TW393578B - A test device and method for the determination of protein in a biological sample - Google Patents

Description

Among them: η is defined as the number of repeated polyethers, which is an integer between 4 and 15, preferably between 4 and 1 1; m is defined as the number of repeated polycarbonates, which is an integer between 1 and 1, preferably between 1 and 7, preferably between 1 and 5. The starting material polyethylene bond L 950 is a difunctional polypropylene glycol having an average molecular weight of about 420 ', which is prepared by polymerizing propylene oxide in the presence of 12-propylene glycol. Each diol residue gives rise to a polymer chain, resulting in a linear polymer ′ whose ends are terminated with secondary radicals (that is, ′ has two functional radicals). The polyether-polycarbonate material obtained from the connection of polyether residues to carbonates has the following ratio: HI- In n ^ il It ^ 〆ei mu mt ^ —HI— \ OJ / St (Please read the precautions on the back first Refill this page) Polyether L 950 Diphenyl Carbonate Ti (OC4H9) 4 amt (g) 4248 1581 0.6 amt (Moore) 10.22 7.39 0.0018 The consumption cooperation of employees of the Central Standards Bureau of the Ministry of Economic Affairs has been found to restrict aliphatics. The amount of polyether-polycarbonate material to 10% by weight or less is important for reducing the specific gravity effect. Regarding the reduction of the filter paper effect, it has been found suitable to reduce the amount of the aliphatic polyether polycarbonate material to 2% by weight or less. The following examples are shown to disclose the preferred specific examples and the application of the present invention, which does not mean to limit the present invention, unless otherwise stated in the scope of the attached patent application. ) -Τστ Consumers' Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs

The scope of the Ming 2 invention of the method for protein determination, in particular, the use of ^ ^ means π agent, buffer sword, and-a kind of aliphatic polycarbonate (its responsibility " 7 or less than ten percent by weight) Novel compositions and methods for measuring proteins in biological samples. The description of the rabbit scene technique The determination of the presence of proteins in biological samples is extremely important in diagnosing several pathological conditions affecting the kidneys, the circulatory system and the central nervous system. Therefore, quantitative and / or qualitative measurement of urine protein (albumin) is often required. This is especially important for the diagnosis of sugar: and kidney disease. The main protein in rheumatic urine measurement is albumin; therefore the model system for the proteinuria test is albumin. Methods for determining the presence of albumin in urine are well known. The most convenient method for measuring albumin is to wet the test strip impregnated with a protein error indicator with > urine. If albumin is present in the urine sample, the test strip will simply change its color display. The color that appears usually depends on the albumin concentration in the sample. Use this variety of color changes to quantify albumin in the sample. Although the above types of test strips are a convenient and fast tool for determining protein at the staining point, one of the problems that has occurred is the so-called gravity effect. The general urine protein test uses a pH indicator dye, which shows a pKa shift when it is complexed with a protein (the "protein error of the pH indicator,"). This method does not provide a sufficient distinction between negative urine and protein-containing urine (approximately 15 micrograms per kilogram), and gives false positive readings in urine with a high specific gravity. It has been found that the addition of certain polycarbonate compounds to the protein f error indicator reduces the initial reactivity of the reagent paper and reduces the

This paper size applies the Chinese national standard (CNS > Λ4 specification (210X29 ^ J " J • 4-A7 B7) Ministry of Economic Affairs t Central Standards Bureau Staff Consumer Cooperative Cooperative Imprint V. Invention Description (2 It is obtained by the tendency to smoke urine samples The positive reading of protein. Another occurrence of this device structure is that if it is an absorbent material, it is usually a piece of paper, and the reagents used in the system are different. There is a constant (ie, uniform) Anti-reversibility. Filter paper often reacts with the protein required for the determination of ^ 'and tincture X, to the extent that the same carrier substrate material reacts with the composition required to measure other components. The problem of adjusting the formulation to obtain a uniform reactivity between the carrier substrate materials used in the reagent composition Xiao is already in this category-a long-standing problem and of great significance in this industry-. At first, Xianxin adjusted The pH of the reagent composition will overcome the problem of reaction variability between the reagent composition and the carrier substrate material. However, adjusting the pH is not effective for all batches of the carrier substrate material. The required results do not achieve one Reagent composition, this composition will cause a lighter negative reaction color and a darker positive reaction color. Triton X-mail, a polyethylene surfactant, was found to have a large effect on reactivity, but Both the negative reaction and the positive reaction end of the color range produce a lighter color. Explore other ways to adjust the reactivity. For example, change all the components of the reagent composition, but this change has not been found to effectively produce a lighter Negative reaction color and darker positive reaction color. Polyethanol was also introduced into the reagent composition and found no reason in controlling the reagent reaction characteristics of different batches of carrier substrate material. Therefore, even proper qualitative control, different batches of carrier substrate Inter-substance attempts to achieve sensible screening and invention to limit the reactivity problem still cannot be effectively controlled. (Please read the precautions on the back before filling this page}

This paper scale applies Chinese national standards (CNS > Λ4 specification (210 X 2W public delay) -5- Α7 Β7 I invention description (3) To overcome the above-mentioned problem of measuring protein reactivity, 'prepare a unique reagent composition' and Prepared by a method that eliminates the reaction variability between the reagent composition and the carrier substrate. In addition, the reduction in urine pH variability on the test results is another benefit seen with this reagent composition. Discussion The present invention provides an analytical test strip for detecting proteins in biological samples, which includes a phenol phthaloprotein error indicator, a buffer, and an aliphatic polycarbonate having an amount equal to or less than ten percent by weight. According to the present invention, the "secondary impregnation reagent composition step using an absorbent carrier" wherein the buffer solution and optionally a dye such as FD & C yellow # 5 and FD & C # 3 are first impregnated into the absorbent carrier from an aqueous solution. The phenol phthaloprotein error indicator and the aliphatic polycarbonate compound are immersed during the second impregnation with a non-aqueous solvent such as an alcohol solution. Another scope of the present invention refers to the detection of A method of analyzing protein in a sample, wherein a test strip is analyzed with a biological sample. The wet test strip contains an absorbent carrier impregnated with the reagent composition as described above, "observing the test strip to detect any color change, which is An indication of the presence and amount of protein in a biological sample. Description of preferred specific examples According to the present invention, it has been found that test strips for the determination of proteins in biological fluids can be prepared without any difference between the carrier substrates of different batches and between the carrier substrate and the reagent composition. Obvious reactivity problem. In addition, it has been found that the influence of urine variability on the detection limit, the visual detection and instrument detection are reduced by the reagent composition of the present invention. The present invention is immersed and absorbed by the reagent composition. The male base material is achieved. The absorptive size of this paper is applicable to China National Standards (CNS) Α4 specifications (210 × 297 mm) -β7 ---,-K ----- Ί-- (Please read the note on the back first (Please fill in this page again for item f.) Order printed by the Central Standards Bureau of the Ministry of Economic Affairs of the Shelling Consumer Cooperative, and print the printed bags of the Central Standards Bureau of the Ministry of Economic Affairs of the Shelling Consumer Cooperative. A7 B7 5-1 Description of the invention Paper Obtained from the Department of Pharmacy F 204, Inc. grade 204 »• It contains a mixture of wood and fiber pulp. The filter paper is different from batch to batch. Screening and invention limits have been ineffective in the past to ensure the maximum efficacy of the reagents. Others can be used as absorption Sexual carriers include felts, perforated ceramic strips, and woven glass fibers (described in US Project No. 3,846,247). Materials suitable for the carrier of test strips such as wood, cloth, sponge materials and clay materials (such as Described in U.S. Patent No. 3,552,928). However, it has been found that filter paper is particularly suitable despite differences between batches. The absorbent carrier substrate impregnates the reagent composition in the manner described above. It has been found that two impregnations are used for impregnation In step, the aqueous solution was immersed for the first time and the non-aqueous soluble back was immersed for the second time to obtain the best results. The absorbent strip is preferably buffered to adjust the pH to between about 1.5 and 4.5. The system is preferably adjusted to a pH of about 3.5, and most preferably about 3.7. However, the nature of the buffer system is not strictly specified. All well-known cushioning materials can be used. The buffer solution preferably includes citric acid and sodium citrate. If the buffer is greater than 9.3 g / dl (g per gram), the operation of the reagent will be a problem. If the buffer is less than 7.1 g / d 1. May be insufficient. A buffer equivalent of 8.7 g / dl. Is most appropriate. In addition to the buffer material, red and yellow dyes are added as needed during the first dip. Any red dye suitable for the reagent composition can be used. However, FD & C Red # 3 is a better material. A preferred yellow dye is FD & C Yellow # 5. Dyes cover urine development at negative reaction color concentrations. Other dyes, surfactants, etc. can also be added as an additional optional component during the first immersion. This paper size is applicable to China National Standard (CNS) Λ4 specification (21〇 × 2mm) (Please read the precautions on the back before filling this page). &Quot;-«I L- ··-· — — I IP · -'9 Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs A7 _ B7 V. Description of Invention (5) Generally, the dye can be used in excess of about 40 g / 100 liters and less than about 60 g / 100 liters. For tetrabromopyrene blue (TBPB), 50 g / 100 liters is most suitable. '' For the second immersion, use a non-aqueous solvent solution, preferably ethanol, to reduce evaporation. A conventional polyhalogenated phenolphthalein protein error indicator may be used as the indicator compound. Suitable substances include: 3,4,5,6, -tetrabromophenolphthalein (TBPSP); 3,, 3 "-dinitro-3,4,5,6, -tetrabromophenol wind phthalide (DNTB) ; 3,, 3 ”-dibromo-3, 4,5,6, -tetrabromophenolphthalein (DBTB); 3 ', 3” -diiodo-3,4,5,6, -tetrabromophenolsulfone Phthalo (〇 汀 6); 3 ', 3 ", 5', 5 " -tetranitro_3,4,5,6, -Sixi test 飒 Europe (-^ 丁 3); 3 ', 3 "- Dinitroso-5 ', 5 " --- bromofen-3,4,5,6, -tetrabromophthalide (DNoHB); 3,, 3 ,,-diiodo-5,5,5, -di Nitrosol-3, 4,5,6, _tetrabromophthalide (DIDNo); 3 ', 3 " -dinitro-5', 5 ", 3,4,5,6, -hexabromo Phenolphthalein (〇ΝΗΒ), · 3 '-Nitro-5', 5 "-Diiodo-3", 3, 4, 5, 6 · Pentabromopyridine (NDIpB); 3, 3 " -Diiododinitro-3, 455,6, _tetrabromofluorphthalide (DIDNTB); 3 ', 3 -dinitro-5' '5 "-dibromo-3,4,5,6,- Tetraiodofluorenphthalide (DNDBTI); 3 ', 3 ”, 5', 5 " -tetrakis-3,4,5,6, -tetrabromosulfone hydrazone (TCTB); 3 ', 3' ', 5 ', 5 " -Tetrabromo-3,4,5,6, _ Tetrahydrophenol sulfone phthalide (TBTI); 3', 3 ”, 5 ', 5- Si-Epan—3,4,5,6, -tetrabromofluorphthalide (butyl ΤΒ); (Please read the precautions on the back before filling in this page)

、 1T This paper size is applicable to the national standard (CNS) A4 specification (210X297 mm) ~-: A7 B7 V. Description of the invention (6), 6, _ tetrabromophthalmide 3, 3 "- Disulfonic acid-5,, 5, __ Digas-3,4,5 (DSDC); 'Qi Zheng (Please read the precautions on the back before filling this page) 3 ,, 3 " -Diphosphonic acid _5,, 5 " -Dibromophenol ~ 3 4) '4,5,6, -Tetrabromo (DSHB) ;,' Tetrabromophenol is preferred. The other essential component of the second impregnation reagent composition is an aliphatic polyether-polycarbonate material. It is prepared using a mixture of diphenyl carbonate and a difunctional hydroxy compound or a difunctional light compound. The description of polycarbonate, its molecular weight and the alcohol used in its preparation are listed in the table below. Table 1 Identification of the numbered alcohol compound_ Proportion molecular weight ΚΟΚ 9209 Tetraethylene glycol / 1 1795 Hexanediol-1, 6 1 ΚΟΚ 10,000 Four Glycol / 1 4187 Hexanediol-1, 6 1 ΚΟΚ 10,001 Triethylene glycol-1972 ΚΟΚ 10,002 Triethylene glycol / 1 1972 Tetraethanol ΚΟΚ 10,071 Polyether L-950 (Polypropylene glycol-1594 Employee Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs The last substance listed in Table 1 is particularly good. This polypropylene oxide carbonic acid polymer is catalyzed by polyether L 950® substance [Bayer AG] with titanium dioxide but with carbonic acid with the following external structure. Diphenyl ester condensed to obtain the paper size applicable to the Chinese National Standard (CNS) Λ4 specification (210X297 mm) -9-

Among them: η is defined as the number of repeated polyethers, which is an integer between 4 and 15, preferably between 4 and 1 1; m is defined as the number of repeated polycarbonates, which is an integer between 1 and 1, preferably between 1 and 7, preferably between 1 and 5. The starting material polyethylene bond L 950 is a difunctional polypropylene glycol having an average molecular weight of about 420 ', which is prepared by polymerizing propylene oxide in the presence of 12-propylene glycol. Each diol residue gives rise to a polymer chain, resulting in a linear polymer ′ whose ends are terminated with secondary radicals (that is, ′ has two functional radicals). The polyether-polycarbonate material obtained from the connection of polyether residues to carbonates has the following ratio: HI- In n ^ il It ^ 〆ei mu mt ^ —HI— \ OJ / St (Please read the precautions on the back first Refill this page) Polyether L 950 Diphenyl Carbonate Ti (OC4H9) 4 amt (g) 4248 1581 0.6 amt (Moore) 10.22 7.39 0.0018 The consumption cooperation of employees of the Central Standards Bureau of the Ministry of Economic Affairs has been found to restrict aliphatics. The amount of polyether-polycarbonate material to 10% by weight or less is important for reducing the specific gravity effect. Regarding the reduction of the filter paper effect, it has been found suitable to reduce the amount of the aliphatic polyether polycarbonate material to 2% by weight or less. The following examples are shown to disclose the preferred specific examples and the application of the present invention, which does not mean to limit the present invention, unless otherwise stated in the scope of the attached patent application. This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm). ) -Τστ A7 B7 Printed by the Shellfish Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (8) Example 1 A. Contains reagents with complete acidity— "and the preparation of polymer-free reagents without buffer Protein reagents are used with buffered test solutions to identify available polymers. Impregnation solution: 0.3 mM tetrabromophenol blue, TBPB, and 0 %% or 100/0 (w / v) solution of polycarbonate in THF. (Control reagents are prepared with the same impregnation solution, which is polymer-free). The impregnation solution was immersed in Whatman CCP-500 paper at a speed of 4 feet per minute and a drying temperature of 60 ° C. The reagent paper is processed into a reagent strip containing a 2 x 02 inch pad. B. Measurement of reagent reactivity 1. Sample. The reagent was immersed in 0.20M potassium citrate buffer, pH 3.5 or 4.0, with or without 20 mg / dL human serum albumin (HSA). 2. The instrument reads the sound. The test strip was immersed in the sample for 20 seconds and the percent reflectance was measured on a CLINTEK® 200 instrument at 630 nanometers (nm) (610 nm can also be used). Calculate K / S according to the equation K / S = (1-R) 2 / 2R. Each number is the average of 10 repeated measurements. C. Results and discussion-1. Data. (Please read the notes on the back before filling this page)

、 1T This paper size applies the Chinese National Standard (CNS) Λ4 specification (210 X 297 mm) -11-V. Description of the invention (9) A7 B7 Table 2: pH 3.5 results at 630nm, 20s K / S (standard deviation) ） Polymer-free HSA 20 mg / dL HSA% decrease in slope blank group ％% decrease in slope (2) None 0.088 0.375 0.0144--(.005) (.020) ΚΟΚ 9209 0.068 0.271 0.0102 56 29 (0.1% W / V) (.007) (.008) ΚΟΚ 10,000 0.079 0.283 0.0102 25 29 (0.1% W / V) (.002) (.008) ΚΟΚ 10,000 0.069 0.229 0.0080 53 44 (1.0% W / V) (.002 ) (.007) ΚΟΚ 10,001 0.077 0.322 0.0123 31 15 (0.1% W / V) (.003) (.018) ΚΟΚ 10,002 0.071 0.327 0.0128 47 11 (0.1% W / V) (.006) (.008) Economy Printed by the Consumer Standards Cooperative of the Central Bureau of Standards (1) K / S of blank paper has been found to be approximately 0.052; therefore. / 〇 Blank group reduced-[U blank group: κ ^ -0.052 without polymer]-(blank group: K / S-0.052 with polymer)} / (blank group: K / S-0.052 without polymer )] X 100. (2)% slope reduction = [(slope without polymer-slope with polymer) / (slope without polymer)] X 100 obtained similar results at pH 4.0. 2. Discussion. In the buffer solution, at pH 3.5 and 4.0, some polycarbonates reduced the white group more than the slope. Since this experiment, it has been thought that adding polycarbonate to the intact protein reagent (both containing buffer and TBPB) will decrease (please read the precautions on the back before filling this page)

This paper size is applicable to Chinese National Standard (CNS) A4 (210X297 mm) -12- Printed by the Ministry of Economic Affairs, Industrial and Consumer Cooperatives A7 B7 V. Description of the invention (10) Initial color (blank group), May reduce the incidence of false positives. Example 2 A. Jggg_i〇〇 / 0 κ〇κ 10.071 protein reagent formulation 1. Buffer stock solution> Dissolve 14.75 grams of citric acid, 10.93 grams of sodium citrate 'and 7.2 mg FD & C yellow # 5 in 154 Ml of distilled water; pH = 3.46. 2 The first dipping solution. ⑻17.94 grams of buffered stock solution; (b) 5.0 grams of ethanol and (c) 2.47 grams of distilled water. 3. The second dipping solution. (A) 75 mg of TPBP; 25 g of KOK 10071 and (c) add an appropriate amount of THF to 25 ml. B. J. The first lysate and the second lysate prepared the control group formulation, which contained no polymer. The impregnation solution was dipped into E & D 237C paper at a rate of 4 feet per minute. The drying temperature for the first impregnation was 80 ° C, and the temperature for the second impregnation was 60 ° C. C. Measurement of reagent reactivity 1. Sample. The test samples were ⑴ negative reaction matrix SG urine pool and HSA was poured into the matrix SG urine pool to obtain a concentration of 15 mg / d L, and (2) three clinically negative 1 response high SG urine samples (SG = A4.028; B, 1.027; and C, 1.027). The reactivity of three samples was measured on a CLINK® 200 instrument (3 replicates for each sample). _ 1 A clinical negative reaction is defined as negative by the Sulfosal test. Contains HSA S2 mg / dL, total protein < 10 mg / dL. The goal is to get a response for clinically negative urine. This urine responded to a small fraction of 15 mg / dL HSA. Standards for this paper Standards for use in poor countries (CNS) A4 (210X297 mm) -1T- (Please read the precautions on the back before filling this page) Order A7 B7 '* V. Description of the invention (11) 2. The following data are shown as K / S 値 for each sample. In addition, the contribution of each high SG urine sample is [(K / S high sg_K / S blank group) / (K / S umg / dL · K / S blank group)] X 100. This continuation is a percentage of 15 mg / dL HSA caused by high SG urine. I l · ——— ^ —n I— Is I I T t (Please read the note on the back before filling this page)

The initial color and responsivity of the printed SL to the HSA by the Central Workstation Consumer Cooperative Bureau of the Ministry of Economic Affairs was reduced by 10% ΚΟΚ 10,071. However, the contribution of high SG urine that also reduces negative reactions is considerable. Therefore, the presence of 10% KOK 10,071 polymer significantly reduces the possibility of false positive results in high-SG urine samples. * Other examples compare TBPB-containing protein reagents with and without 4% KOK 10,071. Example 3 ΑΒΤΒΒ-ΚΟΚ protein reagent formulation (the amount is sufficient to prepare 100 ml infusion) 1_ the first infusion (0.45M sodium citrate-pH 3.70, 0.03mM FD & C yellow; aqueous solution) . (A) 50 ml of 0.9M sodium citrate-pH 3.70; Table 3 Effect of 10% KOK10,071 on protein reagent efficacy at K / S (standard deviation) of 630mn Matrix SG urine high SG urine high SG urine Liquid contribution

Reagent 15 mg / dL without HSA HSA ABCA denier C without polymer 0.165 (.016) 0.29 (.013) 0.258 (.012) 0.238 (.017) 0.240 (019) 74 58 60 10% κοκ 10,071 0.073 (005) 0.137 (.002) 0.088 (013) 0.092 (.004) 0.086 (OH) 23 30 20 Paper New Zealand t HS Jia Liang Zhun (〇 ^) 戍 4 «1 ^ (210 father 297 mm) " 14 'Economy Ministry of Standards Bureau Consumers' Co-operative Printing A7 B7 V. Invention Description (12) (b) 3.0 ml ImM FD & C Yellow # 5; and (c) 47 ml distilled water 〇-2. The second dipping solution (0.30 mM TBPB, 4% (W / V) KOK 10,071; in THF). ⑷ 30 ml of a solution of 1 mM TBPB in THF; (b) 40 ml of a 10% solution of KOKK 10,071 in THF; and (c) 30 ml of THF. Control group formulations were prepared using the first impregnation solution and the second impregnation solution, which contained no polymer. The impregnating solution was immersed in E & D 237C paper as in Example π. Β. Measurement of reagent reactivity 1. Sample. Samples of matrix SG urine were poured into HSA as described above. Highly SG urine with clinical negative response (A; SG = 1.030; pH = 5.6) and a designed alkaline high SG urine pool (B; SG = 1.024; pH = 8.7) are tested for possible false positive readings . High SG urine "B" was used as an example of "bad case" to test the limitation of the KOK polymer effect. The reactivity of each sample was measured 6 times on a CLINITEK® 200+ instrument. The data are shown below: Table 4 Effect of 4% ΚΟΚ 10,071 on the efficacy of protein reagents at K / S at 630 nm (standard deviation with 90% confidence ) (Please read the notes on the back before filling this page)

， 1T matrix SG urine high SG urine high SG urine contribution 値 reagent 15mg / dL_ no HSA HSA A denier A denier polymer 0.268 (.012) 0.458 (.023) 0.379 (〇Π) 0.430 (.023) 58 85 4% ΚΟΚ 10,071 0.110 (.002) 0.241 (.007) 0.145 (.004) 0177 (.004) 26 51 This paper size applies to the Chinese National Standard (CNS) Λ4 specification (210X297 mm) -15- Ministry of Economic Affairs Printed by the Consumer Standards Cooperative of the Central Bureau of Standards A7 B7 V. Description of Invention (13) In the first example, the initial color and reactivity to HSA were all reduced by κοκ 10,071. Similarly-compared to reagents without polycarbonate, it also reduces negative reactions with high SG urine f contribution. This confirms that polycarbonate reduces the likelihood of false positive results in high-SG urine samples. 'Example 4 Aqueous solution (10 liters), adjusted to pH 3 7, with 417 grams of sodium citrate' 455 grams of lemon so '〇29 grams of fd & C yellow # 5 and 0.5 grams of FD & C red well 3 and using 24 grams PVA impregnated filter paper grade 204C. After the first impregnation (temperature 80-100. (:), the filter paper was impregnated with a second solution. The second solution (10 liters) was a solution of 5 grams of tetrabromophenolphthalein in ethanol and 50 grams of citric acid. And 150 grams of propylene oxide carbonate copolymer [from L 95〇 @ Bayer (AG) catalyzed by osmium (IV) butyl oxide] and diphenyl carbonate. Then the obtained material is at a temperature of 35 to 10. 5. (: Drying. Example 5 Only in different pulp pulp batches, when used with KOK-free protein reagents, it will cause a large difference in reactivity (Table 5). It has been proposed that dyes and proteins bind to paper fibers Differences between batches lead to differences between batches. In addition, it has been proposed that an increase in the binding of dye to paper will increase the background of the test, and an increase in the binding of protein to paper will reduce the protein response of the test. Table 5 uses a central prescription similar to Example 4 Available without ΟΟΚ. The paper size is generally made by the country's national structure (CNS) Λ4 size (2) 〇χ 297 mm -16- ---- „----- ^-rL--r- ----------- Order ---------- Γ Mn f. (Please read the notes on the back before filling in this page) V. Invention (彳 4) A7 B7 Table 5 Employees ’Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs of the People ’s Republic of China CLINITEX-10 CLINITEX-200 Grade Batch Negative Average 値 Positive Average 値 Negative Average 値 Positive Average 値 237 7775 672 525 695 479 Background minimum 237 8079 745 549 824 511 237 8137 753 560 839 529 237 14014 706 551 743 510 237 8977 743 586 800 572 237 9241 740 595 787 555 237 9041--761 570 204 331 749 555 838 525 204 225 772 578 866 545 background maximum 204 342 760 562 850 523 204 14011 755 575 818 548 204 14013 759 584 823 555 204 306 740 563 805 537 204 238 767 595 841. 564 204 9115 _ one 839 572 For two different Miles' instruments, CLINITEK -10 and CLINITEK -200, the reagent reactivity is represented by the decoded number. The decoded number is determined by the percent reflectance of the maximum absorption at 610 to 660nm. A low decoded number indicates high reactivity and a large background color. The pH of these reagents was 3.46. This paper size applies to Chinese National Standard (CNS) Λ4 specification (210 × 297 male sauce) -17- (Please read the precautions on the back before filling this page)

, 1T Consumer Policy Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs of the PRC 393578 A7 _ B7 V. Description of the invention (15) 237 and 204 The grade of filter paper is Alhstrom Filtration of Mt. Holly Springs, O, Pennsylvania. Analyze the test strips with urine and observe the resulting color changes. I has been recorded as an indicator of the protein in the urine biological sample. As can be seen in Table 6, the protein 4 reagent " Pr04 " (based on Example 4) has greatly reduced the furnace paper batch effect. This reduction effect increases manufacturability and makes the reagent results consistent. Polymers are considered due to Destroy the combination of dye and wood fiber to reduce filter paper variability. As a result, protein reagent reactivity can be consistent on one target. Table 6 Reactivity

Prescription negative 30 mg / dL

Pro 4 883 570 863 570 904 586 has been found that even when using different batches of filter paper, the reactivity between the reagent composition and the filter paper is not a factor when determining protein. In addition, eye and instrument detection limits on different fecal fluid samples showed reduced test strip variability. This is in contrast to other formulations used to test proteins, in which the detection limit of the matrix and the high specific gravity urine, vision and instrument readings are often changed. Although the present invention allows various corrections and another form, the application examples have shown that the paper size is applicable to the Chinese National Standard (CNS) Λ4 specification (210X297 mm) -18- (Please read the precautions on the back before filling this page) ------ Order ----------- Γ-. 393578 at B7 V. Description of the Invention (16) shows specific examples and has been detailed. However, it should be understood that these examples are not intended to limit the invention to the particular form described, but on the contrary, the present invention covers all amendments, equivalents, and alternatives falling within the spirit and scope defined by the scope of the appended patent application. . (Please read the precautions on the back before filling out this page) Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs -19- This paper size applies to China National Standard (CNS) A4 (210X297 mm)

Claims (1)

10. Patent Application No. 84100273 1 Amendment to the Chinese Patent Application, (8! F0 $$ Announcement VI. Application Patent 393578 1. A detection device for measuring protein in biological samples, including the following immersion Absorptive carrier substrate: —— a kind of polydentified phenol sulfone phthaloprotein error indicator, a kind of buffer, which is added to adjust the ρ ρ value to between 1.5 and 4.5, and an aliphatic polyacid-polycarbonate Esters in an amount of 0.001% to 10% (weight / volume ratio) reducing specific gravity effect and having the following structure: Where: η is an integer between 4 and 15; and m is an integer between 1 and 10. 2. According to the test device in the scope of patent application No. 1, in which the donanized peptone S too protein error indicator is tetrabromopansulfan. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling out this page) 3. According to the testing device of the scope of application for patent i, the aliphatic polymystery and the amount of polycarbonate 0% to 2% (weight / volume ratio) 4. According to the test device of the patent claim, wherein η is between * and I 1 'm is between 1 and 7. 5 · According to The detection device under the scope of patent application No. 4 wherein n is between 4 and II ′ and m is between 1 and 5. -1-(CNS) (2l0x297 ^) -------- 10. Article 84100273 Patent Application No. 1 Chinese Application for Patent Garden Amendment, (8! F0 $$ Announcement 6. Application Patent Range 393578 1. A detection device for measuring protein in biological samples, including the following immersed absorbent carriers Substrate: —— a kind of polydentified phenol sulfone phthaloprotein protein error indicator, a kind of buffer, which is added to adjust the ρ Η value to between 1.5 and 4.5, and an aliphatic polyacid-polycarbonate, the amount It is 0.001% to 10% (weight / volume ratio) to reduce the specific gravity effect, and has the following structure: Where: η is an integer between 4 and 15; and m is an integer between 1 and 10. 2. According to the test device in the scope of patent application No. 1, in which the donanized peptone S too protein error indicator is tetrabromopansulfan. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling out this page) 3. According to the testing device of the scope of application for patent i, the aliphatic polymystery and the amount of polycarbonate 0% to 2% (weight / volume ratio) 4. According to the test device of the patent claim, wherein η is between * and I 1 'm is between 1 and 7. 5 · According to The detection device under the scope of patent application No. 4 wherein n is between 4 and II ′ and m is between 1 and 5. -1-(CNS) (2l0x297 ^) -------- A8 B8 C8 D8 Patent application scope 6 'Method for detecting protein in a biological sample, this method includes the following steps: Wetting the analysis test strip with a biological sample' The test strip includes an absorbent impregnated with a first aqueous infusion and a buffer The carrier substrate is then impregnated with a non-aqueous solvent solution of a polyphenol-polycarbonate, a polyhydric phenolic protein error indicator, and 0.01% to 0% (weight // volume ratio). (1) any color change of the recorded test strip, where the color change may be As an indicator of the protein in the biological sample, the addition of the buffer is to adjust the pH value to between 5 and 4.5, and the presence of the Indian 4 ether-polycarbonate is 0.001% to 1 〇% (weight, volume ratio) to reduce the effect of specific gravity, and has the following structure: --.---- β —— (please read the first $ entry on the back and write θ education) π is an integer between 4 and 5; and m is an integer between 1 and 10. Printed by the Shellfish Consumer Cooperative of the Central Bureau of Economic Affairs of the Ministry of Economics Among them: According to the method in the scope of patent application No. 6, wherein the polydentate phenol phthalate protein error indicator is tetrabromo phenol phthalide. The method according to item 6 of the application, wherein the aliphatic polyether-polycarbonate is present in an amount of from 0.01% to 2% (weight / volume ratio). (21. Reading public works-393578 i D8 VI. Application for patent application scope and data, according to, Gen 1 Gen 1 11 ιί. 1 Α .0 · On 1 Fan to Fan Di encircled law to 4 Yu Jiewei The m in η is the item from the 5th to the French side to 4 and the 4 in Jiewei η --------- ^ —— * * (Please read the precautions on the back before this page) The standard of printed paper waves printed by employees of the Central Standards Bureau of the Ministry of Economic Affairs of the Consumer Cooperative is applicable to the Chinese National Standard (CNS) A4 (210X297 mm)