CN102749414A - 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 a kind of method of quality control 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) announces a kind ofly to have nourishing heart, calm the nerves, strengthen stomach function effect; Be used to treat the pine benefit power preparation of diseases such as palpitaition, neurasthenia, stomachache, stomach trouble, this medicine is to process oral solutions by single medicinal material rhizoma nardostachyos, yet; Only there is 1 to differentiate the experiment that possibly contain terpenoid according to color reaction to its quality control; Still the qualitative and quantitative analysis method of the qualitative identification method of monomer-free effective constituent, classification composition and limit requirement, the also qualitative and quantitative analysis method of non-activity monomeric compound and limit requirement, so proper mass control specific aim is not strong; Method is imperfect; Be difficult to effectively reach the purpose of stable control finished product pharmacy quality, this has brought difficulty for normal production and operation, has brought hidden danger also for clinical use.
Summary of the invention
The objective of the invention is provides a kind of practicality, effective, perfect qualitative, quantitative to control the method for the Uygur medicine pine benefit power quality of the pharmaceutical preparations to the existing means that the Uygur medicine pine benefit power quality of the pharmaceutical preparations is control effectively that lack.
The present invention realizes that the technical scheme that above-mentioned purpose adopts is following,
A kind of method of quality control of Uygur medicine pine benefit power preparation:
(1) nardosinone and the forulic acid in the employing thin-layered chromatography difference qualitative identification pine benefit power preparation;
(2) adopt general flavone content, total polyphenols content and total polysaccharides content in the AAS difference detection by quantitative pine benefit power preparation;
(3) adopt nardosinone content and ferulaic acid content in the high performance liquid chromatography difference detection by quantitative pine benefit power preparation.
Further, the step of thin-layered chromatography qualitative identification nardosinone is:
Pine benefit power preparation contacts with sherwood oil, and nardosinone is transferred in the sherwood oil, obtains A1 solution; Reference substance is the petroleum ether solution of nardosinone standard items, then respectively with A1 solution and reference substance point on the silica G plate, launch; It is (6～10) that developping agent adopts volume ratio: the petroleum ether-ethyl acetate of (1～2); Dry, spray concentration is the vanillic aldehyde concentrated sulfuric acid solution of 5～10g/ml, and it is clear to be heated to colour developing under 100～110 ℃; Pine benefit power preparation with the corresponding position of reference substance chromatogram on, show the spot of same color.
The vanillic aldehyde concentrated sulfuric acid solution is 5～10g vanillic aldehyde to be added in 100ml 98% concentrated sulphuric acid obtain.
Further, the step of thin-layered chromatography qualitative identification forulic acid is:
Pine benefit power preparation contacts with ether, and forulic acid is transferred in the ether, obtains B1 solution; Evaporate to dryness, residue is used dissolve with methanol, gets B2 solution; Reference substance is the methanol solution of forulic acid standard items, and B2 solution and reference substance are put respectively on the silica G plate, launches; It is (6～12) that developping agent adopts volume ratio: (5～10): (2～4): the toluene-methenyl choloride of (1～2)-ethyl acetate-glacial acetic acid, dry, and under ultraviolet light, observe; Pine benefit power preparation with the corresponding position of reference substance chromatogram on, show the spot of same color.
Further, the step of AAS detection by quantitative general flavone content is:
The drafting of typical curve: control substance of Rutin is used dissolve with ethanol, be mixed with standard solution, draw standard solution, add NaNO successively
2Solution, Al (NO
3)
3Solution and NaOH solution add ethanol again, constant volume, serial contrast solution, measure the absorbance of serial contrast solution with ultraviolet-visible spectrophotometer, obtain the typical curve of concentration-absorbance;
Mend the power preparation to pine and add NaNO successively
2Solution, Al (NO
3)
3Solution and NaOH solution are used the ethanol constant volume again, measure solution absorbency with ultraviolet-visible spectrophotometer, according to the typical curve of concentration-absorbance, calculate content of total flavone in the loose benefit power preparation.
More concrete operating process is following:
The drafting of typical curve: with mass concentration is that 50～70% ethanol joins in the control substance of Rutin; Be mixed with the standard solution that rutin concentration is 0.15～0.25mg/ml; Respectively with 0,0.8,1.2,1.6,2.0, the standard solution of 2.4ml places the 10ml volumetric flask, adds 0.2～0.6ml mass concentration and be 3～7% NaNO
2Solution shakes up, and places 4～8min, adds 0.2～0.6ml mass concentration then and be 8～12% Al (NO
3)
3Solution shakes up, and places 4～8min; Add 2～6ml mass concentration again and be 2～6% NaOH solution; Shake up, using mass concentration is that 50～70% ethanol is settled to scale, thereby obtains the serial contrast solution of variable concentrations; Measure the absorbance of serial contrast solution with ultraviolet-visible spectrophotometer, draw the typical curve of concentration-absorbance at 450～550nm place;
With pine mend power preparation water dissolve sample solution, draw the sample solution decide volume, place the 10ml volumetric flask, adding 0.2～0.6ml mass concentration is 3～7% NaNO
2Solution shakes up, and places 4～8min, adds 0.2～0.6ml mass concentration then and be 8～12% Al (NO
3)
3Solution shakes up, and places 4～8min; Add 2～6ml mass concentration again and be 2～6% NaOH solution; Shake up, using mass concentration is that 50～70% ethanol is settled to scale, measures the absorbance of solution at 450～550nm place with ultraviolet-visible spectrophotometer; According to the typical curve of concentration-absorbance, calculate content of total flavone in the loose benefit power preparation.
Further, the step of AAS detection by quantitative total polyphenols content is:
The drafting of typical curve: with the gallic acid is reference substance, is dissolved in water, and obtains the gallic acid standard solution, draws the gallic acid standard solution, adds forint phenol reagent (Folin-Ciocalteu's phenol reagent) and Na successively
2CO
3Solution adds the water constant volume, gets contrast solution, with the absorbance of ultraviolet-visible spectrophotometer mensuration contrast solution, obtains the typical curve of concentration-absorbance;
Pine is mended the dissolving of power preparation water, obtain sample solution, absorb sample solution, add forint phenol reagent and Na successively
2CO
3Solution adds the water constant volume, measures solution absorbency, according to the typical curve of concentration-absorbance, calculates the content of total polyphenols in the loose benefit power preparation.
Operating process is following more specifically:
The drafting of typical curve: with the gallic acid is 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 places the 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 successively are 15～25% Na
2CO
3Solution adds water and is settled to scale, the serial contrast solution of variable concentrations, measure the absorbance of serial contrast solution with ultraviolet-visible spectrophotometer, obtain the typical curve of concentration-absorbance;
Pine is mended power preparation water dissolving, obtain sample solution, absorb sample solution and place the 10ml volumetric flask, the forint phenol reagent and the 1.0～2.0ml mass concentration that add 0.3～0.7ml, 1～3mol/L successively are 15～25% Na
2CO
3Solution adds water and is settled to scale, measures solution absorbency, according to the typical curve of concentration-absorbance, calculates the content of total polyphenols in the loose benefit power preparation.
Further, the step of AAS detection by quantitative total polysaccharides content is:
The drafting of typical curve: with the dissolving of glucose reference substance water, be mixed with standard solution, draw standard solution; Add sulfuric acid anthrone solution, boil, get contrast solution; With the absorbance of ultraviolet-visible spectrophotometer mensuration contrast solution, obtain the typical curve of concentration-absorbance;
Pine benefit power preparation is used alcohol immersion, filters, and filter residue is respectively with ethanol, acetone, ether flushing; Filter residue water dissolving then gets sample solution, draws sample solution; Add sulfuric acid anthrone solution, after the water-bath heating, measure solution absorbency; According to the typical curve of concentration-absorbance, calculate the content of total polysaccharides in the loose benefit power preparation.
More concrete testing process is following:
The drafting of typical curve: the glucose reference substance is dissolved in water, and constant volume obtains the standard solution that concentration is 0.05～0.15mg/ml; The standard solution of absorption 0,0.2,0.4,0.6,0.8,1.0,1.2ml places tool plug test tube respectively; Add water and be settled to 2ml, quantitatively add the sulfuric acid anthrone solution of 4～8ml, shake up; Boiled 10～20 minutes; Obtain the serial contrast solution of variable concentrations, measure the absorbance of solution, draw the typical curve of concentration-absorbance at 700～800nm place with ultraviolet-visible spectrophotometer;
Pine benefit power formulation samples is used alcohol immersion earlier, spends the night, and filters; The gained filter residue washes 2～4 times with ethanol, acetone, ether respectively, and the water dissolving again of the filter residue after washing promptly gets sample solution; Draw sample solution and place tool plug test tube, add water and be settled to 2ml, quantitatively add the sulfuric acid anthrone solution of 4～8ml; Shake up, boiled 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 polysaccharides in the loose benefit power preparation.
Said sulfuric acid anthrone solution is that 50～150mg anthrone is dissolved in 50～150ml mass concentration is 70～90% the concentrated sulphuric acid.
Further, the step of high performance liquid chromatography detection by quantitative nardosinone content is:
With the nardosinone standard items is 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, and the nardosinone in the loose benefit power preparation is transferred in the sherwood oil, and petroleum ether layer is crossed high performance liquid chromatograph; Measure chromatographic peak area; Calculate the content of nardosinone in the loose benefit power preparation, chromatographic condition is: filling agent be octadecylsilane chemically bonded silica, moving phase be methanol-water, detect wavelength 220～280nm, flowing velocity is 0.8～2.0ml/min, theoretical cam curve must not calculate by nardosinone and is lower than 2000.
Further, the volume ratio of methyl alcohol and water is (75～25): (25～75).
Further, the step of high performance liquid chromatography detection by quantitative ferulaic acid content is:
With the forulic acid standard items is 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, loose benefit power preparation contacts with ether; Nardosinone in the loose benefit power preparation is transferred in the ether; Ether layer is crossed high performance liquid chromatograph, measures chromatographic peak area, calculates the content of nardosinone in the loose benefit power preparation; Chromatographic condition is: filling agent be octadecylsilane chemically bonded silica, moving phase be methyl alcohol-mass concentration be 0.5～1.5% acetic acid aqueous solution, detect wavelength 300～350nm, flowing velocity is 0.8～2.0ml/min, theoretical cam curve must not calculate by forulic acid and is lower than 3000.
Further, methyl alcohol and mass concentration are that the volume ratio of 0.5～1.5% acetic acid aqueous solution is (15.4～28.6): (84.6～71.4).
The above loose benefit power preparation is syrup, mixture, injection, tablet, granule, powder, lozenge, pill, pill, soft extract, gel, plaster, emplastrum, liquid extract and extract, vina, tincture, capsule, ointment, distillate medicinal water, medicinal tea, liniment, lotion, plastics, suppository, aerosol, spray, nasal formulations, eye-drops preparations or the aural preparations that makes with single rhizoma nardostachyos medicinal material.
The present invention adopts above technical scheme to be based on: (1) big quantity research shows that nardosinone is one of effective constituent of rhizoma nardostachyos; Have certain calm trend and antidepressant effect, meet the effect of preparation mental-tranquilization basically, (2) are carried out the cycle chemistry composition to existing loose benefit power preparation and are differentiated; The result shows and contains the plain class of hydroxyl phenols, anthraquinone class, phenylpropyl alcohol, glucide compounds etc. in the preparation; Existing research shows that this classification chemical constitution has strong pharmacologically active effect usually, and (3) forulic acid and sodium salt thereof have certain calmness, inhibiting effect to central nervous system, have the effect that improves sleep quality; And this effect with the preparation mental-tranquilization is corresponding basically; Therefore, through measuring nardosinone, general flavone, total polyphenols, total polysaccharides and content of ferulic acid in the loose benefit power preparation, can control the quality of finished product preparation effectively, all sidedly; Guarantee that the drug quality of producing in batches is stable, using for clinical drug provides reliable basis.
That method of the present invention has is highly sensitive, favorable reproducibility, stability wait characteristics by force, accurately and reliably.
Embodiment
Below in conjunction with embodiment the present invention is further specified.
Embodiment 1
Mending the power oral liquid with pine as follows is that example detects respectively wherein contained nardosinone, forulic acid, general flavone, total polyphenols and total polysaccharides.
(1) nardosinone and the forulic acid in thin-layered chromatography (TLC) the qualitative identification pine benefit power preparation
The discriminating of nardosinone: get loose benefit power oral liquid 5ml, use the 5ml petroleum ether extraction, collect petroleum ether layer; Be concentrated into 1ml and obtain need testing solution, it is the petroleum ether solution of the nardosinone standard items of 1mg/ml that reference substance adopts concentration, and need testing solution and each 10 μ l of reference substance are put respectively on same silica G plate; The silica G plate is a bonding agent with the sodium carboxymethyl cellulose, uses the petroleum ether-ethyl acetate expansion of volume ratio as 4:1, dries; The spray mass concentration is 5% vanillic aldehyde concentrated sulfuric acid solution; It is clear under 105 ℃, to be heated to colour developing, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on show identical spot.
The discriminating of forulic acid: get 20ml pine benefit power oral liquid, use the 20ml extracted with diethyl ether, re-extract once; Combined ether layer, evaporate to dryness obtains powder; Powder obtains need testing solution with the 1ml dissolve with methanol again, and it is the methanol solution of the forulic acid standard items of 1mg/ml that reference substance adopts concentration, and need testing solution and each 10 μ l of reference substance are put respectively on same silica G plate; The silica G plate is a bonding agent with the sodium carboxymethyl cellulose, uses the toluene-methenyl choloride-ethyl acetate-glacial acetic acid expansion of volume ratio as 6:5:2:1, dries; Place the ultraviolet light of 365nm to observe down, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on show identical spot.
(2) general flavone, total polyphenols and the total polysaccharides in the spectrophotometry pine benefit power oral liquid
The mensuration of general flavone
(1) standard solution of preparation reference substance: precision takes by weighing 10 mg rutin standard items (lot numbers: 100080-200707; Nat'l Pharmaceutical & Biological Products Control Institute) in 50 mL volumetric flask; Using mass content is the dissolve with ethanol of 60 %; And be settled to scale, promptly process the standard solution that every 1ml contains rutin 0.2mg;
(2) drawing standard curve: respectively 0,0.8,1.2,1.6,2.0,2.4 ml standard solution are placed 10 ml volumetric flasks, adding mass concentration is the NaNO of 5 %
2WS 0.4ml shakes up, place 6 min after, adding mass concentration is 10% Al (NO
3)
3WS 0.4ml shakes up, place 6 min after, adding mass concentration is the 4% NaOH WS, 4 ml, shakes up; Using mass content at last is that the ethanol of 60 % is settled to 10 ml, measures solution absorbency with ultraviolet-visible spectrophotometer respectively at the 510nm place behind 15 min; (C mg/mL) is horizontal ordinate, and absorbance (A) is an ordinate, and the drawing standard curve gets regression equation: A=-0.0051514+24.301C, (r=0.9992) with rutin concentration.
(3) working sample absorbance: 1ml pine benefit power oral liquid is placed 10 ml volumetric flasks, and adding mass concentration is the NaNO of 5 %
2WS 0.4ml shakes up, place 6 min after, adding mass concentration is 10% Al (NO
3)
3WS 0.4ml shakes up, place 6 min after, adding mass concentration is the 4% NaOH WS, 4 ml, shakes up; Using mass content at last is that the ethanol of 60 % is settled to 10 ml, and the mensuration solution absorbency is 0.656 at 510nm place to use ultraviolet-visible spectrophotometer behind 15 min.
(4) calculate general flavone content in the loose benefit power oral liquid sample: 0.27 mg/ml.
The mensuration of total polyphenols
(1) standard solution of preparation reference substance: precision takes by weighing 10 mg gallic acid standard items in 100 ml volumetric flasks, the adding distil water dissolving, and be settled to scale, promptly getting concentration is the gallic acid standard solution of 0.1 mg/ml;
(2) drawing standard curve: the gallic acid standard solution with 0,0.2,0.4,0.6,0.8,1.0,1.2 ml places 10 ml volumetric flasks respectively, adds the forint phenol reagent of 0.5ml 2 mol/L, shakes up; After placing 1 min, adding mass concentration is the Na of 20 %
2CO
3The WS 1.5 ml shake up, and are settled to 10 ml with distilled water, measure the absorbance of solution at 760 nm places respectively with ultraviolet-visible spectrophotometer after placing 10 min; With gallic acid concentration (C mg/mL) is horizontal ordinate, and absorbance (A) is an ordinate, the drawing standard curve, regression equation: A=0.00565+104.32 C, (r=0.9997);
(3) working sample absorbance: 1ml pine benefit power oral liquid sample is placed 10 ml volumetric flasks, add the forint phenol reagent of 0.5ml 2 mol/L, shake up; After placing 1 min, adding mass concentration is the Na of 20 %
2CO
3The WS 1.5 ml shake up, and are settled to 10 ml with distilled water, place that to use behind 10 min ultraviolet-visible spectrophotometer to measure the absorbance of solution at 760 nm places be 0.428.
(4) calculate total polyphenols content: 0.40mg/ml in the loose benefit power oral liquid sample.
The mensuration of total polysaccharides
(1) preparation reference substance solution: the dissolving of 10mg glucose reference substance adding distil water, be settled to 100 m1, promptly getting concentration is the glucose standard solution of 0.10 mg/ml;
(2) drawing standard curve: the glucose standard solution of 0,0.2,0.4,0.6,0.8,1.0,1.2 ml is placed 10 ml tool plug test tubes respectively, add water to 2.0ml, (preparation: precision takes by weighing anthrone 100mg to accurate adding 6 ml sulfuric acid anthrone solution; Adding 100ml mass concentration is 80% sulfuric acid solution, makes dissolving, shakes up); Shake up, put and boil 15 minutes, take out; Place ice bath cooling 15 minutes, measure solution absorbency at 760 nm places with ultraviolet-visible spectrophotometer, with concentration of glucose (C; Mg/mL) be horizontal ordinate, absorbance (A) is an ordinate, the drawing standard curve; Get regression equation: A=53.466 C+0.0112, (r=0.9996);
(3) preparation sample solution: 95% ethanol with 3 times of amounts of 50ml pine benefit power oral liquid adding, spend the night suction filtration; Filter residue washes 3 times with absolute ethyl alcohol, acetone, the absolute ether of 2 times of amounts (quality) respectively; The filter residue water is settled to 100ml, therefrom gets 2ml and is settled to 25ml, promptly gets sample solution;
(4) working sample solution absorbance: accurate extracting sample solution 2.0ml is in 10 ml tool plug test tubes; The accurate 6 ml sulfuric acid anthrone solution that add (are prepared: step (2)); Shake up, put and boil 15 minutes in the water-bath, take out; Place ice bath cooling 15 minutes, measuring solution absorbency is 0.288.
(5) total polysaccharides content: 0.67mg/ml in the calculation sample.
(3) high performance liquid chromatography detection by quantitative nardosinone content and ferulaic acid content
The mensuration of nardosinone
Chromatographic condition: with the octadecylsilane chemically bonded silica is filling agent, and methyl alcohol (A)-water (B) is moving phase (A:B=60:40), detects wavelength 250nm, flowing velocity 1.0ml/min, and theoretical cam curve must not calculate by nardosinone and is lower than 2000.
(1) preparation reference substance solution: precision takes by weighing 5 mg nardosinone standard items; Add dissolve with methanol and be made into the nardosinone standard solution that concentration is 0.5 mg/ml, use methyl alcohol that 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: the reference substance solution of getting concentration respectively and be 0.00,0.01,0.02,0.03,0.04,0.05,0.06 mg/ml is crossed the miillpore filter of 0.45 μ m, respectively gets filtrating 20 μ l and injects high performance liquid chromatograph and measure chromatographic peak area; (C mg/mL) is horizontal ordinate, and peak area (S) is an ordinate, and the drawing standard curve gets regression equation: S=3 * 10 with nardosinone concentration
7C-10445, (r=0.9992);
(3) preparation sample solution: get loose benefit power oral liquid 20ml; With twice of 40 ml petroleum ether extraction; Merge petroleum ether extraction liquid twice, be concentrated into 2 ml, cross the miillpore filter of 0.45 μ m; 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 the calculation sample: 3.7 g/ml.
The mensuration of forulic acid
Chromatographic condition: with the octadecylsilane chemically bonded silica is filling agent; (A:B volume ratio=22:78) is a moving phase with methyl alcohol (A)-1% (quality) acetic acid aqueous solution (B); The detection wavelength is 320nm, flowing velocity 1.0ml/min, and theoretical cam curve must not calculate by forulic acid and is lower than 3000.
(1) preparation reference substance solution: precision takes by weighing 10 mg forulic acid standard items; Use dissolve with methanol to be made into the forulic acid standard solution that concentration is 0.10 mg/ml, use methyl alcohol standard solution to be diluted to respectively concentration is 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 concentration respectively and 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 μ m; Respectively get filtrating 20ul injecting chromatograph and measure chromatographic peak area, (C mg/mL) is horizontal ordinate with forulic acid concentration; Peak area (S) is an ordinate; The drawing standard curve gets regression equation: S=116321+24912 C, (r=0.9991);
(3) preparation sample solution: get loose benefit power oral liquid 50ml, with 50ml extracted with diethyl ether twice, merge ether extraction liquid twice, evaporate to dryness adds the 3ml dissolve with methanol, crosses the miillpore filter of 0.45 μ m, is sample solution;
(4) getting 20 μ l sample solution injecting chromatographs mensuration chromatographic peak area is 12274152;
(5) content of ferulic acid in the calculation sample: 29 g/ml.
Each item testing result of the present invention is estimated
(1) precision test
(2) stability test
(3) recovery test
Claims (9)
1. the method for quality control of Uygur medicine pine benefit power preparation is characterized in that:
(1) nardosinone and the forulic acid in the employing thin-layered chromatography difference qualitative identification pine benefit power preparation;
(2) adopt general flavone content, total polyphenols content and total polysaccharides content in the AAS difference detection by quantitative pine benefit power preparation;
(3) adopt nardosinone content and ferulaic acid content in the high performance liquid chromatography difference detection by quantitative pine benefit power preparation.
2. according to the method for quality control of the said Uygur medicine pine of claim 1 benefit power preparation, it is characterized in that the step of thin-layered chromatography qualitative identification nardosinone is:
Pine benefit power preparation contacts with sherwood oil, and nardosinone is transferred in the sherwood oil, obtains A1 solution; Reference substance is the petroleum ether solution of nardosinone standard items, then respectively with A1 solution and reference substance point on the silica G plate, launch; It is (6～10) that developping agent adopts volume ratio: the petroleum ether-ethyl acetate of (1～2); Dry, spray concentration is the vanillic aldehyde concentrated sulfuric acid solution of 5～10g/ml, and it is clear to be heated to colour developing under 100～110 ℃; Pine benefit power preparation with the corresponding position of reference substance chromatogram on, show the spot of same color.
3. according to the method for quality control of the said Uygur medicine pine of claim 1 benefit power preparation, it is characterized in that the step of thin-layered chromatography qualitative identification forulic acid is:
Pine benefit power preparation contacts with ether, and forulic acid is transferred in the ether, obtains B1 solution; Evaporate to dryness, residue is used dissolve with methanol, gets B2 solution; Reference substance is the methanol solution of forulic acid standard items, and B2 solution and reference substance are put respectively on the silica G plate, launches; It is (6～12) that developping agent adopts volume ratio: (5～10): (2～4): the toluene-methenyl choloride of (1～2)-ethyl acetate-glacial acetic acid, dry, and under ultraviolet light, observe; Pine benefit power preparation with the corresponding position of reference substance chromatogram on, show the spot of same color.
4. according to the method for quality control of the said Uygur medicine pine of claim 1 benefit power preparation, it is characterized in that the step of AAS detection by quantitative general flavone content is:
The drafting of typical curve: control substance of Rutin is used dissolve with ethanol, be mixed with standard solution, draw standard solution, add NaNO successively
2Solution, Al (NO
3)
3Solution and NaOH solution add ethanol again, constant volume, serial contrast solution, measure the absorbance of serial contrast solution with ultraviolet-visible spectrophotometer, obtain the typical curve of concentration-absorbance;
Mend the power preparation to pine and add NaNO successively
2Solution, Al (NO
3)
3Solution and NaOH solution are used the ethanol constant volume again, measure solution absorbency with ultraviolet-visible spectrophotometer, according to the typical curve of concentration-absorbance, calculate content of total flavone in the loose benefit power preparation.
5. according to the method for quality control of the said Uygur medicine pine of claim 1 benefit power preparation, it is characterized in that the step of AAS detection by quantitative total polyphenols content is:
The drafting of typical curve: with the gallic acid is reference substance, is dissolved in water, and obtains the gallic acid standard solution, draws the gallic acid standard solution, adds forint phenol reagent and Na successively
2CO
3Solution adds the water constant volume, gets contrast solution, with the absorbance of ultraviolet-visible spectrophotometer mensuration contrast solution, obtains the typical curve of concentration-absorbance;
Pine is mended the dissolving of power preparation water, obtain sample solution, absorb sample solution, add forint phenol reagent and Na successively
2CO
3Solution adds the water constant volume, measures solution absorbency, according to the typical curve of concentration-absorbance, calculates the content of total polyphenols in the loose benefit power preparation.
6. according to the method for quality control of the said Uygur medicine pine of claim 1 benefit power preparation, it is characterized in that the step of AAS detection by quantitative total polysaccharides content is:
The drafting of typical curve: with the dissolving of glucose reference substance water, be mixed with standard solution, draw standard solution; Add sulfuric acid anthrone solution, boil, get contrast solution; With the absorbance of ultraviolet-visible spectrophotometer mensuration contrast solution, obtain the typical curve of concentration-absorbance;
Pine benefit power preparation is used alcohol immersion, filters, and filter residue is respectively with ethanol, acetone, ether flushing; Filter residue water dissolving then gets sample solution, draws sample solution; Add sulfuric acid anthrone solution, after the water-bath heating, measure solution absorbency; According to the typical curve of concentration-absorbance, calculate the content of total polysaccharides in the loose benefit power preparation.
7. according to the method for quality control of the said Uygur medicine pine of claim 1 benefit power preparation, it is characterized in that the step of high performance liquid chromatography detection by quantitative nardosinone content is:
With the nardosinone standard items is 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; Nardosinone in the loose benefit power preparation is transferred in the sherwood oil, and petroleum ether layer is crossed high performance liquid chromatograph, measures chromatographic peak area; Calculate the content of nardosinone in the loose benefit power preparation, chromatographic condition is: filling agent is that octadecylsilane chemically bonded silica, moving phase are methanol-water, detection wavelength 220～280nm.
8. according to the method for quality control of the said Uygur medicine pine of claim 1 benefit power preparation, it is characterized in that the step of high performance liquid chromatography detection by quantitative ferulaic acid content is:
With the forulic acid standard items is 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; Nardosinone in the loose benefit power preparation is transferred in the ether, and ether layer is crossed high performance liquid chromatograph, measures chromatographic peak area; Calculate the content of nardosinone in the loose benefit power preparation, chromatographic condition is: filling agent is that octadecylsilane chemically bonded silica, moving phase are that methyl alcohol-mass concentration is 0.5～1.5% acetic acid aqueous solution, detection wavelength 300～350nm.
9. according to the method for quality control of the arbitrary said Uygur medicine pine benefit power preparation of claim 1 to 8, it is characterized in that said loose benefit power preparation is syrup, mixture, injection, tablet, granule, powder, lozenge, pill, pill, soft extract, gel, plaster, emplastrum, liquid extract and extract, vina, tincture, capsule, ointment, distillate medicinal water, medicinal tea, liniment, lotion, plastics, suppository, aerosol, spray, nasal formulations, eye-drops preparations or the aural preparations that makes with single rhizoma nardostachyos medicinal material.
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