CN103424395A - Method for detecting medicine components in plasma - Google Patents
Method for detecting medicine components in plasma Download PDFInfo
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- CN103424395A CN103424395A CN201310409045XA CN201310409045A CN103424395A CN 103424395 A CN103424395 A CN 103424395A CN 201310409045X A CN201310409045X A CN 201310409045XA CN 201310409045 A CN201310409045 A CN 201310409045A CN 103424395 A CN103424395 A CN 103424395A
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Abstract
The invention discloses a method for detecting medicine components in plasma. The method comprises the steps that a. medicine water solution and substrate materials are evenly mixed and subjected to standing, the pH value of the mixture is adjusted, the surface enhancing Raman spectrum of medicine is detected; b. blank plasma without proteins and medicine is evenly mixed with the substrate materials, standing is carried out, the pH value of the mixture is adjusted to the pH value in the step a, the surface enhancing Raman spectrum of the blank plasma is detected; c. plasma without proteins and with medicine is evenly mixed with the substrate materials, standing is carried out, the pH value of the mixture is adjusted to the pH value in the step a, the surface enhancing Raman spectrum of the plasma with the medicine is detected; d. the surface enhancing Raman spectrum obtained in the step b is removed from the surface enhancing Raman spectrum obtained in the step c, then the surface enhancing Raman spectrum obtained in the step c is compared with the surface enhancing Raman spectrum obtained in the step a, and whether the medicine is contained in the plasma is judged. According to the method, only the simple protein-removing process is carried out on a plasma sample, other preprocessing processes are of no need, and detecting time is short.
Description
Technical field
The present invention relates to the Pharmaceutical Analysis field, in particular to the method that detects blood plasma Chinese traditional medicine composition.
Background technology
The detection of blood plasma Chinese traditional medicine composition is the common technology carried out in the researchs such as medication effect and drug toxicity.In correlation technique, the drug ingedient detected in blood plasma mainly adopts liquid chromatography technology or LC-MS-MS.Liquid chromatographic detection or liquid chromatograph mass spectrography detect and carry out in laboratory, at first need to remove the protein in plasma sample, then the sample that does not contain protein is carried out that heating water bath, nitrogen stream dry up, the pre-treatment such as dissolving again, re-use liquid chromatograph or liquid chromatograph-mass spectrometer completes.
Adopt liquid chromatography technology or LC-MS-MS to detect the drug ingedient in blood plasma, the complex pretreatment of plasma sample, liquid chromatographic detection or liquid chromatograph mass spectrography are long detection time.
Summary of the invention
The object of the present invention is to provide a kind of method that detects blood plasma Chinese traditional medicine composition, to solve the above problems.A kind of method that detects blood plasma Chinese traditional medicine composition is provided in an embodiment of the present invention, has been operated in accordance with the following steps:
A, by pharmaceutical aqueous solution with backing material, mix, standing, regulate the pH value of gained potpourri to the 1-14 scope, the Surface enhanced raman spectroscopy of this medicine of test under exciting light, wherein, the concentration of pharmaceutical aqueous solution is: 10
-3-10
-8Mol/L;
B, the blank plasma that does not contain medicine that will remove protein mix with backing material, standing, the pH value of regulating the gained potpourri is identical with the pH value in step a, and under described exciting light, test does not contain the Surface enhanced raman spectroscopy of the blank plasma of medicine;
C, the blood plasma that contains medicine that will remove protein mixes with backing material, standing, the pH value of regulating the gained potpourri is identical with the pH value in step a, the Surface enhanced raman spectroscopy of the blood plasma that test contains medicine under described exciting light, the concentration of the blood plasma that wherein, contains medicine is: 0.02-50ug/ml;
D, by after the Surface enhanced raman spectroscopy of the blank plasma that does not contain medicine that obtains in the Surface enhanced raman spectroscopy deduction step b of the blood plasma that contains medicine that obtains in step c, compare with the Surface enhanced raman spectroscopy of the medicine obtained in step a, by whether having corresponding characteristic peak to judge in described blood plasma, whether contain described medicine.
The method of the detection blood plasma Chinese traditional medicine composition of the above embodiment of the present invention, only need to simply remove the processing of protein to plasma sample, do not need other pretreatment process; Detection time is short, usually only needs can complete in 2 minutes; In addition, above-mentioned detection can be used portable Raman wide spectrum instrument, and this instrument volume is little, and low price is easy to carry, can realize Site Detection.
The accompanying drawing explanation
The Surface enhanced raman spectroscopy of the aqueous solution that Fig. 1 is embodiment 1 Chinese traditional medicine atenolol under different pH value conditions;
Surface enhanced raman spectroscopy under the condition that the aqueous solution that Fig. 2 is embodiment 1 Chinese traditional medicine atenolol and the plasma sample that contains the medicine atenolol of variable concentrations are 2 in the pH value;
The Surface enhanced raman spectroscopy of the aqueous solution that Fig. 3 is embodiment 2 Chinese traditional medicine sibutramines under different pH value conditions;
Surface enhanced raman spectroscopy under the condition that the aqueous solution that Fig. 4 is embodiment 2 Chinese traditional medicine sibutramines and the plasma sample that contains the medicine sibutramine of variable concentrations are 2 in the pH value;
The Surface enhanced raman spectroscopy of the aqueous solution that Fig. 5 is embodiment 3 Chinese traditional medicine Rosiglitazone Maleates under different pH value conditions;
Surface enhanced raman spectroscopy under the condition that the aqueous solution that Fig. 6 is embodiment 3 Chinese traditional medicine Rosiglitazone Maleates and the plasma sample that contains the medicine Rosiglitazone Maleate of variable concentrations are 2 in the pH value;
The Surface enhanced raman spectroscopy of the aqueous solution that Fig. 7 is embodiment 4 Chinese traditional medicine DB2s under different pH value conditions;
Surface enhanced raman spectroscopy under the condition that the aqueous solution that Fig. 8 is embodiment 4 Chinese traditional medicine DB2s and the plasma sample that contains the medicine DB2 of variable concentrations are 12 in the pH value;
Fig. 9 shows the Surface enhanced raman spectroscopy of blank plasma under different pH value conditions that does not contain medicine in embodiment 1-4.
Embodiment
Below by specific embodiment, also by reference to the accompanying drawings the present invention is described in further detail.
The embodiment of the present invention provides a kind of method that detects blood plasma Chinese traditional medicine composition, specifically utilizes Surface enhanced raman spectroscopy (Surface Enhanced Raman Scattering, be abbreviated as SERS) to detect the composition of blood plasma Chinese traditional medicine.
Surface enhanced raman spectroscopy refers to that their Raman scattering intensity can increase by 10 when some molecules are adsorbed on some coarse metal (as silver, copper, gold etc.) surface
4-10
6Doubly.Chemical compound lot can produce the SERS effect, and hitherto reported has the molecule of Raman scattering enhancement effect that good hundreds of kind is arranged, and existing inorganic molecule has again the even large molecule of organic molecule, and its main condition precedent is that molecular energy is adsorbed onto substrate material surface.Can't determine now which quasi-molecule is definitely there is no enhancement effect, the SERS effect that can only determine some molecules is apparent in view and other are difficult to observe.Than the molecule major part that is easier to detect enhancement effect, be those molecules that contain two keys, lone electron pair and ring structure, those molecules with saturated structures are difficult to observe the SERS effect.
In the embodiment of the present invention, detect the method for blood plasma Chinese traditional medicine composition, operated in accordance with the following steps:
A, by pharmaceutical aqueous solution with backing material, mix, standing, regulate the pH value of gained potpourri to the 1-14 scope, the Surface enhanced raman spectroscopy of this medicine of test under exciting light, wherein, the concentration of pharmaceutical aqueous solution is: 10
-3-10
-8Mol/L;
B, the blank plasma that does not contain medicine that will remove protein mix with backing material, standing, the pH value of regulating the gained potpourri is identical with the pH value in step a, and under described exciting light, test does not contain the Surface enhanced raman spectroscopy of the blank plasma of medicine;
C, the blood plasma that contains medicine that will remove protein mixes with backing material, standing, the pH value of regulating the gained potpourri is identical with the pH value in step a, the Surface enhanced raman spectroscopy of the blood plasma that test contains medicine under described exciting light, the concentration of the blood plasma that wherein, contains medicine is: 0.02-50ug/ml;
D, by after the Surface enhanced raman spectroscopy of the blank plasma that does not contain medicine that obtains in the Surface enhanced raman spectroscopy deduction step b of the blood plasma that contains medicine that obtains in step c, compare with the Surface enhanced raman spectroscopy of the medicine obtained in step a, by whether having corresponding characteristic peak to judge in described blood plasma, whether contain described medicine.
Utilize said method to detect the medicine in blood plasma, only need to simply remove the processing of protein to plasma sample, do not need other pretreatment process; Detection time is short, usually only needs to complete in 2 minutes; In addition, above-mentioned detection can be used portable Raman wide spectrum instrument, and this instrument volume is little, and low price is easy to carry, can realize Site Detection.
The detection method that the embodiment of the present invention provides, be not particularly limited the concentration of drug ingedient, as long as can carry out the test of Surface enhanced raman spectroscopy.Yet this drug ingedient has the concentration range an of the best, concentration is excessive, does not meet the best scattering condition of monomolecular adsorption layer, and drug molecule can not form mono layer adsorption or combination at nano-particle surface just, affects the absorption orientation of drug molecule; Simultaneously, due to drug molecule, to assemble rapidly the particle diameter made with the backing material nano particle of scattering molecular action excessive, surpassed its effective range, thereby can not produce stronger enhancement effect; Concentration is too small, and owing to not having abundant drug molecule to be adsorbed onto the nano-particle surface in backing material, the scattered signal of generation is more weak, and test is less than stronger SERS signal.
Simultaneously, after clinical application, the concentration range of conventional blood plasma Chinese traditional medicine is 0.02-50ug/ml.
Therefore, the method that the embodiment of the present invention provides, the concentration of pharmaceutical aqueous solution is: 10
-3-10
-8Mol/L, the concentration of the blood plasma that contains medicine is: 0.02-50ug/ml.
When detect certain medicine in blood plasma have situation the time, only need, according to above-mentioned steps a-d operation, can obtain conclusion.
The detection method of the blood plasma Chinese traditional medicine that the embodiment of the present invention provides, described medicine is Western medicine, can be the Western medicine of the various molecules that contain two keys, lone electron pair and ring structure, such as Rosiglitazone Maleate, the two flesh of hydrochloric acid benzene second, the two flesh of hydrochloric acid diformazan, hydrochloric acid, adjoin and cough up row ketone, sibutramine etc.
Method provided by the invention, it is usually relevant with energy that test surfaces strengthens the excitation wavelength that Raman spectrum adopts, and in general, the shorter luminous energy of wavelength is stronger, Raman scattering also should be stronger, and excitation wavelength has selective excitation to different functional groups.Such as, under some excitation wavelengths excite, the SERS effect of group A is the strongest, group B a little less than, but under another excitation wavelength excites, situation may be contrary.Therefore, when detecting different medicines, can be according to the selected situation selective excitation light wavelength excited of functional group.
The method of detection blood plasma Chinese traditional medicine composition provided by the invention, can adopt Portable Raman spectrometer, and the light wavelength that excites that Portable Raman spectrometer adopts is preferably 785nm.The Portable Raman spectrometer volume is little, and price is low, easy to carry, detects fast, only needs within 2 minutes, to complete whole testing process, can realize Site Detection, and accuracy of detection can reach the scope of conventional blood concentration simultaneously.
The method of the detection blood plasma Chinese traditional medicine composition that the embodiment of the present invention provides, under normal conditions, the mass ratio that pharmaceutical aqueous solution or the blood plasma that contains medicine are mixed with backing material is not particularly limited, and usually depends on the adsorptive power of medicine on backing material.Generally, the mass ratio of pharmaceutical aqueous solution and backing material solution is 10:1-10:100, and the middle blood plasma that contains medicine and the mass ratio of backing material solution are 10:1-10:100.Between the two, not contact, can be the same or different.In testing process, the mass ratio that pharmaceutical aqueous solution or the blood plasma that contains medicine mix with backing material is generally 1:1.
In the process of carrying out the Surface enhanced raman spectroscopy test, the change of backing material-drug molecule system pH, to cause that being adsorbed group changes with suction type, relative distance, relative how much factors such as state between the backing material metal surface, thereby appear at the Raman scattering enhancing effect difference of detected molecule under different pH condition.For detected molecule, there is the pH value an of the best, under this optimal pH condition, it is best that the Raman scattering obtained strengthens effect.
Therefore, pass through the experiment in testing process in the embodiment of the present invention, at first under different pH value conditions, under the condition that in the present embodiment, preferably the pH value is 2,7,12, the Surface enhanced raman spectroscopy of testing drug molecule, then select this drug molecule Raman scattering to strengthen the best wide spectrum of effect, and the drug molecule Raman scattering is strengthened to the best pH value of effect as testing the pH value that contains medicine blood plasma and do not contain the Surface enhanced raman spectroscopy of medicine blood plasma.While making to detect Surface enhanced raman spectroscopy, can obtain Raman scattering and strengthen the best wide spectrum of effect, contribute to judge accurately with analysed for plasma in drug ingedient.
Wherein, the pH value can be by acid, alkali be regulated, and acid can be one or more in hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid, and alkali can be one or more in NaOH, potassium hydroxide and sodium carbonate.Generally, regulating the acid of pH value or the concentration of alkali is 0.1mol/L.
The method of the detection blood plasma Chinese traditional medicine composition that the embodiment of the present invention provides, only need to remove the protein containing in medicine blood plasma, will discharge with the drug ingedient of protein bound, just can use Raman spectrometer to be detected.The removal method of protein is generally first precipitation and refilters, and the intermediate processing of protein can adopt enzyme hydrolysis method, acid-base method, neutral sulfity process and heating etc.Wherein, the protein in strong acid method removal system more up hill and dale and do not introduce the material that disturbs Raman detection.When the pH value that adds strong acid to make solution, during lower than the isoelectric point of protein, protein exists with cationic form, then with system in acid radical anion form the insoluble salt precipitation.
In the embodiment of the present invention, remove specifically operation in accordance with the following steps of protein: the ratio that is 1:0.6-1:1 according to the mass ratio of the perchloric acid containing medicine blood plasma and 6% massfraction, to containing in medicine blood plasma, adding perchloric acid, mix, centrifugal 10-15min under the condition of 10000-12000r/min, get supernatant, obtain not containing protein containing medicine blood plasma.
The method of the detection blood plasma Chinese traditional medicine composition that the embodiment of the present invention provides, only need carry out the processing of simple protein removal, compare with correlation technique, not need blood plasma is carried out to complicated pre-treatment, operate simplyr, make testing result reduced by the influence degree of pre-treatment.
The detection method of the blood plasma Chinese traditional medicine composition that the embodiment of the present invention provides, backing material can be the various backing materials that can be used for carrying out the Surface enhanced raman spectroscopy test, can be for example silver sol, aurosol or silver-colored billon colloidal sol.Consider the absorption of medicine in backing material, the present invention preferably adopts silver sol.
The present invention adopts following method to prepare silver sol: silver nitrate aqueous solution is heated to boiling, under agitation dropwise adds trisodium citrate aqueous solution, keep boiling to obtain silver sol, and cooling.
In addition, silver sol also can, by commercially available, can also prepare by the method for this area routine.
Concrete, by the liquor argenti nitratis ophthalmicus 400-500ml of 0.15-0.20mg/ml, be heated to boiling, then the citric acid three sodium solution 10-20ml that is 0.5-1% by massfraction dropwise adds, constantly stir during this time, drip rear continuation and keep boiling within 1-5 hour, to obtain fulmargin, cooling rear standby.
Wherein, heating can adopt the method for this area routine, as long as can be heated to boiling.
The embodiment of the present invention provides a kind of method that detects the medicine atenolol in blood plasma, is operated in accordance with the following steps:
1, the plasma sample that preparation contains the medicine atenolol
Precision takes medicine atenolol standard items 2 ㎎ in the 10ml volumetric flask, adds ultrapure water to scale mark, and dissolving shakes up, and obtains the medicine atenolol storing solution of 200ug/ml.By this storing solution ultrapure water stepwise dilution, obtain the medicine atenolol working fluid that concentration is respectively 200ug/ml, 20ug/ml, 2ug/ml, 0.2ug/ml.
The above-mentioned working fluid 0.5ml of accurate absorption, add 4.5ml not contain the plasma sample of medicine atenolol, and vortex mixes 30s, obtains concentration and is respectively 20ug/ml, 2ug/ml, 0.2ug/ml, the plasma sample that contains the medicine atenolol of 0.02ug/ml.
2, remove the protein in the plasma sample that contains the medicine atenolol
In the plasma sample that contains the medicine atenolol at 5ml, precision adds the perchloric acid 3ml that massfraction is 6%, and vortex mixes 5min, and standing 10min, with the centrifugal 10min of 12000r/min, gets supernatant as the plasma sample detected.
3, the aqueous solution for preparing the medicine atenolol
The medicine atenolol is soluble in water, and being mixed with concentration is 10
-5The aqueous solution of mol/L.
4, prepare silver sol
Get the liquor argenti nitratis ophthalmicus 500ml of 0.15mg/ml, be heated to boiling, then dropwise adding massfraction is citric acid three sodium solution 1%, 10ml, during constantly stir, drip rear continuation and keep boiling within 1～5 hour, to obtain silver sol, cooling rear standby.
5, the Surface enhanced raman spectroscopy of testing drug atenolol
The aqueous solution of medicine atenolol is mixed with silver sol, the pH value that the NaOH aqueous solution that the aqueous hydrochloric acid solution that is 0.1mol/L by concentration and concentration are 0.1mol/L is regulated resulting atenolol-silver sol mixed system is 2,7 and 12, acid, neutral and alkaline, then under the condition that is 2,7 and 12 in the pH value, under the exciting light that is 785nm at wavelength, the Surface enhanced raman spectroscopy of testing drug atenolol.The results are shown in Figure 1, wherein, a is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 2 in the pH value; B is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 12 in the pH value; C is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 7 in the pH value.As seen from Figure 1, at wavelength, be under the 785nm exciting light, under the condition that the pH value is 2, the surface-enhanced Raman effects of medicine atenolol is the most obvious.Therefore, the pH value that in following steps, test surfaces enhancing Raman spectrum is selected is 2.
6, test does not contain the Surface enhanced raman spectroscopy of the blank plasma sample of medicine
The blank plasma sample that does not contain medicine is mixed with silver sol, and the pH value of regulating the gained potpourri is 2 and 12.Under the exciting light that is 785nm at wavelength, test does not contain the Surface enhanced raman spectroscopy of the blank plasma sample of medicine.The results are shown in Figure 9, wherein, the Surface enhanced raman spectroscopy that a is blank plasma sample under the pH condition that is 2, the Surface enhanced raman spectroscopy that b is blank plasma sample under the pH condition that is 12.As can be seen from Figure 9, do not contain the blank plasma of any medicine under different potential of hydrogen conditions, the equal no signal of its Surface enhanced raman spectroscopy.
The Surface enhanced raman spectroscopy of the plasma sample that 7, test contains the medicine atenolol
The plasma sample to be measured prepared is mixed with silver sol, and the pH value of regulating the gained potpourri is 2.Under the exciting light that is 785nm at wavelength, the Surface enhanced raman spectroscopy of the plasma sample that test contains the medicine atenolol.The results are shown in Figure 2, wherein, the Surface enhanced raman spectroscopy that obtains of test under the condition that the aqueous solution that a is the medicine atenolol is 2 in the pH value, it is to test the Surface enhanced raman spectroscopy obtained under 200ug/ml, 20ug/ml, 2ug/ml, the 0.2ug/ml plasma sample condition that is 2 in pH value that b-e is followed successively by concentration.To in Fig. 2, containing the Surface enhanced raman spectroscopy b-e of the plasma sample of the medicine atenolol of variable concentrations and the aqueous solution Surface enhanced raman spectroscopy a under the same conditions of Fig. 2 Chinese traditional medicine atenolol, compare, can find out b-e with have corresponding characteristic peak in a, therefore, can reach a conclusion: contain the medicine atenolol in plasma sample.In addition, from b-e, it can also be seen that, relative intensity and the drug concentration in blood plasma of characteristic peak have certain correlativity, can compare the size of different pharmaceutical blood plasma Chinese traditional medicine concentration by the relative intensity of characteristic peak.
In the present embodiment, use the Portable Raman spectrometer test surfaces to strengthen Raman spectrum.
Visible, the method for the detection blood plasma Chinese traditional medicine atenolol that adopts the embodiment of the present invention to provide can effectively detect the medicine atenolol in blood plasma, and the time of the present embodiment test is total up to 2 minutes.
Embodiment 2
The embodiment of the present invention provides a kind of method that detects the medicine sibutramine in blood plasma, and its operation steps is with embodiment 1, different, and the concentration of medicine sibutramine aqueous solution is 10
-4Mol/L, the concentration of the plasma sample that contains the medicine sibutramine is 500ug/ml, 50ug/ml, 5ug/ml, 0.5ug/ml.
The testing result of medicine sibutramine as shown in Figure 3-4.
In Fig. 3, a is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 2 in the pH value; B is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 12 in the pH value; C is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 7 in the pH value.As can be seen from Figure 3, at wavelength, be under the 785nm exciting light, under the condition that the pH value is 2, the surface-enhanced Raman effects of medicine sibutramine is the most obvious.
In Fig. 4, the Surface enhanced raman spectroscopy that obtains of test under the condition that the aqueous solution that a is the medicine sibutramine is 2 in the pH value, it is to test the Surface enhanced raman spectroscopy obtained under 500ug/ml, 50ug/ml, 5ug/ml, the 0.5ug/ml plasma sample condition that is 2 in pH value that b-e is followed successively by concentration.To in Fig. 4, containing the Surface enhanced raman spectroscopy b-e of the plasma sample of the medicine sibutramine of variable concentrations and the aqueous solution Surface enhanced raman spectroscopy a under the same conditions of Fig. 4 Chinese traditional medicine sibutramine, compare, can find out b-e with have corresponding characteristic peak in a, therefore, can reach a conclusion: contain the medicine sibutramine in plasma sample.In addition, from b-e, it can also be seen that, relative intensity and the drug concentration in blood plasma of characteristic peak have certain correlativity, can compare the size of different pharmaceutical blood plasma Chinese traditional medicine concentration by the relative intensity of characteristic peak.
In the present embodiment, use the Portable Raman spectrometer test surfaces to strengthen Raman spectrum.
Visible, the method for the detection blood plasma Chinese traditional medicine sibutramine that adopts the embodiment of the present invention to provide can effectively detect the medicine sibutramine in blood plasma, and the time of the present embodiment test is total up to 2 minutes.
Embodiment 3
The embodiment of the present invention provides a kind of method that detects the medicine Rosiglitazone Maleate in blood plasma, and its operation steps is with embodiment 1, different, and the concentration of medicine Rosiglitazone Maleate aqueous solution is 10
-8Mol/L, the concentration of the plasma sample that contains the medicine Rosiglitazone Maleate is 300ug/ml, 30ug/ml, 3ug/ml, 0.3ug/ml.
The testing result of medicine Rosiglitazone Maleate is as shown in Fig. 5-6.
In Fig. 5, a is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 2 in the pH value; B is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 12 in the pH value; C is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 7 in the pH value.As can be seen from Figure 5, at wavelength, be under the 785nm exciting light, under the condition that the pH value is 2, the surface-enhanced Raman effects of medicine Rosiglitazone Maleate is the most obvious.
In Fig. 6, the Surface enhanced raman spectroscopy that obtains of test under the condition that the aqueous solution that a is the medicine Rosiglitazone Maleate is 2 in the pH value, it is to test the Surface enhanced raman spectroscopy obtained under 300ug/ml, 30ug/ml, 3ug/ml, the 0.3ug/ml plasma sample condition that is 2 in pH value that b-e is followed successively by concentration.To in Fig. 6, containing the Surface enhanced raman spectroscopy b-e of the plasma sample of the medicine Rosiglitazone Maleate of variable concentrations and the aqueous solution Surface enhanced raman spectroscopy a under the same conditions of Fig. 6 Chinese traditional medicine Rosiglitazone Maleate, compare, can find out b-e with have corresponding characteristic peak in a, therefore, can reach a conclusion: contain the medicine Rosiglitazone Maleate in plasma sample.In addition, from b-e, it can also be seen that, relative intensity and the drug concentration in blood plasma of characteristic peak have certain correlativity, can compare the size of different pharmaceutical blood plasma Chinese traditional medicine concentration by the relative intensity of characteristic peak.
In the present embodiment, use the Portable Raman spectrometer test surfaces to strengthen Raman spectrum.
Visible, the method for the detection blood plasma Chinese traditional medicine Rosiglitazone Maleate that adopts the embodiment of the present invention to provide can effectively detect the medicine Rosiglitazone Maleate in blood plasma, and the time of the present embodiment test is total up to 2 minutes.
Embodiment 4
The embodiment of the present invention provides a kind of method that detects the medicine DB2 in blood plasma, and its operation steps is with embodiment 1, different, and the concentration of medicine DB2 aqueous solution is 10
-3Mol/L, the concentration of the plasma sample that contains the medicine DB2 is 400ug/ml, 40ug/ml, 4ug/ml, 0.4ug/ml.
The testing result of medicine DB2 as Figure 7-8.
In Fig. 7, a is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 2 in the pH value; B is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 12 in the pH value; C is for testing the Surface enhanced raman spectroscopy obtained under the condition that is 7 in the pH value.As can be seen from Figure 7, at wavelength, be under the 785nm exciting light, under the condition that the pH value is 12, the surface-enhanced Raman effects of medicine DB2 is the most obvious.
In Fig. 8, the Surface enhanced raman spectroscopy that obtains of test under the condition that the aqueous solution that a is the medicine DB2 is 12 in the pH value, it is to test the Surface enhanced raman spectroscopy obtained under 300ug/ml, 30ug/ml, 3ug/ml, the 0.3ug/ml plasma sample condition that is 12 in pH value that b-e is followed successively by concentration.To in Fig. 8, containing the Surface enhanced raman spectroscopy b-e of the plasma sample of the medicine DB2 of variable concentrations and the aqueous solution Surface enhanced raman spectroscopy a under the same conditions of Fig. 8 Chinese traditional medicine DB2, compare, can find out b-e with have corresponding characteristic peak in a, therefore, can reach a conclusion: contain the medicine DB2 in plasma sample.In addition, from b-e, it can also be seen that, relative intensity and the drug concentration in blood plasma of characteristic peak have certain correlativity, can compare the size of different pharmaceutical blood plasma Chinese traditional medicine concentration by the relative intensity of characteristic peak.
In the present embodiment, use the Portable Raman spectrometer test surfaces to strengthen Raman spectrum.
Visible, the method for the detection blood plasma Chinese traditional medicine DB2 that adopts the embodiment of the present invention to provide can effectively detect the medicine DB2 in blood plasma, and the time of the present embodiment test is total up to 2 minutes.
The foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, for a person skilled in the art, the present invention can have various modifications and variations.Within the spirit and principles in the present invention all, any modification of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (10)
1. detect the method for blood plasma Chinese traditional medicine composition, it is characterized in that, described method is operated in accordance with the following steps:
A, by pharmaceutical aqueous solution with backing material, mix, standing, regulate the pH value of gained potpourri to the 1-14 scope, the Surface enhanced raman spectroscopy of this medicine of test under exciting light, wherein, the concentration of pharmaceutical aqueous solution is: 10
-3-10
-8Mol/L;
B, the blank plasma that does not contain medicine that will remove protein mix with backing material, standing, the pH value of regulating the gained potpourri is identical with the pH value in step a, and under described exciting light, test does not contain the Surface enhanced raman spectroscopy of the blank plasma of medicine;
C, the blood plasma that contains medicine that will remove protein mixes with backing material, standing, the pH value of regulating the gained potpourri is identical with the pH value in step a, the Surface enhanced raman spectroscopy of the blood plasma that test contains medicine under described exciting light, the concentration of the blood plasma that wherein, contains medicine is: 0.02-50ug/ml;
D, by after the Surface enhanced raman spectroscopy of the blank plasma that does not contain medicine that obtains in the Surface enhanced raman spectroscopy deduction step b of the blood plasma that contains medicine that obtains in step c, compare with the Surface enhanced raman spectroscopy of the medicine obtained in step a, by whether having corresponding characteristic peak to judge in described blood plasma, whether contain described medicine.
2. method according to claim 1, is characterized in that, the described light wavelength that excites is 785nm.
3. method according to claim 1, is characterized in that, the mass ratio of described pharmaceutical aqueous solution and backing material is: 10:1-10:100.
4. method according to claim 1, is characterized in that, the blood plasma that contains medicine of described removal protein and the mass ratio of backing material are: 10:1-10:100.
5. method according to claim 1, it is characterized in that, before step b, also comprise: the Surface enhanced raman spectroscopy of the described medicine that comparison step a obtains under different pH values, select Surface enhanced raman spectroscopy and the corresponding pH value thereof of Raman scattering strength-enhanced effect maximum, the corresponding pH value pH value used in step a, b and c using the Surface enhanced raman spectroscopy of the Raman scattering strength-enhanced effect maximum of described medicine, the Surface enhanced raman spectroscopy obtained under this pH value condition is the Surface enhanced raman spectroscopy that step a, b and c obtain.
6. method according to claim 5, is characterized in that, described pH value is regulated by acid, alkali, and described acid is one or more in hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid, and described alkali is one or more in NaOH, potassium hydroxide and sodium carbonate.
7. method according to claim 5, is characterized in that, described pH value is adjusted to respectively 2,7,12.
8. method according to claim 1, it is characterized in that, remove specifically operation in accordance with the following steps of protein described in step c: the ratio that is 1:0.6-1:1 according to the mass ratio of the perchloric acid containing medicine blood plasma and 6% massfraction, to containing in medicine blood plasma, adding perchloric acid, mix, under the condition of 10000-12000r/min, centrifugal 10-15min, get supernatant, obtain not containing protein containing medicine blood plasma.
9. method according to claim 1, is characterized in that, described backing material is selected from silver sol, aurosol or silver-colored billon colloidal sol.
10. method according to claim 9, it is characterized in that, described silver sol is prepared as follows: by the liquor argenti nitratis ophthalmicus 400-500ml of 0.15-0.20mg/ml, be heated to boiling, then the citric acid three sodium solution 10-20ml that is 0.5-1% by massfraction dropwise adds, constantly stir during this time, drip rear continuation and keep boiling within 1-5 hour, to obtain fulmargin, cooling rear standby.
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| CN109060766A (en) * | 2018-09-27 | 2018-12-21 | 珠海彩晶光谱科技有限公司 | A kind of measurement method and detection device of the concentration in body fluid based on Surface enhanced Raman spectroscopy |
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