8076 J. Agric. Food Chem.

2009, 57, 8076–8088

DOI:10.1021/jf900189s

One Hundred Years of Progress in Food Analysis

ROBERT J. MCGORRIN*
Department of Food Science and Technology Oregon State University, Corvallis, Oregon 97331-6602

Food and agricultural products comprise complex and diverse chemical mixtures that historically
have presented challenges for assessing food safety, nutrient content, stability, and sensory
qualities. The analysis of food composition has significantly evolved over the past 100 years,
progressing from reliance on predominantly “wet chemistry” laboratory methods from the early to
mid-20th century to their gradual replacement by modern instrumental techniques. Pioneering
developments in pH instruments, spectrophotometry, chromatography/separations, and spectro-
metry often had immediate applications to food analysis. Continuous improvements in methodology
over this period have led to significant enhancements in analytical accuracy, precision, detection
limits, and sample throughput, thereby expanding the practical range of food applications. The
growth and infrastructure of the modern global food distribution system heavily relies on food
analysis;beyond simple characterization;as a tool for new product development, quality control,
regulatory enforcement, and problem-solving.

KEYWORDS: Analytical methods; characterization techniques; food analysis; food products; separa-
tion techniques; chromatography; mass spectrometry; spectroscopy

INTRODUCTION The chemical analysis of food composition has significantly

From a chemical perspective, food is an intricate, heteroge- advanced over the period from 1908 to the present and is the
neous mixture of diverse biochemical components. Analytical subject of recent comprehensive reviews and compendia of
food chemists are concerned with the composition and properties methods and instrumentation (1-5). As anaytical chemistry
of food products, beverages, confections, and flavors including evolved over the same era (6), classical monographs and textbooks
the chemical and physical changes they undergo during post- devoted to food analysis underwent multiple editions and revi-
harvest handling, processing, and storage. Measuring the quality, sions, many spanning several decades (7-9). In the United States,
safety, sensory properties, nutrition, and stability of food pro- classical methods of food analysis were initially developed to
ducts is the primary concern of analytical chemists working provide reliable and reproducible information about food com-
within the food-processing industry and in academic and govern- position due to frequent incidents of food adulteration and the
ment laboratories affiliated with food science or testing programs. need for consumer protection. The early focus of food analysis
Food assays have been developed to enable measurement of was to differentiate levels of food components, assess purity, and
food product quality attributes (appearance, color, flavor, expose economic fraud. As the science of food components
texture), depending on the specific product application. Alterna- evolved from 1900 to 1940 through subsequent discoveries and
tively, the instability of foods, which limits their shelf life, can be further understandings of the roles of vitamins, minerals, proteins,
initiated by chemical reactions, enzymatic reactions, microbial lipids, and other essential nutrients, the need arose for the develop-
tranformations, or physical forces. Analysis of the degree of ment of analytical methods that could verify nutrient content.
emulsification or moisture content, for example, may allow a Early agricultural chemists were interested in providing links
prediction of the shelf-life limit for a salad dressing or a snack between food composition and nutrition, just as they are today.
cracker, respectively. Analysis of food pigments concerns darken- New federal regulations were enacted in countries around the
ing, bleaching, or development of desirable colors. Flavor assays world, requiring commercially processed foods to be labeled to
measure oxidative rancidity, undesirable browning reactions, or declare their ingredients (for example, in the United States with
desirable compounds produced through fermentation or thermal the Federal Food Drug and Cosmetic (FD&C) Act of 1938) and,
reactions. Texture assays evaluate toughening or softening, loss subsequently, their nutritional value (U.S. Nutrition Labeling
or gain in solubility or water-holding capacity. Besides quality and Education Act of 1990.) Protein, vitamin A, and vitamin C
assurance, the goals of food analysis may be directed toward basic are among the list of 14 nutrients required to be disclosed on the
research (healthy food ingredients, bioactive compounds for package label since 1993 (10), and this new regulation generated
disease prevention) or product development (convenient prepara- the need for more accurate analytical tests for trace nutrients in
tion, improved sensory attributes). foods. In addition, more rapid analytical methods needed to be
developed to allow food processors to declare levels of cholesterol
*Telephone (541) 737-8737; fax (541) 737-1877; e-mail robert. and saturated fat in foods, rather than reporting total fat content.
[email protected]. The requirement to declare trans-fat content on packaged food

pubs.acs.org/JAFC Published on Web 08/31/2009 © 2009 American Chemical Society

Article J. Agric. Food Chem., Vol. 57, No. 18, 2009 8077
Table 1. Sources of Official Analytical Methods for Food Analysis
standard analytical methods organization/publisher food matrix/analyte

Official Methods of Analysis AOAC International foods, food additives, trace nutrients, animal feed
Approved Methods American Association of Cereal Chemists (AACC) flour, gluten, dough strength, starch, fiber
Official Methods and Recommended American Oil Chemists’ Society (AOCS) vegetable oil source materials: fatty acids, glycerides,
Practices glycerin, lecithin, trans-fats
Standard Methods for the Examination American Public Health Association acidity, fat, lactose, moisture, protein, rancidity
of Dairy Products
Food Chemicals Codex U.S. Pharmacopeia preservatives, flavorings, colors, vitamins
TTB Methods U.S. Alcohol Tobacco Tax and Trade alcohol, calories, protein, carbohydrates in wine, malt,
Bureau (TTB) distilled beverages

labels was added in 2006 to allow consumers to make healthier pesticide, antibiotic, and mycotoxin (e.g., aflatoxin, fumonisin,
food choices to reduce their risk of coronary heart disease (11). ochratoxin) residues in animal feeds and foods. Because of
Mandatory food nutrition labeling provided the impetus to increased incidences of foodborne allergies, the U.S. Food Aller-
develop new standardized methods for food analysis that could gen Labeling and Consumer Protection Act of 2004 required
measure a broader group of components, particularly for complex mandatory labeling of the eight major food groups that comprise
matrices in multicomponent “prepared” food products. These 90% of allergenic responses (milk, eggs, fish, crustacean shellfish,
new methods were essential tools for food processors in manu- tree nuts, peanuts, wheat, soybeans) (18). The need for fast, high-
facturing and for government regulatory agencies in enforcement. volume screening has driven the development of rapid analytical
Standardized food analytical methods enable consistency of test kits. Two examples are those that employ enzyme-linked
results among different laboratories that follow the same proce- immunosorbent assay (ELISA) techniques (e.g., detection of
dures and serve as a benchmark for comparing results in the wheat gluten and peanut protein allergens in processed foods)
development of new methods. Several nonprofit scientific orga- and monoclonal antibody-based affinity chromatography meth-
nizations have compiled and published standard methods of ods (e.g., assay of aflatoxins in peanuts, cereal grains, and milk).
analysis for diverse food products, which have been collabora- Today, as in the past, regulatory and food safety concerns compel
tively studied and validated (Table 1). These include Official the ongoing development of analytical testing methods to ensure
Methods of Analysis (12) published by AOAC International, an food authenticity, monitor economic adulteration, and screen for
organization founded in 1884 to initially provide analytical the presence of chemical toxins and food additives to ensure
methods for fertilizers, feeds, and dairy products to serve food compliance with local government regulations. Sophisticated
chemists in government and regulatory agencies. Current meth- analytical tools such as HPLC, near-infrared spectroscopy, site-
ods cover a range of analytical tests across foods, food additives, specific natural isotope fractionation (SNIF)-NMR, and 13C
beverages, and agricultural materials. The American Association stable isotope ratio analysis-mass spectrometry are frequently
of Cereal Chemists (AACC) publishes Approved Methods (13) for used to verify authenticity of fruit juices, natural flavors, spices,
cereal products relating to baking quality, dough strength, stal- essential oils, and other expensive food ingredients. (19-23).
ing, starch, fiber, and proximate tests (e.g., mosture, crude fat,
protein). Official Methods and Recommended Practices (14) relat- FEDERAL REGULATION DRIVES NEED FOR FOOD
ing to fat and oil analysis are published by the American Oil ANALYSIS
Chemists’ Society (AOCS) for oilseed components, lipid materi- Much of today’s analytical food testing has its roots in food
als, and finished food products, in addition to soaps and safety and the prevalence of economic adulteration in the early
detergents. The American Public Health Association publishes 20th century that ultimately led to the creation of the U.S. Food
Standard Methods for the Examination of Dairy Products (15), and Drug Administration (FDA) (24). A large part of the
which include methods for the chemical analysis of milk and dairy motivation for performing early food analysis was to protect
products (acidity, fat, lactose, protein, water content). Food consumers against fraud, such as unscrupulous food manufac-
Chemicals Codex (16) is a compendium of standardized analytical turers adding alum or gypsum to flour, cottonseed oil to olive oil,
methods and tests for the purity and quality of food ingredients or dried leaves to pepper (25, 26). The FDA’s origins can be
including preservatives, flavorings, colors, and nutrients. In the correlated with the 1906 publication of Upton Sinclair’s best-
United States, the Alcohol Tobacco Tax and Trade Bureau selling novel The Jungle, which described in graphic detail the filth
(TTB) has developed official analytical methods (17) to measure and unsanitary practices then prevailing at the Chicago stock-
alcoholic content, calories, protein, and carbohydrates in wines, yards. The public’s outraged response prompted the U.S. Con-
malt beverages, and distilled alcoholic beverages. gress to propose new legislation. In June 1906, President
As the global food industry continues to expand, food analysis Theodore Roosevelt signed into law the Food and Drug Act,
plays an essential role in understanding and utilizing chemistry to also known as the “Wiley Act” after its chief advocate, Dr.
measure food product stability, quantifying changes in flavor Harvey W. Wiley (27). This was the first federal law aimed at
chemistry associated with sensory characteristics, determining the regulating interstate commerce of misbranded or adulterated
end of product shelf life, developing ingredient specifications, and foods, beverages, and drugs.
problem solving in product development, production, and quality The Act prohibited, under penalty of seizure of goods, the
control. In addition, ensuring the safety of foods has become an interstate transport of food which had been “adulterated”, with
international mandate for food processors and governmental that term referring to the addition of fillers of reduced “quality or
agencies. In the United States, food safety implies the absence of strength”, coloring to conceal “damage or inferiority,” formula-
pathogenic microbiological organisms in foods; however, it tion with additives “injurious to health,” or the use of “filthy,
broadly encompasses the control and chemical measurement of decomposed, or putrid” substances. The Act applied similar
8078 J. Agric. Food Chem., Vol. 57, No. 18, 2009 McGorrin
penalties to the interstate marketing of “adulterated” drugs, in applied to the study of food composition. In 1842, Liebig
which the “standard of strength, quality, or purity” of the active attempted to classify foods on the basis of their chemical
ingredient was not either stated clearly on the label or listed in the constituents and nitrogen content, whereas Henneberg devised
United States Pharmacopoeia or the National Formulary (27). an analysis scheme in the 1850s to determine proximate composi-
The responsibility for examining foods and drugs for such tion in a manner still used today (30). Moisture was determined by
adulteration or misbranding as part of the enforcement of this heat-drying; fat was extracted with diethyl ether; nitrogen content
Act was entrusted to the Bureau of Chemistry headed by Wiley in was converted into protein by applying a factor of 6.25; crude
the U.S. Department of Agriculture (USDA). fiber was measured by its insolubility in dilute acid and alkali,
Interestingly, the foundation for the Bureau been laid 22 years from which residual ash and fat was subtracted; total carbohy-
earlier with the establishment in 1884 of the Association of drates were calculated by difference. The 6.25 conversion factor
Official Agricultural Chemists (AOAC) under the auspices of was based on an assumption that animal proteins contain 16%
the USDA (28). Its key role at that time was to adopt uniform nitrogen, of which all is derived from protein (which we now
methods of analysis for fertilizers. In 1885, a convention establish- know to be false). Overlaps between the nutritional sciences and
ing AOAC as an independent organization was held in Philadel- agricultural chemistry provided early research on food composi-
phia, PA. Its membership was restricted to analytical chemists in tion, energy values of foods by calorimetry, and the digestibility
state and federal government positions, a membership require- and availability of food macronutrients from notable contribu-
ment that remained in effect nearly 100 years later. tions by Rubner in Berlin and Atwater at the Storrs Agricultural
The early years of AOAC were strongly influenced by Wiley, a Experiment Station in Connecticut in the early 1900s (30). Of
founder of the association who served as its president and historical interest is that a preponderance of synthetic organic and
secretary. In 1885, Wiley oversaw the publication of the AOAC analytical chemistry advancements occurred in Germany prior to
Methods of Analysis, a 49 page bulletin of chemical methods for World War I, and early chemists received their training in
the analysis of fertilizers. By 1887, the publication had grown to Germany to learn these techniques up until World War II (31).
include methods for feeds and dairy products as well as fertilizers. Several examples are representative of the development of early
Dr. Wiley’s interest and concern for the adulteration of foods and classical wet chemistry methods. In terms of its relative significance
drugs led to the study and adoption of official methods for foods and impact on quality and safety, moisture determination is one of
in 1889, and by 1912 AOAC was publishing the Official and the most widely used measurements in the processing and testing
Provisional Methods of Analysis. This became the impetus for of foods. However, accurate and precise moisture measurements
development of standardized and collaboratively tested analytical are difficult to obtain, because the degree of water binding within a
methods for food substances and ingredients, which were peer- food product influences its ease of removal (2). Early techniques
reviewed at multiple stages in the methods development process. involved codistillation with a high boiling point solvent or use of
In 1927, the Bureau of Chemistry’s regulatory powers were drying ovens. The Karl Fischer titration (iodine reduction by SO2
reorganized under a new USDA body, the Food, Drug, and in the presence of water) developed early as the preferred method
Insecticide Organization. This name was shortened to the Food for the determination of water in low-moisture foods such as dried
and Drug Administration in 1930. Following the groundwork fruits and vegetables or foods high in sugar or protein.
laid during the first part of the 20th century, food regulation in the The Kjeldahl measurement of protein (digestion with sulfuric
areas of labeling, standard specifications, nutritional claims, and acid and copper catalyst; boric acid titration of NH3) became a
approved additives continues to influence and direct analytical standard food analysis technique for over 100 years (2, 3). The
methods development. Kjeldahl method’s universality, precision, and reproducibility
made it the internationally recognized standard method for
EARLY PERIOD (1908-1950): WET CHEMISTRY REIGNS estimating the protein content in foods. However, the Kjeldahl
The typical food analysis laboratory in the early 20th century assay does not exclusively measure protein content. The assay’s
(Figure 1) appeared very different from modern food research and inability to differentiate protein from nonprotein nitrogen was
testing facilities. During this period, nearly all food analyses were exploited in two adulteration incidents in China (pet food, 2007;
performed using “wet chemistry” methods. A majority of these milk powder, 2008) when melamine, a nitrogen-rich chemical
methods were designed to utilize chemicals and equipment that used in plastics and fertilizer, was added to the diluted food
were readily available in typical chemical laboratories, such as material to falsify high protein contents. Additional disadvan-
glassware, weighing balances, Bunsen burners, and ovens. Typi- tages of the Kjeldahl method, such as the need to use concentrated
cally, classical methods involved combinations of elaborate sulfuric acid at high temperature with a relatively long digestion
analytical procedures such as weighing, mixing, filtering, eva- time, led to the development of the Dumas nitrogen combustion
poration, distillation, or solvent extraction (29). The traditional method for measuring crude protein content. The Dumas method
practices of these wet chemical analyses were highly deductive. was developed in the 1940s, but it initially was not a practical
Clues to identifying substances and their levels arose from volume routine analytical tool until the instrumentation was further
determinations, titrations, or precipitations, with the results developed and made more user-friendly in the 1990s. Instrumen-
judged by the subjective, if trained, eye of the food chemist. tal nitrogen combustion methods (e.g., LECO) are becoming
Physical methods of analysis;measuring light absorption or more frequently utilized to determine protein content, even
electromagnetic properties of food;almost inevitably found though the microscale Kjeldahl method continues to be routinely
the food chemist connecting optical and electrical components used in food analytical testing laboratories at the present time.
with traditional glassware, ceramics, and rubber tubing. Often, the reliability of classical food analysis methods is
The foundation for these methods had been laid in the 1800s directly related to the skill of the analyst, an example being the
with the development of assays used with synthetic organic determination of vitamin A and β-carotene in foods. The official
chemistry and for characterization of natural products in food analysis method is the Carr-Price assay, which was developed in
and feed. The contributions of early German and other European 1947 and utilizes a colorimetric measurement at 620 nm to
chemists to food analysis have been reviewed (26). Many of these determine the analyte concentration (32, 33). The reaction of
early scientists developed chemical procedures for elemental antimony trichloride with vitamin A in chloroform yields an
analysis and isolation of organic substances that were later intense but unstable blue color. The reproducibility and accuracy
Article J. Agric. Food Chem., Vol. 57, No. 18, 2009 8079

Figure 1. Analytical chemistry laboratory at the USDA Fixed Nitrogen Research Laboratory, near Washington, DC, in the 1920s (courtesy Chemical Heritage
Foundation).
of the assay is highly dependent on the subjective interpretation of Bishop electrotitrimeter was a device invented in the early 1900s
the color change by the analyst and the time interval at which the for measuring the concentration and percentages of acid in
titration is performed after the sample extraction (34). In addi- substances (37). These early instruments were composed of fragile
tion, the relatively strong acidic conditions can promote cis/trans glassware, wires, galvanometers, and elaborate components, and
isomerization of the native retinols. Because of other colored considerable analyst time was expended in the construction and
chemical interferences present in many types of food samples that maintenance of these devices in the laboratory. The practical
can influence the accuracy of the results, colorimetric methods utility of glass pH electrodes would require advances in electro-
such as Carr-Price were eventually replaced in the 1980s by faster, nics capable of measuring high resistance values. In 1934, a
more accurate high-performance liquid chromatography (HPLC) significant milestone was achieved when Arnold O. Beckman
methods (35). In most cases, analytical food testing laboratories designed his first electronic pH-meter, the “Acidimeter” (38).
no longer perform vitamin assays using the earlier colorimetric Beckman built the instrument as a personal favor for a former
methods, even though they remain the official methods. classmate who was working as a food chemist at the California
The analyses of trace minerals (calcium, copper, iron, manga- Fruit Growers Exchange on a lemon byproduct. To maintain
nese, zinc) and toxic heavy metals (lead, cadmium, mercury) in uniform product quality, it was necessary to measure the acidity
foods were initially performed by wet chemical titration assays of of lemon juice, which correlated with its relative sourness.
precipitation complexes or reactions with chromogens to form
However, the chemist had considerable difficulty measuring the
colored products that can be quantified by light absorption (2).
pH using a fragile, thin-glass silver/silver chloride electrode (to
These techniques initially were replaced by the commercialization
reduce resistance to small electrical currents) connected to a
of atomic absorption spectrometry in 1963, followed later by
delicate galvanometer. One of these components was frequently
inductively coupled plasma (ICP)-atomic emission spectrometry
breaking, and the electrical signals were weak. Beckman offered
in the 1980s and ICP-mass spectrometry in the 1990s (2, 36).
However, even at the present time, the analysis of table salt to fabricate the system himself, incorporating a thicker walled,
(sodium chloride) in foods is routinely determined as chloride by more rugged glass electrode with an electronic amplifier using
titration with silver ions. vacuum tubes to enhance the signal strength. The Acidimeter was
In the latter half of the 20th century, the time spent on such such a success that other colleagues learned about it and asked if
laborious and difficult wet chemical analyses began to decline. Beckman could build a few more. He immediately recognized the
Although many classical methods are still widely used today, they utility for food scientists to have a compact and portable instru-
were eventually substituted with instrumental methods that ment enclosed in a “black box” that was designed to be rugged in
provided lowered detection limits, increased analyte specificity, laboratory and processing environments. With that vision, Beck-
simplified use, reduced cost, higher sample throughput, and man ushered in a new era by forming a new company, National
automation capabilities. Technical Laboratories, to market the pH-meter, thus initiating
the modern instrumentation industry and the early stages of the
EARLY INSTRUMENTAL DEVELOPMENTS (1930-1950) transformation of food analysis. The Beckman model G pH-
The instruments utilized for food chemical analysis in the early meter introduced in 1937 following two earlier prototyes was the
part of the 20th century were quite crude. As an example, the first commercial laboratory instrument to combine electronics
8080 J. Agric. Food Chem., Vol. 57, No. 18, 2009 McGorrin
followed in 1957 by the model 137, a popular low-cost laboratory
instrument. Infrared analysis proved to be more powerful than
UV spectroscopy, because it provided chemical information
about organic functional groups and structure elucidation. IR
enabled new food components to be rapidly identified by com-
parison with known standards and to be analyzed for purity.
Because IR anaysis is able to differentiate cis- versus trans-
isomers, it is particularly useful in food lipid applications to
assess the trans-content of unsaturated fatty acids in hydroge-
nated oils. IR (followed later by near-IR) analysis provided an
early role in driving the shift from traditional wet methods to
modern instrumental analysis. It demonstrated that direct objec-
tive measurement could be simpler, faster, less expensive, and
considerably more accurate and precise.
PIONEERING ADVANCEMENTS IN CHROMATOGRAPHY
The development of chromatography within the past century
has provided the broadest impact and largest application to the
discipline of food analysis. Just over 100 years ago, a Russian
botanist, Mikhail Tswett, invented liquid chromatography as
part of his investigations on plant leaf pigments and first
presented his findings in 1901 as the XI Congress of Naturalists
and Physicians in St. Petersburg, Russia (40). Tswett’s research
led to the discovery of a liquid adsorption technique that
permitted the separation of chorophylls and xanthophylls on a
calcium carbonate adsorbent, with elution by petroleum ether/
ethanol mixtures. In subsequent years, he further refined the
technique, which in a 1906 publication he termed chromatogra-
phy, or “color-writing”, derived from the Greek words chroma
and graphikos (41). In his application, the separation of chloro-
phyll leaf pigments was clearly visible as colored bands on the
adsorbent column. Because his findings were published in the
Russian technical literature, their applications were largely un-
Figure 2. Beckman model G pH-meter (1937) (courtesy Chemical Heri-
known by Western scientists and fell into obscurity until nearly
tage Foundation).
20 years after his death. By that time, Archer J. P. Martin and
with direct chemical measurement (Figure 2). It demonstrated for Richard L. M. Synge were exploring an alternative to counterflow
the first time that instruments could eliminate the subjectivity, extraction to separate acetylated amino acids in the wool indus-
fragility, and complexity of earlier methods. To recognize its try. Eventually they devised the idea of partitioning one phase
significance, the American Chemical Society designated the while holding the other one stationary, and the result was the
development of the Beckman pH-meter a National Historic invention of liquid-liquid partition chromatography in
Chemical Landmark in 2004 (39). 1941 (42). This led to their receiving the 1952 Nobel Prize in
The next significant analytical instrument that Beckman Chemistry for their invention of partition chromatography.
developed of importance to food analysis was the model DU In his Nobel lecture, Martin casually revealed that he, in
ultraviolet (UV) spectrophotometer (38). It used similar electro- collaboration with A. T. James, had devised a mechanism for
nic technology that was designed for the pH-meter and was first gas-liquid chromatography using a liquid stationary phase
marketed in 1941 by National Technical Laboratories (now coated onto a solid packing, in combination with a gas mobile
Beckman Coulter). With the availability of commercial in- phase. A landmark publication followed shortly thereafter in
strments, UV measurements became an essential component of 1952 describing the invention of modern gas-liquid chromatog-
every food analysis laboratory for various colorimetric assays, raphy (GC) and its application to the separation and quantitation
which related the intensity of solution color as a way to quantify of C1-C12 volatile fatty acids (43). The first commercial GC
concentration of a specific analyte. As previously discussed, instrument manufactured was the Perkin-Elmer model 154 Vapor
colorimetric end points were typically used in vitamin assays. Fractometer in 1955. Early GC column packings included ground
Immediately before World War II, infrared (IR) spectrophoto- firebrick, laundry detergent, and whatever else food chemists
meters were under development by Beckman and others for the could experiment with and were used in combination with early
classified development of synthetic polybutadiene rubber and thermal conductivity detectors.
higher octane aviation fuel. IR spectrophotometers relied on an GC expanded rapidly over the three decades since its invention
optical source, prisms, and electronics to detect the characteristic in 1952. Major milestones in GC development came with column
stretching vibration “fingerprints” of molecules produced by their phase technology, flame ionization and electron capture detec-
interaction with IR light. The Perkin-Elmer Co. developed an tors, cold on-column sample inlet systems, and headspace con-
early prototype model 12 IR spectrophotometer in the 1930s, centrators (44). The introduction of robust, efficient, and
which was further perfected before being sold in 1944 as the first reproducible fused-silica capillary columns in the late 1970s by
IR spectrophotometer made commercially available to the scien- Hewlett-Packard and J&W Scientific propelled GC as a ubiqui-
tific community (29). However, in 1951 Perkin-Elmer introduced tous tool in food analysis laboratories (45). An advantage of
its highly successful infrared spectrophotometer, model 21, which capillary columns over packed columns is their greatly increased
made IR a routine laboratory tool for food analysis. It was separation efficiency, lower temperature, chemical inertness, and
Article J. Agric. Food Chem., Vol. 57, No. 18, 2009 8081
information-rich without the application of HPLC. New phases
continue to be developed that offer expanded resolution or speed
of analysis. Similarly, fatty acids, lipid components, flavors,
pesticide residues, steroids (cholesterol), and other food compo-
nents are routinely analyzed by capillary GC. Prior to the
development of chromatography, food chemical analysis
required repetitive crystallizations, fractional distillation, or che-
mical derivatization techniques.
DAWN OF THE MASSIVE INSTRUMENTS (1940-1980)
The history of mass spectrometry dates back more than
100 years and has its roots in physical and chemical studies
regarding the nature of stable elemental isotopes. Mass spectro-
metry is an analytical technique that identifies chemical composi-
tion based on the mass-to-charge ratio of charged particles as they
undergo chemical fragmentation and pass through electric and
magnetic fields. J. J. Thomson was the first to measure the m/z
values of positive ions, and he can rightfully be called the father of
mass spectrometry. In 1919, Francis Aston constructed the first
velocity focusing mass spectrograph with a mass resolving power
of 130, for which he won the Nobel Prize in 1922 for mass
Figure 3. Comparison of capillary column (A) versus packed column (B) spectrometry. Aston’s design improvements of the early mass
GC separation of calmus oil. Reprinted with permission from ref 44. spectrometer (MS) continued through the 1930s, with resolving
Copyright 2002 Elsevier. power increasing to 2000. The first commercial mass spectrometer
was the model 21-101 constructed in 1942 by the Consolidated
resistance to column bleed. Many trends in current progress Engineering Co. These early instruments were generally large,
can be seen to originate in the first two decades of the history of expensive, difficult to work with, and more suited to solving
GC, but the invention of fused-silica capillary columns greatly research problems. They were do-it-yourself, room-sized instru-
increased the application of high-resolution GC for flavor and ments that had to be used with vacuum pumps and pneumatic
fatty acid analysis. Figure 3 shows a typical comparison of the devices and were driven by large racks of analogue electronics (47).
significantly enhanced resolution power of capillary column GC In 1963, Associated Electrical Industries (now Kratos Ana-
over packed column GC, using the same stationary phase for lytical) introduced the MS-9 as the first high-resolution mass
distilled calmus oil, which has a “spicy-aromatic” fragrance spectrometer sold in the United States, which was applied by the
character (44). By the end of the century, GC had become the flavor and fragrance industry for accurate mass determinations.
single most widely used analytical tool in chemistry, despite the The concept was that a high-resolution mass spectrometer could
restriction that analytes needed to be volatile. measure mass with sufficient accuracy to deduce the elemental
A significant milestone in liquid chromatography was the composition of organic compounds (48). These MS instruments,
development of high-performance liquid chromatography nicknamed “workhorses”, led to significant advances in the
(HPLC) by James Waters (46). Commercial instruments intro- knowledge of flavors and identification of food components.
duced in 1967 by Waters Corp. made HPLC a valuable, widely A breakthrough development occurred in 1960 with the inter-
used tool for food chemists and led the way toward solving facing of a mass spectrometer as the detector for gas chromatog-
difficult separation problems by providing enhanced speed, raphy by Roland Gohlke and Fred McLafferty (49, 50). Through
sensitivity, and resolution. In 1972 Waters introduced the model this powerful combination, GC provided the separation of
6000 with the first dual-reciprocating piston pump (6000 psi), an volatile compounds and MS yielded unambiguous chemical
innovation in precision solvent delivery that delivered nearly identifications. Subsequently, major advances in flavor chemistry
pulseless column flow to increase the signal-to-noise ratio at the and flavor analysis were enabled by the GC-MS identification of
detector. Subsequently, HPLC evolved into an important food trace-level volatile components as commercial instruments were
analysis tool for separation of mixtures of compounds and their made available in the late 1960s (Figure 4).
quantitation, initially using refractive index and ultraviolet detec- One of the major developments that led to wider distribution
tors. The 1978 introduction of the first commercial, disposable and adoption of GC-MS instruments in food analysis labora-
miniature silica-based solid phase extraction (Sep-Pak) cartridges tories was the development of commercial quadrupole MS
by Waters became the predominant technique for food sample technology by Finnigan Instruments that replaced large, expen-
enrichment and purification by removing interfering substances sive, capricious magnetic sector instruments (51). Hewlett-Pack-
prior to HPLC analysis. ard licensed the technology, which eventually led to the
Other new chromatographic and electrophoretic techniques development of their computerized 5970 Mass-Selective Detec-
were introduced for application to food analysis, including thin- tors for GC;reliable and relatively simple instruments that could
layer chromatography (1938), paper chromatography (1944), and be routinely used by food chemists. GC-MS expanded rapidly
more recently sodium dodecyl sulfate-polyacrylamide gel elec- over the remaining half of the 20th century after the introduction
trophoresis (SDS-PAGE, 1967), two-dimensional PAGE (1975), of relatively low-cost but reliable benchtop instruments in the
ion chromatography (1975), capillary electrophoresis (1989), and 1980s. Progress in food chemistry continues to be rapid in
supercritical fluid chromatography (1995). comprehensive two-dimensional 2D-GC, fast analysis, detection
GC and HPLC have the broadest applications to food analysis. by atomic emission, and time-of-flight mass spectrometry and
Current analyses of sugars, polysaccharides, amino acids, pro- with applications to process analysis.
teins, vitamin, colors, herbicide and antibiotic residues, and other Early MS instruments in the 1960s had oscillographic trace
food constituents would not be as be as rapid, productive, or outputs that spewed-out bar graphs of mass spectra on a 3 ft
8082 J. Agric. Food Chem., Vol. 57, No. 18, 2009 McGorrin

Figure 4. Early Hitachi RMU-6E GC-MS instrument employed to identify volatile compounds in flavors (1969) (courtesy International Flavors & Fragrances).

Figure 5. Author using a Hewlett-Packard 5985 GC-MS instrument for identification of dairy flavor volatiles (1982) (courtesy Kraft Foods).
length of photosensitive paper with the push of a button! These sample preparation, provides rapid analysis, and offers the
were gradually obsolesced in the 1980s by computerized graphic potential to run multiple tests on a single sample. The early
displays of MS spectra, and the ability to match unknown commercial NMR instruments of the 1960s were 60 and 100 MHz
compounds with MS library databases (Figure 5). proton and 13C instruments manufactured by Varian Associates,
Nuclear magnetic resonance spectroscopy (NMR) was being which were applied to aroma compound identifications by the
developed initially as a tool for analysis of pure organic com- flavor industry (Figure 6). Later in the 20th century, high-
pounds in the late 1940s. NMR spectroscopy of a bulk material resolution spectrometers were constructed with 300-800 MHz
was first demonstrated independently by Felix Bloch and Edward fields that were coupled to computers with Fourier transform
M. Purcell in 1945. NMR is the most powerful analytical pulse sequences. In food applications, NMR can be used to
technique, because it provides unambiguous structure identifica- measure moisture content, and it can be applied to food lipids to
tion of organic molecules. Whereas NMR is not nearly as measure the degree of conjugated double bonds in fatty acid
sensitive an analytical technique as MS, it requires minimal mixtures in an easy and nondestructive manner.
Article J. Agric. Food Chem., Vol. 57, No. 18, 2009 8083

Figure 6. Varian HA-100 (100 MHz) NMR applied for identification of flavor compounds (1968) (courtesy International Flavors & Fragrances).

By the late 1980s, all-digital instruments incorporated improve- Laser Technology. When the first working laser was reported in
ments developed in both commercial and academic laboratories 1960, it was described as “a solution looking for a problem”.
that allowed them to be more affordable and much smaller, Theodore Maiman created the first operating laser in May 1960 at
occupying several linear feet on a laboratory bench rather than the Hughes Research Laboratory in California, by shining a high-
considerable floor space. During this period, newly designed power flash lamp on a ruby rod with silver-coated surfaces (53).
instruments provided the ability to measure food components Twenty-five years later, the laser’s distinctive qualities;its ability
that were different from or beyond what was previously possible. to generate an intense, very narrow beam of light of a single
These new tools were a source of major innovations in food wavelength;were being harnessed by analytical chemistry for
technology and were able to elucidate many issues about the detection systems in spectrometers and chromatographic instru-
nature of modern chemistry of foods, such as the correct assign- ments. Computer optical disk drives for mainframe and personal
ment of sensory attributes to specific flavor molecutles. computers were also developed that incorporated laser compact
disks for storage and retreival of large mass spectrometry and
KEY ENABLING TECHNOLOGIES chromatography data files.
Three significant converging technological advances occurred The parallel developments of the laser, microprocessor, and
during this period which contributed to significant advances in personal computer industries and the convergence of these
food analysis capabilities. technologies with food chemistry provided phenomenal advan-
Semiconductor Technology. Intel Inc. introduced its first 8-bit tages for further miniaturization of food analysis, accuracy,
microprocessor, the 4004 chip, in 1971. The 4004 was capable of speed, and limits of detection.
addressing up to 1 kB of program memory and up to 4 kB of data
memory. In 1979, Intel developed a new 16-bit microprocessor, MODERN AGE OF FOOD ANALYSIS
the 8088, and IBM engineers used it for the first personal Food analysis is often the beneficiary of technical advance-
computer. The combination of the new 16-bit microprocessor ments (chromatography, separations, MS, FTIR, and NMR
and the name IBM shifted the personal computer to a mainstream spectroscopy) that were initially developed for the petroleum,
business tool. pharmaceutical, chemical manufacturing, environmental, and
Computing Technology. In 1981, IBM Corp. introduced the natural products chemistry areas. Beginning with the commer-
personal computer (PC), viewed as the most ubiquitous labora- cialization of the Beckman Acidimeter in 1934 through the
tory-enabling technology. The affordable and miniaturized PC Caliper LabChip microchip for fluid handling in 1996, the
became an integral component for data processing and graphics Chemical Heritage Foundation has identified “50 chemical
display as part of laboratory instruments such as IR and UV laboratory instruments that changed the world in the 20th
spectrophotometers, chromatography data systems, and mass century” (54). These key instruments were recognized as having
spectrometers. a significant impact across a broad spectrum of chemical research
The impact of modern electronics is global in scope, especially and analysis. Through technology adaptation, many of these key
during the past 50 years since semiconductor technology emerged instrumental capabilities were applied by food chemists to devel-
and began to transform the world. Thirty-five years earlier, John op new analytical methods and procedures for measuring food
Mauchly and J. Presper Eckert developed the ENIAC I (electrical components. For example, the pharmaceutical industry drove the
numerical integrator and calculator). Through research spon- need for chiral separations of asymmetric organic compounds.
sored by the U.S. military, the new computer used vacuum tubes, Whereas one enantiomeric isomer may exhibit the desired pha-
instead of switches and relays, to speed calculations. The only macological activity, the opposite enantiomer could potentially
drawback was that it required 17468 vacuum tubes, which cause undesired side effects (e.g., (S)-thalidomide). Chiral chro-
emitted considerable heat and needed frequent replacement. It matography was readily adapted for authentication of natural
covered 1800 square feet (167 m2) of floor space, weighed 30 tons, flavors to determine the most sensory-active enantiomers during
and consumed 160 kW of electrical power (52). discovery of new flavor compounds. Capillary electrophoresis,
8084 J. Agric. Food Chem., Vol. 57, No. 18, 2009 McGorrin
which was initially introduced by Beckman in 1989 for biochem- The requirement for nutrition labeling of foods juxtaposed with
ical separations, was readily applied to the analysis of pea parallel advances in the fields of moleular biology, biochemistry,
oligosaccharides, citrus pectins, casein aand whey proteins, and genetics has spawned the development of new chemical
gliadins and glutenins in wheat, and myoglobin proteins and methods to measure carbohydrates, fat, essential fatty acids,
peptides in foods (55). protein, fiber, macrominerals (Ca, Mg, K, Na, P), trace minerals
Multidimensional or hyphenated instruments employ two or (Co, Cu, Fe, Cr, Mn, Se, Zn), and vitamins in a variety of food
more techniques either sequentially (such as GC-MS and GC- and beverage matrices.
MS/MS) or in parallel (GC-IR-MS). Fragment ions can only be Sampling. Perhaps one of the most critical aspects of the
used for structure determination, and a tandem instrument with analytical process is obtaining a representative food sample for
two quadrupole mass analyzers allows an analyst to “piece analysis. Consideration for how to conduct appropriate sampling
together the structural puzzle” to deduce the origin of fragment can occur on two levels: (1) developing a protocol for sample
ions. The high-performance liquid chromatography-tandem collection (e.g., field crop location, manufacturing, marketplace,
mass spectrometry (LC-MS/MS) technique has been useful for or home), assessing the statistical number of samples required,
sequencing food peptides and proteins such as caseins. and determining whether the food should be sampled in raw,
Similarly, comprehensive two-dimensional GCGC and cooked, or processed form; (2) compositing the sample and
LCLC instruments became available within the past five years selecting a representative aliquot from a homogenized multi-
that are capable of significantly enhanced separation of complex component food prior to analysis. A homogeneous sample is
mixtures (56-58). These instruments rely on combinations of necessary prior to the performance of chemical analysis to ensure
different chemical selectivities to achieve separations of flavors or that the result represents the composition of the overall food
phenolic antioxidants in beverages. Time-of-flight (TOF) tandem sample (e.g., a pizza that contains bread crust, cheese, tomato,
mass spectrometers with matrix-assisted laser desoprtion ioniza- mushroom, sausage, and green pepper components). Care must
tion (MALDI) are now used routinely for rapid sequencing of be taken to avoid contaminating the sample or exposing
amino acids in proteins. it to excessive heat, which can cause loss of volatile flavor
Simultaneous with the progressive development of new instru- components or accelerate decomposition of labile food compo-
ments with increased analyte specificity over the past 60 years was nents. The food chemist may need to consider the chemical
the ongoing trend toward lowered limits of detection in food stability of the targeted analyte to appropriately blend a
analysis. Relative sensitivities of analytical tests for minerals and food under frozen, inert atmosphere, or low-light conditions
elements over the early period have been tabulated (6). New and (e.g., for vitamin analyses).
improved detectors for GC (flame ionization, electron capture, Sample Preparation. Challenges remain for the analysis of
chemiluminescence, pulsed flame photometric, MS) and HPLC individual chemical species in food systems because of their
(refractive index, ultraviolet, fluorescent, diode array, evapora- complex multicomponent structure. Despite the availability of
tive light-scattering, MS) provided micromole to picomole en- modern techniques of separation and identification, such as GC-
hancements in sensitivity depending on the analyte/application. MS and LC-MS/MS, rarely is it possible to load a syringe with a
Spectroscopy and spectrometry instruments (MS, IR, UV, NMR, food sample and directly inject it into an analytical instrument to
atomic emission) afforded a reduction in detection limits from obtain a sensible result! Sample preparation and extraction
nanomole to picomole. As analytical instruments became increas- remain among the most time-consuming and error-prone steps
ingly sensitive, additional components in foods that were pre- in the food analysis process (61). However, they are critical
viously undetectable could be quantified. Furthermore, procedures because food scientists need to isolate and concentrate
carcinogenic compounds that formerly had been below detection a wide range of analytes from complex and varied food matrices.
limits could now be measured. This had significant implications in Sample preparation typically involves pH adjustment, extraction
the area of food law and food regulation. Perhaps the best-known with organic solvents, solid phase extraction, cleanup (filtration,
example is the 1958 amendment to the U.S. FD&C Act of 1938, liquid-liquid partitioning), and concentration for subsequent
known as the “Delaney clause”, named after the Congressman analysis via chromatography or other methods.
who chaired the Congressional subcommittee (59). The Delaney Recently developed automated sample preparation techniques
clause prohibits the use of food additives that have been shown to that can be easily integrated with other analytical systems offer
cause cancer in humans or animals, without consideration of the productivity improvements for many food analysis laboratories.
concentration level at which a toxic effect is induced. Virtually all Automated preparation techniques incorporate either low-
foods contain traces of carcinogenic substances from environ- volume solvent extraction or solvent-free thermal desorption,
mental and natural sources (e.g., safrole in sassafras and and in most cases they can increase the efficiency relative to
cinnamon), and this law had broad ramifications (59). It required conventional solvent extraction. For example, the Mojonnier
processed foods to be reformulated to eliminate any additives ether extraction method (AOAC 922.06) typically requires 2-3 h
(e.g., those containing trace impurities) that now had measurable and 110 mL of solvent for determining the fat content in
amounts of carcinogens, even though they probably did not chocolate, whereas pressurized fluid extraction (accelerated sol-
consitute a health risk. The Delaney clause was first invoked in vent extraction (ASE), Dionex) reduces the extraction time to
1959 when traces of the cancer-causing herbicide aminotriazole 18 min and solvent use to less than 20 mL (62). Other examples
were discovered on cranberry plants from Oregon and Washing- include supercritical fluid extraction, thermal desorption, solid
ton, causing sales of cranberries to plummet during the week of phase extraction, solid phase microextraction (SPME), and stir-
Thanksgiving. Pesticide use was removed from the Delaney bar sorptive extraction (SBSE). Modular systems are now readily
clause in 1996 by an amendment to the U.S. Food Quality and available that automate these procedures and interface them to
Protection Act (60). analytical instruments, such as GC, GC-MS or HPLC.
Specific measurements of food quality include food safety Macrocomponent Analyses. Proximate analyses (crude fat,
(microbiological) testing, nutrient content, pesticide residue protein, moisture, soluble solids, ash) have been typically con-
testing, and biotechnology applications (use of genetically ducted by classical wet chemistry methods that were tailored for
modified organisms, GMOs). The latter three areas typically in- analyses of food products (Table 2) (12). New developments and
volve development and application of new chemical analyses. improvements in instrumental techniques are providing rapid,
Article J. Agric. Food Chem., Vol. 57, No. 18, 2009 8085
a
Table 2. Comparison of Representative Food Analysis Methods organic acids, ionic species, peptides, and proteins. Highly
analyte classical “wet” method instrumental method sensitive fluorescence detectors for HPLC instruments enable
lower limits of detection for vitamins, proteins, aspartame, and
moisture Karl Fischer near-infrared mycotoxins.
dry fruits/vegetables dry fruits/vegetables Spectroscopic techniques are highly desirable for analysis of
967.19 E-G 967.19 B-D food macrocomponents because they often require minimal
cocoa 977.10
sample preparation, provide rapid analysis, and have the
vacuum oven microwave oven
meat/poultry 950.46 meat/poultry 985.14
potential to run multiple tests on a single sample. These
advantages particularly apply to nuclear magnetic resonance
sugars total dextrins 988.12 HPLC (NMR), infrared (IR), and near-infrared (NIR) spectro-
Munson-Walker 906.03 milk chocolate 980.13 scopy. The latter technique is routinely used as a quality
sucrose (cocoa) 920.82 ion chromatography assurance tool to determine compositional and functional
analysis of food ingredients, process intermediates, and fin-
total dietary fiber enzymatic-gravimetric 985.29 ished products.
Microcomponent Analyses. Volatile compounds are most often
protein Kjeldahl 955.04; 988.05 Dumas combustion/GC analyzed in foods, flavors, and aroma systems. Since the 1960s,
milk 991.23 meat 992.15
the combination of gas chromatography and mass spectrometry
capillary electrophoresis
has become an essential tool to identify flavors (63, 64). More
fat total (crude): total (crude): recent developments in capillary column technology and the use
Mojonnier ether extraction microwave-solvent extraction of various headspace concentrators (equilibrium, dynamic/
flour 922.06 meat 985.15 Tenax, vacuum, solvent-assisted flavor evaporation, solid phase
milk 989.05 accelerated solvent extraction microextraction, stir-bar sorptive extraction) have shortened the
Soxhlet ether extraction fat as fatty acid glycerides: assay time and increased limits of detection for potent flavor
meat 960.39 gas chromatography components (65). GC-olfactometry as a sensory-directed tech-
Roese-Gottlieb (milk) 905.02 fish oil fatty acids 991.39 nique has significantly advanced the field of flavor chemistry
during the analysis of complex flavor mixtures (66). It enables the
cholesterol titrimetric 941.09 gas chromatography 994.10
relative contributions of individual flavor components to be
vitamin A Carr-Price 974.29 HPLC 2001.13; 2002.06 (milk) assessed in real time as they are being separated while allowing
the food chemist to assign aroma descriptors (“fruity”, “green”,
minerals (Ca, Cu, atomic absorption ICP-atomic emission “roasted”) to chromatographic peaks that have sensory impact.
Fe, Mn, Zn) spectrometry 991.25; spectrometry Additional trace analytes in foods include colorants, pigments,
999.10 vitamins, and minerals. Research on bioactive “functional” food
compounds relies on the development of new assays with in-
metals (Pb, Cd, mercury 971.21 ICP-mass spectrometry
creased sensitivity and specificity to measure efficacious phyto-
Hg)
chemical components (anthocyanins, polyphenolic compounds),
a
AOAC Official Analytical Method indicated in bold. which are frequently colored. Atomic spectrometry methods
including atomic absorption, atomic emission, and elemental
automated alternatives to the classical food analysis methods. mass spectrometry are routinely used to measure minerals (e.g.,
Current regulatory requirements for food manufacture obligate calcium, zinc) and heavy metals (e.g., lead, cadmium, mercury) in
not just the analysis of total fat content (e.g., Mojonnier) but also food products (67). Inductively coupled plasma atomic emission
the characteristics of the fat, that is, saturated, monounsaturated, spectrometry (ICP-AES) and inductively coupled plasma mass
polyunsaturated, monoglycerides, diglycerides, cholesterol, etc. spectrometry (ICP-MS) offer the advantage of providing simul-
Classical total protein determinations were historically based on taneous multielement measurement in food samples. In these
nitrogen content (the Kjeldahl technique); however, modern techniques, samples are vaporized, atomized, and excited in a
electrophoretic and chromatographic methods were developed high-temperature argon plasma. Detection limits are 1-100 ppb
that enable rapid and specific amino acid and peptide assays. for ICP-AES and 1-100 ppt for ICP-MS. Microelemental
Total carbohydrate values are required to be listed as specific analysis by ICP-AES has been used to differentiate country of
components: sugars, sugar alcohols, dietary fiber, soluble fiber, origin and geographic growing conditions for potatoes, coffee,
and insoluble fiber. Moisture content has been redefined to pistachios, and fresh strawberry, blueberry, and pear (68), by
specify the states of water as being “free”, “bound” (e.g., hydro- simultaneously measuring relative differences in elemental con-
gen bonding to sugars, salts, proteins), or “adsorbed” (e.g., with centrations. Recently developed micro X-ray fluorescence spec-
phospholipid layers, starch), and each of these modes requires a trometry instruments provide a nondestructive technique for
specific analytical measurement. food quality control applications to rapidly analyze metallic
HPLC is a very powerful and versatile technique for analyzing elements, minerals, and heavy metals at parts per million sensi-
and purifying biomolecules and, consequently, is a well-estab- tivity limits. Analytical measurement of toxicological residues in
lished mode for food and beverage analysis. Ongoing advances in foods including pesticides, aflatoxins, and other mycotoxins have
column supports, bonding chemistry, porous particles, and been devised by the development of chromatographic, spectro-
packing materials have enabled increased speeds and efficient graphic, and immunochemical methods for assaying these com-
separations of a wide range of organic and inorganic food pound classes.
analytes from low parts per million to high parts per billion Enzymes, microbes, and antibodies have been exploited as
levels. Related column techniques that involve separation by biosensors to rapidly measure the presence of amylases or
either ionic mobile phases or charged fields are ion chro- proteases in foods, which can cause starch thinning or bitter
matography and capillary electrophoresis, respectively. Typical flavors, respectively. Enzyme-linked assays have been used to
food analytes quantified by these techniques include mono- measure vitamins, antibiotic residues, microbial toxins, various
and disaccharides, aspartame, phospholipids, vitamins, caffeine, sugars, ethanol, and carbon dioxide (69). In certain cases, enzyme
8086 J. Agric. Food Chem., Vol. 57, No. 18, 2009 McGorrin
activity assays are practical food safety probes to measure disposal. Flexible laboratory designs, moveable casework, and
whether adequate processing of a food product has been overhead utility hook-ups (gases, water, electricity, vacuum) will
achieved. provide the ability to quickly change from one analysis project to
Electrophoretic methods have been applied to food analysis, another as new applications dictate.
initially using SDS-PAGE for separation of milk proteins (caseins, The needs of food-processing and agricultural production
whey) and cereal proteins (glutenin, gliadin, zein) (55, 70). Varia- will continue to create demand for improved analytical methods
tions include isoelectric focusing, 2-D gel elctrophoresis, and that can measure new attributes, features, or molecular compo-
isotachophoresis. Analyte separations are a function of their nents in food products or ingredients while providing more
mass-to-charge ratios in an externally applied electric field. Gel analyte information at lower detection limits. Instrumental
electrophoresis was demonstrated to be especially useful in protein improvements in mass spectrometers enabled state-of-the-art
fingerprinting for the authentication of specific animal species to limits of detection that evolved over the 1970-2000 period
detect labeling fraud. Applications include the identification of from picomole (10-12) to femtomole (10-15). The progression
specific proteins in red snapper, buttermilk powder, egg pasta, over the next couple of decades is toward attomole (10-18) limits
citrus isozymes, and cow’s milk in goat cheese (55,70,71). Follow- for the ability to detect thousands of individual molecules.
ing its commercial introduction, capillary electrophoresis (CE) Ion detectors for mass spectrometry applications are currently
was readily substituted for SDS-PAGE in the 1990s to develop being developed at the frontiers of analytical capability using
new methods for food protein analysis (55, 72-75). CE provides electrode arrays (82). The convergence of biomedical advances,
analytical separation and reproducibility that are superior to those health and nutrition needs, and food choices will likely evolve
of gel electrophoresis and are comparable to those of HPLC, with into the need for information-rich, real-time analytical mea-
a range of detector options including UV, diode array, fluores- surements that address bioavailability and delivery. As the
cence, conductivity, and MS (76). CE applications include protein competitive marketplace evolves, new regulatory requirements
authentication to identify specialty cheeses produced with vege- for food analysis will surface, driven by governmental and
table coagulant instead of rennet (77), monitoring specific flavo- political forces.
noids and amino acids in orange juice for freshness (78), Continued research in the areas of health-promoting con-
determining D-isocitric acid as a marker for authenticity of orange stituents of foods (e.g., carotenoid phytochemicals and poly-
juice (79), and measuring phenolic acids in extra virgin olive phenolic antioxidants) will spawn the development of new
oil (80). assays to measure their presence in foods and changes occur-
Combinations of chromatographic, NMR, isotopic, and en- ring during processing. Flavonoids, isoflavones, isothio-
zyme-linked methods have been developed to screen and dete- cyanates, organosulfur compounds, carotenoids (lycopene,
mine the authenticity of suspect food products and ingredients, as lutein), saponins, and capsaicinoids are active areas of re-
prompted by economic fraud (19-23). Ironically, adulteration search related to potential benefits for cancer prevention, anti-
occasionally persists 100 years after creation of the Food and inflammation, antioxidant effects, and assistance to the immune
Drug Act in Dr. Wiley’s era. Modern examples include partial system (83, 84).
substitution of corn syrup for honey; organic acids and sugars for Rapid methods for testing raw ingredients, product quality,
apple juice; and benzaldehyde for almond extract. and process monitoring will undoubtedly experience continued
growth. The percentage of rapid tests should increase dramati-
FUTURE DEVELOPMENTS AND HORIZONS cally in the coming years, as diagnostic assay companies provide
Looking toward the next century, any prediction about new performance improvements. Food-processing companies will
developments in food analysis must consider the convergence of readily adopt rapid analytical methods to screen raw materials
multiple influences. A first consideration involves the natural to diminish the risk of food safety incidents and ensure compli-
evolution of scientific discoveries, which proceed at their own ance with regulatory standards. Near-infrared (NIR) and mid-
pace, but are accelerating due to rapid sharing of information via infrared (MIR) spectroscopy applications are currently being
the Internet and electronic media. New technological advances examined for online quality monitoring in food processing
will emerge, driven by old-fashioned Edisonian experimentation plants (85). ATP bioluminescence assays using the firefly enzyme
and innovation. The current desire for smaller analytical instru- luciferase are commercially available as test kits to monitor
ments, increased speed of results, lowered detection limits, easier food residues and microorganisms during the sanitation of food
operation, and portable applications of analytical measurements processing equipment (86). Fiber optic biosensors using differ-
in food production or field crop environments other than in the ent surface chemistries and antibody complexes are being
traditional laboratory will continue. explored for the detection of foodborne pathogens (Escherichia
The food analytical laboratory of the future will likely have coli, Listeria, Salmonella) (87). Nanoparticle sensors are under
glassware and equipment that resemble those of current labora- development as DNA probes for the detection of microbial
tories; however, the progression to smaller instruments, compu- pathogens in foods using semiconductor, metallic, metal oxide,
ters, and data collection devices is imminent (81). Laptop or polymeric materials (88). Aroma sensor arrays are being
computers now reside in most laboratories, but miniaturization designed using chemically responsive dyes that provide unique
of electronic hardware and more powerful software systems will colorimetric patterns for individual odorants (89). Future tech-
likely drive integrated instrument control and data collection nology refinements of electronic noses (90, 91) and electronic
systems that incorporate electronic notebooks. Bar-coding tongues (92) will likely find process monitoring applications for
systems will evolve to track and monitor large numbers of flavor quality.
samples. Radio frequency identification (RFID) systems are Trends are emerging through the miniaturization and port-
starting to be used in the commercial and industrial sectors that ability of analytical instruments. Cutting-edge chromatography
have yet to be integrated into laboratory environments. Auto- technologies utilize credit-card-sized polyimide-based fluidic
mation of instruments and analytical measurements is a likely chips in nanoflow channel HPLC-MS instruments to quantitate
expectation for the future of routine food assays. Smaller instru- peptides (93). Field-portable instruments are continuing to be
ments will integrate microfluidic designs that use smaller sample developed, which could be applied to monitor crop ripening and
volumes and fewer solvents and require less cleanup and waste assess fruit and vegetable quality.
Article J. Agric. Food Chem., Vol. 57, No. 18, 2009 8087
Pesticide residue testing of foods is routinely performed to (14) American Oil Chemists’ Society. Official Methods and Recommended
comply with various international food regulations that vary by Practices, 6th ed.; Champaign, IL, 2009.
country. Pesticide testing relies on analytical methods as screen- (15) Wehr, H. M., Frank, J. F., Eds. Standard Methods for the Examina-
ing tools to measure whether an approved pesticide is being tion of Dairy Products, 17th ed.; American Public Health Association:
correctly applied for specific food crops. As future pesticides are Washington, DC, 2002
(16) U.S. Pharmacopeia. Food Chemicals Codex, 6th ed.; Rockville, MD,
developed and approved for food use, new analytical methods
2008.
(usually GC or HPLC) will need to be subsequently developed to (17) TTB official methods, http://www.ttb.gov/ssd/labeling_methods.
quantify their presence and levels in foods. shtml accessed Jan 4, 2009.
The engineering of new genetically modified food crops has (18) U.S. FDA Food Allergen Act of 2004, http://www.cfsan.fda.gov/
and will continue to provide impetus for advancement of new ∼dms/alrgact.html, accessed Jan 4, 2009.
analytical methods to screen for compositional or antinutritive (19) Martin, G. G.; Martin, Y.-L.; Naulet, N.; McManus, H. J. D.
differences compared with conventional food crops. HPLC-MS Application of 2H SNIF-NMR and 13C SIRA-MS analyses to maple
methods have been developed for sequencing food proteins in syrup: detection of added sugars. J. Agric. Food Chem. 1996, 44,
genetically modified crops to characterize peptide fragments and 3206–3213.
other molecular features that correspond with those of known (20) Remaud, G.; Debon, A. A.; Martin, Y.; Martin, G. G.; Martin, G. J.
Authentication of bitter almond oil and cinnamon oil. Application of
food allergens (94).
the SNIF-NMR method to benzaldehyde. J. Agric. Food Chem.
The high-throughput testing demands of the food industry 1997, 45, 4042–4048.
will likely create financial incentives for new generations of (21) Cordella, C.; Moussa, I.; Martel, A.-C.; Sbirrazzuoli, N.; Lizzani-
analytical instruments that can make specialized measure- Cuvelier, L. Recent developments in food characterization and
ments. As the need for global food supply traceability grows, adulteration detection: technique-oriented perspectives. J. Agric.
increasing numbers of food products and ingredients will need Food Chem. 2002, 50, 1751–1764.
to be routinely tested. Requirements for new analytical labora- (22) Jamin, E.; Martin, F.; Santamaria-Fernandez, R.; Lees, M. Detec-
tory instruments will emphasize performance, sensitivity, re- tion of exogenous citric acid in fruit juices by stable isotope ratio
liability, simplified use, and low-cost for high-volume, routine analysis. J. Agric. Food Chem. 2005, 53, 5130–5133.
assays. The transformative dynamics of the revolutionary (23) Wilhelmsen, E. C. Adulteration determination. In ref 1, pp 3862-
3888.
years from 1930 to 1950 are still at work at the beginning of
(24) Anderson, O. E. The Health of a Nation: Harvey W. Wiley
the 21st century. No doubt, analytical instruments have and and the Fight for Pure Food; University of Chicago Press: Chicago,
will continue to profoundly shape the future of food chemistry IL, 1958.
and the food industry. (25) Wiley, H. W. Foods and Their Adulteration, 3rd ed.; P. Blakiston's Son:
Philadelphia, PA, 1917.
ACKNOWLEDGMENT (26) Fennema, O. R.; Tannenbaum, S. R. Introduction to food chemistry.
In Food Chemistry, 3rd ed.; Fennema, O. R., Ed.; Dekker: New York,
I am grateful for the assistance of the Chemical Heritage
1996; pp 1-15.
Foundation, Philadelphia, PA, for valuable background materi- (27) Swann, J. P. How chemists pushed for consumer protection: the
als and resources used in the preparation of this paper. Food and Drugs Act of 1906. Chem. Heritage 2006, 24 (2), 6–11.
(28) The history of AOAC. www.aoac.org/about/aoac.htm, accessed Jan
LITERATURE CITED 4, 2009.
(1) McGorrin, R. J., Section Ed. Food Analysis. In Encyclopedia of (29) Brock, D. Revolutionary tools. Chem. Heritage 2002, 20 (3), 39.
Analytical Chemistry: Instrumentation and Applications; Meyers, R. (30) Koivistoinen, P. E. Introduction: the early history of food com-
A., Ed.; Wiley: New York, 2000; Vol. 5, pp 3857-4332. position analysis;source of artifacts until now. Food Chem. 1996,
(2) Nielsen, S. S., Ed. Food Analysis, 3rd ed.; Kluwer Academic/Plenum 57, 5–6.
Publishers: New York, 2003. (31) Lesch, J. E. The German Chemical Industry in the Twentieth Century;
(3) Nollet, L. M. L., Ed. Handbook of Food Analysis, 2nd ed.; Dekker: Springer: New York, 2000.
New York, 2004; Vol. 1-3. (32) Carr, F. H.; Price, E. A. Color reactions attributed to vitamin A.
(4) Wrolstad, R. E., Acree, T. E., Decker, E. A., Penner, M. H., Reid, D. Biochem. J. 1926, 31, 497–501.
S., Schwartz, S. J., Shoemaker, C. F., Smith, D., Sporns, P., Eds. (33) Dann, W. J.; Evelyn, K. A. The determination of vitamin A with the
Handbook of Food Analytical Chemistry: Vol. 1, Water, Proteins, photoelectric colorimeter. Biochem. J. 1938, 32, 1008–1017.
Enzymes, Lipids, and Carbohydrates; Wiley, Hoboken, NJ, 2005. (34) Augustin, J., Klein, B. P., Becker, D., Venugopal, P. B., Eds.
(5) Wrolstad, R. E., Acree, T. E., Decker, E. A., Penner, M. H., Reid, Methods of Vitamin Assay, 4th ed; Wiley: New York, 1985.
D. S., Schwartz, S. J., Shoemaker, C. F., Smith, D., Sporns, P., Eds. (35) De Leenheer, A. P., Lambert, W. E., Van Bocxlaer, J. F., Eds.
Handbook of Food Analytical Chemistry: Vol. 2, Pigments, Colorants, Modern Chromatographic Analysis of Vitamins, 3rd ed.; Dekker:
Flavors, Texture, and Bioactive Food Components; Wiley, Hoboken, New York, 2000.
NJ, 2005. (36) Baker, S. A.; Miller-Ihli, N. J. Atomic spectroscopy in food analysis.
(6) Ewing, G. W. Analytical chemistry: the past 100 years. Chem. Eng. In ref 1, pp 3888-3912.
News 1976, 54 (15), 128-134, 139-142. (37) Papers of Howard B. Bishop, http://othmerlib.chemheritage.org,
(7) Pearson, D. The Chemical Analysis of Foods, 7th ed.; Chemical accessed Jan 4, 2009.
Publishing: New York, 1976. (38) Bowden, M. E. Beckman pH meter, first and last. Chem. Heritage
(8) Joslyn, M. A., Ed. Methods in Food Analysis, 2nd ed.; Academic Press: 2004, 22 (3), 6–7.
New York, 1970. (39) http://acswebcontent.acs.org/landmarks/landmarks/phmeter/
(9) Pomeranz, Y.; Meloan, C. E. Food Analysis: Theory and Practice, 3rd phmeter.html, accessed Jan 6, 2009.
ed.; Chapman and Hall: New York, 1994. (40) Ettre, L. S.; Tswett, M. S. The invention of chromatography. LC-
(10) Fed. Regist. 1993, 58 (3) 632-690, 2066-2964. GC North Am. 2003, 21 (5), 458–467.
(11) Food labeling: trans fatty acids in nutrition labeling, 21 CFR Part (41) Tswett, M. S. Physical chemical studies on chlorophyl adsorptions.
101. Fed. Regist., 2003, 68 (133), 41433-41506. Ber. Dtsch. Bot. Ges. 1906, 24, 316–323.
(12) Latimer, G. W.; Horwitz, W. Official Methods of Analysis, 18th ed., (42) Martin, A. J. P.; Synge, R. L. M. A new form of chromatogram
2nd rev.; AOAC International: Gaithersburg, MD, 2007. employing two liquid phases: a theory of chromatography. 2.
(13) Grami, B., Ed. Approved Methods of Analysis, 10th ed.; AACC Application to the micro-determination of the higher monoamino-
International: St. Paul, MN, 2000. acids in proteins. Biochem. J. 1941, 35, 1358-1366.
8088 J. Agric. Food Chem., Vol. 57, No. 18, 2009 McGorrin
(43) James, A. T.; Martin, A. J. P. Gas-liquid chromatography: the (71) Malin, E. L.; Basch, J. J.; Shieh, J. J.; Sullivan, B. C.; Holsinger,
separation and microestimation of volatile fatty acids from formic V. H. Detection of adulteration of buttermik powder by gel electro-
acid to dodecanoic acid. Biochem. J. 1952, 50, 679. phoresis. J. Dairy Sci. 1994, 77, 2199–2206.
(44) Bartle, K. D.; Myers, P. History of gas chromatography. TrAc, (72) Lindeberg, J. Capillary electrophoresis in food analysis. Food Chem.
Trends Anal. Chem. 2002, 21 (9-10), 547–557. 1996, 55 (73-94), 95–101.
(45) Freeman, R. R., Ed. High Resolution Gas Chromatography, 2nd ed.; (73) Swedberg, S. Capillary electrophoresis: principles and applications.
Hewlett-Packard: Palo Alto, CA, 1981. In Instrumental Methods in Food Analysis; Pare, J. R. J., Belanger,
(46) McDonald, P. D. Waters Corporation: fifty years of innovation in J. M. R., Eds.; Elsevier: Amsterdam, The Netherlands, 1997; pp 367-
analysis and purification. Chem. Heritage 2008, 26 (2), 32–37. 394.
(47) Judson, C.; Grayson, M. A workhorse comes to CHF. Chem. (74) Frazier, R. A.; Ames, J. M.; Nursten, H. E. The development and
Heritage 1998, 16 (2), 20–21. application of capillary electrophoresis methods for food analysis.
(48) Grayson, M. The MS-9: an icon of mass spectrometry. Chem. Electrophoresis 1999, 20, 3156–3180.
Heritage 2007, 25 (1), 18. (75) Frazier, R. A.; Ames, J. M.; Nursten, H. E. Capillary Electrophoresis
(49) Gohlke, R. S. Time-of-flight mass spectrometry and gas-liquid for Food Analysis: Method Development; Royal Society of Chemistry:
partition chromatography. Anal. Chem. 1959, 31, 535–541. London, U.K., 2000.
(50) Gohlke, R. S.; McLafferty, F. W. Early gas chromatography/mass (76) Simo, C.; Barbas, C.; Cifuentes, A. Capillary electrophoresis-mass
spectrometry. J. Am. Soc. Mass Spectrom. 1993, 4 (5), 367–371. spectrometry in food analysis. Electrophoresis 2005, 26, 1306–1318.
(51) Webb-Halpern, L. Detecting success. Chem. Heritage 2008, 26 (2), 31. (77) Roseiro, L. B.; Gomez-Ruiz, J. A.; Garcia-Risco, M.; Molina, E.
(52) McCartney, S. ENIAC: The Triumphs and Tragedies of the World’s Vegetable coagulant (Cyanara cardunculus) use evidenced by capil-
First Computer; Waller: New York, 1999. lary electrophoresis permits PDO cheese certification. Lait 2003, 83,
(53) Maiman, T. H. Stimulated optical radiation in ruby. Nature 1960, 18 343–350.
(4736) 7, 493-494. (78) Cancalon, P. F. Analytical monitoring of citrus juices by using
(54) Chemical Heritage Foundation. Fifty chemical laboratory instru- capillary electrophoresis. J. AOAC Int. 1999, 82, 95–106.
ments that changed the world, www.chemheritage.org/exhibits/ (79) Jezek, J.; Suhaj, M. Application of capillary isotachophoresis for
50list.htm, accessed Jan 6, 2009. fruit juice authentication. J. Chromatogr., A 2001, 916, 185–189.
(55) Cancalon, P. F. Electrophoresis and isoelectric focusing in food (80) Pancorbo, A. C.; Cruces-Blanco, C.; Carretero, A. S.; Gutierrez,
analysis. In ref 1, pp 3929-3955. A. F. Sensitive determination of phenolic acids in extra-virgin olive
(56) Marriott, P.; Shellie, R. Principles and applications of comprehen- oil by capillary zone electrophoresis. J. Agric. Food Chem. 2004, 52,
sive two-dimensional gas chromatography. TrAC, Trends Anal. 6687–6693.
Chem. 2002, 21, 573. (81) Anonymous. Lab Equipment. R&D 2006, 48, F18-F20.
(57) Adahchour, M.; van Stee, L. L. P.; Beens, J.; Vreuls, R. J. J.; (82) Arnaud, C. H. Advanced detectors take the stage. Chem. Eng. News
Batenburg, M. A.; Brinkman, U. A. Comprehensive two-dimen- 2009, 87 (13), 42–43.
sional gas chromatography with time-of-flight mass spectrometric (83) Chaillou, L. L.; Nazarenko, M. A. New method to determine
detection for the trace analysis of flavor compounds in food. antioxidant activity of polyphenols. J. Agric. Food Chem. 2006, 54,
J. Chromatogr., A 2003, 1019, 157–172. 8397–8402.
(58) Schoenmakers, P.; Majors, R. E. LCxLC: comprehensive two- (84) Li, N.; Liu, J.-H.; Zhang, J.; Yu, B.-Y. Comparative evaluation of
dimensional liquid chromatography. LC-GC North Am. 2008, 26 cytotoxicity and antioxidative activity of 20 flavonoids. J. Agric.
(7), 600–608. Food Chem. 2008, 56, 3876–3883.
(59) Pariza, M. W. Toxic substances. In Food Chemistry, 3rd ed.; Fennema (85) Irudayaraj, J.; Reh, C. Nondestructive Testing of Food Quality;
O. R., Ed.; Dekker: New York, 1996; pp 825-840. Blackwell Publishing/IFT Press: Ames, IA, 2008.
(60) Merrill, R. A. Food safety regulation: reforming the Delaney clause. (86) Kyriakides, A. L.; Patel, P. D. Luminescence techniques for micro-
Annu. Rev. Public Health 1997, 18, 313–340. biological analysis of foods. In Rapid Analysis Techniques in Food
(61) Lichon, M. L. Sample preparation. In Nollet, L. M. L., Ed.; Hand- Microbiology; Patel, P. D., Ed.; Blackie Academic and Professional:
book of Food Analysis. Vol. 3, Methods, Instruments and Applications, London, U.K., 1994; pp 196-231.
2nd ed.; Dekker: New York, 2004; pp 1741-1756. (87) Geng, T.; Bhunia, A. K. Optical biosensors in foodborne pathogen
(62) Majors, R. E. Modern techniques for the extraction of solid detection. In Smart Biosensor Technology; Knopf, G. K., Bassi, A. S.,
materials;an update. LC-GC North Am. 2006, 24 (7), 648–660. Eds.; CRC Press/Taylor and Francis Group: Boca Raton, FL, 2007; pp
(63) Reineccius, G. A. Flavor analysis. In Flavor Chemistry and Technol- 505-519.
ogy, 2nd ed.; CRC Press: Boca Raton, FL, 2006; pp 33-72. (88) Viswanathan, S.; Radecki, J. Nanomaterials in electrochemical
(64) Acree, T. E., Section Ed. Flavors. In Handbook of Food Analytical biosensors for food analysis;a review. Polish J. Food Nutr. Sci.
Chemistry: Vol. 2 Pigments, Colorants, Flavors, Texture, and Bioac- 2008, 58, 157–164.
tive Food Components; Wrolstad, R. E., Acree, T. E., Decker, E. A., (89) Zhang, C.; Suslick, K. S. Colorimetric sensor array for soft drink
Penner, M. H., Reid, D. S., Schwartz, S. J., Shoemaker, C. F., Smith, D., analysis. J. Agric. Food Chem. 2007, 55, 237–242.
Sporns, P., Eds.; Wiley: Hoboken, NJ, 2005; pp 219-362. (90) Berna, A. Z.; Trowell, S.; Cynkar, W.; Cozzolino, D. Comparison of
(65) Marsili, R., Ed. Flavor, Fragrance and Odor Analysis; Dekker: metal oxide-based electronic nose and mass spectrometry-based
New York, 2002. electonic nose for the prediction of red wine spoilage. J. Agric. Food
(66) Marsili, R., Ed. Sensory-Directed Flavor Analysis, CRC Press/Taylor Chem. 2008, 56, 3238–3244.
and Francis Group: Boca Raton, FL, 2007. (91) Deisingh, A. K.; Stone, D. C.; Thompson, M. Applications of
(67) Baker, S. A.; Miller-Ihli, N. J. Atomic spectroscopy in food analysis. electronic noses and tongues in food analysis. Int. J. Food Sci.
In ref 1, pp 3888-3912. Technol. 2004, 39, 587–604.
(68) Perez, A. L.; Smith, B. W.; Anderson, K. A. Stable isotope and trace (92) Ciosek, P.; Wroblewski, W. Sensor arrays for liquid sensing;
element profiling combined with classification models to differenti- electronic tongue systems. Analyst 2007, 132, 963–978.
ate geographic growing origin for three fruits: effects of subregion (93) Ritter, S. New and notable at Pittcon. Chem. Eng. News, 2009, 87
and variety. J. Agric. Food Chem. 2006, 54, 4506–4516. (13), 46-48, 50.
(69) Powers, J. R. Enzyme analysis and bioassays in food analysis. In (94) Koppelman, S. J., Hefle, S. L., Eds. Detecting Allergens in Food; CRC
ref 1, pp 3955-3971. Press/Taylor and Francis Group: Boca Raton, FL, 2006.
(70) Strange, E. D.; Malin, E. L.; Van Hekken, D. L.; Basch, J. J.
Chromatographic and electrophoretic methods used for analysis of Received for review January 17, 2009. Revised manuscript received
mik proteins. J. Chromatogr. 1992, 624, 81–102. April 23, 2009. Accepted July 29, 2009.