U.S. patent number 3,667,607 [Application Number 05/093,299] was granted by the patent office on 1972-06-06 for chromatographic material.
This patent grant is currently assigned to J. T. Baker Chemical Company. Invention is credited to Marcus K. Brandt.
United States Patent |
3,667,607 |
Brandt |
June 6, 1972 |
CHROMATOGRAPHIC MATERIAL
Abstract
A thin layer chromatographic strip in the form of an elongated
tape carrying indicia related to chromatographic test areas, the
areas being in contact with a reservoir for solvent, the
combination of test areas, indicia, and reservoir being
periodically reproduced on the length of the tape, whereby the
indicia may be used to initiate sampling, solvent application, and
control passage through development and drying stages, and
initiation and termination of scanning stages, thereby to make
possible a sequence of unrelated or related analyses automatically
on said tape.
Inventors: |
Brandt; Marcus K. (Easton,
PA) |
Assignee: |
J. T. Baker Chemical Company
(Phillipsburg, PA)
|
Family
ID: |
22238193 |
Appl.
No.: |
05/093,299 |
Filed: |
November 27, 1970 |
Current U.S.
Class: |
210/198.3 |
Current CPC
Class: |
G01N
30/92 (20130101); G01N 35/00009 (20130101) |
Current International
Class: |
G01N
30/92 (20060101); G01N 30/00 (20060101); G01N
35/00 (20060101); B01d 015/08 () |
Field of
Search: |
;210/31,198 |
References Cited
[Referenced By]
U.S. Patent Documents
Primary Examiner: DeCesare; J. L.
Claims
I claim:
1. A thin layer chromatographic adsorption element in the form of
an elongated strip suitable for use in sequential testing of
specimens which comprises, in combination, an inert, non-adsorptive
flexible substrate, a layer of chromatographic adsorbent material
as a test area on a face of said substrate, a solvent reservoir on
the face of said substrate and in abutting contact with the layer
of adsorbent material, and indicia means along said substrate, said
indicia means adapted to provide signals to initiate sample
application to the adsorbent material layer and solvent application
to the solvent reservoir, said substrate including a repetitive
sequence of said layers of chromatographic adsorbent materials,
solvent reservoirs and indicia means spaced thereon.
2. A chromatographic adsorption element in accordance with claim 1
in which the substrate is a flexible plastic tape.
3. A chromatographic adsorption element in accordance with claim 1
in which the substrate is a monofilamentous fabric base.
4. A chromatographic adsorption element in accordance with claim 2
wherein a plurality of layers of chromatographic adsorbent material
are arranged in parallel relationship on said flexible plastic tape
substrate.
5. A chromatographic adsorption element in accordance with claim 1
wherein said layer of chromatographic adsorption material is
selected from the group consisting of silica gel, alumina,
cellulose, derivatized cellulose, microcrystalline cellulose,
kieselguhr, ion-exchange resins, polyamide powders, hydroxapatite,
diatomaceous earth, polysaccharide dextran derived powder, zinc
carbonate, polyethylene powder, calcium sulfate, magnesium
silicate, calcium hydroxide, and magnesia-silica gel.
6. A chromatographic adsorption element in accordance with claim 5
wherein said substrate is formed from a material selected from the
group consisting of monofilamentous fabric of synthetic fiber,
synthetic plastic, synthetic plastic-coated paper, synthetic
plastic-coated fabric and metal foil.
7. A chromatographic adsorption element in accordance with claim 1
wherein the element is in the form of a reel of elongated tape
having a sequence of chromatographic adsorptive material layers
spaced along the length of the tape.
8. A chromatographic adsorption element of claim 2 wherein said
indicia means comprises perforations spaced along the flexible
plastic tape.
9. A chromatographic adsorption element in accordance with claim 2
wherein the indicia means comprises conductive spots spaced along
the flexible tape.
10. A chromatographic adsorption element in accordance with claim 2
wherein the solvent reservoir comprises a pad of inert, open
cellular structure material.
Description
BACKGROUND OF THE INVENTION
The theory and practice of thin layer chromatography, column
chromatography, and the theory of having a solvent move components
of a mixture at different rates through an adsorptive medium is
well documented. In thin layer chromatography, the fundamental
technique is to apply a thin layer of adsorbent material to a
substrate, usually a glass slide, or a large plastic sheet. The
technique of preparing the thin layer involves the use of various
mechanical coaters which will apply an appropriate layer of
adsorptive material to this substrate, generally 50 microns to 300
microns. The material thus coated serves as an adsorption medium
and the test technique involves placing a drop of the material to
be separated at one point on the adsorptive surface, applying
solvent continuously to the base of that adsorptive surface and
allowing it to advance the spot by capillary action, with the
entire test specimen being enclosed in an atmosphere of solvent
vapor enclosed within a chamber. In the course of a few minutes the
sample being separated is transported, through the adsorbent layer
or alternately adsorbed and desorbed from the adsorbent, and its
components separated into distinct areas depending upon the
partition coefficient of each compound. The chromatogram then can
be allowed to react with a chemical or with other mixtures of
chemicals to produce color, if they have none, and scanned to
relate intensity of color to the amount of material in the spot.
Analysis becomes possible when the technique is carefully applied.
The technique is of particular applicability in installations where
many tests of a repetitive type are required, for example, the
testing of body fluids in hospital laboratories. It is also in wide
use in biomedical laboratories as a standard research tool.
It is, accordingly, a fundamental object of this invention to
provide a chromatographic adsorptive test material formed on a
flexible back material in the form of a continuous strip or tape to
permit a continuous moving analysis of a large number of specimens
from related, identical, or diverse sources, the tape carrying
indicia rendering it possible to relate specimens to test results,
through use of an automatic thin layer chromatography
instrument.
Other objects and advantages of the invention will in part be
apparent from the following description and drawings and in part
appear hereinafter.
DETAILED DESCRIPTION OF THE INVENTION
This invention, is, accordingly, embodied in a tape, for example, a
plastic tape as a base, the tape being characterized by its having
mechanical indicia along its edges to permit precise timed movement
of the tape over a timing sprocket, the tap having mounted on a
face thereof a strip or strips of chromatographic adsorbent of
predetermined width, length, and shape, coordinated with a
reservoir for holding a chromatographic solvent, or solvent
mixture, the tape being further coordinated with an indicium
related to the placement of said reservoir and adsorptive surfaces,
the several parts being coordinated so that in forward movement on
a timed basis, a test specimen for chromatography can be applied to
the surface of the adsorbent in response to one of said indicia, be
dried, and travel through a development zone and drying zone, and
also proceed in response to said indicia to a final stage of
inspection, such as, for example, by visual or by optical means,
for the identification of each of the zones in the
chromatogram.
Thus, the invention broadly is embodied in the combination
consisting of a base tape, in the form of reels of any size, which
serves as a substrate and spaced strips of chromatographic
adsorptive material thereon in spaced sequence, said strips
individually being in contact with solvent reservoir pads. The
adsorbent coating may be any material, with or without indicating
dyes, such as silica gel, alumina, cellulose, derivatized
cellulose, microcrystalline cellulose, kieselguhr, ion-exchange
resins, polyamide powders, hydroxapatite, "Celite" diatomaceous
earth, "Sephadex" polysaccharide dextran derived powder, zinc
carbonate, polyethylene powder, calcium sulfate, magnesium
silicate, calcium hydroxide, "Florisil," magnesia-silica gel, and
the like, and it is coated on the base in any manner such as by
dropping, pouring, dipping, spraying or spreading, to form a thin
layer on the substrate upon which mixtures of materials may be
separated when the layer is dried.
The base, or tape, may be nylon, Dacron polyester, fiber made from
polyethylene terephthalate or other monofilamentous fiber, the only
requirement being that it be non-adsorptive and inert. It may vary
in thickness, in fiber type, and class of weight. Metal foil is
also useful as a substrate. The adsorbents may or may not require
the use of a binder such as polyvinylpyrrolidone, polysaccharides,
polyethers or calcium sulfate hemihydrate for extra adhesion. A
preferred form of film base is the Mylar polyester tape commonly
used in photography, and, in particular, in a specific form of our
invention in Mylar, film, 35 millimeters in width, as used for
motion picture film, the adsorptive strips, reservoirs and indicia
being applied thereto.
The timing indicia used in the film may be the conventional
perforations along the edges thereof, alone, or they may be an
additional perforation located in appropriate relationship to the
reservoir, or they may be in the form of conductive spots applied
to the film in appropriate relation to the reservoir. The advantage
of the perforation as an indicium is that in a traverse in an
automatic apparatus the leading and lagging edges of the
perforation are mechanically detectable with microswitches. The
conductive spot is very useful, and can be sensed with electrical
contacts and generally conventional microswitches. The magnetic
spot is similarly readily detectable.
The construction and arrangement of the invention herein may be
better understood by reference to the drawings and detailed
description:
FIG. 1 is a side view exaggerated in thickness to show the
relationship of chromatographic layer to the film;
FIG. 2 represents a plan view of a single chromatographic
combination applied to a tape;
FIG. 3 is a general plan view of the length of tape showing a
plurality of these chromatographic test strips mounted thereon;
FIG. 4 is a perspective view of a reel of tape showing a sequence
of the chromatographic strips;
FIG. 5, a, b, c, d indicates the applicability of shaped strips to
the invention.
Referring to FIG. 1, 10 represents a base of monofilamentous
material or tape having an upper face, 11, a lower face, 12 and
edges 13 and 14. Near each edge are perforations in precisely
spaced sequence, namely, perforations 15, 16, 17, etc.
In the clear area between the rows of perforations 15 and 16 are
mounted the chromatographic adsorptive areas 18 and 19 which are of
a pre-selected length, carefully placed on the strip in relation to
indicium 20. A preferred form of indicium is a perforation a few
millimeters in width or diameter. Also included in specific
relation to indicium 20 is the solvent reservoir or pad 21.
The combination of adsorptive area, indicium, and solvent reservoir
is repeated at spaced intervals, as frequently as the length of
tape being used will permit or less frequently as desired. The tape
may further provide at each end a lead area for the indication
thereon of data concerning the date, time, operator,
chromatographic conditions and nature of the test. By relating the
indicium to the test area the application of sample to adsorptive
area and solvent to the reservoir can be coordinated with movement
of the tape through test apparatus.
It is apparent that while the invention is illustrated in terms of
two parallel test areas each about 5 to 8 millimeters in width, and
as much as 80 to 100 millimeters in length, that the test area
could be a single area approximately 20 millimeters in width or it
could be made 3 or 4 chromatographic areas or the like. For general
use, it has been found that the parallel pair of areas represents a
preferred form. They permit analysis of an unknown sample in one
area and a known sample in the adjacent area for comparison. Wedge
shaped areas as indicated in FIGS. 5, a, b, c, d are useful.
In use the tape can be manually or automatically driven through an
analyzing machine which will automatically cause a specimen to be
applied to each test area, and also initiate the application of
solvent to the reservoir pad. In traversing through the development
chamber of test apparatus which is saturated with solvent vapor,
the solvent is given time to pass completely through the adsorbent
layer and thereby effect separation of a mixture of compounds.
The chromatographic strips 18 and 19, may be of any of a variety of
compositions suitable for performing chromatographic separations,
or any of a variety of shapes as indicated in FIG. 5. It is
understood that the use of the appropriate adsorbent will depend
upon the nature of the compounds which will be separated. Necessary
to the development of the chromatogram is a solvent, or mixture of
solvents, which will perform the separation in combination with the
absorbent coating in each strip. The reservoir pad 21 has been
designated as the material onto which the solvent will be
deposited. As indicated, the reservoir pad is squared off and thus
provides a base line from which the solvent proceeds to migrate
along the chromatographic strip. The material of which the
reservoir is made and the adhesive holding it in place should be
inert to the particular solvents to be used. Generally, it has been
found the polyurethane foams or sponges are particularly useful as
reservoir pad material. As solvent pads or reservoirs any material
having an open cellular structure, e.g., cullulose, felt, cloth,
inert to the solvents and capable of retaining solvent in the
amount of 2 1/2 times the volume of the adsorbent layer is useful.
They are simply lightly glued in direct contact with the base of
each of the chromatographic adsorption strips. Thus, in developing
a chromatogram, solvent is applied to the sponge 21 which serves as
a solvent reservoir, in contact with the strips 18 and 19 and
provides an accurately-defined base line, and solvent can then
migrate along the strips 18 and 19 at a rate determined by its
physical properties and the properties of particles making up the
strips and by the solvent polarity. It contacts the origin of the
chromatogram at positions 22 and 23 and in the course of a few
minutes migrates through most of the length of 18 and 19 in order
to complete the separation of the mixture of compounds originally
deposited at the origin.
Virtually any of the various adsorbents which have been mentioned
can be slurried and placed on the monofilament cloth or plastic
tape as is done routinely in the usual thin layer preparation
methods. Depending upon the adsorbent which is placed on the tape
alkaloids, amines, amino acids, peptides, proteins, steroids,
lipids, carotenoids, chlorophylls, vitamins, quinones, drugs,
poisons, antibiotics, phenols, nucleic acids, nucleotide coenzymes,
sugars, insecticides, dyes, miscellaneous organic compounds,
inorganic ions, and other compounds in need of separation can be
separated with this technique.
Specifically, in practicing the invention any of a variety of
combinations of substrate with adsorbent can be used, but the
following should be considered illustrative:
EXAMPLE I
A monofilament nylon strip, 25 millimeters wide, 0.1 millimeter
thick, was coated in pairs of strips with a silica gel slurry, in
sequence as shown in FIG. 2, containing a calcium sulfate binder.
The strip was hung up to dry. In 15 minutes the strip was
completely dry. The coated tape was hung in an oven at 110.degree.
C. for 10 minutes and removed, allowed to cool slightly in the air,
and was spotted with 4 microliters of Stahl dye mixture. Solvent
was applied to sponge 21 and the strip was then placed in a large
volume tank saturated with solvent vapor and was developed to 8
centimeters from the origin. The separation of the three dyes was
complete and took 6 minutes, the butter yellow, indophenol blue,
and Sudan Red G formed distinct spots, well separated.
Depending upon the thickness of the slurry used to coat these nylon
strips, the time for development will vary. The thicker the layer
on the strip, the longer will be the development time.
EXAMPLE II
A length of monofilament cloth 25 millimeters wide, 100 centimeters
long, and 0.1 millimeter thick was pulled through a silica gel
slurry, as in Example I, and was wiped off with a doctor blade to
remove excess slurry. With this method, a continuous strip of
evenly coated monofilament nylon was made. This strip was air-dried
and then cut into sections corresponding to 18 and 19 of FIG. 2 for
individual chromatographic separations. The sections are glued on
tape. A sponge reservoir then is glued in contact therewith,
solvent is applied to the sponge and the operation completed as
described in connection with Example I.
EXAMPLE III
A 35 millimeter Mylar type, 0.1 millimeter thick, was masked to
leave parallel spaced strips uncovered, and sprayed with a silica
gel slurry containing 1 percent NP-90 Polyvinylpyrrolidone as
binder. The tape was allowed to air dry for 30 minutes, and sponges
were applied as indicated in FIG. 2. The coated strips were
activated for 10 minutes at 110.degree. C. The coated strips were
then spotted with 4 microliters of 0.1 molar solutions of
o-nitroaniline, m-nitroaniline and p-nitroaniline in benzene. The
coated strips were then passed in a chromatographic chamber
saturated with solvent vapor, [phenol/water (8:3)], the sponge
activated with 0.5 milliliter phenol/water (8:3) solvent and the
strips developed to 10 centimeters from the origin. The 3
components separated completely, to give R.sub.f values of 0.55,
0.44 and 0.37 respectively.
In recapitulation it is to be understood that the chromatographic
adsorption device constituting this aspect of the invention
consists of the flexible non-absorptive and non-adsorptive
substrate serving as a carrier for adsorptive areas arranged to
receive samples and to be in contact with a solvent reservoir, the
combination being oriented in a fixed defined relationship with an
indicium, preferably a perforation in the substrate, by arranging
the combination in linear repetitious sequence along a tape. The
several indicia can serve to provide together signals for sample
application, solvent application, etc. and other intelligence built
into a sequence where the operation involves a sequence of tests.
The hole is incorporated in the tape to be used in combination with
an automatic chromatographic apparatus, but it should be clear that
the tape can be manipulated manually through an apparatus. A
suitable apparatus for conducting tests using a substrate in
chromatographic areas made in accordance with this invention is
described in copending application filed of even date herewith,
which application should be read as fully incorporated herein, Ser.
No. 93,064, dated Nov. 27, 1970.
The adsorptive materials which may be used in the chromatographic
layers are subject to virtually infinite variation and will vary
depending upon the requirements of the tests to be conducted. For
example, U.S. Pat. No. 3,418,152, W. S. Staudenmayer et al., issued
Dec. 24, 1968, discloses a large group of specific exemplary
formulations for laying down chromatographically active layers on a
substrate and that disclosure may be considered fully incorporated
herein. Similarly, U.S. Pat. No. 3,415,158, E. S. Perry et al.,
issued Dec. 24, 1968, also discloses a number of specific
formulations and it may be considered fully incorporated
herein.
* * * * *