DU spectrophotometer

DU spectrophotometer

DU Spectrophotometer, National Technical

Laboratories, 1947
measuring ultraviolet light absorbed
Inventor(s) Arnold Orville Beckman
Developed National Technical Laboratories
External video
"The Instrument
that Made the
Ultraviolet Spectrum
Visible to Scientists
Everywhere", Chemical
Heritage Foundation
The DU spectrophotometer or Beckman DU was the first commercially
viable scientific instrument for measuring the amount of ultraviolet
light absorbed by a substance. This model of spectrophotometer enabled
scientists to easily examine and identify a given substance based on
its absorption spectrum, the pattern of light absorbed at different wavelengths.
[1]:148
National Technical Laboratories (later Beckman Instruments) developed
three in-house prototype models (A, B, C) and one limited distribution model (D)
before moving to full production with the DU. Introduced in 1941, approximately
30,000 DU spectrophotometers were manufactured and sold between 1941 and
1976.[2][3]

Measuring both the visible and ultraviolet spectra, [4] the model DU
spectrophotometer yielded more accurate results, and substantially reduced the
time needed to accurately determine the chemical composition of a complex
substance from weeks or hours to minutes.[5] The Beckman Ultraviolet-Visible
(UV-Vis) DU spectrophotometer was essential to several critical secret research
projects during World War II.[6][7] Schmidt credits it with having "brought about a
breakthrough in optical spectroscopy".[2] It has been identified as one of the
"most essential instruments" of postwar science, [8] and "probably the most
important instrument ever developed towards the advancement of bioscience". [9]
Contents
[hide]

1Development
o

1.1Model A

1.2Model B

1.3Model C

1.4Model D

1.5Model DU

2Design

3Use

4Impact
o

4.1Vitamins

4.2Penicillin

4.3Hydrocarbons

4.4Enzyme assays and DNA research

4.5Biotechnology

5Later models

6References

7External links

Development[edit]
The DU was developed at National Technical Laboratories (later Beckman
Instruments) under the direction of Arnold Orville Beckman, an American chemist
and inventor.[1][10][11][12] Beckman's research team was led by Howard Cary, who

went on to co-found Applied Physics Corporation (later Cary Instruments) and

become one of Beckman's strongest competitors. Other scientists included
Roland Hawes and Kenyon George.[13]
Spectroscopic methods for observing absorption in the visible spectrum were
used as early as the 1860s.[14] In 1940, the equipment needed to measure light
energy in the visible spectrum could cost a laboratory as much as $3,000, a huge
amount.[1]:149 Complicated equipment had to be assembled, and test samples
were run through awkward and time-consuming processes to separate them into
analyzable components.[11]Further, the spectrum of visible light was not broad
enough to enable scientists to examine substances such as vitamin A.[15]
Beckman had already developed a successful pH meter for measuring acidity of
solutions. Coleman Instruments had recently coupled a pH meter with an
optical phototube unit to examine the visual spectrum (the Coleman Model DM).
Rather than depending on a human observer's visual ability, or the development
of a photographic plate, to detect wavelengths, phototubes could be used to
register and report specific wavelengths. Beckman made it a goal to create an
easy-to-use instrument extending into the ultraviolet range. [1]:149151
Model A[edit]
The first prototype Beckman spectrophotometer, the Model A, was created at
National Technologies Laboratories in 1940. It used a tungsten light source with a
glass Fery prism as a monochromator. An external amplifier from the Beckman
pH meter and a vacuum tube photocell were used to detect wavelengths. [11]
Model B[edit]

Optical quality quartz crystals

It was quickly realized that the glass prism was not suitable for use with the
ultraviolet spectrum, and a quartz prism was substituted instead, resulting in the
Model B. In the model B, a tangent bar mechanism was used to adjust the
monochromator. The mechanism was highly sensitive and required a skilled
operator.[11]
Model C[edit]
Three Model C instruments were then built, improving the instrument's
wavelength resolution. The Model B's rotary cell compartment was replaced with
a linear sample chamber. The tangent bar mechanism was replaced by a scroll
drive mechanism,[11] which could be more precisely controlled to reset the quartz
prism and select the desired wavelength. [16] With this new mechanism, results
could be more easily and reliably obtained, without requiring a highly skilled
operator. This set the pattern for all of Beckman's later quartz prism instruments.
[11]

Model D[edit]
The A, B, and C models all coupled an external Beckman pH meter to the optical
component to obtain readouts. In developing the Model D, Beckman took the DC
amplifier circuit from the pH meter and combined the optical and electronic
components in a single housing, making it more economical. [16] The model D also
used a hydrogen lamp as a light source rather than tungsten. [11]
Moving from a prototype to production of the Model D involved challenges.
Beckman originally approached Bausch and Lomb about making quartz prisms
for the spectrophotometer. When they turned down the opportunity, National
Technical Laboratories designed its own optical system, including both a control
mechanism and a quartz prism. Large, high optical quality quartz suitable for
creating prisms was difficult to obtain. It came from Brazil, and was in demand
for wartime radio oscillators. Beckman had to obtain a wartime priority listing for
the spectrophotometer to get access to suitable quartz supplies. [11]
The company designed its own hydrogen lamp for the Model D, enclosing
an anode in a thin blown-glass window. The instrument's design also required a
more sensitive phototube than was commercially available at that time.
Beckman was able to obtain small batches of an experimental phototube
from RCA for the first Model D instruments. The Model D spectrophotometer,
using the experimental RCA phototube, was shown at MIT's Summer Conference
on Spectroscopy in July 1941. It was the first model to enter actual production,
and only a small number of Model D instruments were sold before it was
superseded by the DU.[11]
Model DU[edit]
When RCA could not meet Beckman's demand for experimental phototubes,
National Technical Laboratories again had to design its own components inhouse. With the incorporation of Beckman's own newly developed UV-sensitive
phototubes, the Model D became the Model DU UV-Vis spectrophotometer. [11] As

he had done with the pH meter, Beckman had replaced an array of complicated
equipment with a single, easy-to-use instrument. One of the first "black boxes"
used in modern chemical laboratories, [17] it sold for $723 in 1941.[9]
Design[edit]
From 1941 until 1976, when it was discontinued, the Model DU
spectrophotometer was built upon what was essentially the same design. [9] It was
a single beam instrument.[13]:11 The DU spectrophotometers used a quartz prism
to separate light into its absorption spectrum and a phototube to electrically
measure the light energy across the spectrum. This allowed the user to plot the
light absorption spectrum of a substance to obtain a standardized "fingerprint"
characteristic of a compound.[1]:151 [18] All modern UV-Vis spectrometers are built
on the same basic principles as the DU spectrophotometer. [4]
Use[edit]

Beckman DU spectrophotometer in use

The model D and DU spectrophotometer was the first easy-to-use single
instrument containing both the optical and electronic components needed
for ultraviolet-absorption spectrophotometry.[1]:153 The user could insert a sample,
dial up the desired wavelength of light, and read the amount of absorption of
that frequency from a simple meter. A series of readings at different wavelengths
could be taken without disturbing the sample. Working in both the ultraviolet and
the visible regions of the spectrum, the model D produced accurate absorption
spectra which could be obtained with relative ease and accurately replicated.
[19]
The National Bureau of Standards ran tests to certify that the DU's results
were accurate and repeatable and recommended its use. [1]:156
The DU spectrophotometer's manual scanning method was extremely fast,
reducing analysis times from weeks or hours to minutes. It was accurate in both
the visible and ultraviolet spectrums.[4] Advantages included its higher resolution
and the minimization of stray light in the ultraviolet region. [9] Although it was not
cheap, its price point made it available to most scientists. [8]
Impact[edit]

World War II poster encouraged researchers to "Give this job Everything You've
got"
Beckman's DU spectrophotometer has been referred to as the "Model T" of
scientific instruments. It enabled researchers to perform easier analysis of
mixtures of chemicals by quickly taking measurements at more than one
wavelength to produce an absorption spectrum describing the complete
substance. "This device forever simplified and streamlined chemical analysis, by
allowing researchers to perform a 99.9% accurate quantitative measurement of a
substance within minutes, as opposed to the weeks required previously for
results of only 25% accuracy." [20] Theodore L. Brown notes that it "revolutionized
the measurement of light signals from samples".[21]:2 Nobel laureate Bruce
Merrifield is quoted as calling the DU spectrophotometer "probably the most
important instrument ever developed towards the advancement of bioscience." [9]
Vitamins[edit]
Development of the spectrophotometer had direct relevance to World War II and
the American war effort. The role of vitamins in health was of significant concern,
as scientists wanted to identify Vitamin A-rich foods to keep soldiers healthy.
Previous methods of assessing Vitamin A levels involved feeding rats a food for
several weeks and then performing a biopsy to estimate ingested Vitamin A
levels. In contrast, examining a food sample with a DU spectrophotometer
yielded better results in a matter of minutes. [22]The DU spectrophotometer could
be used to study both vitamin A and its precursor carotenoids,[23] and rapidly
became the preferred method of spectrophotometric analysis. [15][24][25]
Penicillin[edit]
The DU spectrophotometer was also an important tool for scientists studying and
producing the new wonder drug penicillin.[16] The development of penicillin was a
secret national mission, involving 17 drug companies, with the goal of providing
penicillin to all U.S. Forces engaged in World War II. [7][26] It was known that

penicillin was more effective than sulfa drugs,[26] and that its use
reduced mortality, severity of long-term wound trauma, and recovery time.
[1]:158
However, its structure was not understood, isolation procedures used to
create pure cultures were primitive, and production using known surface culture
techniques was slow.[26]
At Northern Regional Research Laboratory in Peoria, Illinois, researchers collected
and examined more than 2,000 specimens of molds (as well as
other microorganisms).[27] An extensive research team included Dr.Robert Coghill,
Dr. Norman Heatley, Dr. Andrew Moyer, lab bacteriologist Mary Hunt,[28][29]
[30]
Frank H. Stodola and Morris E. Friedkin. Friedkin recalls that an early model of
the Beckman DU spectrophotometer was used by the penicillin researchers in
Peoria.[26] The Peoria lab was successful in isolating and commercially producing
superior strains of the mold, which were 200 times more effective than the
original forms discovered by Alexander Fleming.[28] By the end of the war,
American pharmaceutical companies were producing 650 billion units of
penicillin each month.[28] Much of the work done in this area during World War II
was kept secret until after the war. [1]:158[7]
Hydrocarbons[edit]
The DU spectrophotometer was also used for critical analysis
of hydrocarbons in crude oil. A number of hydrocarbons were of interest to the
war effort. Toluene, a hydrocarbon in crude oil, was used in production ofTNT for
military use.[1]:158159[11] Benzene and butadienes were used in the production of
synthetic rubber.[31] Rubber, used in tires for jeeps, airplanes and tanks, was in
critically short supply because the United States was cut off from foreign supplies
of natural rubber.[1]:158159[32] The Office of Rubber Reserve organized researchers
at universities and in industry to secretly work on the problem. [6] The demand for
synthetic rubber caused Beckman Instruments to develop infrared
spectrophotometers, which were better suited to measuring wavelengths of
hydrocarbons.[1]:159[14]
Enzyme assays and DNA research[edit]
Gerty Cori and her husband Carl Ferdinand Cori won the Nobel Prize in Physiology
or Medicine in 1947 in recognition of their work on enzymes. They made several
discoveries critical to understanding carbohydrate metabolism, including the
isolation and discovery of the Cori ester, glucose-1 phosphate, and the
understanding of the Cori cycle. They determined that the
enzyme phosphorylase catalyzes formation of glucose 1-phosphate, which is the
beginning and ending step in the conversions of glycogen into glucose and blood
glucose to glycogen. Gerty Cori was also the first to show that a defect in an
enzyme can be the cause of a human genetic disease. [33] The Beckman DU
spectrophotometer was used in the Cori laboratory to calculate enzyme
concentrations, including phosphorylase.[34]
Arthur Kornberg worked with Severo Ochoa, learning the process of enzyme
purification of aconitase, and then spent six months in 1947 at the Cori

laboratory, "the most vibrant place in biochemistry at that time", before

returning to the National Institutes of Health (NIH) in 1948. He too used the DU
spectrophotometer.[35]
"The enzyme could be assayed in a few minutes by coupling it to isocitrate
dehydrogenase and in measuring the NADH formed using the Beckman DU
spectrophotometer, an instrument that transformed biochemistry." [36]
Kornberg and Bernard L. Horecker used the Beckman DU spectrophotometer for
enzyme assays measuring NADH and NADPH. They determined their extinction
coefficients, establishing a basis for quantitative measurements in reactions
involving nucleotides. This work became one of the most cited papers in
biochemistry.[36]:115 Kornberg went on to study nucleotides in DNA synthesis,
isolating the first DNA polymerizing enzyme (DNA polymerase I) in 1956 and
receiving the Nobel Prize in Physiology or Medicine with Severo Ochoa in 1959.[37]
The bases of DNA absorbed ultraviolet light near 260 nm.[16] Inspired by the work
of Oswald Avery[38] on DNA, Erwin Chargaff used a DU spectrophotometer in the
1940s in measuring the relative concentrations of bases in DNA. [39]:260, 290
302
Based on this research, he formulated Chargaff's rules.[40] In the first complete
quantitative analysis of DNA, he reported the near-equal correspondence of pairs
of bases in DNA, with the number of guanine units equaling the number
of cytosine units, and the number of adenine units equaling the number
of thymine units. He further demonstrated that the relative amounts of guanine,
cytosine, adenine and thymine varied between species. In 1952, Chargaff
met Francis Crick and James D. Watson, discussing his findings with them.
Watson and Crick built upon his ideas in their determination of the structure of
DNA.[40]
Biotechnology[edit]
Ultraviolet spectroscopy has wide applicability in molecular biology, particularly
the study of photosynthesis.[41] It has been used to study a wide variety of
flowering plants and ferns[42] by researchers in departments of biology, biology,
plant physiology and agriculture science as well as molecular genetics. [43]
Particularly useful in detecting conjugated double bonds, the new technology
made it possible for researchers like Ralph Holman and George O. Burr to study
dietary fats, work with significant implications for human diet. [44] The DU
spectrophotometer was also used in the study of steroids[23][45] by researchers
like Alejandro Zaffaroni,[46] who helped to develop the birth control pill,
the nicotine patch, and corticosteroids.[47]
Later models[edit]

Beckman Model DK1 Ultraviolet Spectrophotometer

The Beckman team eventually developed additional models, as well as a number
of accessories or attachments which could be used to modify the DU for different
types of work. One of the first accessories was a flame attachment with a more
powerful photo multiplier to enable the user to examine flames such
as potassium, sodium and cesium(1947).[13]:11[48]
In the 1950s, Beckman Industries developed the DR and the DK, both of which
were double-beam ultraviolet spectrophotometers. The DK was named for Wilbur
I. Kaye, who developed it by modifying the DU to expand its range into the nearinfrared.[13] He did the initial work while at Tennessee Eastman Kodak, and later
was hired by Beckman Instruments.[49] The DKs introduced an automatic
recording feature. The DK-1 used a non-linear scroll, and the DK-2 used a linear
scroll to automatically record the spectra.[49]:21
The DR incorporated a "robot operator" which would reset the knobs on the DU
to complete a sequence of measurements at different wavelengths, just like a
human operator would to generate results for a full spectrum. It used a linear
shuttle with four positions, and a superstructure to change the knobs. It had a
moving chart recorder to plot results, with red, green and black dots. [13] The cost
of recording spectrophotometers was substantially higher than non-recording
machines.[23]
The DK was ten times faster than the DR, but not quite as accurate. [13] It used a
photomultiplier, which had introduced a source of error. [49]:21 The DK's speed
made it preferred to the DR.[13] Kaye eventually developed the DKU, combining
infrared and ultraviolet features in one instrument, but it was more expensive
than other models.[49]
The last DU spectrophotometer was produced on July 6, 1976. [50]