REAGENT, TEST PAPER, REAGENT KIT, AND TEST PAPER KIT FOR DETECTING SULFHYDRYL COMPOUND, AND PREPARATION METHOD THEREOF

Abstract

A reagent, test paper, a reagent kit, and a test paper kit for detecting a sulfhydryl compound, and a preparation method thereof are provided. The reagent is obtained by mixing a phosphotungstic acid (PTA) reagent, an acetate buffer, and a shield reagent in a specified ratio. The test paper is composed of porous water-absorbent paper and a dry detection reagent dispersed thereon. The reagent is based on principle innovation. When in use, the reagent is directly mixed with urine to be tested in a volume ratio of 2:1, a resulting mixture is allowed to stand for 10 min to 15 min, and then a result can be directly determined by naked eyes. The reagent overcomes the shortcoming of requiring a control sample to assist in the determination of a result in existing methods, and has the advantages of simple operation, safety and non-toxicity, and rapid detection.

Claims

1. A sulfhydryl compound detection reagent, wherein the sulfhydryl compound detection reagent is composed of a phosphotungstic acid (PTA) reagent, an acetate buffer, and a shield reagent; and the shield reagent is oxidative, transparent, and colorless, and the shield reagent remains colorless after being reduced under specified conditions.

2. The sulfhydryl compound detection reagent according to claim 1, wherein the shield reagent is an aqueous solution of sodium iodate or an aqueous solution of sodium periodate.

3. The sulfhydryl compound detection reagent according to claim 2, wherein the shield reagent is the aqueous solution of sodium periodate.

4. The sulfhydryl compound detection reagent according to claim 1, wherein a preparation method of the PTA reagent is as follows: weighing 5±0.5 g of sodium tungstate, dissolving the sodium tungstate in 40±5 ml of deionized water, and adding 4±0.5 ml of 85±0.5% concentrated phosphoric acid and zeolite to obtain a resulting mixture; heating the resulting mixture to reflux, and continuously heating the resulting mixture under reflux for 2±0.5 h; and stopping heating, cooling to room temperature, diluting the resulting mixture to 100±0.5 ml to obtain a first resulting product, and storing the first resulting product in a first brown reagent bottle.

5. The sulfhydryl compound detection reagent according to claim 4, wherein a preparation method of the acetate buffer is as follows: preparing a 2±0.5 M sodium acetate solution and a 2±0.5 M acetic acid solution separately, and mixing the 2±0.5 M sodium acetate solution and the 2±0.5 M acetic acid solution in a volume ratio of 5:1.

6. The sulfhydryl compound detection reagent according to claim 5, wherein a preparation method of the shield reagent is as follows: weighing 25 mg to 100 mg of a shield agent, dissolving the shield agent in 100±0.5 ml of deionized water to obtain a second resulting product, and storing the second resulting product in a second brown reagent bottle.

7. The sulfhydryl compound detection reagent according to claim 6, wherein the preparation method of the shield reagent is as follows: weighing 50 mg to 100 mg of the shield agent, dissolving the shield agent in 100±0.5 ml of deionized water to obtain the second resulting product, and storing the second resulting product in the second brown reagent bottle.

8. The sulfhydryl compound detection reagent according to claim 6, wherein the sulfhydryl compound detection reagent is obtained by thoroughly mixing the PTA reagent, the acetate buffer, and the shield reagent in a volume ratio of 2:1:1.

9. A preparation method of a sulfhydryl compound detection reagent, comprising the following steps: S1: weighing 5±0.5 g of sodium tungstate, dissolving the sodium tungstate in 40±5 ml of deionized water, and adding 4±0.5 ml of 85±0.5% concentrated phosphoric acid and zeolite to obtain a resulting mixture; heating the resulting mixture to reflux, and continuously heating the resulting mixture under reflux for 2±0.5 h; and stopping heating, cooling to room temperature, diluting the resulting mixture to 100±0.5 ml to obtain a PTA reagent, and storing the PTA reagent in a first brown reagent bottle; S2: preparing a 2±0.5 M sodium acetate solution and a 2±0.5 M acetic acid solution separately, and mixing the 2±0.5 M sodium acetate solution and the 2±0.5 M acetic acid solution in a volume ratio of 5:1 to obtain an acetate buffer; S3: weighing 50 mg to 100 mg of a shield agent, dissolving the shield agent in 100±0.5 ml of deionized water to obtain a shield reagent, and storing the shield reagent in a second brown reagent bottle; and S4: thoroughly mixing the PTA reagent, the acetate buffer, and the shield reagent in a volume ratio of 2:1:1.

10. A sulfhydryl compound detection reagent kit, comprising a sampler, a test reagent, and a test tube, wherein the test reagent is the sulfhydryl compound detection reagent according to claim 1.

11. A urine sulfhydryl compound detection test paper, wherein the urine sulfhydryl compound detection test paper is composed of a porous water-absorbent paper and a dry detection reagent dispersed on the porous water-absorbent paper, and the dry detection reagent is the sulfhydryl compound detection reagent according to claim 1.

12. A preparation method of a urine sulfhydryl compound detection test paper, comprising the following steps: step 1: a preparation of a detection reagent comprising: S1: weighing 5±0.5 g of sodium tungstate, dissolving the sodium tungstate in 40±5 ml of deionized water, and adding 4±0.5 ml of 85±0.5% concentrated phosphoric acid and zeolite to obtain a resulting mixture; heating the resulting mixture to reflux, and continuously heating the resulting mixture under reflux for 2±0.5 h; and stopping heating, cooling to room temperature, diluting the resulting mixture to 100±0.5 ml to obtain a PTA reagent, and storing the PTA reagent in a first brown reagent bottle; S2: preparing a 2±0.5 M sodium acetate solution and a 2±0.5 M acetic acid solution separately, and mixing the 2±0.5 M sodium acetate solution and the 2±0.5 M acetic acid solution in a volume ratio of 5:1 to obtain an acetate buffer; S3: weighing 50 mg to 100 mg of a shield agent, dissolving the shield agent in 100±0.5 ml of deionized water to obtain a shield reagent, and storing the shield reagent in a second brown reagent bottle; and S4: thoroughly mixing the PTA reagent, the acetate buffer, and the shield reagent in a volume ratio of 2:1:0.5; step 2: immersing a porous water-absorbent paper in the detection reagent; and after the porous water-absorbent paper fully absorbs the detection reagent, taking the porous water-absorbent paper out, and draining and drying the porous water-absorbent paper to obtain the urine sulfhydryl compound detection test paper.

13. The preparation method of the urine sulfhydryl compound detection test paper according to claim 12, wherein the porous water-absorbent paper is a filter paper; the porous water-absorbent paper is immersed at room temperature for no less than 5 seconds; and the porous water-absorbent paper is vacuum-dried at 40° C. to 60° C.

14. The preparation method of the urine sulfhydryl compound detection test paper according to claim 13, wherein when the porous water-absorbent paper is immersed, an ultrasonic-assisted treatment is conducted for no less than 10 seconds.

15. A sulfhydryl compound detection test paper kit, comprising a urine sampler, a detection test paper, and a desiccant, wherein the detection test paper is the urine sulfhydryl compound detection test paper according to claim 11.

16. The sulfhydryl compound detection reagent kit according to claim 10, wherein the shield reagent is an aqueous solution of sodium iodate or an aqueous solution of sodium periodate.

17. The sulfhydryl compound detection reagent kit according to claim 16, wherein the shield reagent is the aqueous solution of sodium periodate.

18. The sulfhydryl compound detection reagent kit according to claim 10, wherein a preparation method of the PTA reagent is as follows: weighing 5±0.5 g of sodium tungstate, dissolving the sodium tungstate in 40±5 ml of deionized water, and adding 4±0.5 ml of 85±0.5% concentrated phosphoric acid and zeolite to obtain a resulting mixture; heating the resulting mixture to reflux, and continuously heating the resulting mixture under reflux for 2±0.5 h; and stopping heating, cooling to room temperature, diluting the resulting mixture to 100±0.5 ml to obtain a first resulting product, and storing the first resulting product in a first brown reagent bottle.

19. The sulfhydryl compound detection reagent kit according to claim 18, wherein a preparation method of the acetate buffer is as follows: preparing a 2±0.5 M sodium acetate solution and a 2±0.5 M acetic acid solution separately, and mixing the 2±0.5 M sodium acetate solution and the 2±0.5 M acetic acid solution in a volume ratio of 5:1.

20. The sulfhydryl compound detection reagent kit according to claim 19, wherein a preparation method of the shield reagent is as follows: weighing 25 mg to 100 mg of a shield agent, dissolving the shield agent in 100±0.5 ml of deionized water to obtain a second resulting product, and storing the second resulting product in a second brown reagent bottle.

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0063] Based on the principles of the present disclosure, the technical solutions of the present disclosure are further described below through specific examples. All reagents used in the examples are purchased from the market, and are analytically pure.

[0064] Specific preparation steps of the sulfhydryl compound detection reagent provided by the present disclosure are as follows:

[0065] S1: preparation of a PTA reagent: weighing 5 g of sodium tungstate, dissolving the sodium tungstate in 40 ml of deionized water, and adding 4 ml of 85% concentrated phosphoric acid and zeolite; heating a resulting mixture to reflux, starting timing, and continuously heating under reflux for 2 h; and stopping heating, cooling to room temperature, diluting to 100 ml, and storing a resulting PTA reagent in a brown reagent bottle;

[0066] S2: preparation of an acetate buffer: preparing a 2 M sodium acetate solution and a 2 M acetic acid solution separately, and mixing the two solutions in a volume ratio of 5:1 to obtain the acetate buffer;

[0067] S3: preparation of a shield reagent: weighing 25 mg to 100 mg (preferably 50 mg to 100 mg) of a shield agent, dissolving the shield agent in 100 ml of deionized water, and storing a resulting shield reagent in a brown reagent bottle; and

[0068] S4: preparation of the sulfhydryl compound detection reagent: thoroughly mixing the PTA reagent, the acetate buffer, and the shield reagent in a volume ratio of 2:1:1.

[0069] The present disclosure is used for detection as follows: urine and a sulfhydryl compound detection reagent are mixed in a volume ratio of 1:2 and a resulting mixture is allowed to stand for 10 min to 15 min, where if the mixture turns blue obviously, it indicates a positive result; otherwise it indicates a negative result.

[0070] As described in S3, the shield agent is an oxidant soluble in water; when the shield agent is dissolved, a resulting solution is transparent and colorless, and remains colorless after the shield agent is reduced under specified conditions; and preferably the shield agent is sodium iodate or sodium periodate.

[0071] In order to make the present disclosure clearer and more comprehensible, the present disclosure will be further described in detail below with reference to the examples. It should be understood that the examples described herein are merely intended to explain the present disclosure, rather than to limit the present disclosure.

Example 1

Preparation of a Sulfhydryl Compound Detection Reagent

[0072] preparation of a PTA reagent: 5 g of sodium tungstate was weighed and dissolved in 40 ml of deionized water, and 4 ml of 85% concentrated phosphoric acid and zeolite were added; a resulting mixture was heated to reflux, timing was started, and the mixture was continuously heated under reflux for 2 h; and the heating was stopped, and a resulting product was cooled to room temperature, diluted to 100 ml, and stored in a brown reagent bottle;

[0073] preparation of an acetate buffer: a 2 M sodium acetate solution and a 2 M acetic acid solution were prepared separately, and mixed in a volume ratio of 5:1;

[0074] preparation of a shield reagent: 50 mg of sodium iodate was weighed and dissolved in 100 ml of deionized water, and a resulting product was stored in a brown reagent bottle; and

[0075] preparation of the sulfhydryl compound detection reagent: the PTA reagent, the acetate buffer, and the shield reagent were thoroughly mixed in a volume ratio of 2:1:1.

Example 2

Preparation of a Sulfhydryl Compound Detection Reagent

[0076] preparation of a shield reagent: 100 mg of sodium iodate was weighed and dissolved in 100 ml of deionized water, and a resulting product was stored in a brown reagent bottle; and the rest of the steps were the same as those in Example 1.

Test Example 1

[0077] The effect of the detection reagent of the present disclosure was tested on simulated urine samples below.

[0078] Preparation of simulated urine sample 1: 500 mg of L-cysteine and 7 mg of vitamin C were weighed and dissolved in 100 ml of water.

[0079] Preparation of simulated urine sample 2: 7 mg of vitamin C was weighed and dissolved in 100 ml of water.

[0080] 100 μl of the simulated urine sample 1 was taken and mixed with 200 μl of the detection reagent in Example 1, a resulting mixture was allowed to stand for 10 min to 15 min, and then the mixture turned blue, indicating a positive result. 100 μl of the simulated urine sample 2 was taken and mixed with 200 μl of the detection reagent in Example 1, a resulting mixture was allowed to stand for 10 min to 15 min, and the color of the mixture did not change, indicating a negative result.

Test Example 2

[0081] The effect of the detection reagent of the present disclosure was tested on normal human urine samples below.

[0082] Normal human urine sample 1: urine excreted for the first time after breakfast was collected and cooled to room temperature by standing.

[0083] Normal human urine sample 2: 25 ml of the normal human urine sample 1 was taken, and 125 mg of L-cysteine was added and dissolved.

[0084] Preparation of a PTA reagent without a shield reagent: the PTA reagent in Example 1 and the acetate buffer were thoroughly mixed in a volume ratio of 2:1.

[0085] 100 μl of the normal human urine sample 1 was taken and mixed with 200 μl of the detection reagent in Example 2, a resulting mixture was allowed to stand for 10 min to 15 min, and a color of the mixture did not change, indicating a negative result; 100 μl of the simulated urine sample 2 was taken and mixed with 200 μl of the detection reagent in Example 2, a resulting mixture was allowed to stand for 10 min to 15 min, and then a color of the mixture turned blue, indicating a positive result; and 100 μl of the normal human urine sample 1 was taken and mixed with 200 μl of the PTA reagent without a shield reagent, a resulting mixture was allowed to stand for 10 min to 15 min, and then a color of the mixture turned blue, indicating a false positive result.

[0086] The preparation of the sulfhydryl compound detection test paper of the present disclosure is described below with reference to some examples:

[0087] Specific preparation steps of the sulfhydryl compound detection test paper provided by the present disclosure were as follows:

[0088] S1: preparation of a PTA reagent: 5 g of sodium tungstate was weighed and dissolved in 40 ml of deionized water, and 4 ml of 85% concentrated phosphoric acid and zeolite were added; a resulting mixture was heated to reflux, timing was started, and the mixture was continuously heated under reflux for 2 h; and heating was stopped, a resulting product was cooled to room temperature, diluted to 100 ml, and stored in a brown reagent bottle;

[0089] S2: preparation of an acetate buffer: a 2 M sodium acetate solution and a 2 M acetic acid solution were prepared separately and mixed in a volume ratio of 5:1;

[0090] S3: preparation of a shield reagent: 25 mg to 100 mg (preferably 50 mg to 100 mg) of a shield agent was weighed and dissolved in 100 ml of deionized water, and a resulting product was stored in a brown reagent bottle;

[0091] S4: preparation of the sulfhydryl compound detection reagent: the PTA reagent, the acetate buffer, and the shield reagent were thoroughly mixed in a volume ratio of 2:1:0.5; and

[0092] filter paper was immersed in the prepared urine sulfhydryl compound detection reagent, and after the filter paper fully absorbed the detection reagent, the filter paper was taken out, drained, and dried at 40° C. under vacuum conditions.

[0093] Urine test: a specified amount of urine was taken and added dropwise on the test paper, and then the test paper stood for air-drying and observed, wherein if a color of the test paper turned blue, it indicated a positive result, otherwise it indicated a negative result.

[0094] In order to make the present disclosure clearer and more comprehensible, the present disclosure will be further described in detail below with reference to the examples. It should be understood that the examples described herein are merely intended to explain the present disclosure, rather than to limit the present disclosure.

Example 3

Preparation of Sulfhydryl Compound Detection Test Paper

[0095] preparation of a PTA reagent: 5 g of sodium tungstate was weighed and dissolved in 40 ml of deionized water, and 4 ml of 85% concentrated phosphoric acid and zeolite were added; a resulting mixture was heated to reflux, timing was started, and the mixture was continuously heated under reflux for 2 h; and the heating was stopped, and a resulting product was cooled to room temperature, diluted to 100 ml, and stored in a brown reagent bottle;

[0096] preparation of an acetate buffer: a 2 M sodium acetate solution and a 2 M acetic acid solution were prepared separately, and mixed in a volume ratio of 5:1;

[0097] preparation of a shield reagent: 50 mg of sodium iodate was weighed and dissolved in 100 ml of deionized water, and a resulting product was stored in a brown reagent bottle;

[0098] preparation of the sulfhydryl compound detection reagent: the PTA reagent, the acetate buffer, and the shield reagent were thoroughly mixed in a volume ratio of 2:1:0.5; and

[0099] filter paper was immersed in the prepared urine sulfhydryl compound detection reagent, an ultrasonic-assisted treatment was conducted for 10 s, and after the filter paper fully absorbed the detection reagent, the filter paper was taken out, drained, and dried at 40° C. under vacuum conditions.

Example 4

Preparation of Sulfhydryl Compound Detection Test Paper

[0100] preparation of a shield reagent: 100 mg of sodium iodate was weighed and dissolved in 100 ml of deionized water, and a resulting product was stored in a brown reagent bottle;

[0101] filter paper was immersed in the prepared urine sulfhydryl compound detection reagent, and after the filter paper fully absorbed the detection reagent, the filter paper was taken out, drained, and dried at 60° C. under vacuum conditions; and

[0102] the rest of the steps were the same as those in Example 3.

Test Example 3

[0103] The effect of the detection reagent of the present disclosure was tested on simulated urine samples below.

[0104] Preparation of simulated urine sample 1: 500 mg of L-cysteine and 7 mg of vitamin C were weighed and dissolved in 100 ml of water.

[0105] Preparation of simulated urine sample 2: 7 mg of vitamin C was weighed and dissolved in 100 ml of water.

[0106] The simulated urine sample 1 was taken and added dropwise on the test paper prepared in Example 3, the test paper stood for air-drying, and then the test paper turned blue, indicating a positive result; and the simulated urine sample 2 was taken and added dropwise on the test paper prepared in Example 3, the test paper stood for air-drying, and then a color of the test paper did not change, indicating a negative result.

Test Example 4

[0107] The effect of the detection reagent of the present disclosure was tested on normal human urine samples below.

[0108] Normal human urine sample 1; urine excreted for the first time after breakfast was collected and cooled to room temperature by standing.

[0109] Normal human urine sample 2: 25 ml of the normal human urine sample 1 was taken, and 125 mg of L-cysteine was added and dissolved.

[0110] Preparation of a PTA reagent without a shield reagent: the PTA reagent in Example 1 and the acetate buffer were thoroughly mixed in a volume ratio of 2:1. Then filter paper was immersed in the reagent, after the filter paper fully absorbed the reagent, the filter paper was taken out, drained, and dried at 60° C.

[0111] The normal human urine sample 1 was taken and added dropwise on the test paper prepared in Example 4, the test paper stood for air-drying, and then the test paper turned slightly yellow, indicating a negative result, the normal human urine sample 2 was taken and added dropwise on the test paper prepared in Example 4, the test paper stood for air-drying, and then the test paper turned blue, indicating a positive result; and the normal human urine sample 1 was taken and added dropwise on the PTA test paper without a shield reagent, the test paper stood for air-drying, and then the test paper turned blue, indicating a false positive result.

[0112] Based on the reagent, test paper, reagent kit, and test paper kit for detecting a sulfhydryl compound of the present disclosure, when a positive test result is preliminarily obtained, in order to allow a self-diagnosed patient to further understand an inducement of their HPV infection before seeking medical treatment and facilitate clinicians to understand clinical symptoms and signs, the present disclosure also designs the following self-diagnosis outline and treatment and prevention measures for the reference of self-diagnosed patients and physicians:

[0113] I. Incentive: Do you have any of the following conditions before feeling discomfort?

[0114] 1. Your spouse has been infected.

[0115] 2. In the past 3 to 6 months, you have had multiple sexual partners or your sexual partner has had multiple sexual partners.

[0116] 3. Whether you have recently suffered from gonorrhea, syphilis, chlamydia infection, trichomonal vaginitis, and the like.

[0117] 4. Do you wear short shorts and go out to public places in summer and do you wash your hands before going to the toilet?

[0118] 5. Do you use public bathtubs, bedpans, bath towels, and bathrobes?

[0119] II. Clinical symptoms: Where do you feel uncomfortable?

[0120] 1. You are not uncomfortable.

[0121] 2. You feel genital itching.

[0122] 3. Your leucorrhea increases.

[0123] 4. You feel genital burning pain.

[0124] III. Clinical signs: What did you see or touch?

[0125] 1. Growth parts: labia majora, labia minora, clitoris, vagina, cervix, or perianal area, two of which commonly have papules at the same time.

[0126] 2. Local manifestations: small reddish or gray papules that are verrucous protrusions arranged in clusters or fused into cockscombs, cauliflower-like vegetations, and increased fragility of skin lesions causing bleeding.

[0127] IV. Laboratory examination: Self-test

[0128] 1. A urine sulfhydryl test is conducted, and a positive result is obtained.

[0129] 2. An acetowhite test is conducted as follows: a gauze soaked in a 3% to 5% acetic acid solution is wrapped around or applied to a suspicious skin or mucous membrane surface, and removed 3 min to 5 min later. Typical condyloma acuminatum lesions will be manifested as white papules or warts, and subclinical infection will be manifested as white patches or spots.

[0130] The acetowhite test is a simple and easy detection method to identify early condyloma acuminatum lesions and subclinical infection.

[0131] V: Treatment

[0132] 1. If HPV infection is discovered, you go to the hospital for confirmed diagnosis.

[0133] 2. If HPV infection is suspected, you go to the hospital for confirmed diagnosis.

[0134] VI: Prevention

[0135] 1. Sexual partners are fixed to each other and pay attention to sexual hygiene.

[0136] 2. Condoms can reduce the probability of obvious HPV infection.

[0137] 3. In summer, you should wear skirts or pants that are long enough to reach knee joints before going to public places.

[0138] 4. You should wash your hands thoroughly before going to the toilet, and do not directly contact the toilet door with your hands (20% of people infected with HPV may carry the virus on their fingers).

[0139] The above examples are merely preferred solutions of the present disclosure and are not intended to limit the present disclosure in any form, and other variations and modifications may be made without departing from the technical solutions of the present disclosure as set forth in the appended claims.