JPS62231167A - Pretreatment kit for analyzing catecholamine - Google Patents

Abstract

PURPOSE:To improve the accuracy of analysis by hermetically sealing a buffer, stabilizer and activated adsorptive carrier into a column having a freely attachable and detachable stopper so that the column can be preserved and allowing a catecholamine to be captured on the adsorptive carrier simply by removing the stopper and injecting a vital sample thereon in the stage of the analysis. CONSTITUTION:A funnel-shaped sample injection port 2 is formed to the top end of the PP column 1 which constitutes a centrifugal sepn. tube and a discharge port 3 is formed to the bottom end thereof. A glass filter 4 is disposed near the discharge port 3 and the column is hermetically plugged by the stopper 5 and a flexible cap 6. A boric acid gel 7 and a drum 8 prepd. from about 0.8mol phosphoric acid buffer soln. and about 0.1mol EDTA are sealed into the column. The stopper 5 is removed and sampled blood or urine is dropped into the column from the injection port 2 in the stage of use. The column is then oscillated for about 10min after hermetic sealing with the stopper 5. The cap 6 is thereafter removed and the column is housed into a test tube T. The column is processed by a centrifugal separator to discharge the internal liquid to the test tube T. The sample of the chromatography is thus obtd.

Description

DETAILED DESCRIPTION OF THE INVENTION A. Field of Industrial Application The present invention relates to a pretreatment kit for extracting catecholamines from biological samples.

Previous Art Catecholamines are a general term for 3,4-dihydroxyphenyl-based amines, and by analyzing them from biological samples such as blood and urine, useful data for disease diagnosis and treatment can be obtained.籟 牛7+t-P＊Col
Hit r + - + t → Te17 Pretreatment is performed to extract only catecholamines from the sample.

Taking blood as an example, this pretreatment involves removing protein from sample plasma, adding a stabilizer to suppress enzyme activity, and then taking the sample into a container, and then adding boric acid gel together with a buffer solution. Catecholamines were adsorbed onto the surface of the boric acid gel, and after centrifugal dehydration, acid was added to remove the catecholamines from the boric acid gel, and the resulting solution was collected for analysis.

However, in order to subject the boric acid gel to pretreatment,
This must be soaked in distilled water for a day and night, and then washed with hydrogen chloride or sodium hydroxide solution, which requires a great deal of time and effort when analyzing a large number of samples such as in mass medical examinations. However, there was a problem in that the expensive boric acid gel disappeared when collecting the supernatant.

C. Purpose Uninvented was made in view of this situation, and its purpose is to easily and quickly extract catecholamines from biological samples without causing the disappearance of the boric acid gel. Our goal is to provide a pretreatment kit that allows for

20 Summary of the Invention, that is, the present invention is characterized in that the buffer, stabilizer, and activated adsorption carrier can be sealed and stored in a column with a removable stopper, and the stopper can be removed during analysis. The point is that catecholamines can be obtained in a state captured on an adsorption carrier by simply injecting a biological sample.

E. Embodiments The details of the present invention will now be explained based on illustrated embodiments.

Fig. 1 shows the external appearance of a kit showing an embodiment of the present invention. The reference numeral in the figure is a polypropylene column forming a centrifuge M tube, and the upper end of the column is connected to the sample injection port 2 on the funnel, or A discharge port 3 is formed at the lower end, and a glass filter 4 is disposed near the discharge port 3, each of which is closed with a common stopper 5 and a visible cap 6, and inside the discharge port 3 is filled with washed boric acid. gel 7, and 0.8 molar phosphate buffer and 0
．． Drug 8 prepared with 1 molar EDTA was encapsulated.

In this example, the stopper 4 of the kit prepared in advance
When the sampled blood or urine is dropped from the injection port 2, sealed with the stopper 5, and then shaken, the phosphate buffer will maintain the pH inside the column at around 7 and transfer to the boric acid gel 7. Selection board@ show your personality. As a result, the boric acid gel 7 adsorbs catecholamines on its surface and at the same time
DTA suppresses enzyme activity. After continuing this shaking for 10 minutes, the stopcock 5 and the can 6 were removed, the drug 8 in the column 1 was discharged with an aspirator, and then distilled water was injected.
After washing the boric acid gel 7, it is directly dehydrated by centrifugation. At this time, the glass filter 4 captures the boric acid gel 7 adsorbing catecholamines and prevents this from flowing out. After dehydration, put the cap 6 back on and fill the inlet 2 with an acid that has a pH of 3 or less, e.g. ○,] specified HC.
When β or 0.4 to 0.8 N acetic acid is injected, the boric acid gel 7 desorbs adsorbed catecholamines. After leaving the snow in this way for 5~]0 minutes, remove the cap 6, put it in a test tube T (Fig. 2), centrifuge it, drain the liquid inside into the test tube T, and perform high-performance liquid chromatography. sample.

After use, the boric acid gel 7 is washed with distilled water, hydrochloric acid, or aqueous sodium hydroxide solution, and then a drug 8 prepared with 0.8 molar phosphate buffer and 0.1 molar EDTA is injected and mixed. Seal with stopper 5 and save for next use.

As explained above, according to the present invention, a buffer, a stabilizer, and a boric acid gel are sealed in an analytical column to form a kit, so a large number of samples can be analyzed quickly and under the same conditions with simple operations. Pretreatment can be performed, which not only improves analytical accuracy, but also allows processing to be completed in one container, resulting in less loss of target components and making it possible to analyze with a trace amount of sample. In addition, since the boric acid gel is stored immersed in a solution that satisfies certain conditions, there is no risk of reducing adsorption to catecholamines.

[Brief explanation of drawings]

FIG. 1 is an external view of a pretreatment kit showing one embodiment of the present invention, and FIG. 2 is an external view showing the state at the time of collecting target components. 1...Column 4...Glass filter 7...Boric acid gel 8
...Drugs

Claims (1)

[Claims] The sample inlet and outlet are sealed with a removable stopper and cap, and a drug prepared with a washed boric acid gel, phosphate buffer, and stabilizer is placed in a container with a filter placed near the outlet. A pretreatment kit for analysis of catecholamines.