Composition for detecting occult blood in feces, its preparation method and use

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 relates to the field of disease detection, and in particular to a composition for detecting occult blood in feces, its preparation method and use, and in particular to a composition for detecting occult blood in feces with high sensitivity, its preparation method and use. [Background technology]

[0002] Blood in the excrement is a common pathological symptom. The causes that may cause blood in the stool include, but are not limited to, hemorrhoids, ulcerative colitis, upper gastrointestinal bleeding, lower gastrointestinal bleeding, blood system disease, anal rupture, gastrointestinal tumors, etc. The causes that may cause blood in the urine include, but are not limited to, urinary mucosal damage, urinary stones, nephritis, urinary tumors, etc. The bleeding in the digestive and urinary systems caused by the above pathologies includes two cases: visible blood and occult blood. Blood visible to the naked eye in the excrement is easy to attract attention, but the initial symptom of most diseases is occult blood in the excrement. When there is visible blood in the excrement, the pathology may already be severe, so regular occult blood detection can play an early screening effect on diseases of the urinary and digestive systems.

[0003] CN114354948A discloses a semi-quantitative method for detecting fecal occult blood, the steps of which are as follows: the diluted sample is dropped into the spotting hole on the gold colloid detection card by a spotting device, and after the qualitative negative and positive results appear initially, a photograph is taken, and the qualitative result is read on the instrument 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, and then a certain amount of hydrogen peroxide solution is dropped, and after the 3,3,5,5-tetramethylbenzidine indicator solution and the hydrogen peroxide solution are dropped and reacted for a while, the gold colloid detection card shows the semi-quantitative result, and a photograph is taken again to read the semi-quantitative result. This invention increases the semi-quantitative result of fecal occult blood detection, reduces manual operations, reduces the amount of manual work, solves the defects of fecal occult blood detection in clinical testing, greatly reduces the work of manual retesting, and ensures the accuracy of the detection result.

[0004] CN103175965A discloses a fecal occult blood detection kit, which includes a fecal occult blood detection test paper strip or test paper card, a sample processing tube or a stool collection mixer and a sampling rod, and the test paper strip or test paper 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 above-mentioned reagent kit includes steps such as compounding the processing solution, manufacturing the reagent kit, preparing the gold colloid complex, spraying metal, cutting, assembling, cutting, packaging, etc. The fecal occult blood detection kit has good specificity, high sensitivity, rapid detection, simple, and no dietary or drug restrictions.

[0005] Detection of occult blood can play an important role in early screening for diseases of the urinary and digestive systems. Therefore, how to provide a method or reagent for detecting occult blood that is easy to detect and has high sensitivity is an issue that needs to be solved immediately. 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 excrement, its preparation method and use, and in particular, a composition for detecting occult blood in excrement with high sensitivity, its preparation method and use. The composition of the present application can quickly and effectively detect the occult blood condition in excrement, has a fast color change speed, 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, the composition comprises 20-80 parts of an organic solvent, 0.01-50 parts of a thickener, 0.01-2 parts of benzidine, 0.01-5 parts of peroxide, 0.001-0.5 parts of EDTA-2Na, and 10-80 parts of water; A composition for detecting occult blood in fecal matter 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., 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 above 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, 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 combinations listed above, and other combinations not listed above within the above combination range 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, such as 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 combinations listed above, 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, or 3,3'-dihydroxybenzidine, such as 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 combinations listed above, 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 specific benzidines as 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 combinations listed above, and other combinations not listed above within the above combination range also apply.

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

[0022] In one embodiment, the organic peroxide includes any one or a combination of at least two of cumene hydroperoxide, diisopropylbenzene hydroperoxide, t-amyl peroxy-2-ethylhexanoate, or t-butyl peroxy-2-ethylhexanoate, such as a combination of cumene hydroperoxide and diisopropylbenzene hydroperoxide, a combination of diisopropylbenzene hydroperoxide and t-amyl peroxy-2-ethylhexanoate, or a combination of t-amyl peroxy-2-ethylhexanoate and t-butyl peroxy-2-ethylhexanoate, but is not limited to the above-listed combinations, and other combinations not listed within the above combination 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 speed, extend the color retention time, and further improve the detection sensitivity.

[0025] In one embodiment, the composition further comprises, by weight, 0.01 to 5 parts of a pH adjuster, for example, 0.01 parts, 0.1 parts, 0.5 parts, 1 parts, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts, or 5 parts, etc., but is not limited to the numerical values ​​recited above, and other unrecited numerical values ​​within the above numerical ranges 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, by weight, 1 to 80 parts of starch, such as, 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 recited values, and other unrecited values ​​within the above numerical ranges also apply.

[0028] In one embodiment, the starch includes any one or at least two combinations of corn starch, pea starch, mung bean starch, gelatinized starch, methylol starch, crosslinked 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 combinations listed above, and other combinations not listed above within the above combination range are also applicable.

[0029] The starch can be added according to the needs of the user, and the composition can be made from liquid to solid to be applicable to different scenes.

[0030] In a second aspect, the present embodiment provides a method for preparing the composition for detecting occult blood in fecal matter described above, the method comprising: (1) mixing benzidine, peroxide and an organic solvent to obtain an organic phase; (2) mixing a thickener, EDTA-2Na, and water to obtain an aqueous phase; (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.

[0031] In one embodiment, the step of mixing the thickening agent, 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 fecal waste as 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. Effect of the Invention

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

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

[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 the 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 mixture was 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 ethanol, 40 parts guar gum, 1.5 parts 4,4'-diaminobiphenyl, 4 parts hydrogen peroxide, 0.3 parts EDTA-2Na, 60 parts water, and 5 parts 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 mixture was 20 parts of ethanol, 0.01 parts of pectin, 0.01 parts of tetramethylbenzidine, 0.01 parts of cumene hydroperoxide, 0.001 parts of EDTA-2Na, 10 parts of water, and 0.1 parts 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 was not added and the reduction was 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 was not added and the reduction was 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 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 in Example 1, except that no hydrogen peroxide was added and the reduced 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 the same as 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 otherwise identical to 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 are identical to those of Example 1, except that they do not contain ethanol and the reduction is 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 are identical to those of Example 1, except that they do not contain acetone and the reduction is 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 are identical to those of Example 1, except that they do not contain ethanol and the reduction is 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 paper, then dried to obtain a detection test paper, and then a sample (a normal human fresh blood diluted 10,000 times with water) was used for detection, and the color change time and color retention time were recorded. The results are as follows:

[0072] [Table 1]

[0073] From the above data, it can be seen that the product according to the present application has the advantages of short discoloration time and long color retention time, and samples can be detected quickly. Comparing Examples 1 to 5, it can be seen that the present application can effectively improve the effect of the product by controlling the amount of each raw material used. Comparing Examples 1, 6 to 8, it can be seen that the present application can effectively shorten the discoloration time and extend the color retention time by selecting a specific combination of benzidines. Comparing Examples 1, 9 to 12, it can be seen 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, 13 to 15, it can be seen that the present application can effectively shorten the discoloration time and extend the color retention time by selecting a specific combination of organic solvents.

[0074] Then, the above test paper was used to detect the test samples with different hemoglobin concentrations, and the detection limit of the test paper was analyzed. The results are as follows:

[0075] [Table 2]

[0076] From the above data, it was found that the composition according to the present application has the characteristic of high sensitivity and can effectively detect hemoglobin with an extremely low content. Comparing Examples 1 to 5, it was found that the present application can effectively improve the effect 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 the 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 the 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 the 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 raw material 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 contents 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 are 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 may be combined in any suitable manner unless there is a contradiction, and in order to avoid unnecessary duplication, the present application does not specifically describe various possible combinations.

Claims

1. 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.

2. The composition comprises, in parts by weight, 40-60 parts of organic solvent, 10-40 parts of thickener, 0.5-1.5 parts of benzidine, 1-4 parts of peroxide, 0.1-0.3 parts of EDTA-2Na, and 30-60 parts of water; The organic solvent includes any one or a combination of at least two of ethanol, benzene, toluene, pentane, hexane, methanol, diethyl ether, ethyl acetate, acetone, or carbon tetrachloride; The composition of claim 1.

3. The organic solvent includes a combination of ethanol and acetone. The composition according to claim 1 or 2.

4. The thickening agent may include 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 according to any one of claims 1 to 3.

5. The benzidine includes any one or a combination of at least two of 4,4'-diaminobiphenyl, tetramethylbenzidine, and 3,3'-dihydroxybenzidine. The composition according to any one of claims 1 to 4.

6. The benzidine includes a combination of 3,3'-dihydroxybenzidine and tetramethylbenzidine. The composition according to any one of claims 1 to 5.

7. The peroxide includes inorganic peroxides and / or organic peroxides. The composition according to any one of claims 1 to 6.

8. The peroxide includes a combination of inorganic and organic peroxides. The composition according to any one of claims 1 to 7.

9. The inorganic peroxide includes any one or a combination of at least two of hydrogen peroxide, sodium percarbonate, potassium peroxide, calcium peroxide, or barium peroxide; 9. The composition according to claim 7 or 8.

10. The inorganic peroxide comprises hydrogen peroxide; The composition according to any one of claims 7 to 9.

11. The organic peroxide includes any one or a combination of at least two of cumene hydroperoxide, diisopropylbenzene hydroperoxide, t-amylperoxy-2-ethylhexanoate, and t-butylperoxy-2-ethylhexanoate; The composition according to any one of claims 7 to 9.

12. The organic peroxide includes cumene hydroperoxide. The composition according to any one of claims 7 to 9 and 11.

13. The composition further comprises, by weight, 0.01 to 5 parts of a pH adjuster; Preferably, the pH adjuster comprises any one of citric acid, acetic acid, sodium hydroxide, and hydrochloric acid; Preferably, the composition further comprises, by weight, 1 to 80 parts starch; Preferably, the starch comprises any one or a combination of at least two of corn starch, pea starch, mung bean starch, gelatinized starch, methylol starch, crosslinked starch, maltodextrin, oxidized starch, esterified starch, or etherified starch; The composition according to any one of claims 1 to 12.

14. A method for preparing a composition for detecting occult blood in feces according to any one of claims 1 to 13, comprising the steps of: (1) mixing benzidine, peroxide and an organic solvent to obtain an organic phase; (2) mixing a thickening agent, EDTA-2Na and water to obtain an aqueous phase; (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.

15. The step of mixing the thickening agent, EDTA-2Na, and water to obtain an aqueous phase in step (2) further comprises mixing with a pH adjuster; Preferably, the step of combining the aqueous phase and the organic phase in step (3) further comprises combining with starch.

15. The method of claim 14.

16. Use of the composition for detecting occult blood in excrement according to any one of claims 1 to 13 in the manufacture of an occult blood detection device.

17. The occult blood detection device includes any one of a detection test paper, a detection plate, a reagent kit, and a detection tube; 17. The use according to claim 16.

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