CN106679812A - Color quantitative detection method for traditional Chinese medicine injection - Google Patents
Color quantitative detection method for traditional Chinese medicine injection Download PDFInfo
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- CN106679812A CN106679812A CN201710034735.XA CN201710034735A CN106679812A CN 106679812 A CN106679812 A CN 106679812A CN 201710034735 A CN201710034735 A CN 201710034735A CN 106679812 A CN106679812 A CN 106679812A
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- 238000002347 injection Methods 0.000 title claims abstract description 117
- 239000007924 injection Substances 0.000 title claims abstract description 117
- 239000003814 drug Substances 0.000 title claims abstract description 108
- 238000001514 detection method Methods 0.000 title claims abstract description 23
- 238000000862 absorption spectrum Methods 0.000 claims abstract description 119
- 239000000243 solution Substances 0.000 claims abstract description 14
- 239000013558 reference substance Substances 0.000 claims abstract description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 14
- 238000010790 dilution Methods 0.000 claims description 12
- 239000012895 dilution Substances 0.000 claims description 12
- 239000000284 extract Substances 0.000 claims description 12
- 238000011002 quantification Methods 0.000 claims description 10
- 150000001875 compounds Chemical class 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 5
- 239000003708 ampul Substances 0.000 claims description 2
- 235000013399 edible fruits Nutrition 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 238000010521 absorption reaction Methods 0.000 abstract description 26
- 238000000034 method Methods 0.000 abstract description 25
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- 230000008859 change Effects 0.000 description 36
- 239000009254 shuang-huang-lian Substances 0.000 description 24
- 239000007788 liquid Substances 0.000 description 14
- 230000000007 visual effect Effects 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 10
- 238000007689 inspection Methods 0.000 description 10
- 230000001429 stepping effect Effects 0.000 description 10
- 238000012360 testing method Methods 0.000 description 10
- 238000001228 spectrum Methods 0.000 description 9
- 235000013305 food Nutrition 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 239000002244 precipitate Substances 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 6
- 230000033228 biological regulation Effects 0.000 description 6
- 238000001647 drug administration Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 239000000178 monomer Substances 0.000 description 5
- 230000003595 spectral effect Effects 0.000 description 5
- 230000002708 enhancing effect Effects 0.000 description 4
- 239000004480 active ingredient Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 229920002521 macromolecule Polymers 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 239000002808 molecular sieve Substances 0.000 description 3
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 3
- 229940126678 chinese medicines Drugs 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000002845 discoloration Methods 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
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- 229920006395 saturated elastomer Polymers 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- UAJTZZNRJCKXJN-UHFFFAOYSA-M sodium;2-dodecoxy-2-oxoethanesulfonate Chemical compound [Na+].CCCCCCCCCCCCOC(=O)CS([O-])(=O)=O UAJTZZNRJCKXJN-UHFFFAOYSA-M 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 239000003390 Chinese drug Substances 0.000 description 1
- 206010034719 Personality change Diseases 0.000 description 1
- 239000008923 Qingkailing Substances 0.000 description 1
- DJSISFGPUUYILV-UHFFFAOYSA-N UNPD161792 Natural products O1C(C(O)=O)C(O)C(O)C(O)C1OC(C(=C1O)O)=CC2=C1C(=O)C=C(C=1C=CC(O)=CC=1)O2 DJSISFGPUUYILV-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/46—Measurement of colour; Colour measuring devices, e.g. colorimeters
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
Abstract
The invention discloses a color quantitative detection method for a traditional Chinese medicine injection. The method comprises a step 1) of preparing a solution from a traditional Chinese medicine injection, and starting with a step (2) directly if the injection is traditional Chinese medicine injecta; the step 2) of adding the traditional Chinese medicine injection solution or the traditional Chinese medicine injecta into a perforated plate or a cuvette which is then placed on a microplate reader or a spectrophotometer for the scanning of an absorption spectrum within a set wavelength range; and a step 3) of calculating the area under an absorption spectrum curve, comparing the area under the curve with the reference substance or a specified value, and judging the result. In the invention, color traits in the traditional Chinese medicine injection are accurately and objectively described, and a judgment can be made both by the area under the absorption spectrum curve of the whole ultraviolet-visible region and by the area under the absorption curve including the partial ultraviolet region. The invention provides an accurate method for inspecting the color traits of traditional Chinese medicine injections, and is particularly suitable for inspecting the color traits of the traditional Chinese medicine injections.
Description
Technical field
The present invention relates to Chinese medicine detection technique field, more particularly to a kind of Chinese medicine color quantification detection method.
Background technology
Chinese medicine results from earliest last century the forties, is started by scholars such as Qian Xinzhong, has been widely used in
It is clinical.Through the development of last 100 yearses, nearly 800 kinds are up to during peak, Jing markets and screening are eliminated, and have certification at present
About 142 kinds of the Chinese medicine that can be listed, are a unique ingredients of China's medical market.China was from 1977
Version《Pharmacopoeia of People's Republic of China》(referred to as《Pharmacopeia》) rise, Chinese medicine kind has just been included always.
Chinese medicine can lift drug effect, the curative effect of some Chinese medicines is preferably played, and send out in propulsion form of Chinese drug
Exhibition, expands the aspects such as the range of application of Chinese medicine, service numerous people health and performs meritorious deeds never to be obliterated really.From first Chinese medicine bavin
So far, Chinese medicine substantially experienced birth, development, rectification, the stage of raising for the invention of Hu injection.Technically come
Say, industry has generally solved the validity problem (although mechanism of action is not completely removed) of Chinese medicine, safety
Property is still an industry difficult problem.
With the extensive application of Chinese medicine, untoward reaction constantly finds, or even lethal untoward reaction also has repeatly report
Road.The explanations such as " the QINKAILING ZHUSHEYE event ", " SHUANGHUANGLIAN ZHUSHEYE event ", " the Herba Houttuyniae injectio event " that several years ago occur
The safety of Chinese medicine can not be ignored, by the industry even extensive concern of the public.At present Chinese medicine is in one
Keep under strict control the stage, State-level progressively starts Chinese medicine and revalues activity, many experts also are appealing energetically also to promote
Chinese medicine " is improved again ".
Chinese medicine presses the complexity of ingredient, can be divided into monomer component injection (such as breviscapine injection
Liquid), effective site injection (such as XUESAITONG ZHUSHEYE and injection of Radix Bupleuri), folk prescription injection (such as Radix Salviae Miltiorrhizae Injection) and compound recipe
Injection (QINKAILING ZHUSHEYE)；Wherein folk prescription injection and compound injection have the feature of Chinese medicine total extract, are both anti-
The representative dosage form of Chinese medicine medication, compatibility theory is reflected, is also the more serious dosage form of more problems.Existing data shows, Chinese medicine
The composition of injection is more complicated, and untoward reaction is also more complicated.For vein Chinese medicine, considerable safety sexual behavior is produced
Part be nearly all complicated component injection, such as Radix Et Caulis Acanthopanacis Senticosi injection, SHUANGHUANGLIAN ZHUSHEYE, Herba Houttuyniae injectio, almost
Have no the serious adverse reaction of monomer component injection.
In real work, also find really, the more complicated Chinese medicine of composition, color is often deeper, and folk prescription and
Often color is deeper for compound preparation, is brownish red or brown color etc., referring to table 1.Color trait is the important interior of Chinese medicine
Hold, but the description of various quality standards is all more macroscopical, it is less accurate.Even if certain formulations have the inspection for the character description
Content, is also that naked eyes comparison is carried out with standard color card or only with instrument Single wavelength inspection is carried out.Therefore, in the reality of quality standard
In execution, or coverage rate is not complete, or there is very big subjective error, this character description is caused to be held almost without substantial
Row power.
Color description (part) in the medicine injection quality standard of table 1
Therefore, the description of Chinese medicine color trait is always the short slab of Chinese medicine quality examination, particularly has
There are folk prescription, the compound preparation of total extract feature, it is also multifarious (being shown in Table 1) that its color trait is expressed in quality standard.
Safety issue has become at present the problem that Chinese medicine society widely pays close attention to, because the color trait of Chinese medicine is interior
In the concentrated expression of material, the quantitative description and objective quantitative inspection for paying attention to the character is conducive to lifting the inherence of Chinese medicine
Quality standard, monitors its inherent quality change.It was verified that the Chinese medicine color of expired failure is deeper, discoloration is (mainly
Color burn) it is also the important eometamorphism performance of Chinese medicine；But this performance is ignored always by academia and technos.
The content of the invention
For the deficiencies in the prior art, the invention provides a kind of Chinese medicine color quantification detection method.The present invention
Both can be judged using area under whole ultraviolet-visible area absorption spectrum curves, may also be employed special comprising part ultra-violet (UV) band
Determine area under the absorption curve of wavelength zone to be judged.The present invention provides accurately side for the inspection of Chinese medicine color trait
Method, is particularly well-suited to the folk prescription with total extract feature, herbal mixture injection color trait inspection.
Technical scheme is as follows：A kind of Chinese medicine color quantification detection method, it is characterised in that the inspection
Survey method is comprised the following steps：
Step 1) Chinese medicine is prepared into into solution, if Chinese medicine injection then directly carries out step (2)；
The Chinese medicine is the folk prescription with total extract feature or herbal mixture injection or powder ampoule agent for injection.
Step 2) Chinese medicine injection agent solution or Chinese medicine injection be added to into porous plate or cuvette；Be placed in microplate reader or
Absorption spectrum in spectrophotometer scanning setting wave-length coverage；In order to avoid saturated absorption, can be by Chinese medicine injection agent solution
Or Chinese medicine injection dilutes 1-128 times.
Described porous plate is the orifice plate of flat UV transparent 96 or 384 orifice plates.
The wave-length coverage simultaneously includes at least part ultra-violet (UV) band wavelength simultaneously for the part or all of wavelength of 200-800nm,
Stepping wavelength is 0.5-2nm.200-800nm includes the wave-length coverage of ultraviolet light and visible ray.Can be carried out according to experiment needs
Chosen wavelength range, such as selects the visible ray of 400-800nm, the ultraviolet light of 200-400nm or not only comprising ultraviolet light but also had included
The 200-600nm of visible ray is more preferable using the wave-length coverage Detection results comprising ultra-violet (UV) band.
Step 3) calculate area under absorption spectrum curve；Area under curve is compared with reference substance or setting, is entered
Row result judges.
As a result in judging, to the Chinese medicine being characterized with total extract, it is stipulated that the absorbing light in wave-length coverage is set a song to music
Area should be judged to undesirable more than reference substance or setting under line.Specific setting is regarded according to Chinese medicine kind
Situation determines.
As a result in judging, to the Chinese medicine being characterized with monomer component or active site, the main uptake zone of active component
Absorption spectrum curve under area should be judged to less than reference substance or setting it is undesirable；And the main uptake zone of non-active ingredient
Absorption spectrum curve under area should be judged to more than reference substance or setting it is undesirable.Setting is by according to Chinese medicine injection
Agent kind situation optionally determines.
《Pharmacopeia》In have regulation using Single wavelength colorimetric method detect Chinese medicine injection color method, due to Chinese medicine note
Penetrate agent ingredient complicated, often lack in visual field wavelength zone and there is distinctive absorbing wavelength, this detection is often compared
It is unilateral.《Pharmacopeia》Also the inspection method that useful standard color card naked eyes are compared, but the subjective error of naked eyes colour chart comparison is larger, as a result
Accuracy rate is not high.To solve the above problems, the present invention utilizes the principle of area under absorption curve, quantitative check Chinese medicine
Color.Chinese medicine is prepared into into the liquid of prescribed volume, is used directly for checking if being Chinese medicine injection.Inspection
When looking into, Chinese medicine injection agent solution is placed in into UV transparent container, in such as 96 orifice plates or cuvette, in 200-800nm extinction is scanned
Degree (when light Jing is 1cm) or optical density (porous plate is detected, when light Jing is not 1cm), the absorption calculated under solution 200-800nm is bent
Area under line.For the folk prescription with total extract feature or herbal mixture injection, color burn is reflection Chinese medicine note
The important indicator that agent stability declines, goes bad is penetrated, therefore can be compared with contrast solution or setting, work as absorption spectrum curve
Lower area can be determined that the color trait of the injection is undesirable when being more than certain value.For with monomer component or effective portion
For the Chinese medicine that position is characterized, when area is less than certain value under the main uptake zone absorption spectrum curve of active component, or
Under the absorption spectrum curve of non-active ingredient uptake zone area beyond certain value then can clearly judge color trait do not meet will
Ask.
Compared with prior art, the invention has the advantages that：The color knot that the method that the present invention is provided is detected
Fruit is more objective, more accurately, is particularly well-suited to Chinese medicine color trait inspection.Due to the method simplicity, also can conduct
The quick screening method of Chinese medicine inherent quality change.
It is contemplated that the present invention may be use with the color trait inspection of liquid preparation of Chinese medicine, after solid preparation of Chinese medicine dissolved dilution
Its color trait can be checked with the present invention.In view of the safety issue of Chinese medicine is more projected, it is the pin for strengthening the present invention
To property, therefore with Chinese medicine as main protection domain.
Description of the drawings
Fig. 1 removes macromole and the Chinese medicine rich in macromole.Q, S, D, Dz are respectively QINKAILING ZHUSHEYE, SHUANGHUANLIAN
Injection, Herba Erigerontis injection and Radix Salviae Miltiorrhizae Injection.1-3 represents respectively what is obtained Jing after the process of 3k, 10k and 30k molecular sieve
Remove macromole injection；4 is Chinese medicine stock solution；5-7 represents respectively the richness obtained Jing after the process of 3k, 10k and 30k molecular sieve
Injection containing macromole, macromolecular substances obtain 10 times of enrichments.
Fig. 2 QINKAILING ZHUSHEYE stock solutions are not toasted and the visible absorption spectra after 60 DEG C of bakings.Q0 is the absorption before baking
Spectrum, Q15 and Q30 are respectively the baking absorption spectrums of 15,30 days.
Fig. 3 QINKAILING ZHUSHEYE stock solutions do not toast (0D) and the visible suction after 60 DEG C of bakings 15 days (15D) and 30 days (30D)
Receive area (400-800nm) under the curve of spectrum.
Fig. 4 SHUANGHUANGLIAN ZHUSHEYE stock solutions are not toasted and the visible absorption spectra after 60 DEG C of bakings.S0 is the absorption before baking
Spectrum, S15 and S30 are respectively the baking absorption spectrums of 15,30 days.
Fig. 5 SHUANGHUANGLIAN ZHUSHEYE stock solutions do not toast (0D) and the visible suction after 60 DEG C of bakings 15 days (15D) and 30 days (30D)
Receive area (400-800nm) under the curve of spectrum.
Fig. 6 Radix Salviae Miltiorrhizae Injection stock solutions are not toasted and the visible absorption spectra after 60 DEG C of bakings.D0 is the absorbing light before baking
Spectrum, D15 and D30 is respectively the absorption spectrum for toasting 15 days and 30 days.
Fig. 7 Radix Salviae Miltiorrhizae Injection stock solutions do not toast (0D) and the visible absorbance after 60 DEG C of bakings 15 days (15D) and 30 days (30D)
Area (400-800nm) under the curve of spectrum.
Fig. 8 Herba Erigerontis injection stock solutions are not toasted and the visible absorption spectra after 60 DEG C of bakings.Dz0 is the suction before baking
Spectrum is received, Dz15 and Dz30 is respectively the absorption spectrum for toasting 15 days and 30 days.
Fig. 9 Herba Erigerontis injection stock solutions do not toast (0D) and 60 DEG C baking 15 days (15D) and 30 days (30D) after it is visible
Area (400-800nm) under absorption spectrum curve.
Figure 10 QINKAILING ZHUSHEYE stock solutions are not toasted and the uv-visible absorption spectra after 60 DEG C of bakings.Q0 is before baking
Absorption spectrum, Q15 be toast 15 days absorption spectrum (230-800nm, dilution 128 times afterwards detect).
Figure 11 QINKAILING ZHUSHEYE stock solutions do not toast (Q) and the uv-visible absorption spectra after 60 DEG C of bakings 15 days (Q15)
Area under curve (230-800nm, 128 times of dilution is detected afterwards).
Figure 12 SHUANGHUANGLIAN ZHUSHEYE stock solutions are not toasted and the uv-visible absorption spectra after 60 DEG C of bakings.S0 is before baking
Absorption spectrum, S15 be toast 15 days absorption spectrum (230-800nm, dilution 128 times afterwards detect).
Figure 13 SHUANGHUANGLIAN ZHUSHEYE stock solutions do not toast (S0) and the ultraviolet-ray visible absorbing light after 60 DEG C of bakings 15 days (S15)
Area under spectral curve (230-800nm, 128 times of dilution is detected afterwards).
Figure 14 Radix Salviae Miltiorrhizae Injection stock solutions are not toasted and the uv-visible absorption spectra after 60 DEG C of bakings.D0 is before baking
Absorption spectrum, D15 is the absorption spectrum for toasting 15 days (230-800nm, 128 times of dilution is detected afterwards).
Figure 15 Radix Salviae Miltiorrhizae Injection stock solutions do not toast (D0) and the uv-visible absorption spectra after 60 DEG C of bakings 15 days (D15)
Area under curve (230-800nm, 128 times of dilution is detected afterwards).
Figure 16 Herba Erigerontis injection stock solutions are not toasted and the uv-visible absorption spectra after 60 DEG C of bakings.Dz0 is baking
Front absorption spectrum, Dz15 is the absorption spectrum for toasting 15 days (230-800nm, 128 times of dilution is detected afterwards).
Figure 17 Herba Erigerontis injection stock solutions do not toast (Dz0) and the ultraviolet-visible after 60 DEG C of bakings 15 days (Dz15) is inhaled
Receive area under the curve of spectrum (230-800nm, 128 times of dilution is detected afterwards).
Specific embodiment
The color of the Chinese medicine of embodiment 1 is relevant with macromole.
QINKAILING ZHUSHEYE, specification 10ml, lot number 1003272；SHUANGHUANGLIAN ZHUSHEYE, specification 20ml, lot number 20100324；
Radix Salviae Miltiorrhizae Injection, specification 10ml, lot number 1005104；Herba Erigerontis injection, specification 10ml, lot number 20130533.Four kinds of injections
Liquid is buied from market, is produced by regular pharmaceutical factory, there is the authentication code of State Food and Drug Administration, meets pharmacopeia rule
It is fixed.
Above-mentioned four kinds of Chinese medicine injections are added separately in 3k, 10k, 30k sterile ultrafiltration centrifuge tube (specification 50ml), in
4 DEG C of 3000rpm centrifugation retentions prepare ultrafiltrate, obtain final product it is various remove macromole injection, while also obtaining rich in macromole injection
Liquid.It is 10 times of enrichments rich in macromole injection, that is, takes 10 parts of injection stock solutions and be prepared into 1 part rich in macromole liquid.In order to just
In comparing, each injection open country liquid is placed in 1.5ml centrifuge tubes is taken pictures.The sample preservation of acquisition is standby at -40 DEG C.
Obtain Jing after different molecular sieves retention it is different remove macromole injection, go macromole injection face with what 3k was retained
Color is shallower, and transparency is higher；Next to that 10k retention and 30k retain obtain remove macromole injection (see Fig. 1).To rich in big
For molecule injection, though have no precipitation color like soy sauce opaque (see Fig. 1).Show that macromolecular substances are the main of color
Source.
The QINKAILING ZHUSHEYE visible absorption spectra area under curve of embodiment 2 and color trait and stability change have
Close, but it is still insensitive.
QINKAILING ZHUSHEYE, specification 10ml, lot number 1003272, purchased from market, is produced by regular pharmaceutical factory, there is state food
The authentication code of Drug Administration, meets States Pharmacopoeia specifications.
QINKAILING ZHUSHEYE stock solution 10ml is taken, in being placed in 15ml sterile capped test tubes, in 60 DEG C of constant temperature ovens 30 days.Determine
400-800nm absorption spectrums (stepping wavelength is 1nm) after normal stock solution and baking 15 days and 30 days, take 50 μ l and add during measure
To in the orifice plate of flat bottom clear 384, it is scanned with Tecan microplate reader.Reflect sample face with the area under curve of absorption spectrum
The entire change of color, area under curve is calculated using trapezoidal method, i.e. be divided into area under curve according to wavelength interval many
It is individual trapezoidal, first calculate each trapezoidal area and then each trapezoidal area is added up.
In the thermally-stabilised experiment, there is enhancing and (do not toast, toast 15 days and 30 days in the absorption spectrum of QINKAILING ZHUSHEYE
Visible absorbance curve referring to Fig. 2), area is referring to Fig. 3 under absorption spectrum curve.Areal calculation under Jing absorption curves, QINGKAILING
Area under curve after injection is toasted 30 days substantially increases, increment rate about 24%, but the area under curve toasted 15 days increases
Rate very little (1.08%), is shown in Table 2.
According to quality criteria requirements, the color trait of QINKAILING ZHUSHEYE is brown color or brownish red.From naked eyes, no matter
It is unbaked stock solution or through 60 DEG C of bakings stock solution of 15 days or 30 days, the color of the injection still closes symbol regulation.
Obviously, QINKAILING ZHUSHEYE is toasted for 15 days or 30 days through 60 DEG C, and inherent quality should have occurred and that great change, or even baking
Also occur in that obvious dark precipitate within 15 days, illustrate that color trait has occurred that a certain degree of change, but still for brownish red, use
The method of macroscopic description can not be distinguished, and be then conducive to reflecting with visible absorption spectra area under curve, but susceptiveness is inclined
Low (contrasting with embodiment 6).
The experiment of the QINKAILING ZHUSHEYE of table 2, SHUANGHUANGLIAN ZHUSHEYE, Radix Salviae Miltiorrhizae Injection and Herba Erigerontis injection heat stability
Visible spectrum area under curve changes
The SHUANGHUANGLIAN ZHUSHEYE visible absorption spectra area under curve of embodiment 3 is relevant with color trait and stability, but not
It is enough sensitive.
SHUANGHUANGLIAN ZHUSHEYE, specification 20ml, lot number 20100324, purchased from market, is produced by regular pharmaceutical factory, there is state food
The authentication code of Drug Administration, meets States Pharmacopoeia specifications.
SHUANGHUANGLIAN ZHUSHEYE stock solution 10ml is taken, in being placed in 15ml sterile capped test tubes, in 60 DEG C of constant temperature ovens 30 days.Determine
400-800nm absorption spectrums (stepping wavelength is 1nm) after normal stock solution and baking 15 days and 30 days, take 50 μ l and add during measure
To in the orifice plate of flat bottom clear 384, it is scanned with Tecan microplate reader.Reflect sample face with the area under curve of absorption spectrum
The entire change of color, area under curve is calculated using trapezoidal method, i.e. be divided into area under curve according to wavelength interval many
It is individual trapezoidal, first calculate each trapezoidal area and then each trapezoidal area is added up.
In the thermally-stabilised experiment, the enhancing that the absorption spectrum of SHUANGHUANGLIAN ZHUSHEYE occurs weak degree (is not toasted, toasted
The visible absorbance curve of 15 days and 30 days is referring to Fig. 4), area is referring to Fig. 5 under absorption spectrum curve.Below Jing absorption spectrum curves
Product is calculated, and the area under curve of SHUANGHUANGLIAN ZHUSHEYE increases, and toasting increases about 8% for 15 days, and 30 days increment rates of baking are about 20%,
It is shown in Table 2.
According to quality criteria requirements, the color trait of SHUANGHUANGLIAN ZHUSHEYE is brownish red.From naked eyes, either without baking
Still through 60 DEG C of bakings stock solution of 15 days or 30 days, the color of SHUANGHUANGLIAN ZHUSHEYE still remains true qualities to roasting stock solution.It is aobvious
So, through the baking of 60 DEG C of 15 days and 30 days, inherent quality should have occurred and that great change to the injection, and be baked to 15 days
It can be seen that a small amount of dark precipitate, but liquid is still brownish red, therefore can not be distinguished with the method for macroscopic description, and with absorption curve
Area is conducive to reflecting that area merely add about 8% under color distinction, but 15 days absorption curves of baking.Although the method has progress
Meaning, but it is still not sensitive enough, because in real work, the scope of the 5-10% that fluctuates usually belongs to normal fluctuation range.
The Radix Salviae Miltiorrhizae Injection visible absorption spectra area under curve of embodiment 4 is relevant with color trait and stability, but not enough
It is sensitive.
Radix Salviae Miltiorrhizae Injection, specification 10ml, lot number 1005104, purchased from market, is produced by regular pharmaceutical factory, there is state food medicine
The authentication code of product Surveillance Authority, meets States Pharmacopoeia specifications.
Radix Salviae Miltiorrhizae Injection stock solution 10ml is taken, in being placed in 15ml sterile capped test tubes, in 60 DEG C of constant temperature ovens 30 days.Determine just
Often the 400-800nm absorption spectrums (stepping wavelength is 1nm) after stock solution and baking 15 days and 30 days, take 50 μ l and are added to during measure
In the orifice plate of flat bottom clear 384, it is scanned with Tecan microplate reader.Color sample is reflected with the area under curve of absorption spectrum
Entire change, area under curve calculated using trapezoidal method, will area under curve be divided into according to wavelength interval it is multiple
It is trapezoidal, first calculate each trapezoidal area and then each trapezoidal area is added up.
In the thermally-stabilised experiment, there is the increase of micro degree and (do not toast stock solution, dry in the absorption spectrum of Radix Salviae Miltiorrhizae Injection
Roasting 15 days and 30 days visible absorbance curve is referring to Fig. 6), area is referring to Fig. 7 under absorption spectrum curve.Area under Jing absorption curves
Calculate, area still increases under the absorption spectrum curve of Radix Salviae Miltiorrhizae Injection, but toast the increment rate of 15 days less than 10%, baking
The increment rate of 30 days is also about 11%, is shown in Table 2.
According to quality criteria requirements, the color trait of Radix Salviae Miltiorrhizae Injection should be brown to brownish red.From naked eyes, either
Still through 60 DEG C of bakings stock solution of 15 days or 30 days, the color of the injection still remains true qualities to unbaked stock solution.
Obviously, the injection is toasted for 15 days and 30 days through 60 DEG C, and inherent quality should have occurred and that change, and visible a small amount of is dark heavy
Form sediment, but liquid color is almost unchanged, is still brownish red, therefore can not be distinguished with the method for macroscopic description, and use absorption spectrum
Area under curve is conducive to reflecting the change that darkens, but under absorption curve area only to increase degree less.
The Herba Erigerontis injection visible absorption spectra area under curve of embodiment 5 has with color trait and stability change
Close, but it is not sensitive enough.
Herba Erigerontis injection, specification 10ml, lot number 20130533, purchased from market, is produced by regular pharmaceutical factory, there is national food
The authentication code of product Drug Administration, meets States Pharmacopoeia specifications.
Herba Erigerontis injection stock solution 10ml is taken, in being placed in 15ml sterile capped test tubes, in 60 DEG C of constant temperature ovens 30 days.Survey
400-800nm absorption spectrums (stepping wavelength is 1nm) after determining normal stock solution and toasting 15 days and 30 days, take 50 μ l and add during measure
Enter in the orifice plate of flat bottom clear 384, be scanned with Tecan microplate reader.Reflected with the area under curve of visible absorption spectra
The entire change of color sample, area under curve is calculated using trapezoidal method, will area under curve according to wavelength interval point
It is cut into multiple trapezoidal, first calculates and each trapezoidal area and then each trapezoidal area is added up.
In the thermally-stabilised experiment, the enhancing that the absorption spectrum of Herba Erigerontis injection occurs weak degree (does not toast original
Liquid, the baking visible absorbance curve of 15 days and 30 days are referring to Fig. 8), area is referring to Fig. 9 under absorption spectrum curve.Jing absorption spectrums
Area under curve is calculated, and the area under curve of the injection is toasted 15 days and increases about 2%, and toasting increases about 3% for 30 days, is shown in Table
2。
According to quality criteria requirements, the color trait of Herba Erigerontis injection is brown.From naked eyes, either without baking
Still through 60 DEG C of bakings stock solution of 30 days, the color of the injection still remains true qualities to roasting stock solution.Obviously, the injection
Toast within 15 days and 30 days through 60 DEG C, inherent quality should have occurred and that great change, also visible a small amount of dark precipitate, but face
Color change is not obvious, is still brown, can not be distinguished with the method for macroscopic description, and is conducive to instead with area under absorption spectrum curve
Reflecting area under the change, but absorption curve only increases less.Therefore, the method is still not sensitive enough, because in real work, fluctuation
The scope of 5-10% usually belongs to normal fluctuation range.
The QINKAILING ZHUSHEYE uv-visible absorption spectra area under curve of embodiment 6 can delicately reflect that color trait becomes
Change.
QINKAILING ZHUSHEYE, specification 10ml, lot number 1003272, purchased from market, is produced by regular pharmaceutical factory, there is state food
The authentication code of Drug Administration, meets States Pharmacopoeia specifications.
QINKAILING ZHUSHEYE stock solution 10ml is taken, in being placed in 15ml sterile capped test tubes, in 60 DEG C of constant temperature ovens 15 days.Determine
230-800nm absorption spectrums (stepping wavelength is 1nm) after normal stock solution and baking 15 days, are first diluted to 128 times again during measure
Take 160 μ l to be added in the orifice plate of flat UV transparent 96, be scanned with Tecan microplate reader.It is ultraviolet that absorption curve is calculated respectively
The area under curve in area (230-400nm), visual field (400-800nm) and ultraviolet-visible area (230-800nm).That is, by curve
Lower area is divided into multiple trapezoidal according to wavelength interval, first calculates and each trapezoidal area and then enters each trapezoidal area
Row is cumulative.
In the thermally-stabilised experiment, there is significantly enhancing and (do not toast and toast 15 in the absorption spectrum of QINKAILING ZHUSHEYE
It ultraviolet-ray visible absorbing curve is referring to Figure 10), area is referring to Figure 11 under absorption spectrum curve.Area meter under Jing absorption curves
Calculate, area increases under the ultra-violet (UV) band of QINKAILING ZHUSHEYE and visual field absorption spectrum curve, but below the absorption curve of ultra-violet (UV) band
Compared with visual field, substantially (ultra-violet (UV) band increases by 82%, and 76%) ultraviolet-visible area comprehensively increases, and is shown in Table 3 for product increase.
According to quality criteria requirements, the color trait of QINKAILING ZHUSHEYE is brown color or brownish red.From naked eyes, no matter
It is unbaked stock solution or through 60 DEG C of bakings stock solution of 15 days, the color of the injection still closes symbol regulation.Obviously, clearly
Open clever injection to toast within 15 days through 60 DEG C, inherent quality should have occurred and that great change, or even occur in that dark precipitate, and
Color trait has occurred that a certain degree of change, but liquid is still brownish red, can not be distinguished with the method for macroscopic description, uses
Visible absorption spectra area under curve is insensitive, and uses uv-visible absorption spectra area under curve then can delicately reflect
Out.Show that color trait change is evaluated with uv-visible absorption spectra area under curve has more the meaning that makes progress.
The QINKAILING ZHUSHEYE of table 3, SHUANGHUANGLIAN ZHUSHEYE, Radix Salviae Miltiorrhizae Injection and Herba Erigerontis injection 15 days stability realities of heat
The rate of change of area under visible, the ultraviolet and ultraviolet-visible area absorption spectrum curve tested.
The SHUANGHUANGLIAN ZHUSHEYE uv-visible absorption spectra area under curve of embodiment 7 can delicately reflect that color trait becomes
Change.
SHUANGHUANGLIAN ZHUSHEYE, specification 20ml, lot number 20100324, purchased from market, is produced by regular pharmaceutical factory, there is state food
The authentication code of Drug Administration, meets States Pharmacopoeia specifications.
SHUANGHUANGLIAN ZHUSHEYE stock solution 10ml is taken, in being placed in 15ml sterile capped test tubes, in 60 DEG C of constant temperature ovens 15 days.Determine
230-800nm absorption spectrums (stepping wavelength is 1nm) after normal stock solution and baking 15 days, are first diluted to 128 times and take during measure
160 μ l are added in the orifice plate of flat UV transparent 96, are scanned with Tecan microplate reader.With the area under curve of absorption spectrum
To reflect the entire change of color sample, ultra-violet (UV) band (230-400nm), visual field (400-800nm) and ultraviolet-can be calculated respectively
See area under area (230-800nm) absorption spectrum curve, i.e. area under curve is divided into according to wavelength interval it is multiple trapezoidal,
First calculate each trapezoidal area and then each trapezoidal area is added up.
In the thermally-stabilised experiment, there is a certain degree of increase in the ultraviolet and visible absorption spectra of SHUANGHUANGLIAN ZHUSHEYE
(not toasting the stock solution and baking ultraviolet-ray visible absorbing curve of 15 days referring to Figure 12), area is referring to Figure 13 under absorption spectrum curve.
The area under curve of areal calculation under Jing absorption spectrum curves, SHUANGHUANGLIAN ZHUSHEYE ultra-violet (UV) band and visual field absorption spectrum increases
Plus, but the value that visual field increases is less, and the value much greater (46%) that ultra-violet (UV) band increases, ultraviolet-visible area comprehensively increases
43%, it is shown in Table 3.
According to quality criteria requirements, the color trait of SHUANGHUANGLIAN ZHUSHEYE is brownish red.From naked eyes, either without baking
Still through 60 DEG C of bakings stock solution of 15 days, the color of SHUANGHUANGLIAN ZHUSHEYE still remains true qualities to roasting stock solution.Obviously, the note
Penetrate liquid to toast within 15 days through 60 DEG C, bottom a small amount of dark precipitate occurs, illustrates that inherent quality has occurred and that great change, but liquid
It is still brownish red, can not be distinguished with the method for macroscopic description, and uses visible absorption spectra area under curve delicately anti-
Mirror come, but with include ultra-violet (UV) band absorption spectrum curve under area can significantly reflect.Obviously, use comprising ultra-violet (UV) band
Area then more makes progress meaning evaluating color trait change under absorption curve.
The Radix Salviae Miltiorrhizae Injection uv-visible absorption spectra area under curve of embodiment 8 can delicately reflect that color trait changes.
Radix Salviae Miltiorrhizae Injection, specification 10ml, lot number 1005104, purchased from market, is produced by regular pharmaceutical factory, there is state food medicine
The authentication code of product Surveillance Authority, meets States Pharmacopoeia specifications.
Radix Salviae Miltiorrhizae Injection stock solution 10ml is taken, in being placed in 15ml sterile capped test tubes, in 60 DEG C of constant temperature ovens 15 days.Determine just
Often the 230-800nm absorption spectrums (stepping wavelength is 1nm) after stock solution and baking 15 days, first dilute 128 times and take 160 again during measure
μ l are added in the orifice plate of flat UV transparent 96, are scanned with Tecan microplate reader.Come anti-with the area under curve of absorption spectrum
The entire change of color sample is reflected, ultra-violet (UV) band (230-400nm), visual field (400-800) and ultraviolet-visible area (230- is calculated
800nm) area under absorption curve, i.e. area under curve is divided into according to wavelength interval it is multiple trapezoidal, first calculate each ladder
Then the area of shape is added up each trapezoidal area.
In the thermally-stabilised experiment, the absorption spectrum of Radix Salviae Miltiorrhizae Injection occurs substantially to increase (50%) in ultra-violet (UV) band, and
Visual field only occurs micro degree to be increased, and ultraviolet-visible area comprehensively increases by 37% and (is shown in Table 3, do not toast stock solution and toast 15 days
Ultraviolet-ray visible absorbing curve is referring to Figure 14), area is referring to Figure 15 under each absorption spectrum curve.
According to quality criteria requirements, the color trait of Radix Salviae Miltiorrhizae Injection should be brown to brownish red.From naked eyes, either
Still through 60 DEG C of bakings stock solution of 15 days, the color of the injection still remains true qualities to unbaked stock solution.Obviously, should
Injection is toasted for 15 days through 60 DEG C, and a small amount of dark precipitate occurs in bottom, but still for brownish red, can not with the method for macroscopic description
Difference, can not delicately be reflected with visible absorption spectra area under curve, and use include ultra-violet (UV) band absorption spectrum curve
Lower area can then reflect the change of color trait.Therefore, with comprising area under the absorption spectrum curve of ultra-violet (UV) band to check this in
The color trait of medicine injection has progressive meaning.
The Herba Erigerontis injection uv-visible absorption spectra area under curve of embodiment 9 can delicately reflect color trait
Change.
Herba Erigerontis injection, specification 10ml, lot number 20130533, purchased from market, is produced by regular pharmaceutical factory, there is national food
The authentication code of product Drug Administration, meets States Pharmacopoeia specifications.
Herba Erigerontis injection stock solution 10ml is taken, in being placed in 15ml sterile capped test tubes, in 60 DEG C of constant temperature ovens 15 days.Survey
230-800nm absorption spectrums (stepping wavelength is 1nm) after determining normal stock solution and toasting 15 days, first dilute 128 times again during measure
Take 160 μ l to be added in the orifice plate of flat UV transparent 96, be scanned with Tecan microplate reader.Below the curve of absorption spectrum
The long-pending entire change to reflect color sample, calculates ultra-violet (UV) band (230-400nm), visual field (400-800) and ultraviolet-visible area
(230-800nm) area under absorption curve, will area under curve be divided into according to wavelength interval multiple trapezoidal, first calculate every
Then individual trapezoidal area is added up each trapezoidal area.
In the thermally-stabilised experiment, there is a certain degree of increase in the uv-visible absorption spectra of Herba Erigerontis injection
(not toasting the stock solution and baking ultraviolet-ray visible absorbing curve of 15 days referring to Figure 16), area is referring to Figure 17 under absorption spectrum curve.
Increased substantially (42%) with area under the absorption spectrum curve of ultra-violet (UV) band, it is seen that area increases less, and ultraviolet-visible area comprehensively increases
Substantially (34%), is shown in Table 3.
According to quality criteria requirements, the color trait of Herba Erigerontis injection is brown.From naked eyes, either without baking
Still through 60 DEG C of bakings stock solution of 15 days, the color of the injection still remains true qualities to roasting stock solution.The injection is passed through
60 DEG C are toasted for 15 days, and a small amount of dark precipitate occurs in bottom, but liquid is still brown.Therefore, it is unable to area with the method for macroscopic description
Not, delicately reflected with visible absorption spectra area under curve is also bad, and use uv-visible absorption spectra area under curve then
Can significantly reflect, show the face that Chinese medicine is checked with area change under the absorption spectrum curve comprising ultra-violet (UV) band
Color character has progressive meaning.
Four kinds of Chinese medicine injections of the above are the more common Chinese medicines in market, two kinds be compound recipe, two kinds be folk prescription,
There is certain representativeness in total extract Chinese medicine.Exist or the new macromolecular substances for producing are Chinese medicine colors
Important sources, it is the important stability change performance of Chinese medicine to darken.According to reason, Chinese medicine should keep under strict control color
Character change.Find in the above-described embodiment, only QINKAILING ZHUSHEYE and SHUANGHUANGLIAN ZHUSHEYE have for 30 days in 60 DEG C of conditions
Visually distinguishable color burn, other two kinds little because of discoloration, i.e., sentence using visible absorption spectra area under curve is also bad
It is disconnected；But then can be judged well with ultraviolet or uv-visible absorption spectra area under curve.Without exception, this four
, in 60 DEG C of bakings 15 days, equal visible precipitate shows that inherent quality there occurs significant change for kind of Chinese medicine, but increase can
See that area is below 10% under absorption spectrum curve, and weighed with area under the absorption spectrum curve comprising ultra-violet (UV) band, increase
Plus rate is more than 30%.This is also illustrated, in the change of Chinese medicine color trait, the change of ultra-violet (UV) band is earlier than visible
Area, the detection comprising ultra-violet (UV) band is sensitiveer.
For big multiple instruments, the theoretical maximum of absorbance or optical density (OD) is 4, and the close value represents appearance
Saturated absorption.In order to be able to truly reflect spectral absorption, had better not occur more close 4 value in spectral absorption.Therefore, have
Necessity carries out appropriate dilution to detection object, and the solution after dilution should there are more than 1 but less than 3 in absorption spectrum
Value, is so conducive to improving the susceptiveness of detection.For the judgement of result, can be with absolute value or relative value.With absolute value, just
It is that area cannot be greater than certain value under the absorption spectrum curve in regulation testing sample certain wave-length coverage, or cannot be greater than control
The value of sample.With relative value, exactly specify that area must not under the absorption spectrum curve in testing sample certain wave-length coverage
It is higher by certain ratio.
For the Chinese medicine with total extract feature, area and visual field under general ultra-violet (UV) band absorption spectrum curve
With preferable dependency.But, if only with area under visual field absorption spectrum curve, the susceptiveness of detection is then poor,
Definitely value changes are little, also little with respect to value changes, cause the bad judgement of result.In order to allow area under absorption curve have it is bigger should
With scope, present invention performing creative labour discovery, detection can be greatly improved if spectrum range includes ultra-violet (UV) band sensitive
Property, or even area also can largely improve the spirit that relative value and absolute value are passed judgment under the absorption spectrum curve of alone ultra-violet (UV) band
Sensitivity, and use the area under curve of uv-visible absorption spectra then can further improve the sensitivity that absolute value is passed judgment on.
The instrument of detection uv-visible absorption spectra is laboratory conventional instrument, widely general in laboratory in addition to microplate reader
And domestic 75-2 spectrophotometers etc. can also conveniently realize detection.Calculate uv-visible absorption spectra area under curve
Easily realize, during spectral scan, if with 1nm steppings, by each absorption value (when light Jing is 1cm) or OD values (when light Jing not
For 1cm when, as detected with porous plate) add up, it is area under absorption spectrum curve that can regard as.
It is reflection Chinese medicine due to darkening for the folk prescription with total extract feature or herbal mixture injection
The general character phenomenon of injection stable problem, but the degree of color burn may be different because of different Chinese medicine kinds.For
Specific Chinese medicine injection agent solution, can specify that area is certain value under the absorption spectrum curve in particular range of wavelengths, greatly
Then can be determined that as against regulation in the value.
Except for the folk prescription or herbal mixture injection of total extract feature, for being characterized with monomer or effective site
Chinese medicine for, absorption spectrum change can also reflect the change of its inherent quality.The active component of this kind of Chinese medicine
It is relatively clear, mostly homologue, with certain general character spectral absorption, these regions then constitute the main uptake zone of active component, this
Absorption beyond a little regions is then non-active ingredient uptake zone.For specific Chinese medicine injection agent solution, can specify that a certain
Area is certain value under absorption spectrum curve in particular range of wavelengths, then be can be determined that as (phase against regulation more than the value
When in the absorption spectrum of monitoring foreign pigment)；Also could dictate that area under the absorption spectrum curve in a certain particular range of wavelengths
For certain value, then can be determined that as unqualified (equivalent to the absorption spectrum of monitoring active component) less than the value.When judging
Corresponding concrete scope can as needed be set.
Claims (6)
1. a kind of Chinese medicine color quantification detection method, it is characterised in that the detection method is comprised the following steps：
Step 1) Chinese medicine is prepared into into Chinese medicine injection agent solution, if Chinese medicine injection then directly carries out step (2)；
Step 2) Chinese medicine injection agent solution or Chinese medicine injection are added to into porous plate or cuvette, it is placed in microplate reader or light splitting
Absorption spectrum in photometer scanning setting wave-length coverage；
Step 3) area under absorption spectrum curve is calculated, area under curve is compared with reference substance or setting, tied
Fruit judges.
2. Chinese medicine color quantification detection method as claimed in claim 1, it is characterised in that step 1) the Chinese medicine note
Agent is penetrated for cmm of simple prescription or the Chinese medicine injection or powder ampoule agent for injection of the preparation of compound recipe total extract.
3. Chinese medicine color quantification detection method as claimed in claim 1, it is characterised in that step 2) described in porous
Plate is the orifice plate of flat UV transparent 96 or 384 orifice plates.
4. Chinese medicine color quantification detection method as claimed in claim 1, it is characterised in that step 2) described in Chinese medicine
Injection agent solution or Chinese medicine injection carry out 1-128 times of dilution.
5. Chinese medicine color quantification detection method as claimed in claim 1, it is characterised in that step 2) the wavelength model
Enclose the part or all of wavelength for 200-800nm and include at least part ultra-violet (UV) band wavelength simultaneously, stepping wavelength is 0.5-2nm.
6. Chinese medicine color quantification detection method as claimed in claim 1, it is characterised in that step 3) result judges
In, to the Chinese medicine being characterized with total extract, it is stipulated that area is more than control under the absorption spectrum curve in wave-length coverage
Product or setting should be judged to undesirable.
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Cited By (1)
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| CN112525760A (en) * | 2020-11-24 | 2021-03-19 | 厦门市健康医疗大数据中心(厦门市医药研究所) | Method for digitally and quantitatively evaluating moisture absorption strength of substance |
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| CN101126713A (en) * | 2007-10-12 | 2008-02-20 | 王承华 | Bipolar ultraviolet spectrum identification method |
| CN101791331A (en) * | 2010-03-16 | 2010-08-04 | 浙江大学 | Method for rapid determination of tannin content in Slvia Miltiorrhiza Bunge extracting solution |
| CN102128796A (en) * | 2010-12-07 | 2011-07-20 | 桂林理工大学 | Method for measuring gentamicin sulfate |
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| JPS6125059A (en) * | 1984-07-14 | 1986-02-03 | Shimadzu Corp | Method and apparatus for analyzing saccharide |
| CN101126713A (en) * | 2007-10-12 | 2008-02-20 | 王承华 | Bipolar ultraviolet spectrum identification method |
| CN101791331A (en) * | 2010-03-16 | 2010-08-04 | 浙江大学 | Method for rapid determination of tannin content in Slvia Miltiorrhiza Bunge extracting solution |
| CN102128796A (en) * | 2010-12-07 | 2011-07-20 | 桂林理工大学 | Method for measuring gentamicin sulfate |
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| CN112525760A (en) * | 2020-11-24 | 2021-03-19 | 厦门市健康医疗大数据中心(厦门市医药研究所) | Method for digitally and quantitatively evaluating moisture absorption strength of substance |
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Application publication date: 20170517 |