CN107515206B - A kind of method of sulfur doping carbon quantum dot fluorescence sensitivity detection Norfloxacin - Google Patents
A kind of method of sulfur doping carbon quantum dot fluorescence sensitivity detection Norfloxacin Download PDFInfo
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Abstract
The invention discloses a kind of methods of sulfur doping carbon quantum dot fluorescence sensitivity detection Norfloxacin, use poly- (4- styrenesulfonic acid-co-maleic acid) sodium for carbon source and sulphur source, and hydro-thermal method synthesizes sulfur doping fluorescent carbon quantum dot；Sulfur doping fluorescent carbon quantum dot can enhance the fluorescence of Norfloxacin, and the fluorescence signal intensity and blood concentration norfloxacin of enhancing are in some linear, establish the fluorescence spectrophotometry of detection Norfloxacin accordingly；Sulfur doping fluorescent carbon quantum dot prepared by the present invention has significant fluorescence sensitivity to act on Norfloxacin, and selectivity is strong, for the fluoremetry of Norfloxacin, has the characteristics that high sensitivity, specificity.
Description
Technical field
The present invention relates to chemical analysis detection technique field, specially a kind of sulfur doping carbon quantum dot fluorescence sensitivity detects promise
The method of Flucloxacillin.
Background technique
Norfloxacin is commonly called as orfloxacin, and medicine antibacterial spectrum is wide, antibacterial activity is strong, is the various infectious diseases for the treatment of
Important synthetic drug, is clinically widely applied.Currently, the analysis method of Norfloxacin drug mainly have flow injection spec-trophotometry,
UV visible spectroscopy, fluorimetry, analysis method with quantum dot (Quantum Dot, QDS) as fluorescence probe etc..
Quantum dot has wide excitation band, half-peak width, symmetrical and adjustable emission band, is not susceptible to photobleaching, fluorescent quantum
The advantages that yield height and quantum size effect, skin effect." CdTe quantum fluorometric determination Norfloxacin contains article
Amount " (such as Sun Xuehua chemical research and application, 2013,25 (3): 344-346), selects thioacetic acid as stabilizer, with water
Solution synthetic method has synthesized CdTe quantum, measures Norfloxacin as fluorescence probe using it.
Carbon quantum dot is the new carbon that a kind of partial size is generally less than 10nm.It has excellent optical property, adjustable
Excitation and transmitting behavior, higher fluorescent stability, lower toxicity and good biocompatibility are more and more being led
It is widely used in domain, a nova being increasingly becoming in nano-carbon material.Currently, people are dedicated to exploring preparation height
The method of quantum yield carbon point.Carbon quantum dot is after overdoping or surface passivation, fluorescence quantum yield and photoelectric properties
The raising that may be significantly.There are the document report much about nitrogen-doping carbon quantum dot, carbon quantum dot recent years
Fluorescence quantum yield be improved.However the carbon quantum dot of element sulphur doping or nitrogen, sulphur codope is also seldom reported
Road, influence of the element sulphur to carbon quantum dot also require further study.
Summary of the invention
The purpose of the present invention is to provide a kind of highly sensitive, highly selective sulfur doping carbon quantum dot fluorescence sensitivities to detect promise
The method of Flucloxacillin.
The method of sulfur doping carbon quantum dot fluorescence sensitivity detection Norfloxacin of the present invention includes the following steps:
(1) drafting of standard working curve: in the range of linearity of 0.01~10 μm of ol/L, using citrate-phosphate hydrogen
Disodium buffer configures the Norfloxacin standard solution of 8 various concentration gradients, then water-soluble sulphur is added in each concentration 5mL
Doping fluorescent carbon quantum dot solution is vortexed after mixing and standing, on sepectrophotofluorometer, excitation wavelength 344nm, and transmitted wave
A length of 413nm measures fluorescence intensity, is mapped with fluorescence enhancement intensity to its respective concentration and carries out regression analysis, standard can be obtained
Working curve, this makes it possible to obtain the equation of linear regression of Norfloxacin coefficients associated therewith；
(2) sample preparation
Blood sample preparation: taking 5 mL new bloods, and in being centrifuged 10-15 min with 10000r/min, removal is precipitated, and
The perchloric acid solution protein precipitation of 1-2mL volumetric concentration 10% is added into upper plasma, 10-are centrifuged under 5000r/min
15min takes supernatant, is kept in dark place in 4 DEG C of refrigerator measurements to be analyzed；
Urine sample preparation: taking 5-10 mL urine, and 1-3g active carbon is added, and vortex mixed 30-60 s stands filtering,
Its filtrate is taken, is kept in dark place in 4 DEG C of refrigerator measurements to be analyzed；
Milk sample preparation: taking 5-10 mL milk, and 10-15 mL methanol-water-formic acid (28:72:0.1, v/v) is added
The trichloroacetic acid acetonitrile solution of extracting solution and l-2 mL mass concentration 4%, vortex 1-3 min, ultrasonic 5-10min, 10000r/
It is centrifuged 10-15 min under min, takes supernatant, is kept in dark place in 4 DEG C of refrigerator measurements to be analyzed；
(3) sample measures: taking step, stepOr stepExtracting solution 1-3mL obtained, with citrate-phosphate hydrogen
Disodium buffer is settled to 5mL, and water-soluble sulfur doping fluorescent carbon quantum dot solution is added, is vortexed after mixing standing, in fluorescence point
On light photometer, excitation wavelength 344nm, launch wavelength is that 413nm measures fluorescence intensity, substitutes into step (1) regression equation, meter
Calculate the content of Norfloxacin in sample.
The preparation method of water-soluble sulfur doping fluorescent carbon quantum dot is to weigh poly- (the 4- styrenesulfonic acid-co-of 2.0-5.0g
Maleic acid) sodium is added in 100mL ultrapure water, and ultrasonic 5-10min forms colourless transparent solution, and it is anti-to be transferred to polytetrafluoroethylene (PTFE)
Kettle is answered, after 200-240 DEG C of heating 5-8h, natural cooling, is first 0.22 μm of membrane filtration with aperture, is with molecular cut off afterwards
The bag filter of 3000-3500Da carries out dialysis treatment for 24 hours, obtains water-soluble sulfur doping fluorescent carbon quantum dot.
The citrate-phosphate disodium hydrogen pH of buffer is 4-5.
The dosage of the water solubility sulfur doping fluorescent carbon quantum dot is 300 μ L.
The vortex, which mixes to stand, refers to that vortex mixes 0.5-2 min, is then allowed to stand 10-20 min.
The present invention has the advantages that
1. the present invention is prepared for a kind of sulfur doping carbon amounts water-soluble, with good fluorescence performance using one step hydro thermal method
Sub-, the fluorescence quantum yield of the sulfur doping carbon quantum dot of preparation is high；
2, the present invention prepares water-soluble sulfur doping fluorescence carbon quantum using poly- (4- styrenesulfonic acid-co-maleic acid) sodium
The fluorescence quantum yield of point, the sulfur doping carbon quantum dot of preparation is up to 35.4%, has strong fluorescence enhanced sensitivity work to Norfloxacin
With using sulfur doping fluorescent carbon quantum dot to detect micro Norfloxacin as fluorescence probe, the high sensitivity of method, detection limit can be with
Reach 1.0 nmol/L, fluorescence sensitivity multiple is up to 15 times, while other quinolone medicines, without this effect, method has preferable
Specificity；
3, the sulfur doping fluorescent carbon quantum dot prepared can highly sensitive and high specificity enhancing Norfloxacin fluorescence, be used for ring
The detection of third husky star, method is simple, high sensitivity, high specificity.
Detailed description of the invention
Fig. 1 is that water-soluble sulfur doping fluorescent carbon quantum dot shows the fluorescence sensitivity exercising result of Norfloxacin in embodiment 1
It is intended to spectrum；
Fig. 2 is the influence result of other flouroquinolone drugs for coexisting to sulfur doping fluorescent carbon quantum dot.
Specific embodiment
Explanation, but this hair are described in further detail to technical solution of the present invention below in conjunction with specific embodiments
Bright protection scope is not limited to that.
Embodiment 1: the assay operating procedure of Norfloxacin is as follows in blood:
(1) preparation of water-soluble sulfur doping fluorescent carbon quantum dot (S-CQDs): weighing 2.0g, poly- (4- styrene sulfonic acid-is altogether
Poly- maleic acid) sodium is added in 100mL ultrapure water, and ultrasonic 5min forms colourless transparent solution, and it is anti-to be transferred to polytetrafluoroethylene (PTFE)
Kettle is answered, after 220 DEG C of 5 h of heating, natural cooling, is first 0.22 μm of membrane filtration with aperture, is with molecular cut off afterwards
The bag filter of 3000Da carries out dialysis treatment for 24 hours, obtains water-soluble sulfur doping fluorescent carbon quantum dot；
(2) drafting of standard working curve: use pH5.0 citrate-phosphate disodium hydrogen buffer configuration concentration for 0.01,
0.05,8 Norfloxacin standard solution of 0.1,0.2,0.5,1,5,10 μm of ol/L, and constant volume is in 5 mL；300 μ L step is added
Suddenly the water-soluble sulfur doping fluorescent carbon quantum dot of (1) preparation is vortexed after mixing 0.5 min standing, 10 min, in fluorescence spectrophotometer light
On degree meter, excitation wavelength 344nm, launch wavelength is that 413nm measures fluorescence intensity, with fluorescent quenching intensity to its respective concentration
Mapping carries out regression analysis to get equation of linear regression △ F=2.1343C+4.3039 of standard working curve, related coefficient is arrived
R2=0.9972；
(3) prepared by blood sample: taking 5 mL new bloods, in being centrifuged 15 min with 10000 r/min revolving speeds, removal is heavy
It forms sediment, and the perchloric acid solution protein precipitation of 1mL volumetric concentration 10% is added into upper plasma, be centrifuged under 5000r/min revolving speed
10 min take supernatant to be kept in dark place in 4 DEG C of refrigerator measurements to be analyzed；
(4) sample measures: taking the supernatant 1mL of above-mentioned steps (3), 5.0 citrate-phosphate disodium hydrogen of pH buffering is added
The water-soluble sulfur doping fluorescent carbon quantum dot of 300 μ L steps (1) preparation is added in 5 mL in liquid constant volume, and being vortexed, it is quiet to mix 0.5 min
After setting 10 min, on sepectrophotofluorometer, excitation wavelength 344nm, launch wavelength is that 413nm measures fluorescence intensity, generation
Enter step (2) regression equation, the content that Norfloxacin is calculated is 2.45 μm of ol/L, and detection is limited to 1.2nmol/L, opposite to mark
Quasi- deviation is 4.5%.
Embodiment 2: the assay step of Norfloxacin in urine sample are as follows:
(1) preparation of water-soluble sulfur doping fluorescent carbon quantum dot: the preparation method of water-soluble sulfur doping fluorescent carbon quantum dot
It is added in 100mL ultrapure water to weigh poly- (4- styrenesulfonic acid-co-maleic acid) sodium of 3.0g, ultrasonic 8min forms colourless
Clear solution, and it is transferred to ptfe autoclave, it is first 0.22 μm of filter with aperture after 200 DEG C of heating 8h, natural cooling
Film filtering, after with the bag filter that molecular cut off is 3500Da carry out dialysis treatment for 24 hours, obtain water-soluble sulfur doping fluorescence carbon amounts
Sub- point；
(2) drafting of standard working curve: with 1 step of embodiment (2)；
(3) prepared by urine sample: taking 5 mL urines, 1g active carbon is added, 1 min of vortex mixed stands filtering, takes its filter
Liquid is kept in dark place in 4 DEG C of refrigerator measurements to be analyzed；
(4) sample measures: taking the filtrate 2mL of above-mentioned steps (3), 4.0 citrate-phosphate disodium hydrogen buffer of pH is added
The water-soluble sulfur doping fluorescent carbon quantum dot of 300 μ L steps (1) preparation is added in 5 mL in constant volume, is vortexed and mixes 1min standing 15
After min, on sepectrophotofluorometer, excitation wavelength 344nm, launch wavelength is that 413nm measures fluorescence intensity, substitutes into step
(2) regression equation show that the content of Norfloxacin is 6.20 μm of ol/L, and detection is limited to 1.5 nmol/L, and relative standard deviation is
5.4%。
Embodiment 3: the assay step of Norfloxacin in milk sample are as follows:
(1) preparation of water-soluble sulfur doping fluorescent carbon quantum dot: the preparation method of water-soluble sulfur doping fluorescent carbon quantum dot
It is added in 100mL ultrapure water to weigh poly- (4- styrenesulfonic acid-co-maleic acid) sodium of 5.0g, ultrasonic 10min forms colourless
Clear solution, and it is transferred to ptfe autoclave, it is first 0.22 μm of filter with aperture after 240 DEG C of heating 6h, natural cooling
Film filtering, after with the bag filter that molecular cut off is 3200Da carry out dialysis treatment for 24 hours, obtain water-soluble sulfur doping fluorescence carbon amounts
Sub- point.
(2) drafting of standard working curve: with 1 step of embodiment (2)；
(3) prepared by milk sample: taking 5 mL milk, 10 mL methanol-waters-formic acid (28:72:0.1, v/v) extracting solution is added
With 4% trichloroacetic acid acetonitrile solution of lmL mass concentration, be vortexed 3 min, is centrifuged 10 min under ultrasound 5 min, 10000r/min,
Supernatant is taken, is kept in dark place in 4 DEG C of refrigerator measurements to be analyzed；
(4) sample measures: taking the supernatant 3mL of above-mentioned steps (3), 4.0 citrate-phosphate disodium hydrogen of pH buffering is added
The water-soluble sulfur doping fluorescent carbon quantum dot of 300 μ L steps (1) preparation is added in 5 mL in liquid constant volume, is vortexed and mixes 2 min standing
After 20 min, on sepectrophotofluorometer, excitation wavelength 344nm, launch wavelength is that 413nm measures fluorescence intensity, is substituted into
Step (2) regression equation show that the content of Norfloxacin is 2.28 μm of ol/L, and detection is limited to 1.0 nmol/L, and relative standard is inclined
Difference is 4.1%.
Embodiment 4: steps are as follows for the assay of Norfloxacin in blood:
(1) preparation of water-soluble sulfur doping fluorescent carbon quantum dot: with 1 step of embodiment (1)；
(2) drafting of standard working curve: with 1 step of embodiment (2)；
(3) prepared by blood sample: 5 mL new bloods are taken, in being centrifuged 10min with 10000r/min, removal is precipitated, and to
The perchloric acid solution protein precipitation of 2mL volumetric concentration 10% is added in upper plasma, is centrifuged 15min under 5000r/min, takes
Clear liquid is kept in dark place in 4 DEG C of refrigerators, measurement to be analyzed；
(4) sample measures: taking the preparation solution 2mL of above-mentioned steps (3), it is slow that 4.0 citrate-phosphate disodium hydrogen of pH is added
The water-soluble sulfur doping fluorescent carbon quantum dot of 300 μ L steps (1) preparation is added in 5mL in fliud flushing constant volume, and being vortexed, it is quiet to mix 1 min
After setting 10 min, on sepectrophotofluorometer, excitation wavelength 344nm, launch wavelength is that 413nm measures fluorescence intensity, generation
Enter step (2) regression equation, show that the content of Norfloxacin is 3.74 μm of ol/L, detection is limited to 1.0nmol/L, and relative standard is inclined
Difference is 4.7%.
From Fig. 1 result: water solubility sulfur doping fluorescent carbon quantum dot manufactured in the present embodiment has Norfloxacin relatively strong
Fluorescence sensitivity effect；It includes Norfloxacin, Ofloxacin, left oxygen fluorine that Fig. 2, which is by other flouroquinolone drugs of same concentrations,
In the water-soluble sulfur doping fluorescent carbon quantum dot that Sha Xing, pefloxacin, Enoxacin, gatifloxacin, Lomefloxacin are added, almost
There is no fluorescence sensitivity phenomenon, show that other coexistent flouroquinolone drugs do not interfere with the measurement of Ciprofloxacin,
Method has preferable specificity.
Claims (4)
1. a kind of method of sulfur doping carbon quantum dot fluorescence sensitivity detection Norfloxacin, which is characterized in that specific step is as follows:
(1) drafting of standard working curve: using citrate-phosphate disodium hydrogen buffer configuration concentration in 0.01~10 μm of ol/
Then water-soluble sulfur doping fluorescent carbon quantum dot solution, whirlpool is added in 8 Norfloxacin standard solution in L, each concentration 5mL
After rotation mixes standing, on sepectrophotofluorometer, excitation wavelength 344nm, launch wavelength is that 413nm measures fluorescence intensity,
It is mapped with fluorescence enhancement intensity to its respective concentration and carries out regression analysis to get standard working curve is arrived, it is husky thus to obtain promise fluorine
The equation of linear regression of star；
(2) sample preparation
Blood sample preparation: taking 5 mL new bloods, is centrifuged 10-15 min with 10000r/min, removal precipitating, and to upper layer
The perchloric acid solution protein precipitation of 1-2mL volumetric concentration 10% is added in blood plasma, 10-15min are centrifuged under 5000r/min, are taken
Supernatant is kept in dark place in 4 DEG C of refrigerators, measurement to be analyzed；
Urine sample preparation: being added 1-3g active carbon in 5-10 mL urine, and vortex mixed 30-60 s stands filtering, takes
Its filtrate is kept in dark place in 4 DEG C of refrigerators, measurement to be analyzed；
Dairy produce sample preparation: 10-15 mL methanol-water-formic acid extracting solution and l-2 mL matter are added in 5-10 mL milk sample
The trichloroacetic acid acetonitrile solution of concentration 4% is measured, is centrifuged 10-15 under vortex 1-3 min, ultrasonic 5-10min, 10000r/min
Min takes supernatant, is kept in dark place in 4 DEG C of refrigerators, measurement to be analyzed, wherein methanol-water-formic acid extracting solution is by volume
Methanol, water, formic acid are mixed and are made by the ratio of 28:72:0.1；
(3) sample measures: taking step, stepOr stepExtracting solution 1-3mL obtained, with citrate-phosphate disodium hydrogen
Buffer is settled to 5mL, and water-soluble sulfur doping fluorescent carbon quantum dot solution is added, is vortexed after mixing standing, in fluorescence spectrophotometer light
On degree meter, excitation wavelength 344nm, launch wavelength is that 413nm measures fluorescence intensity, substitutes into step (1) regression equation, calculates
The content of Norfloxacin in sample；
The preparation method of the water solubility sulfur doping fluorescent carbon quantum dot is to weigh poly- (the 4- styrenesulfonic acid-co-of 2.0-5.0g
Maleic acid) sodium is added in 100mL ultrapure water, and ultrasonic 5-10min forms colourless transparent solution, and it is anti-to be transferred to polytetrafluoroethylene (PTFE)
Kettle is answered, after 200-240 DEG C of heating 5-8h, natural cooling, is first 0.22 μm of membrane filtration with aperture, is with molecular cut off afterwards
The bag filter of 3000-3500Da carries out dialysis treatment for 24 hours, obtains water-soluble sulfur doping fluorescent carbon quantum dot.
2. the method for sulfur doping carbon quantum dot fluorescence sensitivity detection Norfloxacin according to claim 1, it is characterised in that:
Citrate-phosphate disodium hydrogen pH of buffer is 4-5.
3. the method for sulfur doping carbon quantum dot fluorescence sensitivity detection Norfloxacin according to claim 1, it is characterised in that:
The dosage of water-soluble sulfur doping fluorescent carbon quantum dot is 300 μ L.
4. the method for sulfur doping carbon quantum dot fluorescence sensitivity detection Norfloxacin according to claim 1, it is characterised in that:
It is vortexed to mix to stand and refers to that vortex mixes 0.5-2 min, be then allowed to stand 10-20 min.
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| CN109142295B (en) * | 2018-08-03 | 2020-10-27 | 咸阳师范学院 | Method for detecting cobalt ions and identifying norfloxacin in relay mode by using sulfur quantum dots |
| CN110398484A (en) * | 2019-09-09 | 2019-11-01 | 云南伦扬科技有限公司 | A kind of method of carbon dots analogue enztme combination fluorescence probe detection dopamine |
| CN112666141B (en) * | 2020-12-14 | 2024-02-27 | 广西大学 | Fluorescent ratio detection method for phosphorus-killing pesticide |
| CN113462377B (en) * | 2021-06-23 | 2023-06-20 | 青岛农业大学 | Preparation method of silicon dioxide coated carbon quantum dot composite material and application of silicon dioxide coated carbon quantum dot composite material in detection of residues of different veterinary drugs |
| CN114958365B (en) * | 2022-06-30 | 2023-08-08 | 北京兴德通医药科技股份有限公司 | Preparation method and application of cicada slough carbon quantum dots |
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