CN105738337A - Application of isorhamnetin to measurement of copper ions - Google Patents
Application of isorhamnetin to measurement of copper ions Download PDFInfo
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- CN105738337A CN105738337A CN201610112140.7A CN201610112140A CN105738337A CN 105738337 A CN105738337 A CN 105738337A CN 201610112140 A CN201610112140 A CN 201610112140A CN 105738337 A CN105738337 A CN 105738337A
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The invention discloses an application of isorhamnetin to measurement of copper ions, and belongs to the technical field of optical analysis and detection. According to the application, a 10<-5>mol/L solution is prepared from isorhamnetin by virtue of a CH3OH-PBS buffer solution with pH of 7.40, copper ions are added to form different concentrations, fluorescence intensity changes are measured by adopting fluorescence spectroscopy, a standard curve of the copper ion concentrations at different fluorescence intensities is drawn, and the copper ion concentration of an unknown sample is calculated according to the standard curve. A measurement process has the characteristics of being simple, sensitive, accurate, environmentally friendly, and the like, and is high in practicability.
Description
Technical field
The invention belongs to light technical field of analysis and detection, be specifically related to isorhamnetin application in measuring copper ion.
Background technology
Copper is widely present in the middle of nature and human body, is the indispensable trace element of human body, meanwhile,
Copper is widely used in the industries closely bound up with human lives such as metal smelt, electronics, packaging, along with people
Class social development speed is accelerated, and mineral resources disorderly exploitation phenomenon is day by day serious, except three industrial wastes, house refuse,
Electronic waste is the most increasing, ultimately results in heavy metal ion and runs off, causes soil, Heavy Metals in Waters ion
Content overproof.Although the copper requisite trace element that is human body, but, if internal copper content mistake
Height, the lighter causes gastrointestinal upset, and severe one can cause nervous lesion, parkinsonism, even life-threatening,
Therefore, in monitoring of environmental, the content of copper ion seems critically important.At present, the method for detection copper ion is a lot, often
That sees has: inductivity coupled plasma mass spectrometry, flame atomic absorption method, electrochemical process etc., but these methods
In will use many precision instruments, and sample pre-treatments is required strict, testing cost is high, inefficient.
For overcoming disadvantage mentioned above, the report of Fluorometric assay copper ion is used to get more and more.Fluorescence method is sensitive except having
Outside degree height, good selective, also have and detect quick, easy, low cost and other advantages.But, mesh
It is to be obtained by organic synthesis approach that front majority is used for detecting the fluorescent probe of copper ion, uses perhaps in experimentation
Many toxic chemical, cause the secondary pollution of environment virtually.
Isorhamnetin (C16H12O7, 3,5,7-trihydroxy-2-(4-hydroxy 3-methoxybenzene base) benzopyran-4-one)
For flavone compound, it is widely present in the plant of nature, such as: Folium Ginkgo, Folium Mori, Fructus Hippophae etc..
Isorhamnetin has the multiple biological activitys such as antioxidation, antiinflammatory, antitumor, but its fluorescence property relevant is studied
Less, not yet report about its research metal ion detected as fluorescent probe.
Summary of the invention
Goal of the invention: for the deficiencies in the prior art, it is an object of the present invention to provide isorhamnetin and is measuring
Application in copper ion, not only has the advantages such as selectivity is good, highly sensitive, quick, convenient, and has
Source is wide, environmental protection, the advantage such as pollution-free.
Technical scheme: in order to realize foregoing invention purpose, the technical solution used in the present invention is:
Isorhamnetin application in measuring copper ion.
Described application, comprises the following steps:
(1) respectively the copper ion solution of different volumes is joined the CH of isorhamnetin3OH-PBS buffering is molten
In liquid, use the fluorescence intensity of each solution of fluorescence spectrophotometer record, draw the fluorescence intensity mark to copper ion concentration
Directrix curve；
(2) liquid to be measured for copper ion is joined the CH of isorhamnetin3In OH-PBS buffer solution, use glimmering
The fluorescence intensity of photothermal spectroscopic analyzer recording solution；
(3) concentration of copper ion in solution to be measured is determined according to standard curve.
Described CH3The pH of OH-PBS buffer solution is 7.40, and methanol is 1 with the V/V of phosphate-buffered salt:
99。
Described fluorescent strength determining condition is that to excite slit be 5nm, and transmitting slit is 15nm, sweep limits
For 450-700nm, excitation wavelength is 390nm, a length of 542nm of transmitted wave.
Described copper ion concentration scope is 1.0 × 10-8-1.9×10-6mol·L-1, detection is limited to 4.0 × 10-9m
ol·L-1, the concentration of isorhamnetin is 1 × 10-5mol·L-1。
The fluorescence intensity standard curve to copper ion concentration, y=-9.57x+234.2, y are fluorescence intensity, x
For copper ion concentration, linearly dependent coefficient is R2=0.997, the range of linearity is 1.0 × 10-8-1.9×10-6mol·
L-1, detection is limited to 4.0 × 10-9mol·L-1。
Beneficial effect: compared with prior art, the present invention utilizes isorhamnetin cheap and easy to get as fluorescent probe
Detect the copper ion in aqueous solution, the Changing Pattern of fluorescence intensity after utilizing isorhamnetin to be coordinated with copper ion,
Draw the fluorescence intensity standard curve to copper ion concentration, obtain copper ion in unknown sample according to standard curve
Concentration.Simple to operate, fast, strong to copper ion identification ability, little by other ion interference, highly sensitive,
The range of linearity is big, and detection limit is low；And in the PBS aqueous solution of pH=7.40, copper ion is detected, have
Hope in the cell of application, by content of copper ion in the technology for detection cells such as cell imaging.
Accompanying drawing explanation
Fig. 1 is CH3In OH-PBS buffer solution, isorhamnetin is to copper ion fluorescence titration spectrogram；
Fig. 2 is the isorhamnetin fluorescence titration curve chart to copper ion；
Fig. 3 is the isorhamnetin selectivity result figure to different metal ion.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described further.
Embodiment 1
Isorhamnetin application in measuring copper ion, comprises the following steps:
(1) accurately weigh 0.0316g isorhamnetin, with in methanol constant volume to 10mL volumetric flask, be made into
Concentration is 1 × 10-2mol·L-1Solution, takes 0.1mL PBS buffer solution (pH=7.40) and is settled to 1
In 00mL volumetric flask, it is made into 1 × 10-5mol·L-1Solution, stand-by；Accurately weigh 0.0200g Cu (AC)2
·H2O water is settled in 10mL volumetric flask, and being made into containing copper ion concentration is 1 × 10-2mol·L-1Molten
Liquid, and it is diluted to 1 × 10 successively-3, 1 × 10-4, 1 × 10-5mol·L-1Solution, stand-by.
(2) prepare the sample cell of some 5mL dried and clean, be separately added into 2mL1 × 10-5mol·L-1
Isorhamnetin solution, and the copper ion solution of different volumes variable concentrations, be made into isorhamnetin concentration and be 1
×10-5mol·L-1, copper ion concentration is respectively 0 × 10-7mol·L-1, 0.1 × 10-7mol·L-1, 0.5 × 10- 7mol·L-1, 1.0 × 10-7mol·L-1, 2.0 × 10-7mol·L-1, 3.0 × 10-7mol·L-1, 4.0 × 10-7
mol·L-1, 5.0 × 10-7mol·L-1, 6.0 × 10-7mol·L-1, 7.0 × 10-7mol·L-1, 8.0 × 10-7
mol·L-1, 9.0 × 10-7mol·L-1, 10.0 × 10-7mol·L-1, 11.0 × 10-7mol·L-1, 12.0 × 1
0-7mol·L-1, 13.0 × 10-7mol·L-1, 14.0 × 10-7mol·L-1, 15.0 × 10-7mol·L-1, 16.0
×10-7mol·L-1, 17.0 × 10-7mol·L-1, 18.0 × 10-7mol·L-1, 19.0 × 10-7mol·L-1, 2
0.0×10-7mol·L-1, 40.0 × 10-7mol·L-1, 60.0 × 10-7mol·L-1, 80.0 × 10-7mol·L-1,
100.0×10-7mol·L-1Solution.
(3) arranging fluorescence spectrophotometer excitation wavelength is 390nm, excites and launches slit width and is respectively 5n
M and 15nm, sweep limits is set to 450nm to 700nm；Above-mentioned solution is tested, respectively
Record above-mentioned solution fluorescence intensity at a length of 542nm of transmitted wave, as it is shown in figure 1, from 1 to 27, copper
Ion concentration is followed successively by 0 × 10-7mol·L-1, 0.1 × 10-7mol·L-1, 0.5 × 10-7mol·L-1, 1.0 × 1
0-7mol·L-1, 2.0 × 10-7mol·L-1, 3.0 × 10-7mol·L-1, 4.0 × 10-7mol·L-1, 5.0 × 10-7mol·L-1, 6.0 × 10-7mol·L-1, 7.0 × 10-7mol·L-1, 8.0 × 10-7mol·L-1, 9.0 × 10-7
mol·L-1, 10.0 × 10-7mol·L-1, 11.0 × 10-7mol·L-1, 12.0 × 10-7mol·L-1, 13.0 × 1
0-7mol·L-1, 14.0 × 10-7mol·L-1, 15.0 × 10-7mol·L-1, 16.0 × 10-7mol·L-1, 17.0
×10-7mol·L-1, 18.0 × 10-7mol·L-1, 19.0 × 10-7mol·L-1, 20.0 × 10-7mol·L-1, 4
0.0×10-7mol·L-1, 60.0 × 10-7mol·L-1, 80.0 × 10-7mol·L-1, 100.0 × 10-7mol·L- 1。
(4) drawing the fluorescence intensity standard curve to copper ion concentration, y=-9.57x+234.2, y are glimmering
Light intensity, x is copper ion concentration, and linearly dependent coefficient is R2=0.997, the range of linearity is 1.0 × 10-8-1.9
×10-6mol·L-1, detection is limited to 4.0 × 10-9mol·L-1, the detection to trace copper ion can be completed, as
Shown in Fig. 2.
(5) mensuration of sample: configuration standard specimen and the copper ion treating test sample (unknown concentration) according to the method described above
Isorhamnetin solution, isorhamnetin concentration is 1 × 10-5mol·L-1, standard specimen copper ion concentration is 5.5 × 10-7m
ol·L-1(standard specimen 1), 10.5 × 10-7mol·L-1(standard specimen 2), 15.5 × 10-7mol·L-1(standard specimen 3)；Adopt
By same procedure, above-mentioned solution is carried out fluorometric investigation, record above-mentioned solution at a length of 538nm of transmitted wave
Fluorescence intensity, repeats to survey three times, according to the fluorescence intensity standard curve to copper ion concentration: y=-9.57x+
234.2, it is calculated the concentration of copper ion, concrete outcome is as shown in table 1.
Table 1 measurement result table.
In table, concentration unit is 10-7mol·L-1。
Embodiment 2
Containing isorhamnetin 1 × 10-5mol·L-1In buffer, it is separately added into containing Li+、Na+、K+、Mg2+、
Ca2+、Cr3+、Mn2+、Fe3+、Co2+、Ni2+、Cu2+、Zn2+、Pb2+、Cd2+、La3+、Ce3+、E
u3+、Gd3+、Dy3+Acetate Solution, wherein Li+、Na+、K+、Mg2+、Ca2+Concentration is copper ion
5 times of concentration, Cr3+、Mn2+、Fe3+、Co2+、Ni2+、Zn2+、Pb2+、Cd2+、La3+、Ce3+、E
u3+、Gd3+、Dy3+Concentration is 2 times of copper ion concentration, records under the identical condition of embodiment 1 and method
Fluorescence intensity (in figure, other metal ions not marked from left to right are followed successively by: Li as shown in Figure 3+、Na+、
K+、Mg2+、Ca2+、Cr3+、Mn2+、Fe3+、Co2+、Ni2+、Zn2+、Pb2+、Cd2+、La3+、Ce3 +、Eu3+、Gd3+、Dy3+), as seen from the figure, the only membership that adds of copper ion causes obvious fluorescence to quench
Going out, other metal ions are less on the impact of isorhamnetin fluorescence intensity, can ignore, demonstrate isorhamnetin
Copper ion is had preferable selectivity.Additionally, from fluorescence titration spectrum and titration curve it can be seen that different Mus
The range of linearity of Li Su detection copper ion crosses over three orders of magnitude, and scope is relatively big, and detection limit can reach 10-9
mol·L-1, detection limit is relatively low, demonstrates that isorhamnetin has preferable susceptiveness to copper ion.It can be said that
Bright, use the method, the copper ion of trace can be detected as fluorescent probe, not only have by isorhamnetin
Have high selectivity and a susceptiveness, and fast, rapidly, environmental protection, result is preferable.
Claims (6)
1. isorhamnetin application in measuring copper ion.
Application the most according to claim 1, it is characterised in that comprise the following steps:
(1) respectively the copper ion solution of different volumes is joined the CH of isorhamnetin3OH-PBS buffering is molten
In liquid, use the fluorescence intensity of each solution of fluorescence spectrophotometer record, draw the fluorescence intensity mark to copper ion concentration
Directrix curve；
(2) liquid to be measured for copper ion is joined the CH of isorhamnetin3In OH-PBS buffer solution, use glimmering
The fluorescence intensity of photothermal spectroscopic analyzer recording solution；
(3) concentration of copper ion in solution to be measured is determined according to standard curve.
Application the most according to claim 2, it is characterised in that: described CH3OH-PBS buffering is molten
The pH of liquid is 7.40, and methanol is 1:99 with the V/V of phosphate-buffered salt.
Application the most according to claim 2, it is characterised in that: described fluorescent strength determining condition is
Exciting slit is 5nm, and transmitting slit is 15nm, and sweep limits is 450-700nm, and excitation wavelength is 390n
M, a length of 542nm of transmitted wave.
Application the most according to claim 2, it is characterised in that: described copper ion concentration scope is 1.
0×10-8-1.9×10-6mol·L-1, detection is limited to 4.0 × 10-9mol·L-1, the concentration of isorhamnetin is 1 ×
10-5mol·L-1。
Application the most according to claim 2, it is characterised in that: the fluorescence intensity mark to copper ion concentration
Directrix curve, y=-9.57x+234.2, y are fluorescence intensity, and x is copper ion concentration, and linearly dependent coefficient is
R2=0.997, the range of linearity is 1.0 × 10-8-1.9×10-6mol·L-1, detection is limited to 4.0 × 10-9mol·L-1。
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| CN104359885A (en) * | 2014-11-21 | 2015-02-18 | 南京林业大学 | Application of quercetin to determination of Cu<2+> as fluorescent probe and method for determining Cu<2+> |
| CN104830315A (en) * | 2015-05-11 | 2015-08-12 | 济南大学 | Bivalent copper ion fluorescent probe and preparation method and application thereof |
| CN104849271A (en) * | 2015-05-26 | 2015-08-19 | 中国科学院烟台海岸带研究所 | Method for detecting trace bivalent copper ions by virtue of cyanine-based probe |
| CN104860879A (en) * | 2015-04-15 | 2015-08-26 | 齐鲁工业大学 | Malononitrile isophorone copper ion fluorescent probe and preparation method thereof |
| US20150276722A1 (en) * | 2012-11-07 | 2015-10-01 | Canox4Drug S.P.A. | Method and kit for determination of free copper in serum |
| CN105352920A (en) * | 2015-10-08 | 2016-02-24 | 河南师范大学 | Method using 1,4-dihydroxy-9,10-anthraquinone thiosemicarbazone compound as fluorescent probe to detect copper ions |
-
2016
- 2016-02-29 CN CN201610112140.7A patent/CN105738337A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150276722A1 (en) * | 2012-11-07 | 2015-10-01 | Canox4Drug S.P.A. | Method and kit for determination of free copper in serum |
| CN104359885A (en) * | 2014-11-21 | 2015-02-18 | 南京林业大学 | Application of quercetin to determination of Cu<2+> as fluorescent probe and method for determining Cu<2+> |
| CN104860879A (en) * | 2015-04-15 | 2015-08-26 | 齐鲁工业大学 | Malononitrile isophorone copper ion fluorescent probe and preparation method thereof |
| CN104830315A (en) * | 2015-05-11 | 2015-08-12 | 济南大学 | Bivalent copper ion fluorescent probe and preparation method and application thereof |
| CN104849271A (en) * | 2015-05-26 | 2015-08-19 | 中国科学院烟台海岸带研究所 | Method for detecting trace bivalent copper ions by virtue of cyanine-based probe |
| CN105352920A (en) * | 2015-10-08 | 2016-02-24 | 河南师范大学 | Method using 1,4-dihydroxy-9,10-anthraquinone thiosemicarbazone compound as fluorescent probe to detect copper ions |
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