.
Skellysolve. Hydrocarbon solvents. Getty Refining and Market-
ing Co., PO Box 1650, Tulsa, OK 74102
Tef/on. Chemically resistant polytetrafluoroethylene. E. I. du
Pont de Nemours & Co.
Tygon. Halogenated vinyl plastic. Norton Co., Plastics & Syn-
thetics Div., 12 E Ave, Tallmadge, OH 44278
(30) The foillowing abbreviations, many of which conform
with those of Chemical Abstracts, are used. In general, principle
governing use of periods after abbreviations is that period is
used where final letter of abbreviation is not the same as final
letter of word it represents. Periods are not used with units,
except inch(es) and gallon(s).
xviii
Abbreviation
a
A
AA
Ac
ACS
addn
addnl
alc.
alk.
alky
amp
amt
anal.
anhyd.
AOCS
app.
approx.
aq.
ASTM
atm.
avo
Be.
bp
Bu
C
ca
calc.
calcd
calcg
calcn
Cat. No.
centrf.
centrfd
centrfg
Chap.
chern.
chromatgc
chromatgd
chromatgy
Ci
CI
CIPAC
cm
compd
com.
conc.
concd
concg
concn
const
contg
cP
cpm
cryst.
crystd
crystg
crystn
cu in.
dc
del.
detd
Word
absorptivity(ies)
absorbance(s) thruout (not restricted to for-
mulas); not absorption. A' is used for std;
Ao for blank; 3 digit subscript numerals
usually denote wavelengths in nm
atomic absorption
CH3CO- (acetyl, not acetate)
American Chemical Society
addition
additional
alcoholic (not alcohol)
alkaline (not alkali)
alkalinity
ampere(s)
amount
analytical(ly)
anhydrous
American Oil Chemists' Society
apparatus
approximate(ly)
aqueous
American Society for Testing and Materials
atmosphere, atmospheric
average (except as verb)
degree Baume
boiling point
butyl
degrees Celsius (Centigrade)
about, approximately
calculate
calculated
calculating
calculation
Catalog Number
centrifuge
centrifuged
centrifuging
Chapter
chemical(ly)
chromatographic
chromatographed
chromatography
curie(s)
Color Index
Collaborative International Pesticides Ana-
lytical Council
centimeter(s)
compound
commercial(ly)
concentrate (as verb or noun)
concentrated
concentrating
concentration
constant
containing
centipoise
counts per minute
crystalline (not crystallize)
crystallized
crystallizing
crystallization
cubic inch(es)
direct current
determine
determined

21. Abbreviation
detg
detn
diam.
diat. earth
dil.
dild
dilg
diln
distd
distg
distn
DMF
DMSO
EDTA
e.g.
elec.
equiv.
est.
estd
estg
estn
Et
EtOH
evap.
evapd
evapg
evapn
ext
extd
extg
extn
F
FAO
Fig.
fl oz
fp
ft
g
g
gal.
GLC
g-s
HCHO
HOAc
HPLC
hr
ht
id
in.
inorg.
insol.
IR
ISO
JAOAC
kg
L
Ib
liq.
m
Word
determining
determination
diameter
diatomaceous earth
dilute
diluted
diluting
dilution
distilled
distilling
distillation
N,N-dimethylformamide
dimethyl sulfoxide
ethylenedinitrilotetraacetic acid (or -tetra-
acetate)
for example
electric(al)
equivalent
estimate
estimated
estimating
estimation
ethyl
ethanol (the chemical entity C2H50H)
evaporate
evaporated
evaporating
evaporation
extract
extracted
extracting
extraction
degrees Fahrenheit (DC = (5/9) x (OF - 32))
Food and Agriculture Organization
Figure (illustration)
fluid ounce(s) (29.57 mL)
freezing point
foot (30.48 cm)
gram(s)
gravity (in centrfg)
gallon(s) (3.785 L)
gas-liquid chromatography
glass-stoppered
formaldehyde
acetic acid (not HAc)
high pressure (or performance) liquid chro-
matography
hourIs)
height
inner diameter (or dimension)
inch(es) (2.54 cm)
inorganic
insoluble
infrared
International Organization for Standardiza-
tion
Journal of the Association of Official Analyt-
ical Chemists (after 1965)
Journal of the Association of Official Agri-
cultural Chemists (before 1966)
kilogram(s)
liter(s)
pound(s) (453.6 g)
liquid
meter(s); milli-as prefix
xix
Abbreviation
m
M
ma
mag.
max.
mech.
Me
MeOH
mg
min
min.
mixt.
mL
mm
mp
mIL
mv
MW
N
N
n
NBS
NCA
neg.
neut.
neutze
neutzd
neutzg
neutzn
NF
ng
nm
No.
-OAc_
-OCN
od
org.
oxidn
oz
p
Pa
par.
pet ether
phthln
pos.
powd
ppb
ppm
ppt
pptd
pptg
pptn
Pr
prep.
prepd
prepg
prepn
psi
psig
pt
Word
molal
molar (as applied to concn). not molal
milliampere (et amp)
magnetic(ally)
maximum
mechanical(ly)
methyl
methyl alcohol
milligram(s)
minute(s)
minimum
mixture
milliliter(s)
millimeter(s)
melting point
millimicron (10-6
mm); use nanometer (nm)
(10-9 m)
millivolt
molecular weight
normal (as applied to concn); in equations.
normality of titrating reagent
Newton (105 dynes)
refractive index
National Bureau of Standards
National Canners Association (now National
Food Processors Associa.tion)
negative
neutral
neutralize
neutralized
neutralizing
neutralization
National Formulary
nanogram (10-9
g)
nanometer (10-9
m); formerly mIL
number
acetate (cf Ac)
cyanate
outer diameter (or dimension)
organic
oxidation
ounce(s) (28.35 g)
pico (10-12
) as prefix
Pascal (1 Newton/m2
; 9.87 x 10-6
atm.; 7.5
x 10-3 mm Hg (torr); 1.45 x 10--4 psi)
paragraph(s)
petroleum ether
phenolphthalein
positive
powdered (as adjective)
parts per billion (1/109
)
parts per million (1/106
)
preci pitate
precipitated
precipitating
precipitation
propyl
prepare
prepared
preparing
preparation
pounds per square inch (absolute)
pounds per square inch gage (atmospheric
pressure = 0)
pint(s) (473 mL)