CN107064090B - A kind of quality determining method of sinew-Soothing Blood-Quickening Tablet - Google Patents
A kind of quality determining method of sinew-Soothing Blood-Quickening Tablet Download PDFInfo
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Abstract
The present invention relates to a kind of quality determining method of sinew-Soothing Blood-Quickening Tablet, assay is carried out including the indentification by TLC to Radix Notoginseng in sinew-Soothing Blood-Quickening Tablet, the rhizome of davallia, and to heavy metal contained by it.Detecting step are as follows: one, using Ginsenoside Rg1 and Rb1 as reference substance, using whether having Radix Notoginseng ingredient in indentification by TLC sinew-Soothing Blood-Quickening Tablet；Two, using aurantiin as reference substance, using whether having rhizome of davallia ingredient in indentification by TLC sinew-Soothing Blood-Quickening Tablet；Three, using lead, cadmium, arsenic, mercury, five heavy metal species unit standard solution of copper as reference substance, whether contain the above 5 heavy metal species ingredient using in atomic absorption spectroscopy determination sinew-Soothing Blood-Quickening Tablet.The invention has the benefit that the quality of sinew-Soothing Blood-Quickening Tablet carry out effectively, reliable control, it is party's forensic science, feasible, reliable.
Description
Technical field
The invention belongs to pharmaceutical fields, and in particular to a kind of quality determining method of sinew-Soothing Blood-Quickening Tablet.
Background technique
Sinew-Soothing Blood-Quickening Tablet is ground bettle, ramulus cinnamomi, safflower, peach kernel, the rhizome of davallia, radix achyranthis bidentatae, cape jasmine, the root of Dahurain angelica, teasel root, Radix Notoginseng, big
The 20 taste pulverizing medicinal materials such as Huang, radix paeoniae rubra, Rehmannia glutinosa, bush, olibanum, catechu, Radix Angelicae Sinensis, vomiting nut, borneol, native copper add at after fine powder
Enter the pill that honey is prepared into；Bitter is puckery.Its major function is channels sootheing and network vessel quickening, promoting blood circulation and stopping pain.For traumatic injury, trouble of spraining one's back
Gas, breaking of muscle and tendon fracture, pain due to blood stasis.
Existing sinew-Soothing Blood-Quickening Tablet has clinically been widely applied, but since native copper is main in sinew-Soothing Blood-Quickening Tablet prescription
Ingredient, the effects of playing the scattered stasis of blood, synthetism, relieve pain, main component is ferrous disulfide (FeS2), separately there is pertinent literature to report nature
Also containing heavy metals such as lead, arsenic, cadmium, copper, arsenic in copper, therefore consider many patients, in order to preferably control the quality of product,
Radix Notoginseng, the rhizome of davallia in sinew-Soothing Blood-Quickening Tablet are carried out to identify research, to five kinds of lead, cadmium, arsenic, mercury, copper huge sum of moneys in sinew-Soothing Blood-Quickening Tablet
Belong to content and carries out content detection and running water.
Summary of the invention
It is of the existing technology in order to solve the problems, such as, the present invention provides a kind of quality determining method of sinew-Soothing Blood-Quickening Tablet,
It has the characteristics that specificity is strong, separating degree is good.
The technical scheme adopted by the invention is as follows:
A kind of quality determining method of sinew-Soothing Blood-Quickening Tablet, including the identification to Radix Notoginseng, the identification to the rhizome of davallia and a huge sum of money
The detection of category, wherein
The identification of Radix Notoginseng the following steps are included:
The preparation of test solution A: taking sinew-Soothing Blood-Quickening Tablet 7g, shreds plus methanol 50ml is heated to reflux 1 hour, let cool, filter
It crosses, filtrate is evaporated, residue is transferred in separatory funnel, first plus the shaking of water saturated n-butanol is extracted 3 times, each 30ml,
Merge n-butanol liquid；It ammoniates test solution again to wash 2 times, each 20ml；Water washing 2 times be saturated again with n-butanol, each 20ml takes
N-butanol liquid, is evaporated, and residue adds methanol 1ml to make to dissolve, and obtains test solution A；
The preparation of reference substance solution A: taking ginsenoside Rb1, ginsenoside Rg1's reference substance, adds methanol that every 1ml is made and contains
The mixed solution of 0.5mg, as reference substance solution A；
It draws test solution A, each 5 μ l of reference substance solution A to be put respectively on same silica gel g thin-layer plate, with chloroform-
Lower layer's solution of 10 DEG C of acetate-methanol-water (15:40:22:10) placements is solvent, is unfolded, colour developing；
The identification of the rhizome of davallia the following steps are included:
The preparation of test solution B: taking sinew-Soothing Blood-Quickening Tablet 30g, diatomite added to disperse, and chloroform 150ml ultrasound is added to mention
It takes 2 times, 30 minutes every time, lets cool, filter, filtrate is evaporated, filter residue is obtained, ethyl acetate 150ml, 30 points of ultrasound are added into residue
Clock, filtration, is evaporated filtrate, makes to dissolve to residue plus methanol 1ml, obtain test solution B；
The preparation of reference substance solution B: taking aurantiin reference substance, adds methanol that solution of every 1ml containing 0.5mg is made, must compare
Product solution B；
4 μ l of reference substance solution B is drawn, 10 μ l of test solution B is put respectively on same silica G prefabricated sheets plate, item
Band application is unfolded using toluene-ethyl acetate-formic acid-water (1:12:2.5:3) upper solution as solvent, colour developing；
The detection of heavy metal the following steps are included:
The preparation of test solution C: taking test sample powder 0.1g, set in kjeldahl flask, adds nitric acid-perchloric acid mixed solution
10ml is mixed, soaked overnight；Heating resolution, digestion solution is transferred in 50ml measuring bottle, washs kjeldahl flask with 2% nitric acid solution,
Washing lotion is incorporated in measuring bottle, and is diluted to scale, is shaken up to get test solution C；
The preparation of test solution D: taking test sample powder 0.1g, set in kjeldahl flask, adds nitric acid-perchloric acid mixed solution
10ml is mixed, soaked overnight；It is heated to clearing up completely, into digestion solution plus 20% sulfuric acid solution 10ml, 5% potassium permanganate are molten
Liquid 0.5ml, shakes up, and just disappears 5% hydroxylamine hydrochloride solution is added dropwise to aubergine, is transferred in 50ml measuring bottle, beginning is washed with water
Flask, washing lotion are incorporated in measuring bottle, are diluted with water to scale to get test solution D.
Further, in the identification step of Radix Notoginseng, further include the preparation of negative test solution A: by recipe quantity, removing three
Other medicinal material powders outside seven mix, add refined honey that Radix Notoginseng negative sample is made, feminine gender is made by the preparation method of test solution A
Contrast solution A.
Further, it in the identification step of Radix Notoginseng, after silica gel g thin-layer plate dries, sprays with 10% ethanol solution of sulfuric acid, In
105 DEG C to be heated to spot development clear.
Further, in the identification step of the rhizome of davallia, further include the preparation of negative test solution B: by recipe quantity, removing
Other medicinal material powders outside the rhizome of davallia mix, add refined honey that rhizome of davallia negative sample is made, by the preparation method system of test solution B
At negative control solution B.
Further, it in the identification step of the rhizome of davallia, after silica gel g thin-layer plate dries, sprays with alchlor test solution, In
It is inspected under 365nm ultraviolet lamp.
Further, further include the preparation of lead standard solution in heavy metal analysis step: precision measures lead single element standard
Solution is diluted with 2% nitric acid solution, the solution of the leaded 1 μ g of every 1ml is made.
Further, further include the preparation of cadmium standard solution in heavy metal analysis step: precision measures lead single element standard
Solution is diluted with 2% nitric acid solution, the solution of every 1ml 1 μ g containing cadmium is made.
Further, further include the preparation of arsenic standard solution in heavy metal analysis step: precision measures lead single element standard
Solution is diluted with 2% nitric acid solution, the solution of every 1ml 1 μ g containing arsenic is made.
Further, further include the preparation of mercury standard solution in heavy metal analysis step: precision measures lead single element standard
Solution is diluted with 2% nitric acid solution, the solution of the mercurous 1 μ g of every 1ml is made.
Further, further include the preparation of copper standard solution in heavy metal analysis step: precision measures lead single element standard
Solution is diluted with 2% nitric acid solution, the solution of every 10 μ g of 1ml cupric is made.
The invention has the benefit that
1. method of the invention can effectively be handled relaxing tendons and activating collaterals pill, it is saturated using methanol, n-butanol
Extraction with aqueous solution, and washed with ammonia solution, it is not only easy to operate, and can effectively extract ginsenoside Rb1 in Radix Notoginseng, ginseng
Saponin(e Rg1, while excluding the interference of other compositions；
2. Radix Notoginseng uses chloroform-acetate-methanol-aqueous systems in identifying, ginsenoside Rb1, ginseng can be made
Two components of saponin(e Rg1 have preferable solute migration distance, obtain ideal Rf value, at the same can guarantee two at
/ good separating degree, and background is noiseless；
3. method of the invention can effectively be handled relaxing tendons and activating collaterals pill, using diatomite dispersion, three chloromethanes
Alkane dissolution is extracted, and successively extracts dissolution using ethyl acetate, methanol, not only easy to operate, and can effectively be extracted
Aurantiin ingredient in the rhizome of davallia, while excluding the interference of other compositions；
4. the rhizome of davallia uses toluene-ethyl acetate-formic acid-aqueous systems in identifying, using its upper solution as solvent, obtain
Ideal Rf value, while background is noiseless；
5. native copper is the main component in the prescription in relaxing tendons and activating collaterals pill, the scattered stasis of blood, synthetism, the work of analgesic are played
With the impurity such as also leaded in native copper, arsenic, cadmium, copper, nickel, arsenic, antimony, to cause in sinew-Soothing Blood-Quickening Tablet (small honey pill) containing weight
Therefore heavy metal analysis is included in quality standard and is of great significance by metal；
6. heavy metal detection method of the invention, is effectively directed to relaxing tendons and activating collaterals pill, repeatability, is loaded back at precision
The results such as yield meet the requirements, and therefore, can accurately detect the content of heavy metal in preparation.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, technical solution of the present invention will be carried out below
Detailed description.Obviously, the described embodiments are only a part of the embodiments of the utility model, rather than whole implementation
Example.Based on the embodiments of the present invention, those of ordinary skill in the art are acquired without making creative work
All other embodiment, belong to the range protected of the present invention.
The identification of 1 Radix Notoginseng of embodiment
1. having carried out the identification of medicinal material: determining the applicability of the presence or absence of target component and method in medicinal material.
The preparation of 1.1 reference substances
Ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1 add methanol to be prepared into every ml respectively containing the mixed liquor of 0.5mg,
As reference substance solution.
1.2. prepared by pseudo-ginseng
Pseudo-ginseng powder 0.5g adds the drop of water 5 to stir evenly, and water-saturated n-butanol 5ml is added to shake 10 minutes, places 2h, is centrifuged,
Supernatant is taken, adds the water of 3 times of amount n-butanol saturations to shake up, placement makes to be layered, and takes n-butanol layer, is evaporated residue and methanol 1ml is added to make
Dissolution, as medicinal material test solution.
1.3 point samples, expansion
Draw each 1ul of pseudo-ginseng test solution and control solution respectively, point on same silica gel g thin-layer plate, with
Chloroform -10 DEG C of acetate-methanol-water (15:40:22:10) lower layer's solution arranged below is solvent, is unfolded, takes
Out, it dries, spray is heated to clear spot in 105 DEG C with sulfuric acid solution (1 → 10), sets daylight and ultraviolet lamp (365nm) respectively
Under inspect.
1.4. as a result, having same color spot and fluorescence spot in medicinal material chromatography and reference substance chromatography in same position.
2. the identification of pseudo-ginseng in sinew-Soothing Blood-Quickening Tablet finished product
The preparation (method one) of 2.1 test samples
Method one takes sinew-Soothing Blood-Quickening Tablet sample 10g, and appropriate diatomite is added to disperse, and takes (30~60 DEG C) of 200ml petroleum ether to surpass
Sound 50 minutes, petroleum ether liquid is discarded, the dregs of a decoction volatilize, and add 200ml chloroform ultrasound 30 minutes, discard chloroform liquid, the dregs of a decoction
It volatilizes, adds 100ml water-saturated n-butanol ultrasound 30 minutes, filtration, filtrate adds 0.5% sodium hydroxide solution shaking 2 times, every time
100ml is washed with water to neutrality, and n-butanol liquid is evaporated, and residue adds methanol 1ml to make to dissolve, as test solution.
2.2 point samples, expansion
Reference substance solution 5ul, medicinal material solution 5ul are drawn, test solution is distinguished 5ul, 10ul, put respectively in same silica gel
It is exhibition with chloroform -10 DEG C of acetate-methanol-water (15:40:22:10) lower layer's solution arranged below on G lamellae
Agent is opened, expansion takes out, dries up immediately, and spray is heated to clear spot in 110 DEG C with 10% sulfuric acid solution, sets daylight and purple respectively
It is inspected under outer smooth lamp (365nm).
2.3. as a result, sample chromatogram in medicinal material chromatography and reference substance chromatography with reference substance Rg1, Re in identical bits
It sets, there is same color spot and fluorescence spot, test sample, without obvious spot, considers the process in decontamination at reference substance Rb1
Middle loss is more, therefore need to change Extraction solvent and decontamination mode.
The preparation (method two) of 2.4 test samples: on one experiment basis of method, removing petroleum ether decontamination, uses ammonia examination instead
Liquid washs decontamination.
Method two takes sinew-Soothing Blood-Quickening Tablet sample 10g, shreds, adds methanol 50ml, be heated to reflux 1h, let cool, filtration, filtrate
It is evaporated, water 20ml is added to make to dissolve, add water-saturated n-butanol shaking to extract 3 times, each 30ml, divide and take n-butanol liquid, use ammonia solution
Washing 2 times, each 200ml, water washing 2 times be saturated with n-butanol, each 20ml takes n-butanol liquid to be evaporated, and residue adds methanol
1ml makes to dissolve, as test solution.
2.5. point sample, expansion
Each 5ul of reference substance, control medicinal material, test solution is taken, is put respectively on same silica gel g thin-layer plate, with three chloromethanes
Alkane -10 DEG C of acetate-methanol-water (15:40:22:10) lower layer's solution arranged below is solvent, is unfolded, takes out, dries in the air
It is dry, it sprays with 10% ethanol solution of sulfuric acid, is heated to clear spot at 105 DEG C, sets examined under daylight and ultraviolet lamp (365nm) respectively
Depending on.
Rf value: Rg1 Rf1=7.5cm/12.2cm=0.615
Rb1 Rf2=2.2cm/12.2cm=0.180
2.6. preparation is extracted to sinew-Soothing Blood-Quickening Tablet Radix Notoginseng negative sample by method two and point sample is unfolded, as a result negative nothing
Interference, specificity are good.Therefore selected second method carries out ginsenoside Rg1 and ginseng soap to the Radix Notoginseng in sinew-Soothing Blood-Quickening Tablet
The identification of glycosides Rb1.
The identification of 2 rhizome of davallia of embodiment
1. having carried out the identification of medicinal material: determining the applicability of the presence or absence of target component and method in medicinal material
The preparation of 1.1 reference substances
Aurantiin reference substance is taken, adds methanol that solution of every 1ml containing 0.5mg is made, as reference substance solution.
1.2. the preparation of medicinal material
Aurantiin medicinal powder 0.5g is taken, methanol 30ml is added, is heated to reflux 1h, is filtered, filtrate is evaporated, and residue adds methanol
1ml makes to dissolve, as medicinal material solution.
1.3. point sample, expansion (method one)
Above-mentioned medicinal material solution and each 4ul of reference substance solution are drawn, is put respectively on same silica gel g thin-layer plate, with toluene-second
Acetoacetic ester-formic acid-water (1:12:0.5:3) upper solution is solvent, is unfolded, and takes out, dries, and is sprayed with alchlor test solution,
It sets and is inspected under ultraviolet lamp (365nm).
1.4. result: medicinal material spot separating effect is undesirable, considers the polarity of solvent and the progress in nature of lamellae
Adjustment.
1.5. point sample, expansion (method two):
Solvent ratio is adjusted on the basis of method one, and uses prefabricated board.
Above-mentioned medicinal material solution and each 4ul of reference substance solution are drawn, is put respectively on same silica G prefabricated board, band is decorateeed
Sample, using toluene-ethyl acetate-formic acid-water (1:12:2.5:3) upper solution as solvent, be unfolded, take out, dry, spray with
Alchlor test solution is set and is inspected under ultraviolet lamp (365nm).
1.6 results: on reference substance chromatography corresponding position in medicinal material chromatography, same color spot is shown.
2. having carried out the identification of the rhizome of davallia in sinew-Soothing Blood-Quickening Tablet: medicinal material method adjusted is directly applied in finished product.
2.1. the preparation of test sample
Sinew-Soothing Blood-Quickening Tablet sample 40g is taken, appropriate diatomite is added to disperse, adds chloroform ultrasound 2 times, 30 minutes every time, often
Secondary 150ml, filtration, discards filtrate, and the dregs of a decoction volatilize, and the dregs of a decoction add ethyl acetate 150ml ultrasound 30 minutes, and filtration, filtrate is evaporated, residual
Slag adds methanol 1ml to make to dissolve, as test solution.
2.2. point sample, expansion
Medicinal material solution, reference substance solution each 4ul, test solution 10ul are drawn, is put respectively in same silica G prefabricated board
On, band decorates sample, and using toluene-ethyl acetate-formic acid-water (1:12:2.5:3) upper solution as solvent, expansion is taken
Out, it dries, sprays with alchlor test solution, set and inspected under ultraviolet lamp (365nm).
Aurantiin Rf value: Rf=7.0cm/11.8cm=0.593
2.3. as a result, in test solution chromatography on reference substance chromatography corresponding position show same color spot, spot
Clearly.
The detection of 3 heavy metal of embodiment
1 instrument and reagent
1.1 instrument Shimadzu AA-6800 atomic absorption spectrophotometers, graphite furnace atomizer, autosampler, hollow yin
Pole lamp, pyrophzing test, hydrogenation generator, thermostatic control oscillator vibration, electric hot plate.
1.2 standard substance
Lead single element standard solution (national non-ferrous metal and material analysis test center GSB04-1742-2004)
Cadmium single element standard solution (national non-ferrous metal and material analysis test center GSB04-1721-2004)
Arsenic single element standard solution (national non-ferrous metal and material analysis test center GSB04-1714-2004)
Mercury single element standard solution (national non-ferrous metal and material analysis test center GSB04-1729-2004)
Copper single element standard solution (national non-ferrous metal and material analysis test center GSB04-1725-2004)
1.3 reagent
Hydrochloric acid is excellent pure grade (Tianjin Yao Hua chemical reagent Co., Ltd, lot number 20121020),
Nitric acid is excellent pure grade (Tianjin Fengchuan Chemical Reagent Science & Technology Co., Ltd., lot number 20130906),
Perchloric acid excellent pure grade (Tianjin moment Miao's chemical reagent development centre is supervised, lot number 20110212),
Sulfuric acid (Tianjin Yao Hua chemical reagent Co., Ltd, lot number 20070921),
Pepsin (Chinese Hui Shi biochemical reagents Co., Ltd, lot number 20120208),
Hydroxylamine hydrochloride (Shantou City's chemical reagent factory, lot number 20090627),
Ascorbic acid (chemical industry subsidiary factory, Tianjin Bazhou City, quartz Clock Factory, lot number 20120429).
1.5 sample
Sinew-Soothing Blood-Quickening Tablet (small honey pill): specification is every 10 ball weight 2.0g；20110402 lot number: 20110401,
20110403。
The research of 2 analysis methods
The preparation of 2.1 test solutions
2.1.1 lead in test sample, cadmium, arsenic, copper sample solution preparation method take test sample powder 0.1g, precision claims
It is fixed, it sets in kjeldahl flask, adds nitric acid-perchloric acid (4:1) mixed solution 10ml, mix, bottleneck adds a small funnel, soaked overnight.
It is placed on electric hot plate and heats resolution, continuous heating continues to be heated to smokeing clogging, until white cigarette dissipates to the clear and bright rear raising temperature of solution
To the greatest extent, digestion solution is in colorless and transparent or yellowish, lets cool, is transferred in 50ml measuring bottle, washs container, washing lotion with 2% nitric acid solution
Be incorporated in measuring bottle, and be diluted to scale, shake up to get.With method while reagent preparation blank solution.
2.1.2 the sample solution preparation method of mercury takes test sample powder 0.1g in test sample, accurately weighed, sets kelvin burning
In bottle, add nitric acid-perchloric acid (4:1) mixed solution 10ml, mix, bottleneck adds a small funnel, soaked overnight.It is placed in electric hot plate
On, in 120~140 heating resolutions, heating resolution lets cool until resolution completely, adds 20% sulfuric acid solution 10ml, 5% permanganic acid
Potassium solution 0.5ml, shakes up, and 5% hydroxylamine hydrochloride solution is added dropwise and just disappears to aubergine, is transferred in 50ml measuring bottle, appearance is washed with water
Device, washing lotion are incorporated in measuring bottle, are diluted with water to scale.With method while reagent preparation blank solution.
The preparation of 2.2 standard solution
2.2.1 the preparation precision measurement lead single element standard solution of lead standard solution is appropriate, is diluted with 2% nitric acid solution,
The solution of 1 μ g of every 1ml leaded (Pb) is made to get (0-5 DEG C of storage).
2.2.2 the preparation precision measurement lead single element standard solution of cadmium standard solution is appropriate, is diluted with 2% nitric acid solution,
Solution of every 1ml containing cadmium (Cd) 1 μ g is made to get (0-5 DEG C of storage).
2.2.3 the preparation precision measurement lead single element standard solution of arsenic standard solution is appropriate, is diluted with 2% nitric acid solution,
Solution of every 1ml containing arsenic (As) 1 μ g is made to get (0-5 DEG C of storage).
2.2.4 the preparation precision measurement lead single element standard solution of mercury standard solution is appropriate, is diluted with 2% nitric acid solution,
The solution of 1 μ g of every 1ml mercurous (Hg) is made to get (0-5 DEG C of storage).
2.2.51 the preparation precision measurement lead single element standard solution of copper standard solution is appropriate, is diluted with 2% nitric acid solution,
The solution of 10 μ g of every 1ml cupric (C μ) is made to get (0-5 DEG C of storage).
2.3 measuring method
It is surveyed according to " Chinese Pharmacopoeia " 2010 editions lead, cadmium, arsenic, mercury, copper measuring method (Ⅸ B atomic absorption spectrophotometry of annex)
It is fixed.
The test of 3 methodology validations
The drafting of 3.1 standard curves
Measuring method: according to " Chinese Pharmacopoeia " 2015 editions the 4th lead, cadmium, arsenic, mercury, copper measuring method (2321) measurement to get.
The drafting of 3.1 standard curves
The Specification Curve of Increasing result of 1 lead of table, cadmium, arsenic, mercury, copper
The investigation of 3.2 sample solution preparation methods
3.2.1 different preparation methods
One simulated gastric fluid method of method:
A) preparation of lead, arsenic, cadmium, copper test liquid takes test sample powder 0.5g, accurately weighed, addition simulated gastric fluid (by " in
State's pharmacopeia " 2010 years versions, one Ⅻ A of annex preparation) 100ml, in 37 DEG C water bath with thermostatic control mechanical shaking extraction 1 hour, be centrifuged (4000r
min-1) 1 hour, it takes supernatant 4ml to set in kjeldahl flask, adds nitric acid-perchloric acid (4:1) mixed solution 10ml, mix, bottleneck adds
One small funnel, soaked overnight.It is placed on electric hot plate and heats resolution, continuous heating continues to heat to the clear and bright rear raising temperature of solution
To smokeing clogging, until white cigarette disperses, digestion solution is in colorless and transparent or yellowish, lets cool, is transferred in 50ml measuring bottle, with 2% nitre
Acid solution wash container, washing lotion are incorporated in measuring bottle, and are diluted to scale, shake up to get.With method, reagent preparation blank is molten simultaneously
Liquid.
B) the test solution preparation of mercury takes test sample powder 0.5g, accurately weighed, and simulated gastric fluid is added and (presses " middle traditional Chinese medicines
Allusion quotation " 2010 years versions, one Ⅻ A of annex preparation) 100ml, in 37 DEG C water bath with thermostatic control mechanical shaking extraction 1 hour, be centrifuged (4000rmin-1) 1 hour, it takes supernatant 4ml to set in kjeldahl flask, adds nitric acid-perchloric acid (4:1) mixed solution 10ml, mix, bottleneck adds one
Small funnel, soaked overnight.It is placed on electric hot plate, in 120~140 heating resolutions, heating resolution lets cool, adds until resolution completely
20% sulfuric acid solution 10ml, 5% liquor potassic permanganate 0.5ml, shake up, and 5% hydroxylamine hydrochloride solution is added dropwise and just disappears to aubergine,
It is transferred in 50ml measuring bottle, container is washed with water, washing lotion is incorporated in measuring bottle, is diluted with water to scale.With method while reagent preparation
Blank solution.
Method 2 2010 editions " Chinese Pharmacopoeia " methods
A) preparation of lead, arsenic, cadmium, copper test liquid takes test sample powder 0.1g, accurately weighed, sets in kjeldahl flask, adds nitre
Acid-perchloric acid (4:1) mixed solution 10ml, mixes, and bottleneck adds a small funnel, soaked overnight.It is placed on electric hot plate and heats resolution,
Continuous heating to solution it is clear and bright after increase temperature, continue to be heated to smokeing clogging, until white cigarette disperses, digestion solution in colorless and transparent or
Yellowish is let cool, and is transferred in 50ml measuring bottle, washs container with 2% nitric acid solution, washing lotion is incorporated in measuring bottle, and is diluted to
Scale, shake up to get.With method while reagent preparation blank solution.
B) the test solution preparation of mercury takes test sample powder 0.1g, accurately weighed, sets in kjeldahl flask, adds nitric acid-height
Chloric acid (4:1) mixed solution 10ml, mixes, and bottleneck adds a small funnel, soaked overnight.It is placed on electric hot plate, adds in 120~140
Heat resolution, heating resolution let cool until resolution completely, add 20% sulfuric acid solution 10ml, 5% liquor potassic permanganate 0.5ml, shake
It is even, 5% hydroxylamine hydrochloride solution is added dropwise and just disappears to aubergine, is transferred in 50ml measuring bottle, container, the washing lotion amount of being incorporated in is washed with water
In bottle, it is diluted with water to scale.With method while reagent preparation blank solution.
2 distinct methods of table prepare the acid- soluble heavy metal content results of test liquid
| Constituent content | Method one | Method two |
| Lead | 10.13μg/g | 12.5μg/g |
| Cadmium | 0.58μg/g | 1.0μg/g |
| Arsenic | 4.52μg/g | 5.06μg/g |
| Mercury | It is not detected | 0.014μg/g |
| Copper | It is not detected | 11μg/g |
Conclusion: it is relatively large using official method heavy metal measured value in the test data measured from two methods, and
Five kinds of acid- soluble heavy metals can be measured, and method is first is that using simulated gastric fluid simulation human gastric juice environment, measured value is opposite
Less than two measured value of method, although the method more meets the absorbent degree of human body institute, the resolution sample used time is also shorter, for examination
Product solution is not sufficiently stable, and bacterium is easily bred after placement, the inspection being not easy in production real process, and cannot be by five kinds of huge sum of moneys
Belong to all detections, therefore, comprehensively consider, method for selecting two is preparation method of test article.
3.2.2 different sampling amounts
Sinew-Soothing Blood-Quickening Tablet finished product is crushed, 0.1g, 0.3g is weighed respectively and is prepared into confession according to preparation method of test article respectively
Test sample solution measures acid- soluble heavy metal content, and when measurement finds, when weighing sample 0.1g, can detecte out each element and contains
Amount, when weighing sample 0.3g, trap is big, and beyond detection limit, therefore sample volume 0.1g is convenient.
3.3 precision test
Accurate sample introduction lead, cadmium, arsenic, mercury, copper single element standard solution repeat respectively according to each element measuring method respectively
Sample introduction 6 times, each element measurement result see the table below.The result shows that precision is preferable.
3 Precision test result of table
The test of 3.4 negative samples
Referring to " Chinese Pharmacopoeia " versions in 2010 and pertinent literature, according to the composition of sinew-Soothing Blood-Quickening Tablet (small honey pill) at
Part, it is known that acid- soluble heavy metal is mainly derived from the native copper in raw material in finished product, and native copper is mineral drug, main component
For ferrous disulfide (FeS2), separately contain iron, gold, mercury, cadmium, arsenic, lead and other elements, therefore, it is living to be prepared for the relaxing muscles and tendons without native copper
Blood ball (small honey pill) negative sample, and it is prepared into test liquid, the assay of acid- soluble heavy metal is carried out, while to finished product
And 2 batches of nature copper raw materials are measured, and the results are shown in Table.The result shows that acid- soluble heavy metal in negative finished product (being free of native copper)
Content is very low or is not detected.
The negative finished acid soluble heavy metal content measurement result of table 4
| Element | Lead | Cadmium | Arsenic | Mercury | Copper |
| Negative sample | 1.277μg/g | It is not detected | It is not detected | It is not detected | It is not detected |
| Finished product | 12.46μg/g | 0.1μg/g | 5.06μg/g | 0.014μg/g | 11μg/g |
| Natural copper raw material 1 | 40.39μg/g | 2.32μg/g | 71.25μg/g | It is not detected | It is not detected |
| Natural copper raw material 2 | 47.56μg/g | 2.12μg/g | 68.17μg/g | It is not detected | It is not detected |
3.5 repetitive test
6 parts of same batch of sample is taken, it is accurately weighed respectively, it is molten that 6 parts of test samples are prepared by the preparation method of test solution
Liquid, sample introduction, measurement respectively, and content average value and RSD (%) are calculated, it the results are shown in Table.The result shows that repeatability is good.
5 repetitive test result of table
The test of 3.6 sample recovery rates
6 parts of sample of known content are weighed, single element standard solution are separately added into, by the preparation method system of test solution
For at 6 parts of test solutions, sample introduction, record is as a result, calculate the rate of recovery and RSD (%).It the results are shown in Table.The result shows that the rate of recovery is good
It is good.
6 recovery test result of table
The measurement of 4 samples
The measurement of 4.1 lab scale samples
According to the prescription ratio of sinew-Soothing Blood-Quickening Tablet, finished product sample 500g is prepared, is prepared by " sample solution preparation method "
At test liquid, the content of acid- soluble heavy metal lead, cadmium, arsenic, mercury, copper is measured, the results are shown in Table 7.
7 lab scale sample measurement result (PPM) of table
The measurement of test agent in 4.2 3 batches
Taking lot number respectively is 20110401,20110402,20110403 sinew-Soothing Blood-Quickening Tablet (small honey pill) finished product 0.1g, essence
It is close weighed, it is prepared into test liquid by " sample solution preparation method ", measurement acid- soluble heavy metal lead, cadmium, arsenic, mercury, copper contain
Amount, the results are shown in Table 8.
8 three batches of finished acid soluble heavy metal content measurement results (PPM) of table
The conclusion of 5 researchs
My company examines center according to State Bureau's medicine and sends out benefit requirement for the first time, has carried out the inspection of heavy metal, arsenic, as a result table
Bright content of beary metal is higher, it is proposed that wouldn't be included in quality standard；Second of center hair, which is examined, according to State Bureau's medicine mends requirement,
We carry out acid- soluble heavy metal (lead, cadmium, arsenic, mercury, copper) content in the sinew-Soothing Blood-Quickening Tablet (small honey pill) declared
Experimental study, the experimental results showed that, content of beary metal is relatively high in the product, and reason is to contain mine in this product prescription
Object medicine native copper passes through the detection to natural copper raw material, the negative sample without native copper and sinew-Soothing Blood-Quickening Tablet (small honey pill), card
Acid- soluble heavy metal in bright preparation is mainly derived from raw material native copper, and native copper is the main component in the prescription, rises
To the scattered stasis of blood, synthetism, the effect of analgesic, (the FeS containing ferrous disulfide based on main component2), according to another document report, gone back in native copper
The impurity such as leaded, arsenic, cadmium, copper, nickel, arsenic, antimony to cause to contain heavy metal in sinew-Soothing Blood-Quickening Tablet (small honey pill), but test knot
Fruit shows the usage and dosage according to the drug, and based on daily maximum dose 8g, the amount of acid- soluble heavy metal is less than daily
The limit regulation of human body intake.
Due in existing pharmacopeia to content of beary metal in Chinese patent drug without clearly stipulate that only with few kind (purple perilla dissipate, the coptis
Supernatant piece) in define content of beary metal, therefore, we combine test result, referring to " Chinese Pharmacopoeia " and pertinent literature, and root
The limit in harmful element limit standard recommended according to FAO/WHO (JECFA) and " food hygiene physical and chemical inspection standard manual " is wanted
It asks and the composite factors such as raw material batch difference considers, by the assay of acid- soluble heavy metal in sinew-Soothing Blood-Quickening Tablet (small honey pill)
25/1000000ths must not be crossed by being defined as lead；Cadmium must not cross hundred a ten thousandths；Arsenic must not cross 10/1000000ths；Mercury must not cross thousand
2/10000ths；Copper must not cross 25/1000000ths.And will test method, bound requirements are included in quality standard.
9 sinew-Soothing Blood-Quickening Tablet of table (small honey pill) acid- soluble heavy metal bound requirements contrast table
Note: 1, day intake limit is to calculate resulting numerical value by adult body weight 60kg；
2, sinew-Soothing Blood-Quickening Tablet day intake maximum is the instructions of taking maximum according to the product, takes 8g within the next few days and calculates institute
The numerical value obtained.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (10)
1. a kind of quality determining method of sinew-Soothing Blood-Quickening Tablet, it is characterised in that: including to Radix Notoginseng identification, to the identification of the rhizome of davallia
And the detection of heavy metal, wherein
The identification of Radix Notoginseng the following steps are included:
The preparation of test solution A: taking sinew-Soothing Blood-Quickening Tablet 7g, shreds plus methanol 50ml is heated to reflux 1 hour, let cool, filter, will
Filtrate is evaporated, and residue is transferred in separatory funnel, and first plus the shaking of water saturated n-butanol is extracted 3 times, each 30ml, merges just
Butanol liquid；It ammoniates test solution again to wash 2 times, each 20ml；Water washing 2 times be saturated again with n-butanol, each 20ml takes n-butanol
Liquid is evaporated, and residue adds methanol 1ml to make to dissolve, and obtains test solution A；
The preparation of reference substance solution A: taking ginsenoside Rb1, ginsenoside Rg1's reference substance, adds methanol that every 1ml is made and contains
The mixed solution of 0.5mg, as reference substance solution A；
It draws test solution A, each 5 μ l of reference substance solution A to be put respectively on same silica gel g thin-layer plate, with chloroform-acetic acid
Lower layer's solution of 10 DEG C of ethyl ester-methanol-water placements is solvent, is unfolded, colour developing；The chloroform, ethyl acetate, methanol with
And the weight ratio between water is 15:40:22:10；
The identification of the rhizome of davallia the following steps are included:
The preparation of test solution B: taking sinew-Soothing Blood-Quickening Tablet 30g, diatomite added to disperse, and adds chloroform 150ml ultrasonic extraction 2
It is secondary, it 30 minutes every time, lets cool, filters, filtrate is evaporated, filter residue is obtained, ethyl acetate 150ml is added into residue, it is 30 minutes ultrasonic,
Filtration, is evaporated filtrate, makes to dissolve to residue plus methanol 1ml, obtain test solution B；
The preparation of reference substance solution B: taking aurantiin reference substance, adds methanol that solution of every 1ml containing 0.5mg is made, it is molten to obtain reference substance
Liquid B；
4 μ l of reference substance solution B is drawn, 10 μ l of test solution B is put respectively on same silica G prefabricated sheets plate, ribbon
Point sample is unfolded using toluene-ethyl acetate-formic acid-water upper solution as solvent, colour developing；The toluene, ethyl acetate, first
Weight ratio between acid and water is 1:12:2.5:3；
The detection of heavy metal the following steps are included:
The preparation of test solution C: taking test sample powder 0.1g, set in kjeldahl flask, adds nitric acid-perchloric acid mixed solution
10ml is mixed, soaked overnight；Heating resolution, digestion solution is transferred in 50ml measuring bottle, washs kjeldahl flask with 2% nitric acid solution,
Washing lotion is incorporated in measuring bottle, and is diluted to scale, is shaken up to get test solution C；
The preparation of test solution D: taking test sample powder 0.1g, set in kjeldahl flask, adds nitric acid-perchloric acid mixed solution
10ml is mixed, soaked overnight；It is heated to clearing up completely, into digestion solution plus 20% sulfuric acid solution 10ml, 5% potassium permanganate are molten
Liquid 0.5ml, shakes up, and just disappears 5% hydroxylamine hydrochloride solution is added dropwise to aubergine, is transferred in 50ml measuring bottle, beginning is washed with water
Flask, washing lotion are incorporated in measuring bottle, are diluted with water to scale to get test solution D.
2. the quality determining method of sinew-Soothing Blood-Quickening Tablet according to claim 1, it is characterised in that: the identification step of Radix Notoginseng
In, further include the preparation of negative test solution A: by recipe quantity, removing other medicinal material powders outside Radix Notoginseng, mixes, add refined honey system
At Radix Notoginseng negative sample, negative control solution A is made by the preparation method of test solution A.
3. the quality determining method of sinew-Soothing Blood-Quickening Tablet according to claim 2, it is characterised in that: the identification step of Radix Notoginseng
In, after silica gel g thin-layer plate dries, sprays with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C.
4. the quality determining method of sinew-Soothing Blood-Quickening Tablet according to claim 1, it is characterised in that: the identification step of the rhizome of davallia
In, further include the preparation of negative test solution B: by recipe quantity, removing other medicinal material powders outside the rhizome of davallia, mixes, add refined honey
Rhizome of davallia negative sample is made, negative control solution B is made by the preparation method of test solution B.
5. the quality determining method of sinew-Soothing Blood-Quickening Tablet according to claim 4, it is characterised in that: the identification step of the rhizome of davallia
In, after silica gel g thin-layer plate dries, sprays with alchlor test solution, inspected under 365nm ultraviolet lamp.
6. the quality determining method of sinew-Soothing Blood-Quickening Tablet according to claim 1, it is characterised in that: heavy metal analysis step
In, further include the preparation of lead standard solution: precision measures lead single element standard solution, is diluted with 2% nitric acid solution, is made every
The solution of the leaded 1 μ g of 1ml.
7. the quality determining method of sinew-Soothing Blood-Quickening Tablet according to claim 1, it is characterised in that: heavy metal analysis step
In, further include the preparation of cadmium standard solution: precision measures cadmium single element standard solution, is diluted with 2% nitric acid solution, is made every
The solution of 1ml 1 μ g containing cadmium.
8. the quality determining method of sinew-Soothing Blood-Quickening Tablet according to claim 1, it is characterised in that: heavy metal analysis step
In, further include the preparation of arsenic standard solution: precision measures arsenic single element standard solution, is diluted with 2% nitric acid solution, is made every
The solution of 1ml 1 μ g containing arsenic.
9. the quality determining method of sinew-Soothing Blood-Quickening Tablet according to claim 1, it is characterised in that: heavy metal analysis step
In, further include the preparation of mercury standard solution: precision measures mercury single element standard solution, is diluted with 2% nitric acid solution, is made every
The solution of the mercurous 1 μ g of 1ml.
10. the quality determining method of sinew-Soothing Blood-Quickening Tablet according to claim 1, it is characterised in that: heavy metal analysis step
In, further include the preparation of copper standard solution: precision measures copper single element standard solution, is diluted with 2% nitric acid solution, is made every
The solution of 10 μ g of 1ml cupric.
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| CN102552616A (en) * | 2010-12-29 | 2012-07-11 | 江西济民可信集团有限公司 | Detection method for tablets capable of relaxing muscles and stimulating blood circulation |
| CN105203469A (en) * | 2015-10-21 | 2015-12-30 | 山西振东开元制药有限公司 | Detection method of medicament for treating femoral head necrosis |
| CN107064090A (en) * | 2017-04-18 | 2017-08-18 | 黑龙江葵花药业股份有限公司 | A kind of quality determining method of sinew-Soothing Blood-Quickening Tablet |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102552616A (en) * | 2010-12-29 | 2012-07-11 | 江西济民可信集团有限公司 | Detection method for tablets capable of relaxing muscles and stimulating blood circulation |
| CN105203469A (en) * | 2015-10-21 | 2015-12-30 | 山西振东开元制药有限公司 | Detection method of medicament for treating femoral head necrosis |
| CN107064090A (en) * | 2017-04-18 | 2017-08-18 | 黑龙江葵花药业股份有限公司 | A kind of quality determining method of sinew-Soothing Blood-Quickening Tablet |
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