DE1767931A1 - Method and diagnostic agent for the detection of urobilinogen bodies - Google Patents

Description

C. F. Soehringer & Soehne

Mannheim 1525a

Method and diagnostic agent for the detection of urobilinogen bodies

_ (Addition to patent application ξ

P s / 4 ty ftf. Γ

The subject of patent application P 16 43 ^ 5? ■ & is an improved method or an improved diagnostic agent for the detection of urobi ^ inogen bodies in body fluids, preferably in urini consisting of an absorbent carrier or a film, a strong solid acid and a Substance which, like the Ehrlioh's sample with urobilinogen bodies, provides a color change, characterized by the content of one or more substances of the formula I

(D, CHO

in which

R, and Rp a halogen atom, a cyano, nitroso, carbalkoxy, carbamido, hydroxy, acyloxy, alkoxy, an open-chain or cyclic, saturated or unsaturated, secondary or tertiary amino group, where R, and R _? also together an oxa, thia, sulfonyl, alkylthionium, iaaino, alkylimino, optionally substituted arylimino or an open-chain, alicyclic or heterocyclic dialkylimino group and one of the radicals H. and Rjj can also be a hydrogen atom, and η and ra are numbers from 0 to 3.

209833/0121

* ' ^U ■ * »BAD ORIGINAL

It has now been found that such substances I ¹ , in which m ■ is 0 and R _{2 is} the radical of a secondary, straight-chain or branched, open-chain, or mono- or polyacrylic alkyl or aralkyl group, are also much more sensitive or less susceptible to interference · Provide diagnostic means for the detection of urobilinogen than were previously known. According to the invention, therefore, particularly suitable derivatives of p-aminobenzaldehyde are substances which have a branched alkyl group on the p-amino group. In addition, the aldehydes I ia benzene ring can be substituted by an alkyl radical E, without thereby losing their positive properties.

A particular advantage of these new substances I · is that they do not display previously unknown metabolic products that occasionally occur in normal urine. It is also surprising to denote that the more basic derivatives of p-aainobenzaldehyde with a branched substituent R_ on the nitrogen atom not with Urea react, although more strongly based on previous observations basic aldehydes also react more strongly with urea.

For carrying out the method according to the invention and for production the diagnostic agents according to the invention use the same methods and agents that have already been described in the main application are*

In the following examples, the inventive method and the diagnostic agents according to the invention explained in more detail, with by Comparative tests with previously known diagnostic agents have demonstrated their superiority over the prior art.

209833/0121

BATH ORIGINAL Example 1

Filter paper (2316 from Schleicher & Gehüll) is mixed with a solution soaked in the following composition:

P- (Cyclohexylamine) benzaldehyde 0.05 g Oxalic acid. 20.0 g

Methanol to 100.0 ml

After drying, a white test paper is obtained, which is filled with urine, the urobilinogen bodies contain one more or less, depending on the content shows intense pink to purple color. TJrobilinogen-body-arae urine or a 3% aqueous urea solution do not result in any color Change of the test paper. Urinary indicator gives no color reaction. The test papers can be read after 1-2 minutes.

example

Filter paper (2316 from Schleicher & Schüll) is mixed with a solution soaked in the following composition:

p- (sec-Butylamino) benzaldehyde 0.05 S

Potassium bisulfate 15.0 g

Polyethylene glycol. 5.0 g

Water to 100.0 ml

After drying, a white test paper is obtained which is the same Has properties like the test paper produced according to Example 1.

ORIGINAL BATHROOM 209833/0121

example J>

Filter paper (2J16 from Schleicher & Schüll) comes with different concentrated solutions of X parts each of p- (isopropylamino) benzaldehyde (iPABA), p- (n-Propylamino) -benzaldehyde (nPABA) or p- (Dimethylamine) -benzaldehyde (DABA) and 20 parts of oxalic acid in 100 parts of methanol as described in Example 1, impregnated. The dry test papers will be immersed in a 5% urea solution. The color changes observed are compiled in the following table:

X iPABA nPABA DABA

0.05 White yellowish yellowish yellow 0.075 White yellowish yellow 0.1 White yellow yellow 0.15 weak yellow strong yellow

As can be clearly seen from this, this is in accordance with the invention. e test paper with iPABA much less sensitive to urea and thus reacts more sensitive and specific to urobilinogen bodies, as the pink color reaction with urobilinogen is not covered by the more or less strong yellow color reaction with urea.

B_ eis ρ ie 1 4

Filter paper (23 S or 23I6 from Schleicher & Sei.till) is impregnated with roethariolic solutions which ^{contain 100 parts of solution 0, OS parts of the following substance I 1} and 20 parts of oxalic acid. White to yellowish test papers are obtained.

The parbo reactions observed when these test papers are immersed: will ' read off after 1-2 minutes and are in the following slot 7usa: n :: 'e: ig:

209833/0121

BATH ORIGINAL

; dance I '

Reaction with

3 pointer urea solution Ubg. -haitigern

or Ubg.-poor urine urine

p- (isopropylamino) benzaldehyde

p- (Isopropylamino) -m-methylbenzaldehyde ■ p- (see-butylamino) -benzaldehyde p- (Pentyl-3-aroino) benzaldehyde

p- (2,4-Dimethylpentyl-3-amino) benzaldehyde

p- (Cyclopentylamino) benzaldehyde

p- (2-Methylcyclopentyl-amino) -benzaldehyde

P- (Cyclohexylamine) benzaldehyde

p- (Cyclohexylamine) -m-methylbene :: aldehyde

p- (Cyclohexylamino) -m-isopropylbenzalde'pyd

p- (2-ITe thy lcyc lohexy 1-amino) benzaldehyde

p-menthylairino-benzaldehyde

p- (1-Cyclohexylpropyl-2-amino) benzaldehyde ? - (Cyclododecylamino) -benzaldehyde p- (Lorbornyl-2-amino) -benzaldehyde p- (α-Hetaylbenzylamino) benzaldehyde

p- (-Isopropylbenzylamino) -bev ./. aldehyde

p- (l-? henyipropyl-2-amino) -i.e »i /, a] deiiyd

V- ( indanyl-2-amino) - "benzaldehyde

p- (N-Diethylaminoäth7l-H-i8O-propylamino) -benzaldehyde

White*

pale yellowish

pink

slightly yellowish ti pale yellowish Il yellowish Il yellow Red pale yellowish pink-purple yellowish II Il nearly white* Il Il

yellowish Il Il yellowish Il Il yellow orange yellowish pink pale yellowish pink yellow pink yellowish pink yellowish pink yellowish pink yellowish pink yellowish purple

*) The strips immersed in the urine show a pale yellow color, which is due to the inherent color of the urine.

BATH ORIGINAL

209833/0121

Example 5_

Λ For the production of a film-like reagent layer for the detection of urobilinogen a mixture of the following Beatand parts is prepared:

p- (Cyclohexylamine) benzaldehyde 0.05 g

colloidal silica ("Aerosil") 1.00 g

Oxalic acid 10.00 g

Polyvinyl butyracetal ("Mowital"). 0.5 g

Polyethylene glycol 6000 0.5 g

Methanol 50.0 ml

The above mixture about 1 mm thick W is applied to a white polyvinyl chloride film and then dried with warm air, a water-insoluble layer being formed. If this is wetted with a drop of urobilinogen-containing urine, a red color appears after 1-2 minutes. With urobiiinogenfreiem urine no staining is observed.

example

25 ml urine are acidified with 12 ml glacial acetic acid and extracted with 40 ml ether. The extract is washed twice with 20 ml of water each time and made up to 40 ml with ether. 20 ml of this are taken, 0.5 ml of a solution of 1 g of p- (cyclopentylamino) benzaldehyde in 5 ml of conc. Hydrochloric acid and shake it up vigorously. Then 2.5 ml of semi-saturated sodium acetate solution and 3 »5 ml of water are added and shaken again, the red dye collecting in the aqueous phase. After separation, the ether is washed again with 2 ml of water. The combined aqueous extracts are made up to 20 ml, measured at 557 nm in the photometer and evaluated by means of a calibration curve drawn up beforehand.

2098 * 3/0121 _{BAD O} r! G, nal

Claims (3)

1 / Diagnostic agent for the detection of urobilinogen bodies in body fluids, preferably in urine, consisting of an absorbent carrier or a film, a strong solid acid and a .Subs Lan · / ,, which look ^{like the Ehrlich 1 sample} provides a color change with urobilinogen bodies, characterized by the content of one or more substances of the formula I ¹ CHO in which R, hydrogen, a halogen atom, a cyano, nitroso, carbalkoxy, Carbamido, hydroxy, acyloxy, alkoxy, an open-chain or cyclic, saturated or unsaturated, secondary or tertiary Amino group means R ^ the remainder of a secondary, straight-chain or branched, open-chain or mono- or polycyclic alkyl or aralkyl group, R represents an alkyl group and η is a number from 0 i> is 3. . 2. A diagnostic Iüttel according to claim 1, characterized in that;, the absorbent carrier is impregnated with a Lösiing which per 100 ml of 0.001 to 1 /> an aldehyde of the general formula I 'and at least 3> preferably 15-30 ^c / ° contains a strong, solid acid. 3. Method for the detection of urobilinogen bodies in body fluids on an absorbent carrier, a reagent film or in solution in the presence of a strong, solid or liquid acid with the help of a uhstan? ,, the kind of honest seeing reagent with urobilinic bodies provides a color scheme, characterized in that. one as color indicators Substances of the formula I ' 209833/0121 BAD ORfGiNAL -N - -CHO in which E, hydrogen, a halogen atom, a cyano, nitroso, Carbalkoxy, carbamido, hydroxy, acyloxy, alkoxy, one open-chain or cyclic, saturated or unsaturated, means secondary or tertiary amino group, R- denotes the radical of a secondary straight-chain or branched, open-chain or mono- or polycyclic alkyl or aralkyl group, ⁷ ^ j
- / R- represents an alkyl group and η is a number from 0 to 5, used. 209833/0121 BATH ORIGINAL