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An improved chemical method for measuring intensity of hair pigmentation

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Title An improved chemical method for measuring intensity of hair pigmentation
Names Costley, Gary Edward (creator)
Stout, Floyd M. (advisor)
Date Issued 1968-04-25 (iso8601)
Note Graduation date: 1968
Abstract Research on mammalian pigmentation has ranged from basic
studies on biochemistry of melanin formation to more practical
studies involving effects of nutrition, environment and genetics on
hair color. In mink such studies have an added significance as fur
color is of considerable economic importance; however, research in
this area has been hampered by lack of aquantitative measure for pigmentation.
Historically, fur color in mink has been evaluated visually;
this has the disadvantage of lacking repeatability, sensitivity and
objectivity. One existing chemical method for measuring hair color
employing spectrophotometric procedures is based on the knowledge
that hair pigmentation is derived from microscopic melanin granules
within the hair, the corollary being that hair color intensity is a
function of melanin granule concentration. The studies reported herein were undertaken to further improve this spectrophotometric
method of quantitating the amount of melanin pigmentation in hair. In
this regard, trials were conducted in the areas of spectrophotometry,
ultrasonic homogenization and sample preparation. Other tests were
made to gain further information on application of the method.
Spectrophotometry. Determining melanin granule concentrations
involves establishment of the turbidity of suspensions of melanin
granules. Hence, trials were conducted to determine effects of light
source wave length, granule concentration and type of spectrophotometer
employed on these turbidimetric measurements. These trials
indicate that wave length must be specified, as although the absorption
curve is linear, optical density values progressively decrease
with longer wave lengths. It was further shown that optical density
values of varying concentrations of melanin granules follow Beer's law
within the tenfold concentration range studied. This strengthens the
fundamental validity of the method since the data indicate optical densities
of melanin granule suspensions are directly related to granule
concentrations. Results further indicate that the technique can be
adapted to any spectrophotometer; however, absorption curves vary
as a result of differences in cuvette size and other differences inherent
to the equipment giving relative but not absolute results. Establishment
of standard curves for particular instruments would render
values directly comparable.
Ultrasonic Homogenization. Investigations in this area indicate
that homogenization of the digested hair by sound waves increases
the optical density of melanin granule suspensions by separating
groups of pigment granules into individual particles. One and one-half minutes of ultrasonic homogenization, at all four power intensity
levels studied, resulted in maximum optical density values indicating
complete granule separation. Homogenization of melanin granule
suspensions increased the repeatability and consequently the sensitivity
of the technique by maximally increasing the surface area of the
suspended particles and by uniformly dispersing the particles.
Sample Preparation. Studies indicate that grinding the hair
sample was not necessary to achieve complete granule liberation when
followed by ultrasonic homogenization. This modification resulted in
a more useable procedure since grinding of hair required considerable
time and effort; however, more difficulty was encountered in
obtaining a representative sample of guard hair and underfur, which
slightly lowered the repeatability of the procedure.
Application of Method. Research indicates that the procedure
of hair color analysis, as modified by spectrophotometry and ultrasonic
homogenization techniques, is sensitive to small differences in
hair pigmentation and can be applied to relatively large numbers of
samples, These trials further indicate that hair pigmentation as
expressed by "melanin value" is definitely correlated to visually appraised intensities of hair pigmentation. The research also suggests
that numerous applications of the technique exist.
The studies reported herein indicate that the hair color analysis,
as modified, is sensitive and repeatable and can be fairly rapidly
applied to relatively large numbers of samples.
Genre Thesis/Dissertation
Topic Melanin
Identifier http://hdl.handle.net/1957/47095

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