JPH0721157B2 - Cleaning agent for constant temperature bath - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Use of the Invention] The present invention relates to an antiseptic and antifungal agent to be added to a reaction thermostatic bath using water as a medium in scientific instruments, particularly in an autoanalyzer. , A novel cleaning agent having an anti-algal action.

BACKGROUND OF THE INVENTION In general, in the field of clinical chemistry, measurement of physiologically active substances such as enzymes, lipids, proteins, etc. in biological samples such as serum, urine and tissue fluid is widely used for the purpose of diagnosing diseases, understanding pathological conditions, etc. Has been done.

The automatic analyzer is fast, highly efficient and highly accurate,
It has recently been widely used in this field because it has various features such as easy operation, small amount of sample and reagent, and labor saving. The measurement is usually a sample collection, reagent addition, mixing, heating, colorimetric (absorbance measurement),
The calculation is performed in order. Among these, there are air bath (air bath) system and water bath or oil bath system as a system for heating, but the system in which the water bath is a constant temperature bath is the most common. Reaction temperature is normal
It is performed below 50 ° C, but most commonly at 37 ° C. Regarding the absorbance measurement, there are a method in which a reaction solution is sucked up from a reaction container into a cell for measurement, and a method in which the reaction container is directly measured as a measurement cell, but the latter method is currently the mainstream. In this method of measurement, when the water bath is a constant temperature bath, the light emitted from the light source installed outside the bath passes through the constant temperature bath.
Further, it is carried out by detecting through a reaction container of a constant temperature bath by a detector installed on the opposite side of the bath. That is, the reaction vessel is also directly measured as a reaction cell. The wavelength used for measurement is usually 340 to 900 nm.

By the way, the water in the constant temperature bath of the automatic analyzer is usually exchanged once to several times a day, but air bubbles often adhere to the outer wall of the reaction vessel during the water exchange. A small amount of detergent is usually added for the purpose of preventing this. Detergents used for this purpose are usually composed of various surfactants as main components, to which chelating agents, pH adjusters, preservatives, etc. are added, and they are of low foaming property, The concentration as a cleaning agent is usually
It is 0.05 to 2.0V / V%. However, the water in a constant temperature bath containing such a detergent, when used for a long period of time, the ingredients of the detergent serve as a nutritional source, promoting the development of algae and the growth of microorganisms (mold etc.) (even with the addition of an antiseptic agent). As a result, the effect was not recognized), and as a result, algae adhered to the reaction vessel and turbidity of water in the thermostat occurred, which caused a large error in the absorbance measurement. Therefore, when using the apparatus, it is a fact that a great deal of labor is required for accuracy control and maintenance control such as frequently monitoring the inside of the tank and periodically cleaning the inside of the tank. As described above, the addition of the cleaning agent to the constant temperature bath is necessary and effective, but on the other hand, it is necessary to sufficiently carry out daily maintenance and management, and therefore improvement in this point has been strongly demanded.

[Object of the Invention] The present invention has been made in view of the above-mentioned situation, and a preservative added to a reaction thermostatic bath using water as a medium in a scientific instrument, especially in an autoanalyzer, It is an object of the present invention to provide a novel detergent having antifungal and antialgal action for a long period of time.

[Constitution of the Invention] The present invention is an invention of a cleaning agent for a constant temperature bath, which comprises a triazine derivative and a surfactant.

That is, the inventors of the present invention have conducted extensive studies to solve the above-mentioned problems, and as a result, the cause of algae development and the growth of microorganisms (bacteria etc.) is that the concentration of a normal detergent used (0.05 to 2.0). V / V
%), The final concentration of the preservative existing as one component in the thermostatic bath is below the effective concentration, and the effect disappears, so we focused on the triazine derivative as a preservative with a low effective concentration. The inventors have found that the use of a detergent containing a surfactant as a component can prevent the generation of algae and the growth of microorganisms (bacteria, etc.) in a constant temperature bath, and have completed the present invention.

As the triazine derivative which is a preservative having a low effective concentration used in the present invention, a 1,3,5-triazine derivative is particularly effective, and specific compounds include cyanuric acid, cyanuric chloride and hexahydro-1. , 3,5-Tris (β
-Hydroxyethyl) triazine, 2-chloro-4,6-dialkylamino-1,3,5-triazine, 2-methylthio-
4,6-dialkyl-1,3,5-triazine, hexahydro-
1,3,5-triethyltriazine and the like can be mentioned. These triazine derivatives may be used alone or in combination of two or more. Also, as the amount of addition,
It is sufficient that the concentration of algae is prevented and the growth of microorganisms (bacteria, etc.) is prevented, and that it does not affect the measurement. For 1,3,5-triazine derivatives, the total concentration in a constant temperature bath is 0.003 to 0.0
About 8W / V%, preferably 0.005-0.05W / V%,
The detergent is used in an amount of about 3 to 80%, preferably 5 to 50% alone or as a mixture of two or more kinds.

Various compounds other than triazine compounds are marketed as preservatives, and typical examples thereof include phenols, cresols, chlorine compounds, salicylic acid compounds, benzoic acid compounds and sodium azide. However,
These are effective in preventing the growth of microorganisms, but when used as a component of a cleaning agent for a constant temperature bath, they may affect the measurement wavelength or damage the metal or plastics that make up the constant temperature bath. I have a problem with. That is, as the preservative that can be used for the purpose of the present invention, the measurement wavelength (34
Has almost no absorption at 0 to 900 nm), is compatible with water or / and a surfactant, does not cause precipitation and turbidity with other components in the cleaning agent, and does not cause glass, plastic, metal, etc. No corrosion, stable quality for a long time,
In addition, it is an essential condition that the generation of algae and the growth of microorganisms (bacteria etc.) can be prevented at a low effective concentration.

As the surfactant used in the present invention, there is no influence on the measurement, and it is almost good as long as it can prevent the adhesion of bubbles to the reaction vessel, but more strictly speaking, there is no mixture of water-insoluble substances, Low foaming, high cloud point and reaction temperature (37
(° C) is clear, does not cause reaction or precipitation with the triazine derivative which is the preservative according to the present invention, has almost no absorption in the range of 340 to 900 nm, and the thermostat and the reaction vessel of the automatic analyzer are It can be used without particular limitation as long as it has properties such as no influence on the constituent glass, metal, plastic, etc., stable quality at all times, no danger and easy handling,
Particularly, nonionic surfactants are preferably used. Examples of nonionic surfactants include fatty acid glyceride, polyoxyethylene fatty acid ester, polyoxyethylene alkyl ether, polyoxyethylene alkylaryl ether, sorbitan fatty acid ester, sucrose fatty acid ester, polyoxyethylene sorbitan fatty acid ester, and polyoxyethylene sorbitan fatty acid ester. Examples thereof include oxyethylene alkylamine, polyoxyethylene fatty acid amide, polyoxyethylene polypropylene glycol ether and the like. Also,
The concentration in the detergent is not particularly limited, but is preferably 1 to 20 W / V%, more preferably 3 to 1
It is selected in the range of 0 W / V%.

Further, the detergent according to the present invention may optionally contain various surfactants, chelating agents, pH adjusters, preservatives and the like as long as they do not impair the object of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto.

[Examples] Experimental Example 1. Measurement of minimum inhibitory concentration of triazine compound against microorganisms (test bacteria) Microorganisms (molds, bacteria, yeasts, algae, etc.) grown in a constant temperature water tank of an automatic analyzer as shown below. ) And JIS-Z
The mold described in the mold resistance test method of No. 2911 was used.

・ Bacteria: Pseudomonas aeruginosa, Bacillus subtilis,
Escherichia coli.

Molds: Aspergillus niger, Fusarium moniliforme, Cl
adosporium cladosprioides, Penicillium citrinum.

-Algae: green algae, cyanobacteria, diatoms.

・ Yeasts: Saccharomyces cerevisiae, Rhodotorula s
p ..

(Culture liquid) The following were used according to the type of microorganism.

・ Bacteria Meat extract 3g Polypeptone 10g Sodium chloride 5g Distilled water Total 1000ml ・ Molds (potato dextrose broth) Potato extract 4g Glucose 20g Distilled water 1000ml ・ Algae (dead melt broth) Ca ( _{No 3) 2 · 4H 2 O} 1g MgSO 4 · 7H 2 O 0.25g KCl 0.25g KH 2 PO 4 0.25g FeCl 3 trace distilled water total amount for 1000 ml, yeasts (malt yeast broth) yeast extract 3g glucose 10g malt Extract 3g Peptone 5g Distilled water Total volume 1000ml (Procedure) After culturing each microorganism in the prescribed culture solution until the number of bacteria exceeds the prescribed number (bacteria / yeast: 10 ⁷ / ml or more, algae:
10 ⁸ pieces / ml or more, molds: 10 ⁶ pieces / ml or more), hexahydro-1,3,5-tris (β-hydroxyethyl) triazine which is a triazine compound according to the present invention (Hereinafter, abbreviated as THT.) Was added to the culture solution to a predetermined concentration.
After the lapse of time, the minimum inhibitory concentration (minimum addition amount required to inhibit the growth) of fungi was determined after 72 hours at 28 ° C. and at 35 ° C. for algae after 168 hours.

The determination was carried out using the number of bacteria per ml as a control, although the same operation was performed in a medium without THT.

(Results) Table 1 shows the minimum inhibitory concentration of THT for each microorganism.

Experimental Example 2. Measurement of the minimum inhibitory concentration of microorganisms of the constant temperature bath cleaning agent according to the present invention (constant temperature bath cleaning agent) THT: polyoxyethylene nonylphenyl ether: distilled water = 10: 2: 88 by weight ratio The mixture thus obtained was used as a cleaning agent for a constant temperature bath.

By replacing THT in the experiment example 1 with the above-mentioned bath for cleaning the constant temperature bath and using the same microorganism and medium as in the experiment example 1, the same operation method as in the experiment example 1 was used to prevent the minimum growth of the present bath cleaning agent against the microorganism. The concentration was measured.

(Results) The results are shown in Table 2.

As is clear from the results shown in Tables 1 and 2, it can be seen that the triazine compound according to the present invention is effective against microorganisms at a low concentration even when used alone or as the detergent according to the present invention. Further, it was also found that the triazine compound according to the present invention is effective at a lower concentration in addition to the use of a surfactant in combination with a surfactant.

Example 1. (Cleaning agent for constant temperature bath) A cleaning agent for constant temperature bath was prepared by mixing THT: polyoxyethylene nonylphenyl ether: distilled water in a weight ratio of 30:10:60.

(Operation method) The above cleaning agent diluted 1000 times with distilled water was stored at 28 ° C in a polyethylene container.

Separately, a detergent without the addition of THT was prepared and stored under the same conditions as a control.

After storage, the presence or absence of algae and the growth of microorganisms (mold, etc.) in the THT-added and non-added detergent-containing liquids were visually examined daily.

In addition, using a Hitachi 736 type automatic analyzer, the above cleaning agent was added to the same constant temperature bath so as to be diluted 1000 times, and the influence on the measured value was examined.

Separately, a cleaning agent containing no THT was prepared and added to a constant temperature bath of Hitachi 736 automatic analyzer under the same conditions as a control.

The effect on the measured values is the daily reproducibility of the simultaneous reproducibility of transaminase (GOT, GPT), which is an inspection item by the UV rate method, in which the effects of water stains in the constant temperature bath and bubbles adhering to the reaction vessel are most noticeable. The changes were tracked. [N = 40, reagent: transaminase-HR (manufactured by Wako Pure Chemical Industries, Ltd.), standard serum: autonorm (manufactured by Daiichi Pure Chemicals Co., Ltd.)] (Result) A detergent containing no THT was used. Observed the growth of microorganisms (mold etc.) on the 7th day, and the measured values were also affected by the test results using an automatic analyzer, but those using the THT-added detergent also developed algae on the 60th day. , No growth of microorganisms (mold etc.) was observed.

In addition, Table 3 shows the results of measuring the viable cell count in the thermostat with the automatic analyzer using the detergent containing THT over time.
Table 4 shows the results obtained by tracing the daily variation in the simultaneous reproducibility of T.

As is clear from the results in Tables 3 and 4, the test results using the automatic analyzer did not show the development of algae or the growth of microorganisms (mold etc.), and did not show any influence on the measured values.

Similar results were obtained when cyanuric acid was used as the triazine compound instead of THT.

Experimental example 3. Stability test of detergent for constant temperature bath (cleaner for constant temperature bath) Preservative: Polyoxyethylene nonylphenyl ether: Distilled water = 30:10:60 The resulting solution was used as a constant temperature cleaning agent stock solution.

(Operation method-1) The above-mentioned stock solution of the cleaning agent for a constant temperature bath was stored in a polyethylene container at 25 ° C for 1 to 6 months. After storage for a predetermined number of days, the presence or absence of precipitation was visually inspected for each stock solution of the cleaning agent for constant temperature baths.

The results are shown in Table 5.

(Operation method-2) A 1000-fold dilution of the above-mentioned stock solution for a constant temperature bath with distilled water was stored in a polyethylene container at 30 to 50 ° C for 1 to 6 months. After storage for a predetermined number of days, the presence or absence of precipitation was examined for each diluted constant temperature bath cleaning agent. In each case, the presence or absence of absorption in the measurement wavelength range (200 to 900 nm) was examined using a spectrophotometer.

The results are also shown in Table 5.

As is clear from Table 5, the constant temperature bath cleaning agent of the present invention is
It can be seen that even if it is stored for 6 months or more in the undiluted state, no precipitation occurs and it is stable. On the other hand, it can be seen that the cleaning agents for constant temperature baths prepared using the conventional preservatives all precipitate after storage for about 1 month.

Further, it can be seen that even when the cleaning agent for constant temperature bath of the present invention is diluted to an actual use state, it does not cause precipitation even when stored for a long time (6 months or more) and is stable. On the other hand, it can be seen that the diluted cleaning agent for the constant temperature bath prepared by using the conventional preservative causes all precipitation after storage for about one month.

Further, the diluted bath for cleaning the thermostatic bath of the present invention has no absorption in the measurement wavelength range, but the diluted bath for cleaning the bath prepared by using the conventional preservative is measured. It can be seen that it has absorption in the wavelength range.

From the above results, it is understood that the constant temperature bath cleaning agent of the present invention has excellent properties as a cleaning agent used for such a purpose.

[Effects of the Invention] As described above, in the present invention, in a scientific instrument having a constant temperature water bath using water as a medium, particularly in an automatic analyzer, generation of microorganisms (mold etc.) generated in water in the constant temperature bath is prevented. The present invention provides a cleaning agent for a constant temperature bath, which prevents the generation of bubbles and adheres to the outer wall of the reaction vessel in the constant temperature bath, which prevents the measurement accuracy from deteriorating. As a result, a remarkable effect is exerted in that the quickness, high efficiency, high accuracy, and simple operation which are the original merits of the automatic analyzer can be utilized more than ever before.

Claims (3)

[Claims] 1. A cleaning agent for a constant temperature bath comprising a triazine derivative and a surfactant. 2. The cleaning agent according to claim 1, wherein the triazine derivative is a 1,3,5-triazine derivative. 3. The cleaning agent according to claim 1, wherein the surfactant is a nonionic surfactant.