CN107643345A - A kind of method of quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract - Google Patents
A kind of method of quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract Download PDFInfo
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Abstract
The present invention relates to a kind of Rhizoma Gastrodiae and its method for quality control of medicine materical crude slice and extract, the present invention is based on high-efficient liquid phase chromatogram technology, using the mark analysis methods surveyed more, pass through relative correction factor, Gastrodin in Gastrodia eleta Bl., p-Hydroxybenzylalcohol, the gloomy glycosides A of Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E6 kinds Gastrodin constituents content of Bali are calculated, establish with cheap, the stable Gastrodin of property is internal standard compound, while determines the multi objective method of quality control of 6 kinds of active ingredient in rhizoma Gastrodiae.This method detection sensitivity is high, analyze speed is fast, stability is good, can with it is objective, comprehensively and accurately evaluate Rhizoma Gastrodiae and its quality of medicine materical crude slice and extract, can solve the problems, such as the factors such as the gloomy methods of glycosides reference substance of Bali is unstable, price is high and Rhizoma Gastrodiae and its medicine materical crude slice and extract quality can not be controlled comprehensively, to ensureing that rhizoma Gastrodiae curative effect is significant.
Description
Technical field
The present invention relates to quality control method, and in particular to the quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract
Method.
Background technology
Rhizoma Gastrodiae is orchid rhizoma Gastrodiae Gastrodia elata BL. dry tuber, has suppressing liver-YANG, dispelling wind and removing obstruction in the meridians
The effect of.At present it is generally acknowledged that Gastrodin is the important active component of rhizoma Gastrodiae.Research in recent years find rhizoma Gastrodiae also have increase intelligence,
The neuroprotections such as brain tonic, there is the effect of certain to senile dementia, modern pharmacological research shows, the gloomy glycosides of the Bali in rhizoma Gastrodiae
C, the composition such as gloomy glycosides B of Bali learning memory disorder etc. to caused by learning memory disorder caused by hyoscine and vascular dementia
With significant improvement result, there is larger researching value and potentiality to be exploited.
One mark methods of surveying are using a certain typical component in sample as internal standard more, are established relative between the component and other components
Correction factor, the content of other components is calculated by relative correction factor, a mark surveys method to be successfully applied to Chinese medicine multicomponent
The quality control of multi objective, high testing cost and cumbersome detection can be avoided to operate.The quality control of rhizoma Gastrodiae at present it is more with
Gastrodin, p-Hydroxybenzylalcohol are index, carry out assay using external standard method, Con trolling index is relatively easy, has no adopt at present
With the mark reports for surveying 6 kinds of active constituent contents such as methods measure Gastrodin in Gastrodia eleta Bl., the gloomy glycosides A of Bali, the present invention solves bar more
The factors such as the gloomy methods of glycosides reference substance of profit is unstable, price is high and Rhizoma Gastrodiae and its medicine materical crude slice and extraction of substance can not be controlled comprehensively
The problem of amount, to control rhizoma Gastrodiae quality and ensure that curative effect is significant.
CN105277642 A disclose method that is a kind of while determining a variety of phenols component contents in rhizoma Gastrodiae, and this method is adopted
A variety of phenols component contents such as Gastrodin in Gastrodia eleta Bl., p-Hydroxybenzylalcohol, parishin B are determined with fluorescence detection, with this
Invention is compared, and it is primarily present following weak point：1. carrying out assay using external standard method, each reference substance quantity consumed is big,
Testing cost is high.2. being detected using fluorescence detector, its popularization, anti-interference are poor, are carried out using UV-detector
Detection, the reappearance and accuracy of testing result are more preferable.
The content of the invention
Present invention aim to address the deficiency of existing rhizoma Gastrodiae Quality Control Technology, using high-efficient liquid phase chromatogram technology, leads to
Cross a survey and comment method more, with cheap, the stable Gastrodin of property is internal standard compound, establishes itself and p-Hydroxybenzylalcohol, Bali
The gloomy glycosides B of gloomy glycosides A, Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali relative correction factor, and the method for calculating each component content.We
Method detection sensitivity is high, and stability is good, can with it is objective, comprehensively and accurately evaluate Rhizoma Gastrodiae and its matter of medicine materical crude slice and extract
Amount, can solve the factors such as the gloomy methods of glycosides reference substance of rhizoma Gastrodiae Bali is unstable, price is high and can not control comprehensively Rhizoma Gastrodiae and
The problem of its medicine materical crude slice and extract quality, to control rhizoma Gastrodiae quality and ensure that curative effect is significant.
Based on this, to achieve the above object, the technical solution adopted by the present invention：
A kind of method of quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract, is mainly included the following steps that：
（1）Prepare rhizoma Gastrodiae need testing solution；
（2）It is single right to prepare the gloomy glycosides A of Gastrodin, p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali
According to product solution and mixed reference substance solution；
（3）Aspiration step respectively（2）Described single reference substance solution and mixed reference substance solution, inject high performance liquid chromatograph
In tested, according to formula Rm/k=tR(m)/tR(k)（TR (m) is other component m retention time in formula, and tR (k) is internal standard
The retention time of thing）, calculate p-Hydroxybenzylalcohol, the gloomy glycosides A of Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali relative to
The relative retention time of internal standard compound Gastrodin；
（4）Aspiration step（2）Described mixed reference substance solution, the μ L of sample introduction 1,2,4,6,8,12, inject high performance liquid chromatography
Instrument, measure, records each Component peak area, according to relative correction factor calculation formula fkm=fk/fm=(Wk×Am)/(Wm×Ak) (formula
Middle AkFor internal standard compound peak area, WkFor internal standard compound quality；AmFor other component m peak areas, WmFor other component m mass), with day
Numb element is internal reference thing, calculates the relative correction factor f of other 5 kinds of compositionskm；
（5）Aspiration step respectively（1）Described need testing solution and step（2）Described Gastrodin reference substance, injects efficient liquid
Chromatography, measure, the gloomy glycosides A of p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali pass through step（3）
Described relative retention time progress is qualitative, passes through step（4）Described relative correction factor calculates respective content.
Preferably, the chromatographic condition of high performance liquid chromatograph described above is：Using C18 chromatographic columns, mobile phase：
Acetonitrile (the A)-aqueous acid of volume fraction 0.02~0.1% (B), using gradient elution：0 min~5 min, 2%A；5 min~20
Min, 2%~30%A, 20 min~25 min, 30%~2%A；Detection wavelength：220 nm；Volume flow：0.8~1.2mL
min-1；Sample size is 5~25 μ L；Column temperature：20~35 DEG C；As more preferably scheme, mobile phase described above is acetonitrile-sour water
Solution.Acid is preferably phosphoric acid, and its volume fraction is 0.05%；Volume flow is 1.0 mL min-1, sample size is 10 μ L, and column temperature is
25℃。
Physicochemical property of the invention according to Rhizoma Gastrodiae and its medicine materical crude slice and each composition in extract, due to Gastrodin class chemical combination
Thing polarity approaches, on a column more difficult separation, and the present invention passes through the composition of substantial amounts of experiment screening mobile phase, experimental result
Show, well can be separated 6 kinds of Gastrodin constituents with other materials using currently preferred gradient elution program, and peak
Type is good, and analysis time is short.
Preferably, above step（3）The gloomy glycosides A of the p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides of Bali
C, the gloomy glycosides E of Bali is respectively relative to the relative retention time of internal standard compound Gastrodin：1.3480、1.8173、1.684 6、
1.7286、1.6443。
Preferably, above step（4）The gloomy glycosides A of the p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides of Bali
C, the relative correction factor of the gloomy glycosides E of Bali and Gastrodin is respectively：0.6632nd, 1.2777,1.4351,1.3885,1.5907.
Preferably, above step（1）The preparation method of the rhizoma Gastrodiae need testing solution is：Precision weighs a day anaesthetic
Material, Processed Product of Rhizoma Gastrodiae or Rhizoma Gastrodiae extract, 15~25 times of volume fractions are added as 60%~80% ethanol, heating and refluxing extraction 40~
60min, filtration, 5 times of amount subsequent filtrates are taken, be concentrated near do and be dissolved in water without alcohol taste, residue, be transferred in 25 mL measuring bottles, use water
Scale is diluted to, is shaken up, is filtered, is taken subsequent filtrate to cross 0.22 μm of filter membrane, obtain need testing solution.
Preferably, above step（2）The gloomy glycosides A of the Gastrodin, p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali,
The gloomy glycosides C of Bali, the single reference substance solutions of the gloomy glycosides E of Bali and mixed reference substance solution, solvent are water, and the mass concentration of each material is
0.01~0.08mg/mL.
Beneficial effect
Compared with prior art, the present invention relates to a kind of method of the quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract to have
Advantages below and marked improvement：The present invention filters out the colors such as optimal flowing phase composition, elution program, flow velocity by many experiments
Spectral condition, and show by many experiments checking.
The present invention is directed to the deficiency of existing rhizoma Gastrodiae Quality Control Technology, more by a mark using high-efficient liquid phase chromatogram technology
Survey method, with cheap, the stable Gastrodin of property is internal standard compound, establish its with p-Hydroxybenzylalcohol, the gloomy glycosides A of Bali, bar
The gloomy glycosides B of profit, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali relative correction factor, and the method for calculating each component content.This method detects
High sensitivity, analyze speed is fast, and stability is good, can be with objective, comprehensively and accurately evaluation Rhizoma Gastrodiae and its medicine materical crude slice and extraction
The quality of thing, can solve the factors such as the gloomy methods of glycosides reference substance of rhizoma Gastrodiae Bali is unstable, price is high and rhizoma Gastrodiae can not be controlled comprehensively
The problem of medicinal material and its medicine materical crude slice and extract quality, to control rhizoma Gastrodiae quality and ensure that curative effect is significant.
Brief description of the drawings：
Fig. 1 is that reference substance (A) and test sample (B) HPLC scheme, and wherein chromatographic peak 1 is Gastrodin, 2 is p-Hydroxybenzylalcohol, 3 is
The gloomy glycosides E of Bali, 4 be the gloomy glycosides B of Bali, 5 be the gloomy glycosides C of Bali, 6 be the gloomy glycosides A of Bali.
Embodiment
Below in conjunction with quality control of the specific embodiment to a kind of Rhizoma Gastrodiae and its medicine materical crude slice and extract of the present invention
Method is described further the present invention.
1 material
The type high performance liquid chromatographs of Aglient 1260 (An Jie Agilent Co., Ltds)；The a ten thousandth day of AB135-S types ten
It is flat（Switzerland's plum Teller-support benefit Instrument Ltd.）；EP214C type a ten thousandth electronic balances（Switzerland's plum Teller-support benefit
Instrument Ltd.）；KQ-250 type ultrasonic cleaners（Kunshan Ultrasonic Instruments Co., Ltd.）.
Gastrodin reference substance（110807-201306）Purchased from Nat'l Pharmaceutical ＆ Biological Products Control Institute；Bali gloomy glycosides A, B, C, E
（Purity>98%）, purchased from Chengdu Man Site Bioisystech Co., Ltd；Acetonitrile, phosphoric acid are chromatographically pure, and water is purified water, other examinations
Agent is that analysis is pure.
15 batch rhizoma Gastrodiaes are collected from all parts of the country, orchid is accredited as through pharmaceutical college of Shandong Traditional Chinese Medicine University professor Li Feng
Rhizoma Gastrodiae Gastrodia elata BL. stem tuber.
2 methods and result
2.1 liquid phase chromatogram condition
Chromatographic column：Agilent Zorbax SB-C18 posts（4.6 mm × 250 mm, 5 μm）；Mobile phase：Acetonitrile (A)-
0.05% phosphate aqueous solution (B), using gradient elution：0 min~5 min, 2%A；5 min~20 min, 2%~30%A, 20
Min~25 min, 30%~2%A；Detection wavelength：220 nm；Volume flow：1.0 mL·min-1；Column temperature：25 ℃；Sample size
10 μL.On this condition, 6 kinds of chemical compositions and the separating degree of adjacent chromatographic peak are all higher than 1.5 in rhizoma Gastrodiae, see Fig. 1.
The preparation of 2.2 need testing solutions
Tall Gastrodis Tuber is taken, takes about 2.0 g, it is accurately weighed, put in conical flask with cover, accurate addition 70% ethanol 50 mL is weighed heavy
Amount, the min of heating and refluxing extraction 40, lets cool, then weighed weight, the weight of less loss is supplied with 70% ethanol, filtration, precision measures continuous
The mL of filtrate 10, it is concentrated near do and adds aqueous to dissolve without alcohol taste, residue, be transferred in 25 mL measuring bottles, be diluted with water to scale, shake
It is even, filtration, take subsequent filtrate to cross 0.22 μm of filter membrane, produce.
The preparation of 2.3 reference substance solutions
Mixed reference substance solution：Take the gloomy glycosides A of Gastrodin, p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, Bali gloomy
Glycosides E reference substances are appropriate, accurately weighed, add aqueous to dissolve and every 1 mL is made containing the mg of Gastrodin 0.051 5, p-Hydroxybenzylalcohol
The gloomy mg of glycosides A 0.043 8 of 0.032 3 mg, Bali, the gloomy mg of glycosides B 0.043 8 of Bali, the gloomy mg of glycosides C 0.054 6 of Bali, bar
The gloomy mg of glycosides E 0.058 6 of profit mixed solution, is produced.
Single reference substance solution：The gloomy glycosides A of Gastrodin, p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides of Bali are taken respectively
C, the gloomy glycosides E reference substances of Bali are appropriate, accurately weighed, add aqueous dissolving that every 1 mL is respectively prepared containing the mg of Gastrodin 0.128 8, right
The gloomy mg of glycosides A 0.087 6 of the mg of salicylic alcohol 0.065 2, Bali, the gloomy mg of glycosides B 0.087 6 of Bali, the gloomy glycosides C0.109 of Bali
The gloomy mg of glycosides E0.117 2 of 2 mg, Bali single solution, is produced.
3 Method validations
3.1 linear relationships are investigated
The single reference substance solution under " 2.3 " item is taken, series concentration is configured to aqueous dilution, according to 2.1 lower chromatographic conditions
It is measured.With Gastrodin, p-Hydroxybenzylalcohol, the gloomy glycosides A of Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E sample sizes of Bali
（μg）For abscissa（X）, corresponding peak area is ordinate（Y）, standard curve is drawn respectively, obtains regression equation, the results are shown in Table 1.
The regression equation and the range of linearity, coefficient correlation of 6 kinds of chemical compositions in the rhizoma Gastrodiae of table 1
3.2 precision test
Precision draws 2.3 lower mixed reference substance solutions, is measured by 2.1 lower chromatographic conditions, replication 6 times, sample introduction
Measure 10 μ L.As a result the gloomy glycosides A of Gastrodin, p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali peak face
Long-pending RSD is respectively 0.79%, 1.18%, 2.10%, 1.10%, 1.29%, 0.14%, shows that instrument precision is good.
3.3 stability test
0,2,4,8,12,24 hs of the need testing solution under 2.2 after preparation are taken to determine Gastrodin, para hydroxybenzene first
The gloomy glycosides A of alcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali peak area, the μ L of sample size 10.As a result corresponding 7 kinds into
The RSD of swarming area is respectively 1.75%, 0.81%, 1.07,1.22%, 0.81%, 1.07,1.22%, shows that need testing solution exists
There is good stability in 24h.
3.4 replica test
Tall Gastrodis Tuber is taken, totally 6 parts, need testing solution is obtained by 2.2 lower section legal systems respectively, is surveyed by 2.1 lower chromatographic conditions
It is fixed, the μ L of sample size 10, peak area is recorded, it is gloomy to calculate Gastrodin, p-Hydroxybenzylalcohol, the gloomy glycosides A of Bali, the gloomy glycosides B of Bali, Bali
The gloomy glycosides E of glycosides C, Bali content, as a result the RSD of 7 kinds of component contents be respectively 1.09%, 1.18%, 1.21%, 1.14%, 2.28%,
0.88%, show that this method repeatability is good.
3.5 sample-adding recovery tests
The Tall Gastrodis Tuber of known content is taken, totally 6 parts, respectively about 1 g, accurately weighed, accurate respectively to add Gastrodin reference substance solution
（0.660 4 mg•mL-1）, citric acid reference substance solution（3.014 2 mg•mL-1）, p-Hydroxybenzylalcohol reference substance solution
（0.210 4mg•mL-1）, the gloomy glycosides A reference substance solutions of Bali（2. 125 0mg•mL-1）, the gloomy glycosides B reference substance solutions of Bali（1.
042 mg•mL-1）, the gloomy glycosides C reference substance solutions of Bali（0. 102 5 mg•mL-1）, the gloomy glycosides E reference substance solutions of Bali（0. 102
5 mg•mL-1）Each 5 mL, then accurate addition each 15 mL of 70% ethanol, weighed weight, remaining is made by 2.2 lower methods operations
Need testing solution, it is measured according to 2.1 lower chromatographic conditions using criterion keying method, the μ L of sample size 10, calculates sample-adding recovery
Rate.The gloomy glycosides A of Gastrodin, p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali average recovery rate（n=
6）Respectively 100.59%, 101.31%, 101.87%, 100.04%, 99.88%, 99.63%, RSD be respectively 1.66%, 2.30%,
1.36%、1.93%、2.07%、1.80%.As a result show that this method has the preferable rate of recovery.
The measure of 4 relative correction factors
The measure of 4.1 composition relative correction factors to be measured
Precision draws 2.3 lower mixed reference substance solutions, respectively the μ L of sample introduction 1,2,4,6,8,12, parallel determination 2 times, records peak
Area, according to relative correction factor calculation formula fkm=fk/fm=(Wk × Am)/(Wm × Ak), (Ak is internal standard compound peak face in formula
Product, Wk is internal standard compound quality；Am is other component m peak areas, and Wm is other component m mass), calculated by internal reference thing of Gastrodin
The relative correction factor fkm of other 5 kinds of compositions, the results are shown in Table 2.
Relative correction factor of the table 2 using Gastrodin as 5 kinds of compositions of internal reference thing（n=2）
Influence of the 4.2 different instruments to relative correction factor
Investigate Agilent 1200, Waters2996,3 brands of Shimadzu 2010A high performance liquid chromatograph to relative correction because
The influence of son, the results are shown in Table 3.As a result show that relative correction factor has good adaptation in different high performance liquid chromatographs
Property.
The relative correction factor result of 5 kinds of compositions in the different Instrument measuring rhizoma Gastrodiaes of table 3（n=2）
Influence of the 4.3 different chromatographic columns to relative correction factor
Investigate SinoChrom ODS posts（4.6 mm × 250 mm, 5 μm）, Kromasil C18 posts（The mm of 4.6 mm × 250,
5 μm）, Merck ODS posts（4.6 mm × 250 mm, 5 μm）Influence of 3 kinds of chromatographic columns to relative correction factor, the results are shown in Table
4.As a result show that relative correction factor has good adaptability in different brands chromatographic column.
The relative correction factor result of 5 kinds of compositions in 4 different chromatographic columns of table measure rhizoma Gastrodiae（n=2）
4.4 influences different in flow rate to relative correction factor
Investigation flow velocity is 0.8,1.0,1.2 mLmin-1When influence to relative correction factor, the results are shown in Table 5.As a result show
Relative correction factor is when different in flow rate with good adaptability.
The relative correction factor result of 5 kinds of compositions in the measure rhizoma Gastrodiae different in flow rate of table 5（n=2）
Influence of the 4.5 different column temperatures to relative correction factor
The influence to relative correction factor when column temperature is 20 DEG C, 25 DEG C, 30 DEG C is investigated, the results are shown in Table 6.As a result relative correction is shown
Factor adaptability under different column temperatures is good.
The relative correction factor result of 5 kinds of compositions in 6 different column temperatures of table measure rhizoma Gastrodiae（n=2）
The positioning of 5 chromatographic peaks to be measured
This research is using relative reservation method Rm/k=tR(m)/tR(k) it is accurately positioned.This research determines relative retention value not
With reappearance when brand instrument, different chromatographic columns, the results showed that, the relative retention value RSD of each composition to be measured is respectively less than 5%, can
Positioned for composition chromatographic peak to be measured, the results are shown in Table 6.
The relative retention value result of 5 kinds of compositions in 6 different instruments of table and chromatographic column measure rhizoma Gastrodiae（n=2）
6 one surveys comments method and external standard method results contrast
Using external standard method（ESM）Assay is carried out to 6 kinds of chemical compositions in rhizoma Gastrodiae, and comments method (QAMS) to calculate with being surveyed with one more
Result be compared, checking one survey comment method be used for 6 in rhizoma Gastrodiae in chemical composition content determine accurate fixed, reliability, knot
Fruit is shown in Table 7.As a result showing the result of 2 kinds of methods measure, there was no significant difference, and one surveys and comments method to be used for 6 kinds of Gastrodin classes in rhizoma Gastrodiae more
The assay of composition is accurate feasible.
The QAMS methods of table 7 and ESM methods determine content results (%, n=2 of 6 kinds of rhizoma Gastrodiae constituents in 15 batches of rhizoma Gastrodiaes）
In summary, the present invention surveys using one comments method gloomy to Gastrodin in Gastrodia eleta Bl., p-Hydroxybenzylalcohol, the gloomy glycosides A of Bali, Bali more
The gloomy glycosides E6 kinds active ingredient of the gloomy glycosides C of glycosides B, Bali, Bali carries out assay, and the one kind for establishing rhizoma Gastrodiae quality control has efficacious prescriptions
Method, the external standard method content of 6 kinds of active ingredient is used again, compare it with using the difference between relative correction factor calculated value
It is different, the results showed that the content that 2 kinds of methods measure does not have significant difference.
Chinese Pharmacopoeia version in 2010 is to rhizoma Gastrodiae assay using Gastrodin single component as index, it is desirable to must not be less than
0.20%, and increase the assay of p-Hydroxybenzylalcohol composition under version rhizoma Gastrodiae assay item in 2015, it is desirable to Gastrodin and right
The total amount of salicylic alcohol must not be less than 0.25%, and the present invention surveys one first comments method to be applied to Gastrodin in Gastrodia eleta Bl., Bali more
The multi objective Quality Evaluation Model of gloomy methods of glycosides, the factors such as the gloomy methods of glycosides reference substance of Bali is unstable, price is high are avoided,
From cheap, the stable Gastrodin of property be internal standard compound, establish one of 6 kinds of active ingredients in rhizoma Gastrodiae survey more comment method, it is and right
The rhizoma Gastrodiae of national different sources and different batches has carried out assay, and measurement result shows 6 kinds of compositions such as Gastrodin in Gastrodia eleta Bl.
Total content in 2.7%-11.3% scopes, be 10-45 times or so of version rhizoma Gastrodiae assay requirement lower bound in 2015, to controlling day
Numb quality and guarantee curative effect are significant.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill of the art
For personnel, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications should
Belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (8)
- A kind of 1. method of quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract, it is characterised in that：Method is surveyed using a mark, with Gastrodin detects Rhizoma Gastrodiae and its medicine materical crude slice glycosides A gloomy with p-Hydroxybenzylalcohol, Bali in extract, Bali is gloomy as internal standard compound The gloomy glycosides C of glycosides B, Bali, the gloomy glycosides E of Bali quality, specifically include following steps：（1）Prepare rhizoma Gastrodiae need testing solution；（2）It is single right to prepare the gloomy glycosides A of Gastrodin, p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali According to product solution and mixed reference substance solution；（3）Aspiration step respectively（2）Described single reference substance solution and mixed reference substance solution, inject high performance liquid chromatograph In tested, according to formula Rm/k=tR(m)/tR(k)（T in formulaR(m) retention time for being other component m, tR (k) is internal standard The retention time of thing）, calculate p-Hydroxybenzylalcohol, the gloomy glycosides A of Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali relative to The relative retention time of internal standard compound Gastrodin；（4）Aspiration step（2）Described mixed reference substance solution, the μ L of sample introduction 1,2,4,6,8,12, inject high performance liquid chromatography Instrument, measure, records each Component peak area, according to relative correction factor calculation formula fkm=fk/fm=(Wk×Am)/(Wm×Ak) (formula Middle AkFor internal standard compound peak area, WkFor internal standard compound quality；AmFor other component m peak areas, WmFor other component m mass), with day Numb element is internal reference thing, calculates the relative correction factor f of other 5 kinds of compositionskm；（5）Aspiration step respectively（1）Described need testing solution and step（2）Described Gastrodin reference substance, injects efficient liquid Chromatography, measure, the gloomy glycosides A of p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali pass through step（3） Described relative retention time progress is qualitative, passes through step（4）Described relative correction factor calculates respective content.
- 2. a kind of Rhizoma Gastrodiae according to claim 1 and its method for quality control of medicine materical crude slice and extract, its feature exists In the chromatographic condition of the high performance liquid chromatograph is：Using C18 chromatographic columns, mobile phase：Acetonitrile (A)-volume fraction 0.02~ 0.1% aqueous acid (B), using gradient elution：0 min~5 min, 2%A；The min of min, 2%~30%A, 20 of 5 min~20~ 25 min, 30%~2%A；Detection wavelength：220 nm；Volume flow：0.8~1.2mL min-1；Sample size is 5~25 μ L；Post Temperature：20~35 DEG C.
- 3. a kind of Rhizoma Gastrodiae according to claim 2 and its method for quality control of medicine materical crude slice and extract, its feature exists In mobile phase is acetonitrile-aqueous acid, and acid is preferably phosphoric acid, and its volume fraction is 0.05%；Volume flow is 1.0 mL min-1, sample size is 10 μ L, and column temperature is 25 DEG C.
- 4. a kind of method of the quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract, its feature exist according to claim 1 In step（3）The gloomy glycosides A of the p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali are relative to internal standard The relative retention time of thing Gastrodin is respectively：1.3480、1.8173、1.684 6、1.7286、1.6443.
- 5. a kind of method of the quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract, its feature exist according to claim 1 In step（4）The gloomy glycosides A of the p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali and Gastrodin Relative correction factor is respectively：0.6632nd, 1.2777,1.4351,1.3885,1.5907.
- 6. a kind of method of the quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract, its feature exist according to claim 1 In step（1）The preparation method of the rhizoma Gastrodiae need testing solution is：Precision weighs Rhizoma Gastrodiae, Processed Product of Rhizoma Gastrodiae or rhizoma Gastrodiae extraction Thing, the ethanol that 15~25 times of volume fractions are 60%~80% is added, 40~60min of heating and refluxing extraction, filtration, takes 10 times of amounts continuous Filtrate, it is concentrated near do and is dissolved in water without alcohol taste, residue, be transferred in 25 mL measuring bottles, be diluted with water to scale, shake up, filter Cross, take subsequent filtrate to cross 0.22 μm of filter membrane, obtain need testing solution.
- 7. a kind of method of the quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract, its feature exist according to claim 1 In step（2）The gloomy glycosides A of the Gastrodin, p-Hydroxybenzylalcohol, Bali, the gloomy glycosides B of Bali, the gloomy glycosides C of Bali, the gloomy glycosides E of Bali are single Reference substance solution and mixed reference substance solution, solvent are water, and the mass concentration of each material is 0.01~0.2mg/mL.
- 8. a kind of method of the quality control of Rhizoma Gastrodiae and its medicine materical crude slice and extract, its feature exist according to claim 1 In described Rhizoma Gastrodiae extract is water extraction or ethanol extract.
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| CN111948305A (en) * | 2020-07-24 | 2020-11-17 | 江阴天江药业有限公司 | Quality control method and preparation method for producing Qingda granules and compound traditional Chinese medicine based on Qingda granules |
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| CN108020615A (en) * | 2018-03-29 | 2018-05-11 | 宁波大学 | A kind of method of total gastrodia elata genin content in quantitative determination rhizoma Gastrodiae |
| CN111257437A (en) * | 2018-11-30 | 2020-06-09 | 华中科技大学 | Method for detecting various components in gastrodia elata, gastrodia elata extract and gastrodia elata-containing preparation |
| CN111257481A (en) * | 2018-11-30 | 2020-06-09 | 华中科技大学 | Method for establishing fingerprint of rhizoma gastrodiae medicinal material and fingerprint thereof |
| CN111929378A (en) * | 2020-07-24 | 2020-11-13 | 江阴天江药业有限公司 | Method for measuring content of 6 index components of gastrodia elata in Qingda granules |
| CN111948305A (en) * | 2020-07-24 | 2020-11-17 | 江阴天江药业有限公司 | Quality control method and preparation method for producing Qingda granules and compound traditional Chinese medicine based on Qingda granules |
| CN111948305B (en) * | 2020-07-24 | 2023-02-03 | 江阴天江药业有限公司 | Quality control method and preparation method for producing Qingda granules and compound traditional Chinese medicine based on Qingda granules |
| CN114062526A (en) * | 2020-08-05 | 2022-02-18 | 汉义生物科技（北京）有限公司 | Detection method for quantifying cannabinoids based on relative correction factor method |
| CN114062526B (en) * | 2020-08-05 | 2024-03-08 | 汉义生物科技（北京）有限公司 | Detection method for quantifying cannabinoid compounds based on relative correction factor method |
| CN115779042A (en) * | 2022-12-21 | 2023-03-14 | 浙江中医药大学 | Rhizoma gastrodiae total polyphenol for preventing and treating migraine |
| CN115779042B (en) * | 2022-12-21 | 2023-12-05 | 浙江中医药大学 | Gastrodia elata total polyphenol for preventing and treating migraine |
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