DE1289668B - Stabilizing agent for enzymatic test reagents - Google Patents

Description

1 2

To enzymatic investigations (ζ. B. the Mes- Die according to the teaching of the present invention

solution of enzyme activities for diagnostically available reagent compositions,

Purposes) to be carried out routinely and quickly, very quickly and easily soluble in water,

it is of great importance for the practitioner - porous powders that can be kept for years. they provide processing, which for the expiry of the relevant enzyme 5 represents a considerable advance in the field of

substances required for the reaction (auxiliary enzymes, reagent combinations for enzymatic

Coenzymes, substrates, buffer substances, etc.) are investigations. The practitioner now has

ready for use - dosed in the right proportion - nothing more to do than the content of the relevant item

available. Container in a certain amount of water

In the interests of rationalization of work, an io should solve to add the substance to be examined

To measure the reagent composition for enzymatic under- and according to instructions (e.g. measurement of the

searches already as many of the extinctions to be carried out in a photometer),

The following are some reagent compositions

one, around time-consuming pipetting and the like, and indicated in which, according to the invention, glutathione

Reduce sources of error to a minimum. 15 is used as a stabilizer.

Diagnostic means are known that all

Examples for the enzymatic detection of a compound

contain required substances. In the German j determination

Auslegeschrift 1,075,869 is a test reagent for _the glutamate-pyruvate transaminase (GPT)

Glucose described, which the components on ao ττργ » ο

comprises an absorbent carrier. The for the test- tf ⁶ § ^ ^a ? ViJ; ⁴ ' ί

Reagents required for the reaction can also be expressed in ϊ'η <ai · ^

^Powder form, the production of the f '«8101;,," ¹ ,'.

dry reagent composition, as in the at- U, 35 g a-ketoglutarate,

gischen patent specification 627 961 described by 25 005ΌΡΝΗ "'

Freeze-drying of the solutions to very active and. '„„ Α _Ύ J ^ ~, _Λ

dissolving preparations quickly. 0.0015 g lactate dehydrogenase,

The U, 3U g Lrlutation required in many test combinations

both coenzymes di- and triphosphopyridine nucleo- are redistilled in 90 ml. Dissolved in water and the pH

In order to be fully active, tid must be set to a reduced 30 value with dilute sodium hydroxide solution to 7.6.

Form (DPNH and TPNH). Unfortunately they are then the solution with bidistilled. Water to 100 ml

not very stable in this form. filled up, then frozen and lyophized.

The object of the present invention is to provide a

Finding stabilizers that stabilize the 2nd determination

this coenzyme causes, but otherwise practically 35, the glutamate, oxaloacetate transaminase (GOT)

no influence, at least not of a negative kind, on 2 43 s Na HPO · 2 HO

carries out the test reactions. 0 ^ 21 g NaH ^ 2 HA

It has now surprisingly been found that 1.80 g of sodium aspartate;

that as a stabilizer for enzymatic test 0 35 ε α-ketoglutarate

reagents containing the di- or triphosphopyri- 40 030 g serum albumin ',

Contains dinnucleotide in reduced form, Glut- 0 05 2 DPNH

athion can be used with success, the ₀ ' ₀₀₁₅ Lact'at dehydrogenase,

Glutathione in reduced form, i.e. with free 0.0015 g malate dehydrogenase,

SM group must be present. By the invention 0 3Oe glutathione

proper use of the glutathione now succeeds 45 '

these coenzymes in mixtures with the other preparation as indicated in example 1,
necessary reagents even after long periods of storage

keep active. 3. Determination of lactate dehydrogenase (LDH)

The preparation of the reagent compositions requires 2.43 g Na ₂ HPO ₄ · 2 H ₂ O,

follows by mixing all of the 50 o, 21 g NaH ₂ PO ₄ · 2 H ₀ O,

intended enzymatic reactions required 0.004 g sodium pyruvate,

Substances such as auxiliary enzymes, coenzymes, substrates, buffers o'3O g serum albumin,

substances, auxiliaries and the o, O5 g DPNH according to the invention,

Stabilizer and optionally fillers in the 0.30 g glutathione,

the optimal 55 '.._, .., _Λ ,

ratio with water to a solution, the volume of which is prepared as in Example 1.

and pH value can be adjusted in the usual way ₄ _ _{Determination of} ^ _{01386 (ALD)}

can. These test solutions are then processed according to the tron

each desired unit amount to suitable r ^? S ^. ^a |? pjr * '; r iprj'

Divided container and freeze-drying 60 £ ² * S NaH ₂ PO ₄ · .2H ₂ C >>

(Lyophilization) of the contents of these containers and 0.14 g of fructose-1,6-diphsophate,

immediate closure of the same in durable test "> ™ ⁵ g iodine acetate,

Prepared transferred. °> 30 g serum albumin,

Such mchtionogenic orga- ^ ™ Ä- ™,, τ ^, j

niche compounds are used, soft 6 ₅ 0.0015 g glycerophosphate dehydrogenase,

- such as gelatin, polyvinylpyrrolidone, albumin 0 _; 0015gtriosephosphate isomerase,

etc. - in a manner known ^{per se the lyophilization υ} '3υ § Wutatnion.

facilitate the process. Preparation as indicated in Example 1.

Claims (1)

Claim: Use of glutathione as a stabilizer for enzymatic test reagents which Contain di- or triphosphopyride nucleotide in reduced form.