CN107121427A - Detection reagent of tyrosine phenolic metabolism thing and preparation method thereof in human urine - Google Patents
Detection reagent of tyrosine phenolic metabolism thing and preparation method thereof in human urine Download PDFInfo
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- CN107121427A CN107121427A CN201710281212.5A CN201710281212A CN107121427A CN 107121427 A CN107121427 A CN 107121427A CN 201710281212 A CN201710281212 A CN 201710281212A CN 107121427 A CN107121427 A CN 107121427A
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Abstract
The present invention relates to external diagnosis reagent field, further detection reagent of tyrosine phenolic metabolism thing and preparation method thereof specifically related in human urine.The detection reagent of tyrosine phenolic metabolism thing, at least includes in the human urine：1~20mol/L of mercurous nitrate；1~5mol/L of mercuric sulfate；0.1~2mol/L of nickel nitrate；3~20mol/L of sulfuric acid；0.01~1g/L of P-Mo-Wo acid chitosan nano silica composite.
Description
Technical field
The present invention relates to external diagnosis reagent field, further tyrosine phenolic metabolism thing specifically related in human urine
Detection reagent and preparation method thereof.
Background technology
In recent years, the incidence of disease of whole world tumor disease is in continuous ascendant trend, according to ACS (American
Cancer Society) statistics of world wide cancer is shown, from 2008 to the year two thousand thirty, the newly-increased cancer occurrence numbers in the whole world
21,400,000 will be increased to from annual 12,700,000, number of cancer deaths will rise to 13,200,000 people by 7,600,000.At the same time, China
In recent years statistics shows, it is contemplated that the year two thousand twenty Cancer in China new cases will be up to 3,490,000, the dead number because of malignant tumour
60% [" 8 will be risen compared with 2002 up to 2,630,000].As can be seen here, no matter in developed country or developing country, malignant tumour
(cancer) oneself substitution cardiovascular disease turns into the most disease of death toll.Due to pathogenesis of cancer concealment, be in progress fast, grade malignancy
Height, therefore, " early detection, early diagnosis, early treatment " are still the reduction topmost measure of cancer mortality.Due to tumour mark
Will thing has important directive significance to the diagnosis of tumour, classification, Index for diagnosis and treatment, therefore, for tumor markers
Research has obtained the extensive concern of domestic and foreign scholars.
Tyrosine (Tyrosine, Tyr) belongs to aromatic amino acid, can be light by phenylalanine (Phenylalaine, Phe)
Change, can also be absorbed directly from food, be the important semi-dispensable amino acid of human body.To through phenyl-lactic acid (p-
Hydroxyphenyllactic acid, PHPLA), DOPA (3,4-dihydroxyphenylalanine, DOPA) and alcapton
(2,5-dihydroxyphenylacetic acid, HGA) are the important metabolites of tyrosine.The metabolic disorder of tyrosine can
Cause the generation of several amino acids metabolic disease, such as tyrosinemia, alkaptonuria, and with liver and kidney disease, nervous system
Degenerative disease, gas malignant tumour etc. has certain correlation.Therefore, the research for tyrosine and its metabolite not only exists
Protein chemistry and particularly significant, and early diagnosis in a variety of diseases, examination of curative effect in terms of evaluating the nutrition condition of patient
With it is significant in terms of Study of Etiology.
Protein present in biological sample can disturb the measure of tyrosine and its metabolite, and then have impact on detection
The degree of accuracy.
The content of the invention
For above-mentioned technical problem, one side of the invention provides tyrosine phenolic metabolism thing in a kind of human urine
Detection reagent, at least include：
In a preferred embodiment, in the human urine tyrosine phenolic metabolism thing detection reagent, at least
Including：
In a preferred embodiment, in the human urine tyrosine phenolic metabolism thing detection reagent, at least
Including：
In a preferred embodiment, in the P-Mo-Wo acid-chitosan-nanometer silica composite, phosphorus molybdenum
Weight ratio between wolframic acid, chitosan, nano silicon is (1~10)：1：(5~30).
In a preferred embodiment, in the P-Mo-Wo acid-chitosan-nanometer silica composite, phosphorus molybdenum
Weight ratio between wolframic acid, chitosan, nano silicon is (3~7)：1：(10~25).
In a preferred embodiment, in the P-Mo-Wo acid-chitosan-nanometer silica composite, phosphorus molybdenum
Weight ratio between wolframic acid, chitosan, nano silicon is 5：1：15.
In a preferred embodiment, the detection reagent of tyrosine phenolic metabolism thing also includes in the human urine
Sodium phosphate.
Second aspect of the present invention provides the preparation method of the detection reagent of tyrosine phenolic metabolism thing in human urine,
At least comprise the following steps：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
Third aspect of the present invention provides the detection kit of tyrosine phenolic metabolism thing in human urine, the human body
The detection kit of tyrosine phenolic metabolism thing includes the detection examination of tyrosine phenolic metabolism thing in above-mentioned human urine in urine
Agent.
The 4th aspect of the present invention provides a kind of detection method of tyrosine phenolic metabolism thing in human urine, at least wraps
Include following steps：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.
Embodiment
The participation in the election detailed description of the invention below for being preferable to carry out method and including embodiment this hair can be more easily understood
Bright content.Unless otherwise defined, all technologies used herein and scientific terminology have common with art of the present invention
The identical implication that technical staff is generally understood that.When there is contradiction, the definition in this specification is defined.
Equivalent, concentration or other values or parameter are excellent with scope, preferred scope or a series of upper limit preferred values and lower limit
During the Range Representation that choosing value is limited, this, which is appreciated that, specifically discloses by any range limit or preferred value and any scope
All scopes that any pairing of lower limit or preferred value is formed, regardless of whether whether the scope separately discloses.For example, when open
During scope " 1 to 5 ", described scope should be interpreted as including scope " 1 to 4 ", " 1 to 3 ", " 1 to 2 ", " 1 to 2 and 4 to
5 ", " 1 to 3 and 5 " etc..When number range is described herein, unless otherwise indicated, otherwise the scope is intended to include its end
Value and all integers and fraction within the range.
Singulative includes plural number and object is discussed, unless the context clearly dictates otherwise." optional " or it is " any
It is a kind of " refer to that the item or event that describe thereafter may or may not occur, and the description include situation that event occurs and
The situation that event does not occur.
Approximate term in specification and claims is used for modifying quantity, represents that the present invention is not limited to this specific
Quantity, includes the part of the amendment of the acceptable change without cause related basic function close to the quantity.Phase
Answer, modify a numerical value with " about ", " about " etc., mean that the invention is not restricted to the exact numerical.In some examples, approximately
Term likely corresponds to the precision of the instrument of measured value.In present specification and claims, scope is limited can be with
Combine and/or exchange, these scopes include all subranges contained therebetween if not stated otherwise.
In addition, the indefinite article " one kind " and " one " before key element of the present invention or component are to key element or the quantitative requirement of component
(i.e. occurrence number) unrestriction.Therefore " one " or " one kind " should be read as including one or at least one, and odd number
The key element or component of form also include plural form, unless the obvious purport of the quantity refers to singulative.
As used herein, each of following term has the implication related in this part to it.Article " one " and "
It is a kind of " to be used for one or the grammar object more than one (i.e. at least one) for referring to the article herein.For example, " key element " means
One key element or more than one key element.
As used herein, term " sensitivity " is defined as determining the statistics of (for example, method, test) performance and measured, and leads to
Cross true positives number divided by true positives and false negative and to calculate.
As used herein, term " specificity " is defined as determining the statistics of (for example, method, test) performance and measured, and leads to
True negative number divided by true negative and false positive sum is crossed to calculate.
For above-mentioned technical problem, one side of the invention provides tyrosine phenolic metabolism thing in a kind of human urine
Detection reagent, at least include：
In a preferred embodiment, in the human urine tyrosine phenolic metabolism thing detection reagent, at least
Including：
In a preferred embodiment, in the human urine tyrosine phenolic metabolism thing detection reagent, at least
Including：
In the application, the main component that mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid develop the color as detection reagent works as detection
When reagent is in contact with tyrosine phenol metabolism thing, occurs chromogenic reaction, the positive is rendered as red；Weakly positive is rendered as pink
Color；Feminine gender is rendered as white or light yellow.
In the application, the content of mercurous nitrate is 1~20mol/L, refers to mercurous nitrate containing in whole detection reagent
Measure as 1~20mol/L, volume calculating benchmark is defined by the volume of detection reagent.The content of mercuric sulfate is 1~5mol/L；Refer to
Mercuric sulfate is 1~5mol/L in the content of whole detection reagent, and volume calculating benchmark is defined by the volume of detection reagent；Nickel nitrate
Content be 0.1~2mol/L；It is 0.1~2mol/L, volume calculating benchmark in the content of whole detection reagent to refer to nickel nitrate
It is defined by the volume of detection reagent；The content of sulfuric acid is 3~20mol/L；Refer to content of the sulfuric acid in whole detection reagent for 3~
20mol/L, volume calculating benchmark is defined by the volume of detection reagent；P-Mo-Wo acid-chitosan-nanometer silica composite
Content is 0.01~1g/L, and volume calculating benchmark is defined by the volume of detection reagent.
In the application, term " P-Mo-Wo acid-chitosan-nanometer silica composite " refers to chitosan and nano-silica
After SiClx is grafted, chitosan-nanometer silica composite is obtained, then again chitosan-nano silicon is combined
Thing, carries out load P-Mo-Wo acid, obtains P-Mo-Wo acid-chitosan-nanometer silica composite.
In the application, P-Mo-Wo acid, chitosan, nano silicon are commercially available acquisition.
The P-Mo-Wo acid is purchased from Xiamen seamark Science and Technology Ltd.；
The chitosan is purchased from Xi'an Rayleigh bio tech ltd；
The nano silicon is purchased from win wound Degussa；
In a preferred embodiment, the preparation side of the P-Mo-Wo acid-chitosan-nanometer silica composite
Method, at least comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
The present inventor is found surprisingly that, by adding very small amount P-Mo-Wo acid-chitosan-nanometer silica composite,
On the one hand can preferably adsorb biological alkali number in the alkaloid in urine, such as urea, urine excessive, can with system
Mercurous ion is combined, and influences chromogenic reaction, and then influence the precision of detection；P-Mo-Wo acid-chitosan-nano silicon is multiple
Not only physisorption had been produced between compound and alkaloid but also chemisorption is produced, precipitation has been eventually formed, very accurately
Eliminate the interference of alkaloid.On the other hand, the addition of P-Mo-Wo acid-chitosan-nanometer silica composite causes colour developing
The color of reaction is more obvious, can accurately judge the positive, weakly positive, feminine gender, and the positive is rendered as red；Weakly positive is presented
For pink；Feminine gender is rendered as white or light yellow, solves the technical problem that people judge by accident for color aberration.
Second aspect of the present invention provides the preparation method of the detection reagent of tyrosine phenolic metabolism thing in human urine,
At least comprise the following steps：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
Third aspect of the present invention provides the detection kit of tyrosine phenolic metabolism thing in human urine, the human body
The detection kit of tyrosine phenolic metabolism thing includes the detection examination of tyrosine phenolic metabolism thing in above-mentioned human urine in urine
Agent.
The 4th aspect of the present invention provides a kind of detection method of tyrosine phenolic metabolism thing in human urine, at least wraps
Include following steps：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.
The present invention is specifically described below by embodiment.It is necessarily pointed out that, following examples are only used
In the invention will be further described, it is impossible to be interpreted as limiting the scope of the invention, professional and technical personnel in the field
Some the nonessential modifications and adaptations made according to the content of the invention described above, still fall within protection scope of the present invention.
In addition, if without other explanations, raw materials used is all commercially available, and number used in following material is weight
Part.
Embodiment 1：
Embodiment 1 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 1：1：5.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：1.
Embodiment 2：
Embodiment 2 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 10：1：30.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：2.
Embodiment 3：
Embodiment 3 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 3：1：25.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：2.
Embodiment 4：
Embodiment 4 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 3：1：25.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：5.
Embodiment 5：
Embodiment 5 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 10：1：30.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：5.
Embodiment 6：
Embodiment 6 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 10：1：5.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：5.
Embodiment 7：
Embodiment 7 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 7：1：25.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：10.
Embodiment 8：
Embodiment 8 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 3：1：10.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：8.
Embodiment 9：
Embodiment 9 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 5：1：15.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：8.
Embodiment 10：
Embodiment 10 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 5：1：15.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：10.
Embodiment 11：
Embodiment 11 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the P-Mo-Wo acid-chitosan-nanometer silica composite, comprises the following steps：
(1) nano silicon is dissolved in ethanol solution, at room temperature after ultrasonic disperse 60min, and adds silane coupler
K560, stirs 3h at 45 DEG C, obtains nanosilica solution；Chitosan is dissolved in acetum again, 3h is stirred at room temperature,
Obtain chitosan solution；Chitosan solution is added in nanosilica solution, after stirring reaction 8h, centrifugation washing 3 times,
Vacuum drying obtains chitosan-nanometer silica composite at ethanol is washed 2 times, 60 DEG C.
(2) first P-Mo-Wo acid is dissolved in deionized water, makes mass fraction for 10% P-Mo-Wo acid solution, add
Chitosan-the nanometer silica composite obtained in step (1), constant temperature return stirring 20h, vacuum filtration is obtained after drying
P-Mo-Wo acid-chitosan-nanometer silica composite.
Wherein, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is 5：1：15.
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, then add P-Mo-Wo acid-chitosan-and receive
Rice silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
The detection method of tyrosine phenolic metabolism thing, at least comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：10.
Comparative example 1：
Comparative example 1 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, tyrosine phenolic metabolism in human urine is obtained
The detection reagent of thing.
The detection method of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：10.
Comparative example 2：
Comparative example 2 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid, P-Mo-Wo acid are well mixed, tyrosine in human urine is obtained
The detection reagent of phenolic metabolism thing.
The detection method of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：10.
Comparative example 3：
Comparative example 3 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid, chitosan are well mixed, tyrosine phenol in human urine is obtained
The detection reagent of metabolite.
The detection method of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：10.
Comparative example 4：
Comparative example 4 provides a kind of detection reagent of tyrosine phenolic metabolism thing in human urine, including：
The preparation method of the detection reagent of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid, nano silicon are well mixed, junket in human urine is obtained
The detection reagent of propylhomoserin phenolic metabolism thing.
The detection method of tyrosine phenolic metabolism thing, comprises the following steps in human urine：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged
Whether chromogenic reaction is occurred.Wherein, the volume ratio of urine and detection reagent is：1：10.
Characterization test
Choose 1000 cancer patient mornings on an empty stomach, urine 10mL is taken respectively, human urine to be measured is used as.
Detection method：
1st, human urine to be measured is contacted into mixing with embodiment 1~11 and comparative example 1~4 respectively, judges whether occur
Chromogenic reaction.
Wherein, the positive is rendered as red；Weakly positive is rendered as pink；Feminine gender is rendered as white or light yellow.
The characterization test of table 1
Respectively from embodiment 5, embodiment 8, embodiment 9, embodiment 10, embodiment 11, comparative example 1, comparative example 2, right
Ratio 3, comparative example 4 are respectively to 100 cervical carcinomas, 100 stomach cancers, 100 breast cancer, 100 liver cancer, 100 patients with lung cancer
Test recall rate is carried out, as a result as shown in table 2.
The cancer recall rate of table 2
The detection reagent of tyrosine phenolic metabolism thing has very in the human urine of the application it can be seen from table 1,2
Good pertinency factor, meanwhile, the applicant is found surprisingly that the product of the application has extraordinary recall rate to cervical carcinoma.
Foregoing example is merely illustrative, some features of the feature for explaining the disclosure.Appended claim
The scope as wide as possible for requiring to be contemplated that is intended to, and embodiments as presented herein is only according to all possible embodiment
Combination selection embodiment explanation.Therefore, the purpose of applicant is appended claim not by the explanation present invention
Feature example selectional restriction.And the progress in science and technology by formed language performance it is inaccurate due to and not
The possible equivalent or son being presently considered are replaced, and these changes should also be interpreted by appended in the conceived case
Claim is covered.
Claims (10)
1. the detection reagent of tyrosine phenolic metabolism thing in a kind of human urine, it is characterised in that at least include：
2. the detection reagent of tyrosine phenolic metabolism thing in human urine as claimed in claim 1, it is characterised in that at least wrap
Include：
3. the detection reagent of tyrosine phenolic metabolism thing in human urine as claimed in claim 2, it is characterised in that at least wrap
Include：
4. the detection reagent of tyrosine phenolic metabolism thing in the human urine as described in claims 1 to 3, it is characterised in that institute
State in P-Mo-Wo acid-chitosan-nanometer silica composite, the weight between P-Mo-Wo acid, chitosan, nano silicon
Than for (1~10)：1：(5~30).
5. the detection reagent of tyrosine phenolic metabolism thing in human urine as claimed in claim 4, it is characterised in that the phosphorus
In molybdenum wolframic acid-chitosan-nanometer silica composite, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is
(3~7)：1：(10~25).
6. the detection reagent of tyrosine phenolic metabolism thing in human urine as claimed in claim 5, it is characterised in that the phosphorus
In molybdenum wolframic acid-chitosan-nanometer silica composite, the weight ratio between P-Mo-Wo acid, chitosan, nano silicon is
5：1：15.
7. the detection reagent of tyrosine phenolic metabolism thing in human urine as claimed in claim 4, it is characterised in that the people
The detection reagent of tyrosine phenolic metabolism thing also includes sodium phosphate in body urine.
8. the preparation method of the detection reagent of tyrosine phenolic metabolism thing, its feature in the human urine described in claim 1~7
It is, at least comprises the following steps：
After mercurous nitrate, mercuric sulfate, nickel nitrate, sulfuric acid are well mixed, P-Mo-Wo acid-chitosan-nanometer two is then added
Silica composite obtains the detection reagent of tyrosine phenolic metabolism thing in human urine after stirring.
9. a kind of detection kit of tyrosine phenolic metabolism thing in human urine, it is characterised in that junket in the human urine
The detection kit of propylhomoserin phenolic metabolism thing includes tyrosine phenolic metabolism thing in the human urine described in claim 1~7
Detection reagent.
10. the detection method of tyrosine phenolic metabolism thing in a kind of human urine, it is characterised in that at least comprise the following steps：
Obtain human urine to be measured；
The detection reagent of tyrosine phenolic metabolism thing in above-mentioned human urine is contacted into mixing with human urine to be measured, judged whether
Generation chromogenic reaction.
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Effective date of registration: 20210118 Address after: 411100 2 / F, building 2, Xiangtan Jiuhua innovation and entrepreneurship service center, No.5 ya'ai Road, Xiangtan Economic Development Zone, Xiangtan City, Hunan Province Patentee after: Hunan aneng Medical Instrument Co.,Ltd. Address before: 310000 715, building 1, 175 Tiancheng Road, Jianggan District, Hangzhou City, Zhejiang Province Patentee before: Zhu Jianhua |