US20020040507A1 - Use of food colours to dye enzyme solutions - Google Patents

Use of food colours to dye enzyme solutions Download PDF

Info

Publication number
US20020040507A1
US20020040507A1 US09/957,658 US95765801A US2002040507A1 US 20020040507 A1 US20020040507 A1 US 20020040507A1 US 95765801 A US95765801 A US 95765801A US 2002040507 A1 US2002040507 A1 US 2002040507A1
Authority
US
United States
Prior art keywords
food colours
colours
food
enzyme solutions
enzyme
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US09/957,658
Inventor
Roger Blum
Sabine Kadow
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Eppendorf SE
Original Assignee
Eppendorf SE
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Eppendorf SE filed Critical Eppendorf SE
Assigned to EPPENDORF AG reassignment EPPENDORF AG ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: BLUM, ROGER, KADOW, SABINE
Publication of US20020040507A1 publication Critical patent/US20020040507A1/en
Priority to US10/624,192 priority Critical patent/US7462473B2/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/40—Colouring or decolouring of foods

Definitions

  • the present invention relates to the use of food colours to dye enzyme solutions, to enzyme solutions provided with food colours as well as their use.
  • DE 197 37 173 A1 relates to a microdispensing system for the dispensing of very small amounts of liquid.
  • the system is used for example in molecular biology to carry out enzymatic reactions.
  • the very small amounts of liquid e.g. enzyme solutions
  • the very small amounts of liquid are fed in measured quantities into a reaction vessel using a free-jet dosing apparatus e.g. electrostatically, piezoelectrically or thermomechanically, the nozzle of the free-jet dosing apparatus having to be positioned exactly centrally over the reaction vessel, in order to guarantee that the dispensed liquid is actually introduced into the reaction mixture and does not remain completely or partly suspended on the wall of the reaction vessel, without coming into contact with the solution.
  • the object of the present invention is thus to create a possibility for the person skilled in the art to test the dispensing of the amounts of liquid using the aforementioned microdispensing system, i.e. in order to be able to judge whether the dispensed sample volumes have also actually been completely added to the reaction mixture.
  • the food colours are selected from the group consisting of Ponceau 4R (E124), Patent Blue (E131) and tartrazin (E102). Practically any shades can be mixed from those three basic colours.
  • the enzyme solution contains one or more enzymes, such as e.g. enzymes from the group consisting of restriction enzymes and polymerases.
  • the invention further relates to the use of the above-named enzyme solutions in a microdispensing system according to DE 197 37 173 A1, which is incorporated herein by reference.
  • the dying of the enzyme solution produces the following advantages as regards handling: The very small drops (up to 10 nl) dispensed using this system can be recognised only with difficulty when uncoloured.
  • the dying of the enzyme solution facilitates the effortless control of the results, namely a) whether the drop was dispensed and b) whether it reached the correct position.
  • the subject-matter of the present invention is thus furthermore the use of the above-named food colours for the preparation of enzyme solutions, preferably for use in a microdispensing system according to DE 197 37 173 A1.
  • the enzyme solutions can contain one or more enzymes, such as e.g. from the group consisting of restriction enzymes and polymerases.
  • Dyed enzyme solutions were thus prepared which contained 500 ppm colour.
  • the enzyme activity was measured by digestion of ⁇ -DNA in the form of a time series.
  • the rate of reaction of the dyed enzyme solution can be compared with the undyed reference from the gelectrophoretic image of the reaction stopped at 8 to 10 successive time points.
  • the rate of reaction is at a first approximation inversely proportional to the time which is required for the digestion of the substrate.
  • FIGS. 1 to 4 show the rates of the digestion of ⁇ -DNA for the 4 enzymes after 1 week's storage at ⁇ 21° C., FIGS. 5 to 8 after almost a half-year's storage at ⁇ 21° C.

Landscapes

  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

The present invention relates to the use of food colours for colouring enzyme solutions, to enzyme solutions provided with food colours as well as their use.

Description

  • The present invention relates to the use of food colours to dye enzyme solutions, to enzyme solutions provided with food colours as well as their use. [0001]
  • DE 197 37 173 A1 relates to a microdispensing system for the dispensing of very small amounts of liquid. The system is used for example in molecular biology to carry out enzymatic reactions. The very small amounts of liquid (e.g. enzyme solutions) are fed in measured quantities into a reaction vessel using a free-jet dosing apparatus e.g. electrostatically, piezoelectrically or thermomechanically, the nozzle of the free-jet dosing apparatus having to be positioned exactly centrally over the reaction vessel, in order to guarantee that the dispensed liquid is actually introduced into the reaction mixture and does not remain completely or partly suspended on the wall of the reaction vessel, without coming into contact with the solution. [0002]
  • The object of the present invention is thus to create a possibility for the person skilled in the art to test the dispensing of the amounts of liquid using the aforementioned microdispensing system, i.e. in order to be able to judge whether the dispensed sample volumes have also actually been completely added to the reaction mixture. [0003]
  • The object is achieved according to the invention by an enzyme solution which contains one or more food colours, preferably one or more food colours according to Annex I for EC directive 94/36 of Jun. 30, 1994 in the version published on Sep. 10, 1994 in the Official Journal No. L 237 (p. 0013-0029) of the European Communities), which is incorporated herein by reference. Among the particularly preferably used food colours are azo dyes and triarylmethane dyes which are soluble in water and distinguish themselves among food colours due to their depth of colour, i.e. the very high molar extinction coefficient. Only small amount of dye are necessary, in order to achieve a sufficient colouring (cf. e.g. Ullmann—Encyklopädie der technischen Chemie, 4[0004] th Edition, Verlag Chemie 1974, Volume 8, pp. 260 ff).
  • According to a particularly preferred version of the invention, the food colours are selected from the group consisting of Ponceau 4R (E124), Patent Blue (E131) and tartrazin (E102). Practically any shades can be mixed from those three basic colours. [0005]
  • There are no limitations whatsoever with regard to the enzyme. The enzyme solution contains one or more enzymes, such as e.g. enzymes from the group consisting of restriction enzymes and polymerases. [0006]
  • The enzyme solutions according to the invention are characterized in particular in that they show no loss of activity even during prolonged storage periods (or only a slight loss of activity, mainly under 20%). [0007]
  • With regard to the amount of colour used, there are no limitations whatsoever according to the invention. For use in conjunction with the microdispensing system of DE 197 37 173 A1, a minimum quantity has proved to be advantageous which allows a colouring of a drop to be just recognisable visually against a white background. It is not necessarily required that a shade can also still be recognised after mixing the dyed enzyme solution with the reaction mixture. [0008]
  • The invention further relates to the use of the above-named enzyme solutions in a microdispensing system according to DE 197 37 173 A1, which is incorporated herein by reference. The dying of the enzyme solution produces the following advantages as regards handling: The very small drops (up to 10 nl) dispensed using this system can be recognised only with difficulty when uncoloured. The dying of the enzyme solution facilitates the effortless control of the results, namely a) whether the drop was dispensed and b) whether it reached the correct position. [0009]
  • The subject-matter of the present invention is thus furthermore the use of the above-named food colours for the preparation of enzyme solutions, preferably for use in a microdispensing system according to DE 197 37 173 A1. The enzyme solutions can contain one or more enzymes, such as e.g. from the group consisting of restriction enzymes and polymerases. [0010]
  • The invention is explained in more detail in the following examples.[0011]
    • EXAMPLES
  • 1) Materials and Methods: [0012]
  • Restriction enzymes (Roche Basel, Switzerland) and dilution buffers: [0013]
  • Bam H1, 10 units/μl (No. 567 604, dilution buffer=P2) [0014]
  • [0015] Cla 2, 10 units/μl (No. 404 217, dilution buffer=P1)
  • [0016] Dra 1, 10 units/μl (No. 779 695, dilution buffer=P1)
  • Eco R1, 10 units/μl (No. 1 175 084, dilution buffer=P3) [0017]
  • Colours: [0018]
  • All colours from Synopharm, 22882 Barsbüttel, Germany. [0019]
  • Ponceau 4R, No. 145 130, batch: 94030770 [0020]
  • Patent blue, No. 700 141, batch: 95060280 [0021]
  • Tartrazine, No. 145 160, batch: 96040020 [0022]
  • Preparation of the Dyed Enzyme Solutions [0023]
  • The three colours red, blue and yellow, and the mixture blue and yellow for the preparation of green were added to the dilution buffers P1, P2 or P3 to be used according to the instructions of the manufacturer Roche to 0.5% (w/v) colour content. [0024]
  • 1 part or corresponding, dyed dilution buffer was added to 9 parts of an enzyme solution. The colour of the dilution buffer corresponded to the code colours for the reaction buffers allocated to the enzymes (blue for Bam H1, red for [0025] Cla 1, green for Dra 1, red for Eco R1).
  • Yellow is thus tested in the combination with blue. [0026]
  • Dyed enzyme solutions were thus prepared which contained 500 ppm colour. [0027]
  • Mixtures of 9 parts of the enzyme solutions used with 1 part of the corresponding undyed buffer were used as reference. [0028]
  • Measurement of the Enzyme Activity [0029]
  • The enzyme activity was measured by digestion of λ-DNA in the form of a time series. [0030]
  • The rate of reaction of the dyed enzyme solution can be compared with the undyed reference from the gelectrophoretic image of the reaction stopped at 8 to 10 successive time points. The rate of reaction is at a first approximation inversely proportional to the time which is required for the digestion of the substrate. [0031]
  • The Roche buffers, modified by an addition of 0.02% [0032] Tween 20 and 2% of the corresponding dilution buffers, were used as reaction buffers.
  • 2) Results: [0033]
  • FIGS. [0034] 1 to 4 show the rates of the digestion of λ-DNA for the 4 enzymes after 1 week's storage at −21° C., FIGS. 5 to 8 after almost a half-year's storage at −21° C.
  • A change in the activity of the enzymes caused by the presence of the colours used cannot be recognised, at both points in time the rate of digestion by dyed and undyed enzyme solution is the same. [0035]

Claims (11)

1. Enzyme solution, characterized in that it contains one or more food colours.
2. Enzyme solution according to claim 1, characterized in that the food colours are food colours according to Annex I to EC directives 94/36 of Jun. 30, 1994.
3. Enzyme solution according to claim 1 or 2, characterized in that the food colours are azo dyes and/or triarylmethane dyes.
4. Enzyme solution according to claim 1 or 2, characterized in that the food colours are selected from the group consisting of ponceau 4R (E124), patent blue (E131) and tartrazine (E102).
5. Enzyme solution according to claims 1 to 4, characterized in that it contains one or more enzymes from the group consisting of restriction enzymes and polymerases.
6. Use of enzyme solutions according to claims 1 to 5 in a microdispensing system according to DE 197 37 173 A1.
7. Use of food colours for the preparation of enzyme solutions for use in a microdispensing system according DE 197 37 173 A1.
8. Use according to claim 7, characterized in that the food colours are food colours according to Annex I to EC directive 94/36 of Jun. 30, 1994.
9. Use according to claim 7 or 8, characterized in that the food colours are also azo dyes and/or triarylmethane dyes.
10. Use according to claim 7 or 8, characterized in that the food colours are selected from the group consisting of ponceau 4R (E124), patent blue (E131) and tartrazine (E102).
11. Use according to claims 7 to 10, characterized in that it contains one or more enzymes from the group consisting of restriction enzymes and polymerases.
US09/957,658 2000-09-18 2001-09-17 Use of food colours to dye enzyme solutions Abandoned US20020040507A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US10/624,192 US7462473B2 (en) 2000-09-18 2003-07-21 Use of food colours to dye enzyme solutions

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10046034A DE10046034B4 (en) 2000-09-18 2000-09-18 Use of food dyes for staining enzyme solutions
DE10046034.8 2000-09-18

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US10/624,192 Continuation US7462473B2 (en) 2000-09-18 2003-07-21 Use of food colours to dye enzyme solutions

Publications (1)

Publication Number Publication Date
US20020040507A1 true US20020040507A1 (en) 2002-04-11

Family

ID=7656573

Family Applications (2)

Application Number Title Priority Date Filing Date
US09/957,658 Abandoned US20020040507A1 (en) 2000-09-18 2001-09-17 Use of food colours to dye enzyme solutions
US10/624,192 Expired - Lifetime US7462473B2 (en) 2000-09-18 2003-07-21 Use of food colours to dye enzyme solutions

Family Applications After (1)

Application Number Title Priority Date Filing Date
US10/624,192 Expired - Lifetime US7462473B2 (en) 2000-09-18 2003-07-21 Use of food colours to dye enzyme solutions

Country Status (2)

Country Link
US (2) US20020040507A1 (en)
DE (1) DE10046034B4 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10204336A1 (en) * 2002-02-01 2003-08-14 Merck Patent Gmbh Use of multi-layer pigments in the food and pharmaceutical sectors
WO2008045984A1 (en) * 2006-10-12 2008-04-17 Suncolor Coporation Polymeric composition

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2774673A (en) * 1954-06-02 1956-12-18 Robert H Young Food additive
DE3631195A1 (en) * 1986-05-16 1987-11-19 Miles Lab METHOD FOR PRODUCING TEST STRIPS BY IMPREGNATING SUCTIONABLE SUBSTRATES
US5428163A (en) 1986-12-31 1995-06-27 Mills; Randell L. Prodrugs for selective drug delivery
US5556761A (en) * 1994-04-26 1996-09-17 Phillips; Kevin J. Test strip for blood glucose testing
US5851817A (en) 1994-07-19 1998-12-22 Board Of Regents, The University Of Texas System Species specific egg-binding proteins of sperm
US6200762B1 (en) 1997-08-01 2001-03-13 Aurora Biosciences Corporation Photon reducing agents and compositions for fluorescence assays
US7485263B2 (en) 1997-08-26 2009-02-03 Eppendorf Ag Microproportioning system
DE19737173B4 (en) * 1997-08-26 2007-04-05 Eppendorf Ag micro-dosing system

Also Published As

Publication number Publication date
DE10046034B4 (en) 2006-07-27
DE10046034A1 (en) 2002-04-04
US7462473B2 (en) 2008-12-09
US20040023352A1 (en) 2004-02-05

Similar Documents

Publication Publication Date Title
Delcour et al. A new colourimetric assay for flavanoids in pilsner beers
KR100516563B1 (en) Non-settling, universal machine dispensable pearlescent pigment dispersions
CN102741361A (en) Architectural paint and stain tinting system with tip drying resistance
HU228746B1 (en) Multiple flavor beverage dispensing air-mix nozzle
EP3942012A1 (en) Device for preparing and dispensing reconstituted beer
CN101861380A (en) Acid cleaning method in the brewing industry
US20020040507A1 (en) Use of food colours to dye enzyme solutions
JPH0653656B2 (en) Package containing two containers of oxidative hair dye composition
CN109394558A (en) A kind of method of preparation and the hair dyeing of imitative melanin hair dyed agent
Düsterhöft et al. Evidence that acidic fibroblast growth factor promotes maturation of rat satellite‐cell‐derived myotubes in vitro
US5934513A (en) Pigment dispensing apparatus with non-settling universal pearlescent pigment dispersions
AU592346B2 (en) Sugar cubes containing aspartame
WO1998057544A1 (en) Improvements in or relating to disinfecting materials
ES2307500T3 (en) SUSPENSION OF ACETOACETILARILIDE IN PIGMENTARY APPLICATIONS.
CN109206942A (en) A kind of carmoisine composition and its tint applications
Norwitz et al. Continued investigation of the diazotization and coupling spectrophotometric technique for the determination of aromatic amines with 8-amino-1-hydroxynaphthalene-3, 6-disulphonic acid and N-(1-naphthyl) ethylenediamine as coupling agents
PT86136B (en) PROCESS OF PRODUCTION OF SPARKLING WINE
JP2002191349A (en) Mixed liqueur
JP3620037B2 (en) Reagent for measuring γ-glutamyltransferase activity
CN108178937A (en) A kind of weaving vegetalitas orchil
US189764A (en) Improvement in flavoring aerated beverages
JPH05236907A (en) Gold colloidal solution for food and food containing the same colloidal solution
RU2069178C1 (en) Method of controlling deep desiliconization of aluminate solution
US3574514A (en) Naphthoic acid arylamide,aminobenzene-azo-naphthalene alkali metal sulfite modified formaldehyde aromatic hydroxy compound and omega-sulfomethyl - hydroxynaphthalenesulfonic acid condensation product for dyeing cellulose acetate and aromatic polyester
SU1601122A1 (en) Method of producing dyes substrate for determining cellulose activity

Legal Events

Date Code Title Description
AS Assignment

Owner name: EPPENDORF AG, GERMANY

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:BLUM, ROGER;KADOW, SABINE;REEL/FRAME:012398/0562

Effective date: 20011025

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION