CN102749414B - Quality control method for Uighur medicine spikenard strength replenishing preparation - Google Patents
Quality control method for Uighur medicine spikenard strength replenishing preparation Download PDFInfo
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Abstract
The invention belongs to the field of quality control of Uighur medicine preparations, and particularly discloses a quality control method for a Uighur medicine spikenard strength replenishing preparation. The method comprises the following steps of: (1) qualitatively identifying nardosinone and asafetida in the spikenard strength replenishing preparation by using thin layer chromatography respectively; (2) quantitatively detecting total flavone content, total polyphenol content and total polysaccharide content in the spikenard strength replenishing preparation by using spectrophotography respectively; and (3) quantitatively detecting nardosinone content and asafetida content in the spikenard strength replenishing preparation by using high performance liquid chromatography respectively. According to the quality control method, the nardosinone, the total flavone, the total polyphenol, the total polysaccharide and the asafetida in the spikenard strength replenishing preparation are determined in a combined qualitative and quantitative mode, so that the quality of the finished preparation can be effectively and completely controlled and a reliable foundation is provided for clinical application of medicaments. The method has the characteristics of high sensitivity, high repeatability, high stability, accuracy, reliability and the like.
Description
Technical field
The invention belongs to the field of quality control of Uygur medicine preparation, be specifically related to the method for quality control of a kind of Uygur medicine pine benefit power preparation.
Background technology
" Drug Standard of Ministry of Public Health of the Peoples Republic of China " (Uygur medicine fascicle) is announced one and is had and nourish heart, calm the nerves, strengthen stomach function effect, be used for the treatment of palpitaition, neurasthenia, stomachache, the pine of the diseases such as stomach trouble mends power preparation, this medicine makes oral solutions by single medicinal material rhizoma nardostachyos, but, 1 experiment differentiating to contain terpenoid according to color reaction is only had to its quality control, there is no the qualitative identification method of monomer effective constituent, the qualitative and quantitative analysis method of class component and bound requirements, also the qualitative and quantitative analysis method of non-activity monomeric compound and bound requirements, therefore proper mass control specific aim is not strong, method is imperfect, be difficult to the object effectively reaching stability contorting finished product pharmacy quality, this brings difficulty to normal production and operation, also hidden danger is brought to Clinical practice.
Summary of the invention
The object of the invention is the means Uygur medicine pine benefit power quality of the pharmaceutical preparations control effectively for existing shortage, a kind of method of practicality, effective, the perfect qualitative, quantitative control Uygur medicine pine benefit power quality of the pharmaceutical preparations is provided.
It is as follows that the present invention realizes the technical scheme that above-mentioned purpose adopts,
A kind of method of quality control of Uygur medicine pine benefit power preparation:
(1) nardosinone in thin-layered chromatography difference Qualitive test pine benefit power preparation and forulic acid is adopted;
(2) spectrophotometric method is adopted quantitatively to detect general flavone content, Determination of Polyphenols and total starches content in loose benefit power preparation respectively;
(3) high performance liquid chromatography is adopted quantitatively to detect nardosinone content and ferulaic acid content in loose benefit power preparation respectively.
Further, the step of thin-layered chromatography Qualitive test nardosinone is:
Pine benefit power preparation contacts with sherwood oil, nardosinone is made to transfer in sherwood oil, obtain A1 solution, reference substance is the petroleum ether solution of nardosinone standard items, then respectively by A1 solution and reference substance point on silica G plate, launch, developping agent adopts volume ratio to be (6 ~ 10): the petroleum ether-ethyl acetate of (1 ~ 2), dry, spray concentration is the vanillic aldehyde concentrated sulfuric acid solution of 5 ~ 10g/ml, be heated to colour developing at 100 ~ 110 DEG C clear, loose benefit power preparation, on the position corresponding to reference substance chromatogram, shows the spot of same color.
Vanillic aldehyde concentrated sulfuric acid solution is added in 100ml 98% concentrated sulphuric acid by 5 ~ 10g vanillic aldehyde to obtain.
Further, the step of thin-layered chromatography Qualitive test forulic acid is:
Pine benefit power preparation contacts with ether, forulic acid is made to transfer in ether, obtain B1 solution, evaporate to dryness, residue methyl alcohol dissolves, obtain B2 solution, reference substance is the methanol solution of forulic acid standard items, B2 solution and reference substance are put respectively on silica G plate, launch, developping agent adopts volume ratio to be (6 ~ 12): (5 ~ 10): (2 ~ 4): the toluene-methenyl choloride-ethyl acetate-glacial acetic acid of (1 ~ 2), dry, observe under ultraviolet light, loose benefit power preparation, on the position corresponding to reference substance chromatogram, shows the spot of same color.
Further, the step of spectrophotometric standard measure detection general flavone content is:
The drafting of typical curve: dissolved by control substance of Rutin ethanol, be mixed with standard solution, draws standard solution, adds NaNO successively
2solution, Al (NO
3)
3solution and NaOH solution, then add ethanol, constant volume, obtain control series solution, measure the absorbance of control series solution with ultraviolet-visible spectrophotometer, obtain the typical curve of concentration-absorbance;
Mend power preparation to pine and add NaNO successively
2solution, Al (NO
3)
3solution and NaOH solution, then use ethanol constant volume, measure the absorbance of solution with ultraviolet-visible spectrophotometer, according to the typical curve of concentration-absorbance, calculate the content of general flavone in loose benefit power preparation.
More specifically operating process is as follows:
The drafting of typical curve: by mass concentration be 50 ~ 70% ethanol join in control substance of Rutin, be mixed with the standard solution that rutin concentration is 0.15 ~ 0.25mg/ml, respectively by 0,0.8,1.2,1.6,2.0, the standard solution of 2.4ml is placed in 10ml volumetric flask, adds the NaNO that 0.2 ~ 0.6ml mass concentration is 3 ~ 7%
2solution, shakes up, and places 4 ~ 8min, then adds the Al (NO that 0.2 ~ 0.6ml mass concentration is 8 ~ 12%
3)
3solution, shake up, place 4 ~ 8min, add the NaOH solution that 2 ~ 6ml mass concentration is 2 ~ 6% again, shake up, be settled to scale with the ethanol that mass concentration is 50 ~ 70%, thus obtain the control series solution of variable concentrations, measure the absorbance of control series solution at 450 ~ 550nm place with ultraviolet-visible spectrophotometer, draw the typical curve of concentration-absorbance;
Pine is mended power preparation and solve sample solution with water-soluble, draw the sample solution determining volume, be placed in 10ml volumetric flask, add the NaNO that 0.2 ~ 0.6ml mass concentration is 3 ~ 7%
2solution, shakes up, and places 4 ~ 8min, then adds the Al (NO that 0.2 ~ 0.6ml mass concentration is 8 ~ 12%
3)
3solution, shake up, place 4 ~ 8min, add the NaOH solution that 2 ~ 6ml mass concentration is 2 ~ 6% again, shake up, be settled to scale with the ethanol that mass concentration is 50 ~ 70%, measure the absorbance of solution at 450 ~ 550nm place with ultraviolet-visible spectrophotometer, according to the typical curve of concentration-absorbance, calculate the content of general flavone in loose benefit power preparation.
Further, the step of spectrophotometric standard measure detection Determination of Polyphenols is:
The drafting of typical curve: be reference substance with gallic acid, is dissolved in water, obtains gallic acid standard solution, draws gallic acid standard solution, adds forint phenol reagent (Folin-Ciocalteu's phenol reagent) and Na successively
2cO
3solution, add water constant volume, obtains contrast solution, measures the absorbance of contrast solution, obtain the typical curve of concentration-absorbance with ultraviolet-visible spectrophotometer;
Pine is mended the water-soluble solution of power preparation, obtain sample solution, absorb sample solution, add forint phenol reagent and Na successively
2cO
3solution, add water constant volume, measures the absorbance of solution, according to the typical curve of concentration-absorbance, calculate the content of total polyphenols in loose benefit power preparation.
Operating process is as follows more specifically:
The drafting of typical curve: take gallic acid as reference substance, be dissolved in water, obtain the gallic acid standard solution of 0.05 ~ 0.15mg/ml, draw 0,0.2,0.4,0.6,0.8,1.0, the gallic acid standard solution of 1.2ml is placed in 10ml volumetric flask respectively, the forint phenol reagent and the 1.0 ~ 2.0ml mass concentration that add 0.3 ~ 0.7ml, 1 ~ 3mol/L are successively the Na of 15 ~ 25%
2cO
3solution, adds water and is settled to scale, obtains the control series solution of variable concentrations, measures the absorbance of control series solution, obtain the typical curve of concentration-absorbance with ultraviolet-visible spectrophotometer;
Pine is mended the water-soluble solution of power preparation, obtain sample solution, absorb sample solution and be placed in 10ml volumetric flask, the forint phenol reagent and the 1.0 ~ 2.0ml mass concentration that add 0.3 ~ 0.7ml, 1 ~ 3mol/L are successively the Na of 15 ~ 25%
2cO
3solution, adds water and is settled to scale, measures the absorbance of solution, according to the typical curve of concentration-absorbance, calculates the content of total polyphenols in loose benefit power preparation.
Further, the step of spectrophotometric standard measure detection total starches content is:
The drafting of typical curve: by the water-soluble solution of glucose control product, be mixed with standard solution, draws standard solution, add sulfuric acid anthrone solution, boil, obtain contrast solution, measure the absorbance of contrast solution with ultraviolet-visible spectrophotometer, obtain the typical curve of concentration-absorbance;
Pine benefit power preparation alcohol immersion, filter, filter residue uses ethanol, acetone, washed with ether respectively, and then the water-soluble solution of filter residue, obtains sample solution, pipette samples solution, add sulfuric acid anthrone solution, after heating water bath, measure the absorbance of solution, according to the typical curve of concentration-absorbance, calculate the content of total starches in loose benefit power preparation.
More specifically testing process is as follows:
The drafting of typical curve: glucose control product are dissolved in water, constant volume, obtain the standard solution that concentration is 0.05 ~ 0.15mg/ml, draw 0,0.2,0.4,0.6,0.8,1.0, the standard solution of 1.2ml is placed in tool plug test tube respectively, add water and be settled to 2ml, quantitatively add the sulfuric acid anthrone solution of 4 ~ 8ml, shake up, boil 10 ~ 20 minutes, obtain the control series solution of variable concentrations, measure the absorbance of solution at 700 ~ 800nm place with ultraviolet-visible spectrophotometer, draw the typical curve of concentration-absorbance;
Pine benefit power formulation samples first uses alcohol immersion, spend the night, filter, gained filter residue uses ethanol, acetone, washed with ether 2 ~ 4 times respectively, filter residue after washing uses water-soluble solution again, obtain sample solution, pipette samples solution is placed in tool plug test tube, add water and be settled to 2ml, quantitatively add the sulfuric acid anthrone solution of 4 ~ 8ml, shake up, boil 10 ~ 20 minutes, measure the absorbance of solution at 700 ~ 800nm place with ultraviolet-visible spectrophotometer, according to the typical curve of concentration-absorbance, calculate the content of total starches in loose benefit power preparation.
Described sulfuric acid anthrone solution is that 50 ~ 150mg anthrone is dissolved in the concentrated sulphuric acid that 50 ~ 150ml mass concentration is 70 ~ 90%.
Further, high performance liquid chromatography quantitatively detects the step of nardosinone content and is:
With nardosinone standard items for reference substance, preparation series standard solution, sampling, cross high performance liquid chromatograph, measure chromatographic peak area, draw the typical curve of concentration-chromatographic peak area, pine benefit power preparation contacts with sherwood oil, the nardosinone in loose benefit power preparation is made to transfer in sherwood oil, petroleum ether layer crosses high performance liquid chromatograph, measure chromatographic peak area, calculate the content of nardosinone in loose benefit power preparation, chromatographic condition is: filling agent is octadecylsilane chemically bonded silica, mobile phase is methanol-water, determined wavelength 220 ~ 280nm, flowing velocity is 0.8 ~ 2.0ml/min, theoretical cam curve is pressed nardosinone and must not calculate lower than 2000.
Further, the volume ratio of methyl alcohol and water is (75 ~ 25): (25 ~ 75).
Further, high performance liquid chromatography quantitatively detects the step of ferulaic acid content and is:
With forulic acid standard items for reference substance, preparation series standard solution, sampling, cross high performance liquid chromatograph, measure chromatographic peak area, draw the typical curve of concentration-chromatographic peak area, pine benefit power preparation contacts with ether, the nardosinone in loose benefit power preparation is made to transfer in ether, ether layer crosses high performance liquid chromatograph, measure chromatographic peak area, calculate the content of nardosinone in loose benefit power preparation, chromatographic condition is: filling agent is octadecylsilane chemically bonded silica, mobile phase to be methyl alcohol-mass concentration be 0.5 ~ 1.5% acetic acid aqueous solution, determined wavelength 300 ~ 350nm, flowing velocity is 0.8 ~ 2.0ml/min, theoretical cam curve is pressed forulic acid and must not calculate lower than 3000.
Further, methyl alcohol and mass concentration are the volume ratio of the acetic acid aqueous solution of 0.5 ~ 1.5% is (15.4 ~ 28.6): (84.6 ~ 71.4).
The above loose benefit power preparation is with obtained syrup, mixture, injection, tablet, granule, powder, lozenge, pill, pill, soft extract, gel, plaster, emplastrum, liquid extract and the extract of single rhizoma nardostachyos medicinal material, vina, tincture, capsule, ointment, distillate medicinal water, medicinal tea, liniment, lotion, plastics, suppository, aerosol, spray, nasal formulations, eye-drops preparations or aural preparations.
The present invention adopt above technical scheme be based on: (1) large quantity research shows that nardosinone is one of effective constituent of rhizoma nardostachyos, there is certain calm trend and antidepressant effect, substantially effect of preparation mental-tranquilization is met, (2) discriminating of cycle chemistry composition is carried out to existing loose benefit power preparation, result shows in preparation containing hydroxyl phenols, Anthraquinones, Phenylpropanoid Glycosides class, glucide compounds etc., now there are some researches show that this classification chemical composition has stronger pharmacologically active effect usually, (3) forulic acid and sodium salt thereof have certain calmness to central nervous system, inhibiting effect, there is the effect of improving water flood quality, and this is substantially corresponding with effect of preparation mental-tranquilization, therefore, by measuring nardosinone in loose benefit power preparation, general flavone, total polyphenols, the content of total starches and forulic acid, can be effective, control the quality of finished product preparation all sidedly, ensure that the drug quality of batch production is stablized, for clinical drug application provides reliable basis.
Method of the present invention have highly sensitive, favorable reproducibility, stability by force, the feature such as accurately and reliably.
Embodiment
Below in conjunction with embodiment, the present invention will be further described.
Embodiment 1
Mend power oral liquid for pine as follows to detect respectively wherein contained nardosinone, forulic acid, general flavone, total polyphenols and total starches.
(1) nardosinone in thin-layered chromatography (TLC) Qualitive test pine benefit power preparation and forulic acid
The discriminating of nardosinone: get loose benefit power oral liquid 5ml, use 5ml petroleum ether extraction, collect petroleum ether layer, be concentrated into 1ml and obtain need testing solution, reference substance employing concentration is the petroleum ether solution of the nardosinone standard items of 1mg/ml, need testing solution and each 10 μ l of reference substance are put respectively on same silica G plate, silica G plate take sodium carboxymethyl cellulose as bonding agent, launch with the petroleum ether-ethyl acetate that volume ratio is 4:1, dry, spray mass concentration is 5% vanillic aldehyde concentrated sulfuric acid solution, colour developing is heated to clear at 105 DEG C, in test sample chromatogram, aobvious identical spot on the position corresponding with reference substance chromatogram.
The discriminating of forulic acid: get 20ml pine benefit power oral liquid, use 20ml extracted with diethyl ether, re-extract once, combined ether layer, evaporate to dryness, obtain powder, powder dissolves with 1ml methyl alcohol and obtains need testing solution, reference substance employing concentration is the methanol solution of the forulic acid standard items of 1mg/ml, need testing solution and each 10 μ l of reference substance are put respectively on same silica G plate, silica G plate take sodium carboxymethyl cellulose as bonding agent, launch with toluene-methenyl choloride-ethyl acetate-glacial acetic acid that volume ratio is 6:5:2:1, dry, observe under being placed in the ultraviolet light of 365nm, in test sample chromatogram, aobvious identical spot on the position corresponding with reference substance chromatogram.
(2) general flavone, total polyphenols and total starches in spectrophotometry pine benefit power oral liquid
the mensuration of general flavone
(1) standard solution of reference substance is prepared: precision takes 10 mg rutin standard items (lot numbers: 100080-200707, Nat'l Pharmaceutical & Biological Products Control Institute) in 50 mL volumetric flasks, dissolve with the ethanol that mass content is 60 %, and be settled to scale, namely make the standard solution of every 1ml containing rutin 0.2mg;
(2) drawing standard curve: respectively 0,0.8,1.2,1.6,2.0,2.4 ml standard solution are placed in 10 ml volumetric flasks, adds the NaNO that mass concentration is 5 %
2aqueous solution 0.4ml, shakes up, and after placing 6 min, adding mass concentration is 10% Al (NO
3)
3aqueous solution 0.4ml, shakes up, and after placing 6 min, adding mass concentration is 4% NaOH aqueous solution 4 ml, shakes up; Finally be settled to 10 ml with the ethanol that mass content is 60 %, after 15 min, measure the absorbance of solution with ultraviolet-visible spectrophotometer at 510nm place respectively; With rutin concentration (C, mg/mL) for horizontal ordinate, absorbance (A) is ordinate, and drawing standard curve, obtains regression equation: A=-0.0051514+24.301C, (r=0.9992).
(3) working sample absorbance: 1ml pine benefit power oral liquid is placed in 10 ml volumetric flasks, adds the NaNO that mass concentration is 5 %
2aqueous solution 0.4ml, shakes up, and after placing 6 min, adding mass concentration is 10% Al (NO
3)
3aqueous solution 0.4ml, shakes up, and after placing 6 min, adding mass concentration is 4% NaOH aqueous solution 4 ml, shakes up; Finally be settled to 10 ml with the ethanol that mass content is 60 %, measuring the absorbance of solution with ultraviolet-visible spectrophotometer at 510nm place after 15 min is 0.656.
(4) general flavone content in loose benefit power oral liquid sample is calculated: 0.27 mg/ml.
the mensuration of total polyphenols
(1) prepare the standard solution of reference substance: precision takes 10 mg gallic acid standard items in 100 ml volumetric flasks, adding distil water dissolves, and is settled to scale, obtains the gallic acid standard solution that concentration is 0.1 mg/ml;
(2) drawing standard curve: respectively the gallic acid standard solution of 0,0.2,0.4,0.6,0.8,1.0,1.2 ml is placed in 10 ml volumetric flasks, adds the forint phenol reagent of 0.5ml 2 mol/L, shakes up; After placing 1 min, add the Na that mass concentration is 20 %
2cO
3aqueous solution 1.5 ml, shakes up, and is settled to 10 ml with distilled water, measures the absorbance of solution at 760 nm places respectively after placing 10 min with ultraviolet-visible spectrophotometer; With gallic acid concentration (C, mg/mL) for horizontal ordinate, absorbance (A) is ordinate, and drawing standard curve, obtains regression equation: A=0.00565+104.32 C, (r=0.9997);
(3) working sample absorbance: 1ml pine benefit power oral liquid sample is placed in 10 ml volumetric flasks, adds the forint phenol reagent of 0.5ml 2 mol/L, shake up; After placing 1 min, add the Na that mass concentration is 20 %
2cO
3aqueous solution 1.5 ml, shakes up, and is settled to 10 ml with distilled water, and measuring solution in the absorbance at 760 nm places with ultraviolet-visible spectrophotometer after placing 10 min is 0.428.
(4) Determination of Polyphenols: 0.40mg/ml in loose benefit power oral liquid sample is calculated.
the mensuration of total starches
(1) reference substance solution is prepared: 10mg glucose control product adding distil water dissolves, and is settled to 100 m1, obtains the Glucose standards solution that concentration is 0.10 mg/ml;
(2) drawing standard curve: by 0, 0.2, 0.4, 0.6, 0.8, 1.0, the Glucose standards solution of 1.2 ml is placed in 10 ml tool plug test tubes respectively, add water to 2.0ml, precision adds the (preparation: precision takes anthrone 100mg of 6 ml sulfuric acid anthrone solution, add the sulfuric acid solution that 100ml mass concentration is 80%, make dissolving, shake up), shake up, put and boil 15 minutes, take out, be placed in ice bath cooling 15 minutes, measure the absorbance of solution at 760 nm places with ultraviolet-visible spectrophotometer, with concentration of glucose (C, mg/mL) be horizontal ordinate, absorbance (A) is ordinate, drawing standard curve, obtain regression equation: A=53.466 C+0.0112, (r=0.9996),
(3) prepare sample solution: 95% ethanol 50ml pine benefit power oral liquid being added 3 times amount, spend the night, suction filtration, filter residue uses the absolute ethyl alcohol of 2 times amount (quality), acetone, absolute ether to rinse 3 times respectively, filter residue water is settled to 100ml, therefrom gets 2ml and is settled to 25ml, obtain sample solution;
(4) working sample solution absorbance: accurate extracting sample solution 2.0ml is in 10 ml tool plug test tubes, precision adds 6 ml sulfuric acid anthrone solution (preparation: step (2)), shake up, put in water-bath and boil 15 minutes, take out, be placed in ice bath cooling 15 minutes, the absorbance measuring solution is 0.288.
(5) total starches content: 0.67mg/ml in calculation sample.
(3) high performance liquid chromatography quantitatively detects nardosinone content and ferulaic acid content
the mensuration of nardosinone
Chromatographic condition: take octadecylsilane chemically bonded silica as filling agent, methyl alcohol (A)-water (B) (A:B=60:40) is mobile phase, determined wavelength 250nm, flowing velocity 1.0ml/min, theoretical cam curve is pressed nardosinone and must not calculate lower than 2000.
(1) reference substance solution is prepared: precision takes 5 mg nardosinone standard items, add methyl alcohol dissolving and be made into the nardosinone standard solution that concentration is 0.5 mg/ml, with methyl alcohol, standard solution is diluted to the reference substance solution that concentration is 0.00,0.01,0.02,0.03,0.04,0.05,0.06 mg/ml respectively;
(2) drawing standard curve: getting concentration is respectively the miillpore filter that the reference substance solution of 0.00,0.01,0.02,0.03,0.04,0.05,0.06 mg/ml crosses 0.45 μm, respectively gets filtrate 20 μ l and injects high performance liquid chromatograph and measure chromatographic peak area; With nardosinone concentration (C, mg/mL) for horizontal ordinate, peak area (S) is ordinate, and drawing standard curve, obtains regression equation: S=3 × 10
7c-10445, (r=0.9992);
(3) sample solution is prepared: get loose benefit power oral liquid 20ml, with 40 ml petroleum ether extraction twice, merge twice petroleum ether extraction liquid, be concentrated into 2 ml, cross the miillpore filter of 0.45 μm, be sample solution, get 20 μ l sample solution liquid and inject high performance liquid chromatograph mensuration chromatographic peak area 1098654;
(4) content of nardosinone in calculation sample: 3.7 μ g/ml.
the mensuration of forulic acid
Chromatographic condition: take octadecylsilane chemically bonded silica as filling agent, with methyl alcohol (A)-1% (quality) acetic acid aqueous solution (B) (A:B volume ratio=22:78) for mobile phase, determined wavelength is 320nm, flowing velocity 1.0ml/min, theoretical cam curve is pressed forulic acid and must not calculate lower than 3000.
(1) reference substance solution is prepared: precision takes 10 mg forulic acid standard items, dissolve with methyl alcohol and be made into the forulic acid standard solution that concentration is 0.10 mg/ml, with methyl alcohol standard solution is diluted to respectively concentration be 0.02,0.04,0.06,0.08,0.10,0.12, the reference substance solution of 0.14mg/ml.
(2) drawing standard curve: get respectively concentration be 0.02,0.04,0.06,0.08,0.10,0.12, the reference substance solution of 0.14mg/ml crosses the miillpore filter of 0.45 μm, respectively get filtrate 20ul injecting chromatograph and measure chromatographic peak area, with forulic acid concentration (C, mg/mL) be horizontal ordinate, peak area (S) is ordinate, drawing standard curve, obtains regression equation: S=116321+24912 C, (r=0.9991); ;
(3) prepare sample solution: get loose benefit power oral liquid 50ml, by 50ml extracted with diethyl ether twice, merge twice ether extraction liquid, evaporate to dryness, add 3ml methyl alcohol and dissolve, cross the miillpore filter of 0.45 μm, be sample solution;
(4) getting 20 μ l sample solution injecting chromatographs mensuration chromatographic peak areas is 12274152;
(5) content of forulic acid in calculation sample: 29 μ g/ml.
the every evaluation of the present invention
(1) precision test
(2) stability test
(3) recovery test
Claims (2)
1. a detection method for Uygur medicine pine benefit power preparation, is characterized in that:
(1) nardosinone in thin-layered chromatography difference Qualitive test pine benefit power preparation and forulic acid is adopted;
(2) spectrophotometric method is adopted quantitatively to detect general flavone content, Determination of Polyphenols and total starches content in loose benefit power preparation respectively;
(3) high performance liquid chromatography is adopted quantitatively to detect nardosinone content and ferulaic acid content in loose benefit power preparation respectively;
Wherein,
The step of thin-layered chromatography Qualitive test nardosinone is:
Pine benefit power preparation contacts with sherwood oil, nardosinone is made to transfer in sherwood oil, obtain A1 solution, reference substance is the petroleum ether solution of nardosinone standard items, then respectively by A1 solution and reference substance point on silica G plate, launch, developping agent adopts volume ratio to be the petroleum ether-ethyl acetate of 4:1, dry, spray concentration is the vanillic aldehyde concentrated sulfuric acid solution of 5 ~ 10g/ml, be heated to colour developing at 100 ~ 110 DEG C clear, loose benefit power preparation, on the position corresponding to reference substance chromatogram, shows the spot of same color;
The step of thin-layered chromatography Qualitive test forulic acid is:
Pine benefit power preparation contacts with ether, makes forulic acid transfer in ether, obtains B1 solution, evaporate to dryness, residue methyl alcohol dissolves, and obtains B2 solution, reference substance is the methanol solution of forulic acid standard items, B2 solution and reference substance are put respectively on silica G plate, launch, developping agent employing volume ratio is the toluene-methenyl choloride-ethyl acetate-glacial acetic acid of 6:5:2:1, dry, observe under ultraviolet light, loose benefit power preparation, on the position corresponding to reference substance chromatogram, shows the spot of same color;
The step that spectrophotometric standard measure detects general flavone content is:
The drafting of typical curve: dissolved by control substance of Rutin ethanol, be mixed with standard solution, draws standard solution, adds NaNO successively
2solution, Al (NO
3)
3solution and NaOH solution, then add ethanol, constant volume, obtain control series solution, measure the absorbance of control series solution with ultraviolet-visible spectrophotometer, obtain the typical curve of concentration-absorbance;
Mend power preparation to pine and add NaNO successively
2solution, Al (NO
3)
3solution and NaOH solution, then use ethanol constant volume, measure the absorbance of solution with ultraviolet-visible spectrophotometer, according to the typical curve of concentration-absorbance, calculate the content of general flavone in loose benefit power preparation;
The step that spectrophotometric standard measure detects Determination of Polyphenols is:
The drafting of typical curve: be reference substance with gallic acid, is dissolved in water, obtains gallic acid standard solution, draws gallic acid standard solution, adds forint phenol reagent and Na successively
2cO
3solution, add water constant volume, obtains contrast solution, measures the absorbance of contrast solution, obtain the typical curve of concentration-absorbance with ultraviolet-visible spectrophotometer;
Pine is mended the water-soluble solution of power preparation, obtain sample solution, absorb sample solution, add forint phenol reagent and Na successively
2cO
3solution, add water constant volume, measures the absorbance of solution, according to the typical curve of concentration-absorbance, calculate the content of total polyphenols in loose benefit power preparation;
The step that spectrophotometric standard measure detects total starches content is:
The drafting of typical curve: by the water-soluble solution of glucose control product, be mixed with standard solution, draws standard solution, add sulfuric acid anthrone solution, boil, obtain contrast solution, measure the absorbance of contrast solution with ultraviolet-visible spectrophotometer, obtain the typical curve of concentration-absorbance;
Pine benefit power preparation alcohol immersion, filter, filter residue uses ethanol, acetone, washed with ether respectively, and then the water-soluble solution of filter residue, obtains sample solution, pipette samples solution, add sulfuric acid anthrone solution, after heating water bath, measure the absorbance of solution, according to the typical curve of concentration-absorbance, calculate the content of total starches in loose benefit power preparation;
The step that high performance liquid chromatography quantitatively detects nardosinone content is:
With nardosinone standard items for reference substance, preparation series standard solution, sampling, cross high performance liquid chromatograph, measure chromatographic peak area, draw the typical curve of concentration-chromatographic peak area, pine benefit power preparation contacts with sherwood oil, the nardosinone in loose benefit power preparation is made to transfer in sherwood oil, petroleum ether layer crosses high performance liquid chromatograph, measure chromatographic peak area, calculate the content of nardosinone in loose benefit power preparation, chromatographic condition is: filling agent is octadecylsilane chemically bonded silica, mobile phase is methanol-water, determined wavelength 220 ~ 280nm, wherein methyl alcohol: water=60:40,
The step that high performance liquid chromatography quantitatively detects ferulaic acid content is:
With forulic acid standard items for reference substance, preparation series standard solution, sampling, cross high performance liquid chromatograph, measure chromatographic peak area, draw the typical curve of concentration-chromatographic peak area, pine benefit power preparation contacts with ether, the nardosinone in loose benefit power preparation is made to transfer in ether, ether layer crosses high performance liquid chromatograph, measure chromatographic peak area, calculate the content of nardosinone in loose benefit power preparation, chromatographic condition is: filling agent is octadecylsilane chemically bonded silica, mobile phase to be methyl alcohol-mass concentration be 1% acetic acid aqueous solution, determined wavelength 300 ~ 350nm, wherein methyl alcohol and mass concentration are the volume ratio=22:78 of the acetic acid aqueous solution of 1%.
2. the detection method of Uygur medicine pine benefit power preparation according to claim 1, is characterized in that described loose benefit power preparation is with obtained syrup, mixture, injection, tablet, granule, powder, lozenge, pill, pill, soft extract, gel, plaster, emplastrum, liquid extract and the extract of single rhizoma nardostachyos medicinal material, vina, tincture, capsule, ointment, distillate medicinal water, medicinal tea, liniment, lotion, plastics, suppository, aerosol, spray, nasal formulations, eye-drops preparations or aural preparations.
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