Composition for detecting occult blood in feces, method for preparing same and use thereof

Abstract

The present application provides compositions for detecting occult blood in fecal waste, methods for their preparation and use. [Solution] The composition contains, by weight, 20 to 80 parts of an organic solvent, 0.01 to 50 parts of a thickener, 0.01 to 2 parts of benzidine, 0.01 to 5 parts of a peroxide, 0.001 to 0.5 parts of EDTA-2Na, and 10 to 80 parts of water. The composition according to the present application can quickly and effectively detect the state of occult blood in excrement, has a fast color change speed, a long color retention time, and high sensitivity.

Description

[Technical Field]

[0001] The present application belongs to the field of disease detection, and specifically relates to a composition for detecting occult blood in feces, its preparation method and use, and more particularly to a composition for detecting occult blood in feces with high sensitivity, its preparation method and use. [Background technology]

[0002] Blood in feces is a common pathological symptom. Potential causes of blood in stool include, but are not limited to, hemorrhoids, ulcerative colitis, upper gastrointestinal bleeding, lower gastrointestinal bleeding, blood-related diseases, anal rupture, and gastrointestinal tumors. Potential causes of blood in urine include, but are not limited to, urinary mucosal damage, urinary stones, nephritis, and urinary tumors. Bleeding in the digestive and urinary systems due to the above conditions includes two types: visible blood and occult blood. While visible blood in feces is easy to notice, occult blood in feces is the initial symptom of most diseases. Visible blood in feces may indicate a serious condition, so regular occult blood detection can serve as an early screening method for urinary and digestive system diseases.

[0003] CN114354948A discloses a semi-quantitative method for detecting fecal occult blood, which involves the following steps: A diluted sample is dropped into a spotting hole on a colloidal gold detection card using a spotting device; after a qualitative negative or positive result initially appears, a photograph is taken and the qualitative result is read on the instrument's reading display; after the qualitative result is read, a certain amount of 3,3,5,5-tetramethylbenzidine indicator solution is dropped into the same spotting hole, followed by a certain amount of hydrogen peroxide solution; after the 3,3,5,5-tetramethylbenzidine indicator solution and hydrogen peroxide solution are dropped and allowed to react for a while, a semi-quantitative result is displayed on the colloidal gold detection card; a photograph is taken and the semi-quantitative result is read. This invention increases the semi-quantitative result of fecal occult blood detection, reduces manual operation and manual workload, solves the deficiencies of fecal occult blood detection in clinical testing, significantly reduces the need for manual retesting, and ensures the accuracy of the detection results.

[0004] CN103175965A discloses a fecal occult blood detection kit, which includes a fecal occult blood detection test strip or test strip card, a sample processing tube or stool mixer, and a sampling rod. The test strip or test strip card includes a detection area (T) coated with an anti-Hb1 monoclonal antibody, a quality control area (C) coated with a sheep anti-mouse polyclonal antibody, and a gold colloid coated with an anti-Hb2 monoclonal antibody. The manufacturing method of the reagent kit includes steps such as blending a processing solution, manufacturing the reagent kit, preparing the gold colloid complex, metal spraying, cutting, assembling, cutting, and packaging. The fecal occult blood detection kit has good specificity, high sensitivity, rapid detection, and is simple, and is not restricted by diet or medication.

[0005] Since the detection of occult blood can play an important role in early screening for diseases of the urinary and digestive systems, it is an urgent issue to provide a method or reagent for detecting occult blood that is easy to detect and highly sensitive. Summary of the Invention [Problem to be solved by the invention]

[0006] In response to the deficiencies of the related art, the present application provides a composition for detecting occult blood in feces, its preparation method and use, and in particular, a composition for detecting occult blood in feces with high sensitivity, its preparation method and use, etc. The composition of the present application can quickly and effectively detect the status of occult blood in feces, has a fast color change rate, a long color retention time, and is highly sensitive. [Means for solving the problem]

[0007] The present application provides the following technical solutions.

[0008] In aspect 1, the present embodiment comprises: In parts by weight, it comprises 20 to 80 parts of an organic solvent, 0.01 to 50 parts of a thickener, 0.01 to 2 parts of benzidine, 0.01 to 5 parts of peroxide, 0.001 to 0.5 parts of EDTA-2Na, and 10 to 80 parts of water; A composition for detecting occult blood in feces is provided.

[0009] Here, the number of parts of the organic solvent may be 20 parts, 30 parts, 40 parts, 50 parts, 60 parts, 70 parts, or 80 parts, etc.; the number of parts of the thickener may be 0.01 parts, 0.1 parts, 0.5 parts, 1 part, 5 parts, 10 parts, 15 parts, 20 parts, 25 parts, 30 parts, 35 parts, 40 parts, 45 parts, or 50 parts, etc.; the number of parts of the benzidine may be 0.01 parts, 0.1 parts, 0.5 parts, 1 part, 1.5 parts, or 2 parts, etc.; and the number of parts of the peroxide may be 0.01 parts, 0.1 parts, 0.5 parts, 1 part, 1.5 parts, or 2 parts, etc. The number of parts of EDTA-2Na may be 0.001, 0.01, 0.1, 0.2, 0.3, 0.4, or 0.5 parts, etc., and the number of parts of water may be 10, 20, 30, 40, 50, 60, 70, or 80 parts, etc., but is not limited to the numerical values ​​recited above, and other unrecited numerical values ​​within the above numerical ranges also apply.

[0010] By selecting specific components, the composition can effectively detect the occult blood condition in excrement, and has a fast color change speed, long color retention time, and high sensitivity.

[0011] In one embodiment, the composition comprises, in parts by weight, 40-60 parts organic solvent, 10-40 parts thickener, 0.5-1.5 parts benzidine, 1-4 parts peroxide, 0.1-0.3 parts EDTA-2Na, and 30-60 parts water.

[0012] In one embodiment, the organic solvent includes one or a combination of at least two of ethanol, benzene, toluene, pentane, hexane, methanol, diethyl ether, ethyl acetate, acetone, or carbon tetrachloride, such as a combination of ethanol and acetone, a combination of ethanol and toluene, or a combination of methanol and ethanol, but is not limited to the above-listed combinations, and other combinations not listed above also apply.

[0013] In one embodiment, the organic solvent comprises a combination of ethanol and acetone.

[0014] In one embodiment, the thickener comprises a combination of one or at least two of xanthan gum, guar gum, pectin, sodium polyacrylate, hydroxypropyl methylcellulose, sodium hydroxymethylcellulose, or hydroxymethylcellulose, for example, a combination of xanthan gum and guar gum, a combination of guar gum and pectin, or a combination of pectin and sodium polyacrylate, but is not limited to the above-listed combinations, and other unlisted combinations within the above range of combinations also apply.

[0015] In one embodiment, the benzidine includes one or a combination of at least two of 4,4'-diaminobiphenyl, tetramethylbenzidine, and 3,3'-dihydroxybenzidine, for example, a combination of 4,4'-diaminobiphenyl and tetramethylbenzidine, a combination of tetramethylbenzidine and 3,3'-dihydroxybenzidine, or a combination of 4,4'-diaminobiphenyl and 3,3'-dihydroxybenzidine, but is not limited to the above-listed combinations, and other unlisted combinations within the above combination range also apply.

[0016] In one embodiment, the benzidine comprises a combination of 3,3'-dihydroxybenzidine and tetramethylbenzidine.

[0017] The combination of the specific benzidines mentioned above can further improve the technical effects of the composition, accelerating the color change rate, extending the color retention time, and further improving the detection sensitivity.

[0018] In one embodiment, the peroxide comprises an inorganic peroxide and / or an organic peroxide.

[0019] In one embodiment, the peroxide comprises a combination of an inorganic peroxide and an organic peroxide.

[0020] In one embodiment, the inorganic peroxide includes one or a combination of at least two of hydrogen peroxide, sodium percarbonate, potassium peroxide, calcium peroxide, or barium peroxide, such as a combination of hydrogen peroxide and sodium percarbonate, a combination of sodium percarbonate and potassium peroxide, or a combination of potassium peroxide and calcium peroxide, but is not limited to the above-listed combinations, and other combinations not listed within the above range also apply.

[0021] In one embodiment, the inorganic peroxide comprises hydrogen peroxide.

[0022] In one embodiment, the organic peroxide includes one or a combination of at least two of cumene hydroperoxide, diisopropylbenzene hydroperoxide, t-amylperoxy-2-ethylhexanoate, and t-butylperoxy-2-ethylhexanoate, for example, a combination of cumene hydroperoxide and diisopropylbenzene hydroperoxide, a combination of diisopropylbenzene hydroperoxide and t-amylperoxy-2-ethylhexanoate, or a combination of t-amylperoxy-2-ethylhexanoate and t-butylperoxy-2-ethylhexanoate, but is not limited to the above-listed combinations, and other combinations not listed within the above range also apply.

[0023] In one embodiment, the organic peroxide comprises cumene hydroperoxide.

[0024] The above-mentioned specific peroxides and their combinations can effectively improve the technical effects of the composition, accelerate the color change rate, extend the color retention time, and further improve the detection sensitivity.

[0025] In one embodiment, the composition further comprises 0.01 to 5 parts by weight of a pH adjuster, for example, 0.01 parts, 0.1 parts, 0.5 parts, 1 part, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts, or 5 parts, but is not limited to the above-listed values, and other unlisted values ​​within the above range also apply.

[0026] In one embodiment, the pH adjuster comprises one of citric acid, acetic acid, sodium hydroxide, or hydrochloric acid.

[0027] In one embodiment, the composition further comprises 1 to 80 parts by weight of starch, for example, 1 part, 10 parts, 20 parts, 30 parts, 40 parts, 50 parts, 60 parts, 70 parts, or 80 parts, but is not limited to the above-listed values, and other unlisted values ​​within the above ranges also apply.

[0028] In one embodiment, the starch includes one or a combination of at least two of corn starch, pea starch, mung bean starch, gelatinized starch, methylol starch, cross-linked starch, maltodextrin, oxidized starch, esterified starch, or etherified starch, such as a combination of corn starch and pea starch, a combination of pea starch and mung bean starch, or a combination of mung bean starch and gelatinized starch, but is not limited to the above-listed combinations, and other combinations not listed above also apply.

[0029] The starch can be added according to the user's needs, and the composition can be made from a liquid to a solid, allowing it to be applied to different occasions.

[0030] In a second aspect, the present application provides a method for preparing the composition for detecting occult blood in fecal excrement described above, the method comprising: Step (1) of mixing benzidine, peroxide and an organic solvent to obtain an organic phase; Step (2) of mixing a thickener, EDTA-2Na and water to obtain an aqueous phase; and (3) mixing the aqueous phase and the organic phase to obtain a composition for detecting occult blood in the feces, Step (1) and step (2) do not distinguish in terms of priority.

[0031] In one embodiment, the step of mixing a thickener, EDTA-2Na, and water to obtain an aqueous phase in step (2) further comprises mixing with a pH adjuster.

[0032] In one embodiment, the step of combining the aqueous phase and the organic phase in step (3) further comprises combining with starch.

[0033] In a third aspect, the present embodiment further provides a use of the composition for detecting occult blood in feces described above in the manufacture of an occult blood detection device.

[0034] In one embodiment, the occult blood detection device includes one of a detection test strip, a detection plate, a reagent kit, or a detection tube. [Effects of the Invention]

[0035] Compared with the related art, the present invention has the following beneficial effects:

[0036] The present application provides a composition for detecting occult blood in feces, and by selecting specific benzidine and peroxide, it can effectively detect occult blood in feces, has a fast color change rate, a long color retention time, and high sensitivity. DETAILED DESCRIPTION OF THE INVENTION

[0037] In order to further explain the technical means adopted in the present application and their effects, the technical solution of the present application will be further described below with reference to preferred embodiments of the present application, but the present application is not limited to the scope of the embodiments.

[0038] [Example 1] This example provides a composition for detecting occult blood in feces, the preparation ingredients of which are, in parts by weight: The mixture was 30 parts of ethanol, 20 parts of acetone, 25 parts of xanthan gum, 0.5 parts of 3,3'-dihydroxybenzidine, 0.5 parts of tetramethylbenzidine, 1 part of hydrogen peroxide, 1 part of cumene hydroperoxide, 0.25 parts of EDTA-2Na, 50 parts of water, and 2.5 parts of citric acid.

[0039] The preparation method is as follows.

[0040] (1) Ethanol, 3,3'-dihydroxybenzidine, tetramethylbenzidine, hydrogen peroxide, and cumene hydroperoxide were mixed and stirred to obtain an organic phase.

[0041] (2) Water, xanthan gum, EDTA-2Na, and citric acid were mixed to obtain an aqueous phase.

[0042] (3) The aqueous phase and the organic phase were mixed and stirred to obtain a composition for detecting occult blood in the excrement.

[0043] [Example 2] This example provides a composition for detecting occult blood in feces, the preparation ingredients of which are, in parts by weight: The ingredients were 40 parts of ethanol, 10 parts of pectin, 0.5 parts of tetramethylbenzidine, 1 part of cumene hydroperoxide, 0.1 part of EDTA-2Na, 30 parts of water, and 0.1 part of citric acid.

[0044] The preparation method was as described in Example 1.

[0045] [Example 3] This example provides a composition for detecting occult blood in feces, the preparation ingredients of which are, in parts by weight: The mixture was 60 parts of ethanol, 40 parts of guar gum, 1.5 parts of 4,4'-diaminobiphenyl, 4 parts of hydrogen peroxide, 0.3 parts of EDTA-2Na, 60 parts of water, and 5 parts of citric acid.

[0046] The preparation method was as described in Example 1.

[0047] [Example 4] This example provides a composition for detecting occult blood in feces, the preparation ingredients of which are, in parts by weight: The ingredients were 20 parts of ethanol, 0.01 part of pectin, 0.01 part of tetramethylbenzidine, 0.01 part of cumene hydroperoxide, 0.001 part of EDTA-2Na, 10 parts of water, and 0.1 part of citric acid.

[0048] The preparation method was as described in Example 1.

[0049] [Example 5] This example provides a composition for detecting occult blood in feces, the preparation ingredients of which are, in parts by weight: The mixture was 80 parts ethanol, 50 parts guar gum, 2 parts 4,4'-diaminobiphenyl, 5 parts hydrogen peroxide, 0.5 parts EDTA-2Na, 80 parts water, and 5 parts citric acid.

[0050] The preparation method was as described in Example 1.

[0051] [Example 6] This example provides a composition for detecting occult blood in feces, and the preparation materials are identical to those in Example 1, except that 3,3'-dihydroxybenzidine is not added and the remaining amount is allocated to tetramethylbenzidine.

[0052] The preparation method was as described in Example 1.

[0053] [Example 7] This example provides a composition for detecting occult blood in feces, and the preparation materials are identical to those in Example 1, except that tetramethylbenzidine is not added and the remaining amount is allocated to 3,3'-dihydroxybenzidine.

[0054] The preparation method was as described in Example 1.

[0055] [Example 8] This example provides a composition for detecting occult blood in feces, and the preparation materials were otherwise identical to those in Example 1, except that 3,3'-dihydroxybenzidine was replaced with an equal amount of 4,4'-diaminobiphenyl.

[0056] The preparation method was as described in Example 1.

[0057] [Example 9] This example provides a composition for detecting occult blood in feces, and the preparation materials are identical to those of Example 1, except that hydrogen peroxide was not added and the remaining amount was allocated to cumene hydroperoxide.

[0058] The preparation method was as described in Example 1.

[0059] [Example 10] This example provides a composition for detecting occult blood in feces, and the preparation materials are identical to those in Example 1, except that cumene hydroperoxide was not added and the reduced amount was allocated to hydrogen peroxide.

[0060] The preparation method was as described in Example 1.

[0061] [Example 11] This example provides a composition for detecting occult blood in feces, and the preparation ingredients were otherwise identical to Example 1, except that hydrogen peroxide was replaced with an equal amount of sodium percarbonate.

[0062] The preparation method was as described in Example 1.

[0063] [Example 12] This example provides a composition for detecting occult blood in feces, and the preparation materials were the same as those in Example 1, except that cumene hydroperoxide was replaced with an equal amount of t-butylperoxy-2-ethylhexanoate.

[0064] The preparation method was as described in Example 1.

[0065] [Example 13] This example provides a composition for detecting occult blood in feces, and the preparation materials were identical to those in Example 1, except that ethanol was omitted and the remaining amount was allocated to acetone.

[0066] The preparation method was as described in Example 1.

[0067] [Example 14] This example provides a composition for detecting occult blood in feces, and the preparation materials were identical to those in Example 1, except that acetone was omitted and the remaining amount was allocated to ethanol.

[0068] The preparation method was as described in Example 1.

[0069] [Example 15] This example provides a composition for detecting occult blood in feces, and the preparation materials were identical to those in Example 1, except that ethanol was omitted and the remaining amount was allocated to toluene.

[0070] The preparation method was as described in Example 1.

[0071] [Effectiveness test] The compositions for detecting occult blood in excrement according to Examples 1 to 15 were impregnated into test strips, which were then dried to obtain detection test strips. Then, samples (fresh normal human blood diluted 10,000 times with water) were used for detection, and the time required for color change and color retention were recorded. The results are as follows:

[0072] [Table 1]

[0073] The above data demonstrate that the product of the present application has the advantages of short discoloration time and long color retention time, allowing for rapid sample detection. Comparing Examples 1 to 5, it was found that the present application can effectively enhance the product's effectiveness by controlling the amount of each ingredient used. Comparing Examples 1 and 6 to 8, it was found that the present application can effectively shorten the discoloration time and extend the color retention time by selecting a specific benzidine combination. Comparing Examples 1 and 9 to 12, it was found that the present application can effectively shorten the discoloration time and extend the color retention time by selecting a specific peroxide and its combination. Comparing Examples 1 and 13 to 15, it was found that the present application can effectively shorten the discoloration time and extend the color retention time by selecting a specific organic solvent combination.

[0074] Then, the test samples with different hemoglobin concentrations were detected using the test paper, and the detection limit of the test paper was analyzed. The results are as follows:

[0075] [Table 2]

[0076] The above data demonstrate that the composition of the present application is characterized by high sensitivity and can effectively detect extremely low levels of hemoglobin. Comparing Examples 1 to 5, it was found that the present application can effectively enhance the effectiveness of the product by controlling the amount of each raw material used. Comparing Examples 1, 6 to 8, it was found that the present application can effectively reduce the detection limit and improve detection sensitivity by selecting a specific combination of benzidines. Comparing Examples 1, 9 to 12, it was found that the present application can effectively reduce the detection limit and improve detection sensitivity by selecting a specific peroxide and its combination. Comparing Examples 1, 13 to 15, it was found that the present application can effectively reduce the detection limit and improve detection sensitivity by selecting a specific combination of organic solvents.

[0077] The present application describes the composition for detecting occult blood in feces, its preparation method and use through the above examples. However, the applicant declares that the present application is not limited to the above examples, that is, it does not mean that the present application must be carried out depending on the above examples. Those skilled in the art should understand that any improvements to the present application, equivalent replacement of each ingredient of the product of the present application, addition of auxiliary ingredients, selection of specific forms, etc. are all within the protection scope and disclosure scope of the present application.

[0078] The above is a detailed description of the preferred embodiments of the present application. However, the present application is not limited to the specific content of the above embodiments. Various simple modifications can be made to the technical solution of the present application within the scope of the technical idea of ​​the present application. All of these simple modifications fall within the scope of the claims of the present application.

[0079] It should be noted that the specific technical features described in the above specific embodiments can be combined in any suitable manner unless there is a contradiction, and in order to avoid unnecessary repetition, various possible combinations will not be specifically described in this application.

Claims

1. In parts by weight, the composition comprises 20 to 60 parts of an organic solvent, 0.5 to 40 parts of a thickener, 0.1 to 1.5 parts of benzidine, 0.1 to 5 parts of a peroxide, 0.01 to 0.3 parts of EDTA-2Na, and 10 to 60 parts of water; The benzidine includes a combination of 3,3'-dihydroxybenzidine and tetramethylbenzidine; The peroxide includes a combination of an inorganic peroxide and an organic peroxide, the organic peroxide includes cumene hydroperoxide, and the inorganic peroxide includes hydrogen peroxide; The organic solvent includes a combination of ethanol and acetone. A composition for detecting occult blood in feces.

2. The composition comprises, in parts by weight, 40 to 60 parts of an organic solvent, 10 to 40 parts of a thickener, 0.5 to 1.5 parts of benzidine, 1 to 4 parts of a peroxide, 0.1 to 0.3 parts of EDTA-2Na, and 30 to 60 parts of water; The composition of claim 1.

3. The thickening agent may comprise any one or a combination of at least two of xanthan gum, guar gum, pectin, sodium polyacrylate, hydroxypropyl methylcellulose, sodium hydroxymethylcellulose, or hydroxymethylcellulose. The composition of claim 1.

4. The composition further comprises 0.01 to 5 parts by weight of a pH adjuster. The composition of claim 1.

5. The composition described in claim 4, wherein the pH adjuster includes one of citric acid, acetic acid, sodium hydroxide, or hydrochloric acid.

6. The composition described in claim 1, further comprising 1 to 80 parts by weight of starch.

7. The starch comprises one or a combination of at least two of corn starch, pea starch, mung bean starch, gelatinized starch, methylol starch, cross-linked starch, maltodextrin, oxidized starch, esterified starch, or etherified starch. The composition of claim 6.

8. A method for preparing a composition for detecting occult blood in feces according to any one of claims 1 to 7, comprising: Step (1) of mixing benzidine, peroxide and an organic solvent to obtain an organic phase; Step (2) of mixing a thickener, EDTA-2Na and water to obtain an aqueous phase; and (3) mixing the aqueous phase and the organic phase to obtain a composition for detecting occult blood in the feces, Step (1) and step (2) do not distinguish between priorities. Preparation method.

9. The step of mixing the thickener, EDTA-2Na, and water to obtain an aqueous phase in step (2) further comprises mixing with a pH adjuster. The preparation method according to claim 8.

10. The step of mixing the aqueous phase and the organic phase in step (3) further comprises mixing with starch. The preparation method according to claim 8.

11. Use of the composition for detecting occult blood in excrement according to any one of claims 1 to 7 in the manufacture of an occult blood detection device.

12. The occult blood detection device includes any one of a detection test paper, a detection plate, a reagent kit, and a detection tube.

12. The use according to claim 11.

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