CN103760293B - The method of 10 kinds of undeclared prescription drugs in rapid screening Small Meridian-Activating Pill - Google Patents
The method of 10 kinds of undeclared prescription drugs in rapid screening Small Meridian-Activating Pill Download PDFInfo
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Abstract
A kind of method of 10 kinds of undeclared prescription drugs in rapid screening Small Meridian-Activating Pill.First reference substance and need testing solution is prepared, then reference substance solution is drawn respectively, need testing solution, point is on same silica gel thin-layer plate, with developping agent ascending development, taking-up is dried, thin-layer chromatography is observed under putting ultraviolet lamp respectively, spray ethanol solution of sulfuric acid again, and it is clear to the spot development on thin layer plate to be placed in baking oven heated at constant temperature, observe thin-layer chromatography under putting daylight and ultraviolet lamp more respectively, if Rf value place identical with reference substance has the fluorescent quenching spot of same color, fluorescence spot or spot to occur simultaneously, then can judge that sample result is as the positive.Obviously, the present invention is easy and simple to handle, fast, cost is low, both the problem that vacation " positive " is wherein disturbed had been solved, it also avoid tradition adopts infrared spectrum, HPLC-MS etc. to measure the expensive expense and plenty of time that produce, makes in effective monitoring Chinese patent drug Small Meridian-Activating Pill whether undeclared prescription drugs becomes possibility.
Description
Technical field
The present invention relates to traditional Chinese medicine quality detection field, be specifically related to the method for 10 kinds of undeclared prescription drugs in a kind of rapid screening Small Meridian-Activating Pill.
Background technology
Small Meridian-Activating Pill is by arisaema cum bile, aconiti preparata,radix, wild aconite root, earthworm, frankincense (system), myrrh (system) Chinese medicine preparation that forms of the totally 6 taste prepared slices of Chinese crude drugs.Primary efficacy is expelling wind and clearing away cold, dehumidifying of reducing phlegm, promoting blood circulation and stopping pain.Sick for the numbness caused by wind-cold damp pathogen impatency, phlegm stagnation in collateral, disease sees limbs joint pain, or crymodynia, or shouting pain, or pain night very, joint joint stuffiness, numbness contracture.Its preparation method is that arisaema cum bile, aconiti preparata,radix, wild aconite root, earthworm, frankincense (system), myrrh (system) are ground into fine powder respectively, sieves, and mixing, every 100g fine powder adds refined honey 120 ~ 130g and makes large honeyed bolus.This Chinese medicine preparation complicated component, the clinical bi Zheng such as pain in the back, skelagia, cervicodynia, brachialgia being used for the treatment of rheumatic arthritis, rheumatoid arthritis and caused by hyperosteogeny, sciatica, duodenal stasis etc.Belong to Anti-rheumatism Chinese patent drugs.In order to pursue economic interests, some lawless persons illegal interpolation in Small Meridian-Activating Pill has antipyretic, anti-inflammatory, analgesic effect cheap chemicals, to reach effect effective fast.And patient after too much these chemicalss of absorption, can cause great harm to health in unwitting situation.
In Anti-rheumatism Chinese patent drugs, undeclared prescription drugs mainly contains two large classes: antipyretic-antalgic class and glucocorticoids.At present, the method detecting undeclared prescription drugs in Anti-rheumatism Chinese patent drugs is the universal method not limiting the concrete kind of Chinese patent drug.This universal method relates to all Anti-rheumatism Chinese patent drugs, and scope is wide, but Anti-rheumatism Chinese patent drugs is of a great variety, and the prescription of each Chinese patent drug is all different, very easily produce vacation " positive " interference, cannot result of determination clearly, therefore universal method only can accomplish scalping.And due to the interference of false " positive ", the chemicals detected is only limitted in antipyretic-antalgic class or glucocorticoids limited several.As " the detection method research about undeclared prescription drugs in antirheumatic Chinese medicine preparation " (Asia-Pacific traditional medicine .2010.6(6) such as Zhang He: 31-32) first adopt thin-layered chromatography to carry out scalping, then must infrared spectrum or high performance liquid chromatography be adopted to verify positive wherein further again.But due to infrared spectrometer, HPLC-MS, GC-MS expensive equipment, and must be manipulated by technical professional, maintenance cost is also high, the object that rapid screening reaches monitoring cannot be realized.Obviously, build for interpolation number of chemical medicine illegal in Small Meridian-Activating Pill the method carrying out rapid screening to be completely very important.Therefore, in order to make the illegal activities of undeclared prescription drugs in Small Meridian-Activating Pill effectively be controlled, ensureing that the people is safe and effective for medication, needing the quick screening method setting up undeclared prescription drugs in Small Meridian-Activating Pill badly.
Summary of the invention
The object of this invention is to provide the method for 10 kinds of undeclared prescription drugs in a kind of rapid screening Small Meridian-Activating Pill, to overcome the deficiencies in the prior art.
Another object of the present invention gets rid of vacation " positive " interference of Small Meridian-Activating Pill and undeclared prescription drugs, realizes the rapid screening of multiple undeclared prescription drugs, reach the universal feasible of monitoring.
Small Meridian-Activating Pill is a Chinese medicine preparation, although prescription flavour of a drug are few, chemical composition is very complicated, in addition undeclared prescription drugs, very easily occurs that vacation " positive " is disturbed during mensuration.Usual vacation " positive " interference must adopt infrared spectrometer, HPLC-MS, GC-MS carry out composition confirmation.Research finds, because 10 kinds of chemicals compositions of examination required in Small Meridian-Activating Pill all have the character being dissolved in methyl alcohol, and methyl alcohol is low to the refined honey solubleness in large honeyed bolus, therefore utilizes this character, selects methyl alcohol as Extraction solvent.Consider that Small Meridian-Activating Pill is large honeyed bolus, if do not disperseed, this Extraction solvent cannot penetrate in sample, can not fully extract chemicals to be measured.Therefore get Small Meridian-Activating Pill one ball (3g), after shredding, add zeyssatite grinding, large honeyed bolus fully can be disperseed.Add methyl alcohol again, sample is all soaked, and adopt ultrasonic process, with effectively by these 10 kinds of constituents extraction out.Both can avoid the stripping of refined honey in Small Meridian-Activating Pill, be convenient to point sample, other compositions in Small Meridian-Activating Pill can have been greatly reduced again to the interference measured.Again because of the illegal difference of chemicals on thin layer plate in adsorptive power of adding, then go out in Chinese patent drug Small Meridian-Activating Pill whether illegally with the addition of the chemicals that 10 kinds are divided into 2 groups with developping agent examination respectively.
Quick screening method of the present invention is as follows
(1) preparation of reference substance solution: get prednisone acetate, dexamethasone acetate, Indomethacin, naproxen and each 10mg of C14H10Cl2NNaO2 reference substance respectively, dissolve with methyl alcohol 10ml, make reference substance solution A; Separately get prednisolone, hydrocortisone, metacortandracin, Triamcinolone acetonide and each 10mg of hydrocortisone acetate reference substance respectively, dissolve with methyl alcohol 10ml, make reference substance solution B.
(2) preparation of need testing solution: get Small Meridian-Activating Pill 3g, shred, adds zeyssatite 2g, grinds well, add methyl alcohol, ultrasonic process, filters, gets filtrate as need testing solution.
(3) examination mensuration is carried out: draw reference substance solution A1 μ l respectively, need testing solution 5 μ l, puts in same silica G F
254on thin layer plate, with developping agent I ascending development, take out, dry, thin-layer chromatography is observed under putting ultraviolet lamp (wavelength 254nm) and ultraviolet lamp (wavelength 365nm) respectively, and then spray 10% ethanol solution of sulfuric acid, and it is clear to the spot development on thin layer plate to be placed in 105 DEG C of baking oven heated at constant temperature, then put observed under daylight thin-layer chromatography.In the chromatogram of above-mentioned three kinds of view modes, Rf value place identical with reference substance occurs if any the fluorescent quenching spot of same color, fluorescence spot or spot simultaneously, then can judge that sample result is as the positive.
The another reference substance solution B1 of absorption respectively μ l, need testing solution 5 μ l, puts in same silica G F
254on thin layer plate, with developping agent II ascending development, take out, dry, observe thin-layer chromatography under putting ultraviolet lamp (wavelength 254nm), and then spray 10% ethanol solution of sulfuric acid, and it is clear to the spot development on thin layer plate to be placed in 105 DEG C of baking oven heated at constant temperature, then put observed under daylight thin-layer chromatography.In the chromatogram of above-mentioned two kinds of view modes, Rf value place identical with reference substance occurs if any the fluorescent quenching spot of same color or spot simultaneously, then can judge that sample result is as the positive.
Above-mentioned developping agent I is n-hexane-ethyl acetate-glacial acetic acid=15: 5: 1(volume ratio).
Above-mentioned developping agent II is methenyl choloride-acetone-methanol-strong ammonia solution=18: 6: 2: 0.1(volume ratio).
The condition of above-mentioned ultrasonic process is 30 minutes, power 250W, frequency 50kHz.
The method judges from being prepared into of reference substance solution sample result, and overall process only needs 1 hour, and can at same silica G F
254the product of the multiple producer of examination on thin layer plate.Obviously, the inventive method is easy and simple to handle, fast, cost is low, both the problem that vacation " positive " that Small Meridian-Activating Pill patent medicine causes because of complicated component is disturbed had been solved, it also avoid and adopt infrared spectrum, HPLC-MS, GC-MS measure the expensive expense and plenty of time that produce, are therefore that in effective monitoring Chinese patent drug Small Meridian-Activating Pill, whether undeclared prescription drugs becomes possibility.
Accompanying drawing explanation
Fig. 1: Small Meridian-Activating Pill of the present invention and prednisone acetate, dexamethasone acetate, Indomethacin, naproxen, the TLC chromatogram of C14H10Cl2NNaO2 reference substance under developping agent I and ultraviolet lamp (wavelength 254nm) condition; Wherein, 1: the need testing solution of Small Meridian-Activating Pill negative sample; 2: the need testing solution of Small Meridian-Activating Pill positive; 3: reference substance solution A(spot a: prednisone acetate; Spot b: dexamethasone acetate; Spot c: Indomethacin; Spot d: naproxen; Spot e: C14H10Cl2NNaO2).
Fig. 2: Small Meridian-Activating Pill of the present invention and prednisone acetate, dexamethasone acetate, Indomethacin, naproxen, the TLC chromatogram of C14H10Cl2NNaO2 reference substance under developping agent I and ultraviolet lamp (wavelength 365nm) condition; Wherein, 1: the need testing solution of Small Meridian-Activating Pill negative sample; 2: the need testing solution of Small Meridian-Activating Pill positive; 3: reference substance solution A(spot d: naproxen).
Fig. 3: Small Meridian-Activating Pill of the present invention and prednisone acetate, dexamethasone acetate, Indomethacin, naproxen, the TLC chromatogram of C14H10Cl2NNaO2 reference substance under developping agent I and sunshine condition; Wherein, 1: the need testing solution of Small Meridian-Activating Pill negative sample; 2: the need testing solution of Small Meridian-Activating Pill positive; 3: reference substance solution A(spot a: prednisone acetate; Spot b: dexamethasone acetate; Spot c: Indomethacin; Spot d: naproxen; Spot e: C14H10Cl2NNaO2).
The need testing solution of the Small Meridian-Activating Pill negative sample in above-mentioned Fig. 1, Fig. 2, Fig. 3 is not containing the need testing solution that the Small Meridian-Activating Pill sample of prednisone acetate, dexamethasone acetate, Indomethacin, naproxen, C14H10Cl2NNaO2 is prepared according to the preparation method of need testing solution.The need testing solution of Small Meridian-Activating Pill positive is the need testing solution that the Small Meridian-Activating Pill sample containing prednisone acetate, dexamethasone acetate, Indomethacin, naproxen, C14H10Cl2NNaO2 is prepared according to the preparation method of need testing solution.
Fig. 4: Small Meridian-Activating Pill of the present invention and prednisolone, hydrocortisone, metacortandracin, Triamcinolone acetonide, the TLC chromatogram of hydrocortisone acetate under developping agent II and ultraviolet lamp (wavelength 254nm) condition; Wherein, 1: the need testing solution of Small Meridian-Activating Pill negative sample; 2: the need testing solution of Small Meridian-Activating Pill positive; 3: reference substance solution B(spot a: prednisolone; Spot b: hydrocortisone; Spot c: metacortandracin; Spot d: Triamcinolone acetonide; Spot e: hydrocortisone acetate).
Fig. 5: Small Meridian-Activating Pill of the present invention and prednisolone, hydrocortisone, metacortandracin, Triamcinolone acetonide, the TLC chromatogram of hydrocortisone acetate under developping agent II and sunshine condition; Wherein, 1: the need testing solution of Small Meridian-Activating Pill negative sample; 2: the need testing solution of Small Meridian-Activating Pill positive; 3: reference substance solution B(spot a: prednisolone; Spot b: hydrocortisone; Spot c: metacortandracin; Spot d: Triamcinolone acetonide; Spot e: hydrocortisone acetate).
The need testing solution of the Small Meridian-Activating Pill negative sample in above-mentioned Fig. 4, Fig. 5 is not containing the need testing solution that the Small Meridian-Activating Pill sample of prednisolone, hydrocortisone, metacortandracin, Triamcinolone acetonide, hydrocortisone acetate is prepared according to the preparation method of need testing solution.The need testing solution of Small Meridian-Activating Pill positive is the need testing solution that the Small Meridian-Activating Pill sample containing prednisolone, hydrocortisone, metacortandracin, Triamcinolone acetonide, hydrocortisone acetate is prepared according to the preparation method of need testing solution.
Embodiment
Provide detailed embodiment below.
The present invention adopts: instrument and reagent: Sartoris BP211D electronic balance, AUTO SCIENCEAS10200BT type ultrasonic cleaner, P/G2007 type constant temperature oven, thin layer plate: silica G F
254thin layer plate (German MERCK lot number: HX929689 specification: 20 × 20cm), it is pure that reagent is analysis, prednisone acetate reference substance (lot number: 100012-200706), dexamethasone acetate reference substance (lot number: 100122-201206), Indomethacin reference substance (lot number: 100258-200904), naproxen reference substance (lot number: 100198-201205), C14H10Cl2NNaO2 reference substance (lot number: 100334-200302), prednisolone reference substance (lot number: 100153-201004), hydrocortisone reference substance (lot number: 100152-200206), metacortandracin reference substance (lot number: 100199-201002), Triamcinolone acetonide reference substance (lot number: 100055-201103), hydrocortisone acetate reference substance (lot number: 100013-200607), above reference substance provides by National Institute for Food and Drugs Control, Small Meridian-Activating Pill (indicates lot number: 130110,20130715,130401,120801,20121201,120012,2111105).
Chromatographic condition: developping agent I: n-hexane-ethyl acetate-glacial acetic acid=15: 5: 1 (volume ratios); Developping agent II: methenyl choloride-acetone-methanol-strong ammonia solution=18: 6: 2: 0.1 (volume ratios); Unfolding condition: temperature: 23 DEG C; Relative humidity: 59%; Exhibition distance: 10cm.
The preparation of reference substance solution: get prednisone acetate, dexamethasone acetate, Indomethacin, naproxen and each 10mg of C14H10Cl2NNaO2 reference substance respectively, dissolve with methyl alcohol 10ml, make reference substance solution A; Separately get prednisolone, hydrocortisone, metacortandracin, Triamcinolone acetonide and each 10mg of hydrocortisone acetate reference substance respectively, dissolve with methyl alcohol 10ml, make reference substance solution B.
The preparation of need testing solution: get Small Meridian-Activating Pill finished product 3g, shred, add zeyssatite 2g, grind well, add methyl alcohol 10ml, carry out ultrasonic process, filters, gets filtrate as need testing solution.
Above-mentioned ultrasonic treatment conditions are 30 minutes, power 250W, frequency 50kHz.
Carry out examination mensuration: draw reference substance solution A1 μ l respectively, need testing solution 5 μ l, puts in same silica G F
254on thin layer plate, with developping agent I ascending development, take out, dry, thin-layer chromatography is observed under putting ultraviolet lamp (wavelength 254nm) and ultraviolet lamp (wavelength 365nm) respectively, and then spray 10% ethanol solution of sulfuric acid, and it is clear to the spot development on thin layer plate to be placed in 105 DEG C of baking oven heated at constant temperature, then put observed under daylight thin-layer chromatography.In the chromatogram of above-mentioned three kinds of view modes, Rf value place identical with reference substance occurs if any the fluorescent quenching spot of same color, fluorescence spot or spot simultaneously, then can judge that sample result is as the positive.Having detected 5 kinds of chemicalss is prednisone acetate, dexamethasone acetate, Indomethacin, naproxen and C14H10Cl2NNaO2.Corresponding chromatogram as shown in Figure 1, Figure 2, Fig. 3.
The another reference substance solution B1 of absorption respectively μ l, need testing solution 5 μ l, puts in same silica G F
254on thin layer plate, with developping agent II ascending development, take out, dry, observe thin-layer chromatography under putting ultraviolet lamp (wavelength 254nm), and then spray 10% ethanol solution of sulfuric acid, and it is clear to the spot development on thin layer plate to be placed in 105 DEG C of baking oven heated at constant temperature, then put observed under daylight thin-layer chromatography.In the chromatogram of above-mentioned two kinds of view modes, Rf value place identical with reference substance occurs if any the fluorescent quenching spot of same color or spot simultaneously, then can judge that sample result is as the positive.Having detected again 5 kinds of chemicalss is prednisolone, hydrocortisone, metacortandracin, Triamcinolone acetonide and hydrocortisone acetate.Corresponding chromatogram is as Fig. 4, Fig. 5.
Result shows, 2 groups of mixing reference substance good separations, and all the components of Small Meridian-Activating Pill self is noiseless to mensuration, and namely at reference substance identical Rf value place, noiseless composition exists test sample.
To sum up, the thin-layered chromatography that the present invention sets up can detect in Small Meridian-Activating Pill whether illegally with the addition of these 10 kinds of chemicalss of prednisone acetate, dexamethasone acetate, Indomethacin, naproxen, C14H10Cl2NNaO2, prednisolone, hydrocortisone, metacortandracin, Triamcinolone acetonide and hydrocortisone acetate fast, have easy and simple to handle, fast, the feature of low cost, can as the universal method of Small Meridian-Activating Pill Quality of Chinese Traditional Proprietary Medicine monitoring.
Claims (6)
1. the method for 10 kinds of undeclared prescription drugs in rapid screening Small Meridian-Activating Pill, is characterized in that preparing each reference substance solution (1) and need testing solution (2) first respectively, then carries out examination mensuration (3):
(1) preparation of reference substance solution: get prednisone acetate, dexamethasone acetate, Indomethacin, naproxen and each 10mg of C14H10Cl2NNaO2 reference substance respectively, dissolve with methyl alcohol 10ml, make reference substance solution A; Separately get prednisolone, hydrocortisone, metacortandracin, Triamcinolone acetonide and each 10mg of hydrocortisone acetate reference substance respectively, dissolve with methyl alcohol 10ml, make reference substance solution B;
(2) preparation of need testing solution: get Small Meridian-Activating Pill one ball, shred, add zeyssatite, grind well, add methyl alcohol, ultrasonic process, filters, gets filtrate as need testing solution;
(3) examination mensuration is carried out: draw reference substance solution A respectively, need testing solution, point sample is in same silica G F
254on thin layer plate, using n-hexane-ethyl acetate-glacial acetic acid as developping agent ascending development, take out, dry, thin-layer chromatography is observed under putting ultraviolet lamp 254nm and 365nm respectively, and then spray 10% ethanol solution of sulfuric acid, and it is clear to the spot development on thin layer plate to be placed in 105 DEG C of baking oven heated at constant temperature, then put observed under daylight thin-layer chromatography; If in the chromatogram of above-mentioned three kinds of view modes, Rf value place identical with reference substance has the fluorescent quenching spot of same color, fluorescence spot or spot to occur simultaneously, then judge that sample result is as the positive, having detected 5 kinds of chemicalss is prednisone acetate, dexamethasone acetate, Indomethacin, naproxen and C14H10Cl2NNaO2;
Separately, draw reference substance solution B respectively, need testing solution, point sample is in same silica G F
254on thin layer plate, using methenyl choloride-acetone-methanol-strong ammonia solution as developping agent ascending development, take out, dry, thin-layer chromatography is observed under putting ultraviolet lamp 254nm, spray again with 10% ethanol solution of sulfuric acid, and it is clear to the spot development on thin layer plate to be placed in 105 DEG C of baking oven heated at constant temperature, then puts observed under daylight thin-layer chromatography; If in the chromatogram of above-mentioned two kinds of view modes, Rf value place identical with reference substance has the fluorescent quenching spot of same color or spot to occur simultaneously, then judge that sample result is as the positive, having detected again 5 kinds of chemicalss is prednisolone, hydrocortisone, metacortandracin, Triamcinolone acetonide and hydrocortisone acetate; The volume ratio of above-mentioned developping agent n-hexane-ethyl acetate-glacial acetic acid is 15: 5: 1; The volume ratio of above-mentioned developping agent methenyl choloride-acetone-methanol-strong ammonia solution is 18: 6: 2: 0.1.
2. the method for 10 kinds of undeclared prescription drugs in rapid screening Small Meridian-Activating Pill as claimed in claim 1, is characterized in that above-mentioned zeyssatite weight is 2g.
3. the method for 10 kinds of undeclared prescription drugs in rapid screening Small Meridian-Activating Pill as claimed in claim 1, it is characterized in that in the preparation of above-mentioned (2) need testing solution, the volume adding methyl alcohol is 10ml.
4. the method for 10 kinds of undeclared prescription drugs in rapid screening Small Meridian-Activating Pill as claimed in claim 1, is characterized in that the point sample volume of above-mentioned each reference substance solution is 1 μ l.
5. the method for 10 kinds of undeclared prescription drugs in rapid screening Small Meridian-Activating Pill as claimed in claim 1, is characterized in that the point sample volume of above-mentioned need testing solution is 5 μ l.
6. the method for 10 kinds of undeclared prescription drugs in rapid screening Small Meridian-Activating Pill as claimed in claim 1, is characterized in that the condition of above-mentioned ultrasonic process is 30 minutes, power 250W, frequency 50kHz.
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