CN104833669A - Rapid detection method of drugs with weak main drug signal - Google Patents
Rapid detection method of drugs with weak main drug signal Download PDFInfo
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Abstract
The invention provides a rapid detection method of drugs with a weak main drug signal. The rapid detection method comprises the following steps: determining a drug with a weak main drug signal; preparing a sample solution, a standard solution, and a structural analog standard solution; carrying out in-situ detection on speckles on a lamella after sample application; and calculating the rf value of TLC (thin layer chromatograph) expansion. The provided rapid detection method can carry out rapid detection on drugs, which have a structural analog and a weak main drug signal, so as to judge authenticity and quality of the drugs.
Description
Technical field
The present invention relates to a kind of method for quick of weak main ingredient signal medicine, belong to drug measurement techniques field.
Background technology
Raman spectroscopy is a kind of medicine Fast Detection Technique emerging in recent years, has the advantages such as easy, quick.The drug content spectral signal that is higher, main ingredient of general commercially available chemicals is stronger, the raman spectral signal of adjuvant is not enough to the characteristic raman spectral signal covering main ingredient, and main ingredient composition does not have analogue, therefore detect fast easily via Portable Raman spectrometer, thus the true and false and the quality of this medicine can be judged.
But some main ingredient signal is weak and have the quick detection difficult of the medicine of analogue, often used by illegal retailer, manufacture and sell pass a fake product off as a genuine one, shoddy counterfeit drug.
This similar drug due to drug content low, auxiliary material content is relatively high, and therefore main ingredient is buried in auxiliary material substantially.When irradiating this similar drug by Raman spectrometer laser, be difficult to be irradiated to main ingredient position.Or because the Raman scattering of main ingredient is more weak, and the raman spectral signal of adjuvant is relatively strong, substantially masks the characteristic raman spectral signal of main ingredient.Thus difficulty is brought to the rapid field detection of authenticity of medicament.
This similar drug analogue that also chemical constitution is similar, chemical property is similar owing to existing, Raman spectral information is similar, although the analogue of these low price also can play result for the treatment of, spinoff is large.Due to the existence of analogue, make this similar drug by Raman signal determination main ingredient composition, also just cannot not realize the Site Detection to this similar drug by Portable Raman spectrometer.
For this kind of main ingredient signal weak (hereinafter referred to as weak main ingredient signal medicine) there is the detection of the medicine of analogue, the realizations such as chromatographic technique, application of gas chromatorgraphy/mass technology, spectral imaging technology can only be passed through at present, but above-mentioned technology instrument is huge and expensive, complicated operation, analysis time is long, and the scene being not suitable for medicine is examined soon.
Therefore, need a kind of new method of exploitation at present badly, weak and there is the medicine of analogue for field quick detection main ingredient signal.
Summary of the invention
The present invention carries out to solve the problem, and object is that providing a kind of can realize weak to main ingredient signal and have the medicine of analogue to detect fast, thus judges the method for authenticity of medicament and quality.
The present invention to achieve these goals, have employed following technical scheme:
The invention provides a kind of method for quick of weak main ingredient signal medicine, for the identification of the main ingredient composition going out weak main ingredient signal medicine, it is characterized in that, comprise the following steps:
Step one, the determination of weak main ingredient signal medicine
Portable Raman spectrometer is utilized to detect medicine to be checked and standard items thereof, laser power is 150mw ~ 300mw, irradiation time is 10s ~ 15s, obtain respective Raman spectrogram respectively, the similarity of both calculating, if the similarity of the two is less than 0.5, then can judge that this medicine to be checked is as weak main ingredient signal medicine.
Step 2, the preparation of sample solution and standard solution thereof, analogue standard solution
Weak main ingredient signal medicine is ground successively, dissolve, ultrasonic process, then centrifuging and taking supernatant, obtain certain density sample solution, the standard items corresponding with main ingredient composition are dissolved, after ultrasonic process, obtain the isocyatic standard solution with sample solution, the analogue standard items corresponding with main ingredient composition are dissolved, after ultrasonic process, obtain the isocyatic analogue standard solution with sample solution, sample solution and standard solution, solvent in analogue standard solution is identical, this solvent does not dissolve the adjunct ingredient in weak main ingredient signal medicine,
Step 3, the in situ detection of the spot of thin-layer sample application
Sample solution and standard solution are distinguished point sample on thin layer plate, obtain two spots, nano-silver colloid solution is dripped on two spots again, Portable Raman spectrometer is utilized to carry out in-situ scanning to two spots, obtain respective Surface enhanced raman spectroscopy figure respectively and the similarity of both calculating, if the similarity of the two is (preferred 0.9-1) between 0.7-1, then judge the composition of main ingredient composition as the composition of standard items or the analogue of standard items;
Step 4, the calculating of the Rf value of thin-layer developing
By sample solution, standard solution, analogue standard solution difference point sample on thin layer plate, developping agent is utilized to launch respectively, taking-up volatilizes, inspect under being placed in uviol lamp, determine to launch the position of rear spot on thin layer plate and obtain Rf value, the Rf value of comparative sample spot and other spots, if sample point is equal with the Rf value of standard items spot, then judges that the main ingredient composition in this weak main ingredient signal medicine is consistent with the composition of standard items; If unequal, then the Rf value of comparative sample spot and analogue spot, if sample point is equal with the Rf value of analogue spot, then judges that the main ingredient composition in this weak main ingredient signal medicine is consistent with the composition of its analogue.
Further, in the method for quick of a kind of weak main ingredient signal medicine of the present invention, can also have such feature: wherein, in described step one, three, Raman spectra pretreatment method comprises spectral coverage and chooses (400cm
-1-1800cm
-1), baseline correction (airPLS method), level and smooth (Sgolay method), similarity calculating method is the Corrcoef function in Matlab.
Further, in the method for quick of a kind of weak main ingredient signal medicine of the present invention, can also have such feature: wherein, in described step 2, the time of described ultrasonic process is 20-30min.
Further, in the method for quick of a kind of weak main ingredient signal medicine of the present invention, such feature can also be had: wherein, in step 3, laser power when described Portable Raman spectrometer scans is 150mw ~ 300mw, and irradiation time is 10s ~ 15s.
Further, in the method for quick of a kind of weak main ingredient signal medicine of the present invention, such feature can also be had: wherein, in step 3, four, the dripping quantity of described sample solution, standard solution and analogue standard solution is 1-2 μ L, and the dripping quantity of described nano-silver colloid solution is 5-10 μ L.
Further, in the method for quick of a kind of weak main ingredient signal medicine of the present invention, can also have such feature: wherein, in described step 3, four, described thin layer plate is thin-layer silicon offset plate.
Further, in the method for quick of a kind of weak main ingredient signal medicine of the present invention, can also have such feature: wherein, in described step 4, the wavelength of described uviol lamp is 254nm.
Further, in the method for quick of a kind of weak main ingredient signal medicine of the present invention, such feature can also be had: wherein, the excitation wavelength 785nm of described Portable Raman spectrometer.
Invention effect and effect
According to the method for quick of a kind of weak main ingredient signal medicine of the present invention, because the sample solution obtained by weak main ingredient signal medicine and standard solution point sample obtain spot on thin layer plate, previously prepared good nano-silver colloid solution is dripped on spot, Portable Raman spectrometer is utilized to detect spot, respective Surface enhanced raman spectroscopy figure can be obtained, and by comparing the similarity of collection of illustrative plates between them, whether the principal ingredient determining this medicine is effective constituent, reaches the object judging authenticity of medicament; Further, sample solution, standard solution and analogue standard solution are carried out thin-layer developing, judge that whether main ingredient composition is consistent with standard items according to Rf value or be the analogue of standard items, reach the object judging drug quality quality; And the nano-silver colloid solution prepared and thin layer plate are all easy to carry about with one, can coordinate with portable Raman spectrometer at Test Field, be easy to the quick detection realized weak main ingredient signal authenticity of medicament and quality.
Accompanying drawing explanation
Fig. 1 is Surface enhanced raman spectroscopy (SERS) figure of the present invention weak main ingredient signal medicine and standard items in embodiment one;
Fig. 2 is the present invention's weak main ingredient signal medicine, standard items and analogue standard items schematic diagram after thin-layer developing in embodiment one;
Fig. 3 is Surface enhanced raman spectroscopy (SERS) figure of the present invention weak main ingredient signal medicine and standard items in embodiment two; And
Fig. 4 is process flow diagram of the present invention.
Embodiment
Below the method for quick of a kind of weak main ingredient signal medicine involved in the present invention is elaborated.
< embodiment one >
In the present embodiment one, for terazosin, specific explanations detects the method for weak main ingredient signal medicine fast.
The method for quick of the weak main ingredient signal medicine of the present embodiment one comprises the following steps,
Step one, the determination of weak main ingredient signal medicine
Utilize Portable Raman spectrometer to detect medicine to be checked and standard items thereof, laser power is 150mw, and irradiation time is 10s, obtains respective Raman spectrogram respectively.
In the present embodiment, the model of Portable Raman spectrometer is BWS415-785H (Bi Da Imtech of the U.S.), excitation wavelength 785nm.
(400cm is chosen by spectral coverage
-1-1800cm
-1), baseline correction, smoothly pre-service is carried out to two Raman spectrums after, the Corrcoef function in Matlab is utilized to calculate the similarity of two above-mentioned Raman spectrograms, found that the two similarity is-0.0525, be less than 0.5, then can judge that this medicine to be checked is as weak main ingredient signal medicine, also namely the Raman signal of auxiliary material masks the Raman signal of main ingredient composition.
Step 2, the preparation of nano-silver colloid solution
Precision takes silver nitrate, the 8.5mgPVP (polyvinylpyrrolidone) of 17mg, is dissolved in 5mL water, obtains the AgNO that mass ratio is 2:1
3/ PVP solution.Measure 50mLDMF (DMF) in the three-necked bottle of 250mL, be heated to boiling.Add rapidly above-mentioned AgNO
3/ PVP solution constantly boiling a period of time, be cooled to room temperature and be placed in brown bottle and keep in Dark Place.
Step 3, the preparation of sample solution and standard solution thereof, analogue standard solution
Get a slice terazosin, the quality of the Terazosin Hydrochloride in this terazosin is 2mg.
After being ground by this terazosin, in the methyl alcohol of 1mL after dissolving, ultrasonic process 20min, centrifuging and taking supernatant, obtains sample solution.Methyl alcohol can not dissolve the adjunct ingredient in terazosin, and after solution is centrifuged, adjunct ingredient can be removed by forming precipitation, therefore only has Terazosin Hydrochloride in sample solution as solute.
Standard items are dissolved, after ultrasonic process 20min, obtain the Terazosin Hydrochloride standard solution of 2mg/mL by the standard items of the Terazosin Hydrochloride of accurately weighed 2.0mg in the methyl alcohol of 1mL.
Standard items are dissolved, after ultrasonic process 20min, obtain the minipress standard solution of 2mg/mL by the standard items of the minipress of accurately weighed 2.0mg in the methyl alcohol of 1mL.
Step 4, the in situ detection of the spot of thin-layer sample application
Sample solution and each 1 μ L of Terazosin Hydrochloride standard solution is pipetted respectively with kapillary, point sample is on silica gel thin-layer plate respectively, obtain two spots, again the nano-silver colloid solution of 5 μ L is dripped on two spots, Portable Raman spectrometer is utilized to carry out in-situ scanning to two spots, obtain respective Surface enhanced raman spectroscopy figure respectively, specifically as shown in Figure 1.
(400cm is chosen by spectral coverage
-1-1800cm
-1), baseline correction, smoothly pre-service is carried out to two Raman spectrums after, the Corrcoef function in Matlab is utilized to calculate the similarity of two above-mentioned each and every one Raman spectrograms, show that sample solution and Terazosin Hydrochloride standard solution similarity are 0.7920, between 0.7-1, then judge that the composition of main ingredient composition and standard items or analogue standard items cannot be distinguished, also the main ingredient composition namely in this terazosin is Terazosin Hydrochloride or minipress, need judge further.
Step 5, the calculating of the Rf value of thin-layer developing
Sample solution and each 1 μ L point sample of standard solution is pipetted respectively in thin layer plate original position with kapillary, with methenyl choloride: normal butyl alcohol: the mixed solvent (methenyl choloride: normal butyl alcohol: the volume ratio of glacial acetic acid is 4:3:0.5) of glacial acetic acid is for after developping agent expansion, taking-up volatilizes, under uviol lamp 254nm, inspect the position of spot on thin layer plate after launching, and calculate Rf value.Wherein, Rf value be in thin-layered chromatography initial point to the distance of spot centers and initial point to the ratio of the distance of solvent front.
As can be seen from Figure 2, Terazosin Hydrochloride sample solution Rf value is 0.575, and Terazosin Hydrochloride standard solution Rf value is 0.225, and both Rf values are unequal.Illustrate that the composition of main ingredient composition in the terazosin in the present embodiment and Terazosin Hydrochloride standard items is inconsistent.
Minipress is the analogue of Terazosin Hydrochloride.Prazosin Hydrochloride Tablets spinoff is very large, and the low price half of cost ratio terazosin, is therefore often used to pretend to be terazosin to carry out market sale.
Then, launch by the standard solution of step identical in step 5 by minipress, and the Rf value after Rf value and sample solution being launched contrasts, found that the Rf value that minipress standard items obtain is 0.575, identical with the Rf value of sample solution.Therefore can judge that the main ingredient composition in the terazosin in the present embodiment is consistent with the composition of minipress standard items, show that the terazosin in the present embodiment exists the situation of pretending to be with less expensive configuration analog.
Embodiment effect and effect
The method for quick of a kind of weak main ingredient signal medicine involved by the present embodiment, because the sample solution made by Terazosin Hydrochloride, standard solution respectively point sample obtain spot on thin layer plate, previously prepared good nano-silver colloid solution is dripped on spot, Portable Raman spectrometer is utilized to detect spot, respective Surface enhanced raman spectroscopy figure can be obtained, and by comparing the similarity of collection of illustrative plates between them, whether the principal ingredient determining this medicine is effective constituent, reaches the object judging authenticity of medicament; Further, the standard solution of sample solution, standard solution and analogue is carried out thin-layer developing, judge that whether main ingredient composition is consistent with standard items according to Rf value or be the analogue of standard items, reach the object judging drug quality quality; And the nano-silver colloid solution prepared and thin layer plate are all easy to carry about with one, can coordinate with portable Raman spectrometer at Test Field, be easy to realize to the terazosin true and false and quality quick detection.
< embodiment two >
In the present embodiment two, for glimepiride tablet, specific explanations detects the method for weak main ingredient signal medicine fast.
The method for quick of the weak main ingredient signal medicine of the present embodiment two comprises the following steps,
Step one, the determination of weak main ingredient signal medicine
Utilize Portable Raman spectrometer to detect medicine to be checked and standard items thereof, laser power is 300mw, and irradiation time is 15s, obtains respective Raman spectrogram respectively.
In the present embodiment, the model of Portable Raman spectrometer is BWS415-785H (Bi Da Imtech of the U.S.), excitation wavelength 785nm.
(400cm is chosen by spectral coverage
-1-1800cm
-1), baseline correction, smoothly pre-service is carried out to two Raman spectrums after, the Corrcoef function in Matlab is utilized to calculate the similarity of two above-mentioned Raman spectrograms, found that the two similarity is 0.1200, be less than 0.5, then can judge that this medicine to be checked is as weak main ingredient signal medicine, also namely the Raman signal of auxiliary material masks the Raman signal of main ingredient composition.
Step 2, the preparation of nano-silver colloid solution
Precision takes silver nitrate, the 8.5mgPVP of 17mg, is dissolved in 5mL water, obtains the AgNO that mass ratio is 2:1
3/ PVP solution.Measure in the three-necked bottle of 50mLDMF to 250mL, be heated to boiling.Add rapidly above-mentioned AgNO
3/ PVP solution constantly boiling a period of time, be cooled to room temperature and be placed in brown bottle and keep in Dark Place.
Step 3, the preparation of sample solution and standard solution thereof
Get a slice glimepiride tablet, the quality of the Glimepiride in this glimepiride tablet is 2mg.
After this glimepiride tablet is ground, be that in the mixed solvent of 1:1 after dissolving, ultrasonic process 30min, centrifuging and taking supernatant, obtains the sample solution of 2mg/mL at the methyl alcohol of 1mL and the volume ratio of chloroform.Mixed solvent can not dissolve the adjunct ingredient in glimepiride tablet, and after solution is centrifuged, adjunct ingredient can be removed by forming precipitation, therefore only has Glimepiride in sample solution as solute.
Standard items are dissolve in the mixed solvent of 1:1, after ultrasonic process 30min, obtain the standard solution of 2mg/mL at the methyl alcohol of 1mL and the volume ratio of chloroform by the standard items of the Glimepiride of accurately weighed 2.0mg.
Step 4, the in situ detection of the spot of thin-layer sample application
Sample solution and each 2 μ L of standard solution are pipetted respectively with kapillary, point sample is on silica gel thin-layer plate respectively, obtain two spots, again the nano-silver colloid solution of 10 μ L is dripped on two spots, Portable Raman spectrometer is utilized to carry out in-situ scanning to two spots, obtain respective Surface enhanced raman spectroscopy figure respectively, specifically as shown in Figure 3.
(400cm is chosen by spectral coverage
-1-1800cm
-1), baseline correction, smoothly pre-service is carried out to two Raman spectrums after, the Corrcoef function in Matlab is utilized to calculate the similarity of two above-mentioned Raman spectrograms, show that the Similarity value of sample solution and Glimepiride standard items is 0.9660,0.9 is greater than between 0.7-1, then judge that main ingredient composition is consistent with the composition of standard items, the main ingredient composition also namely in this glimepiride tablet is Glimepiride.
Embodiment effect and effect
The method for quick of a kind of weak main ingredient signal medicine involved by the present embodiment, because the sample solution made by Glimepiride and standard solution respectively point sample obtain spot on thin layer plate, previously prepared good nano-silver colloid solution is dripped on spot, Portable Raman spectrometer is utilized to detect spot, respective Surface enhanced raman spectroscopy figure can be obtained, and by comparing the similarity of collection of illustrative plates between them, whether the principal ingredient determining this medicine is effective constituent, reaches the object judging authenticity of medicament; And the nano-silver colloid solution prepared and thin layer plate are all easy to carry about with one, can coordinate with portable Raman spectrometer at Test Field, be easy to realize the quick detection to the glimepiride tablet true and false.
Certainly, the method for quick of a kind of weak main ingredient signal medicine that the present invention relates to not merely is defined in the description in above embodiment.
Claims (8)
1. a method for quick for weak main ingredient signal medicine, for the identification of the main ingredient composition going out weak main ingredient signal medicine, is characterized in that, comprise the following steps:
Step one, the determination of weak main ingredient signal medicine
Portable Raman spectrometer is utilized to detect medicine to be checked and standard items thereof, laser power is 150mw ~ 300mw, irradiation time is 10s ~ 15s, obtain respective Raman spectrogram respectively, the similarity of both calculating, if the similarity of the two is less than 0.5, then can judge that this medicine to be checked is as weak main ingredient signal medicine.
Step 2, the preparation of sample solution and standard solution thereof, analogue standard solution
Weak main ingredient signal medicine is ground successively, dissolve, ultrasonic process, then centrifuging and taking supernatant, obtain certain density sample solution, the standard items corresponding with main ingredient composition are dissolved, after ultrasonic process, obtain the isocyatic standard solution with sample solution, the analogue standard items corresponding with main ingredient composition are dissolved, after ultrasonic process, obtain the isocyatic analogue standard solution with sample solution, sample solution and standard solution, solvent in analogue standard solution is identical, this solvent does not dissolve the adjunct ingredient in weak main ingredient signal medicine,
Step 3, the in situ detection of the spot of thin-layer sample application
Sample solution and standard solution are distinguished point sample on thin layer plate, obtain two spots, nano-silver colloid solution is dripped on two spots again, Portable Raman spectrometer is utilized to carry out in-situ scanning to two spots, obtain respective Surface enhanced raman spectroscopy figure respectively and the similarity of both calculating, if the similarity of the two is (preferred 0.9-1) between 0.7-1, then judge the composition of main ingredient composition as the composition of standard items or the analogue of standard items;
Step 4, the calculating of the Rf value of thin-layer developing
By sample solution, standard solution, analogue standard solution difference point sample on thin layer plate, developping agent is utilized to launch respectively, taking-up volatilizes, inspect under being placed in uviol lamp, determine to launch the position of rear spot on thin layer plate and obtain Rf value, the Rf value of comparative sample spot and other spots, if sample point is equal with the Rf value of standard items spot, then judges that the main ingredient composition in this weak main ingredient signal medicine is consistent with the composition of standard items; If unequal, then the Rf value of comparative sample spot and analogue spot, if sample point is equal with the Rf value of analogue spot, then judges that the main ingredient composition in this weak main ingredient signal medicine is consistent with the composition of its analogue.
2. the method for quick of a kind of weak main ingredient signal medicine according to claim 1, is characterized in that:
Wherein, in described step one, three, Raman spectra pretreatment method comprises spectral coverage and chooses (400cm
-1-1800cm
-1), baseline correction (airPLS method), level and smooth (Sgolay method), similarity calculating method is the Corrcoef function in Matlab.
3. the method for quick of a kind of weak main ingredient signal medicine according to claim 1, is characterized in that:
Wherein, in described step 2, the time of described ultrasonic process is 20-30min.
4. the method for quick of a kind of weak main ingredient signal medicine according to claim 1, is characterized in that:
Wherein, in step 3, laser power when described Portable Raman spectrometer scans is 150mw ~ 300mw, and irradiation time is 10s ~ 15s.
5. the method for quick of a kind of weak main ingredient signal medicine according to claim 1, is characterized in that:
Wherein, in step 3, four, the dripping quantity of described sample solution, standard solution and analogue standard solution is 1-2 μ L, and the dripping quantity of described nano-silver colloid solution is 5-10 μ L.
6. the method for quick of a kind of weak main ingredient signal medicine according to claim 1, is characterized in that:
Wherein, in described step 3, four, described thin layer plate is thin-layer silicon offset plate.
7. the method for quick of a kind of weak main ingredient signal medicine according to claim 1, is characterized in that:
Wherein, in described step 4, the wavelength of described uviol lamp is 254nm.
8. the method for quick of a kind of weak main ingredient signal medicine according to claim 1, is characterized in that:
Wherein, the excitation wavelength 785nm of described Portable Raman spectrometer.
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