CN106841470B - A kind of the efficient liquid phase characteristic spectrum and its acquisition methods of dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae - Google Patents
A kind of the efficient liquid phase characteristic spectrum and its acquisition methods of dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae Download PDFInfo
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Abstract
The invention discloses a kind of methods of efficient liquid phase characteristic spectrum for obtaining dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae, include the following steps: a, prepare test solution: taking dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae, methanol extracts, and concentration is spare；B, following chromatographic condition is used to detect test solution: stationary phase: using octadecylsilane chemically bonded silica as filler；Mobile phase: methanol-water solution gradient elution, methanol percent by volume are 30~80%；Wherein, water phase contains 0.1%w/v lauryl sodium sulfate and 0.1%w/v phosphoric acid.High performance liquid chromatography is applied to the quality testing of dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae by the present invention, can effectively control the quality of dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae, provides scientific basis for effectively control medicinal material inherent quality standard.
Description
Technical field
The present invention relates to a kind of efficient liquid phase characteristic spectrum of dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae and its acquisition methods, belong to medicine
Field.
Background technique
Rhizoma Atractylodis Macrocephalae be compositae plant Rhizoma Atractylodis Macrocephalae (Atractylodes macroephala Koidz.) dry rhizome, main product in
The ground such as Zhejiang, Anhui, Hunan.Its is warm-natured, bitter, sweet, returns spleen, stomach meridian, has strengthening the spleen and replenishing qi, eliminate dampness and have diuretic effect, hidroschesis, miscarriage prevention etc.
Effect cures mainly deficiency-weakness of spleen-QI, anorexia and loose stool caused by dysfunction of the spleen in transportation and transformation, abdominal fullness and distention, fatigue and weakness, phlegm retention oedema, spontaneous perspiration, movement of the foetus
The card such as uneasiness.
The concocting method of Rhizoma Atractylodis Macrocephalae is numerous, ancient literature textual criticism, the processing of Rhizoma Atractylodis Macrocephalae is endured first recorded in the Tang Dynasty supplement to Thousand Golden Prescriptions
Yellow, native stir-fry method.Song Dynasty have fry after big gun, stir-bake to yellowish, the leaching of rice swill, water in which rice has been washed leaching, vinegar leaching stir-frys, roasting, simmer make, soil the methods of boils.The Qing Dynasty
It increases the dried immature fruit of citron orange and decocts water stain stir-fry, " book on Chinese herbal medicine state hook member " is described as that " spleen deficiency and stagnation of the circulation of vital energy person, the dried immature fruit of citron orange is fried or rhizoma cyperi about the processing of Rhizoma Atractylodis Macrocephalae
It fries ".In the record of numerous processed products of Rhizoma Atractylodis Macrocephalae, shown unique characteristics especially with decoction (dried immature fruit of citron orange juice) Rhizoma Atractylodis Macrocephalae processed, deepened processing intension -- it entangles
Bias and synergy.
However, few for the research of dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae so far, this concocting method is detected in thick in ancient literature
It slightly records, is not studied and dissected by modern means, more have no the relevant report for carrying out quality control to dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae.
Therefore, in order to effectively control processing quality, the clinical value of dried immature fruit of citron orange atractylodes slice is improved, needs to establish its spy
Sign property chromatographic fingerprint spectrogram, to provide a kind of stabilization, reliable, comprehensive method of quality control.
Summary of the invention
The purpose of the present invention is to provide a kind of efficient liquid phase characteristic spectrum of dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae and its acquisition sides
Method.
The present invention provides a kind of methods for obtaining dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae efficient liquid phase characteristic spectrum, including walk as follows
It is rapid:
A, it prepares test solution: taking dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae, methanol extracts, and concentration is spare；
B, test solution is detected using following chromatographic condition:
Stationary phase: using octadecylsilane chemically bonded silica as filler；
Mobile phase: methanol-water solution gradient elution, methanol percent by volume are 30~80%；Wherein, water phase contains
0.1%w/v lauryl sodium sulfate and 0.1%w/v phosphoric acid.
Further, eluent gradient elution requirement is as follows:
Further, the stationary phase is XAquaC18 chromatographic column；Chromatographic column specification: column length 25cm, internal diameter are
4.6mm, partial size are 5 μm.
Further, column temperature is 30 DEG C, and flow velocity is 1.0ml per minute.
Further, Detection wavelength 225nm.
Further, the every 1ml of the test solution corresponds to dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae 0.4g.
Further, a step methanol extraction conditions are as follows: dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae is taken, 20 times of volume methanol are added, in
80 DEG C are extracted 2 hours.
Further, the sample volume of the test solution is 10 μ L.
The present invention provides it is a kind of according to the method obtain dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae efficient liquid phase characteristic spectrum,
Including 5 characteristic peaks, retention time is identical as aurantiamarin, synephrine, atractylodes lactone III, atractylodes lactone I, atractylone respectively.
The present invention provides it is a kind of according to the method obtain dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae efficient liquid phase characteristic spectrum,
Including 5 characteristic peaks, retention time be respectively as follows: 28.588~29.524min, 40.784~41.16min, 59.329~
59.594min, 81.249~81.564min, 109.774~110.386min.
Dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae of the present invention is the bulk pharmaceutical chemicals by following weight proportion through processing: the dried immature fruit of citron orange 1~
10 parts, 100 parts of Rhizoma Atractylodis Macrocephalae.
Further, it is the bulk pharmaceutical chemicals by following weight proportion through processing: 10 parts of the dried immature fruit of citron orange, 100 parts of Rhizoma Atractylodis Macrocephalae.
Further, it is extract and Rhizoma Atractylodis Macrocephalae by the weight proportion dried immature fruit of citron orange through processing；The dried immature fruit of citron orange mentions
It takes object to be prepared by the following method to obtain: taking the dried immature fruit of citron orange, add 6~16 times of volume water, impregnate 0~2.0h, boil 0.5~3h, filter,
The dregs of a decoction repeat extract 1~2 time, filtration, merging filtrate, concentration to get.
Further, soaking time is 0~1.0h.
Further, each boiling time is 1~1.5h.
Further, it extracts every time and adds 8~14 times of volume water.
Further, the Bitter Orang P.E is prepared by the following method to obtain: taking the dried immature fruit of citron orange, adds 12 times of volume water, soaks
Steep 0.5h, boil 1.5h, filter, the dregs of a decoction repeat extract 1 time, filtration, merging filtrate, concentration to get.
Further, it is concentrated into 1mL Bitter Orang P.E and is equivalent to 0.2g Fructus Aurantii Immaturus.
Further, it is processed by following methods: Rhizoma Atractylodis Macrocephalae is taken, Bitter Orang P.E is added, impregnates 12~36h, in
100~150 DEG C of 4~12min of frying, in 30~70 DEG C of 24~72h of baking to get.
Further, it is processed by following methods: taking Rhizoma Atractylodis Macrocephalae, Bitter Orang P.E is added, impregnate for 24 hours, in 110 DEG C
Frying 6min, in 40 DEG C bake for 24 hours to get.
Further, the content of atractylodes lactone III is 0.035~0.054% in the processed products of Rhizoma Atractylodis Macrocephalae, synephrine
Content is 0.045~0.053%.
HPLC chromatogram method is applied to the quality testing of dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae, the dried immature fruit of citron orange of 10 batches of different sources by the present invention
The chromatogram entirety appearance of processed products of Rhizoma Atractylodis Macrocephalae processed is essentially identical, and separation analysis efficiency is high, the relative retention time one of each characteristic peak
It causes.High performance liquid chromatography is applied to inside dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae, the quality of dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae can be effectively controlled, to be
Effective control medicinal material inherent quality standard provides scientific basis.
Obviously, above content according to the present invention is not being departed from according to the ordinary technical knowledge and customary means of this field
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The specific embodiment of form by the following examples remakes further specifically above content of the invention
It is bright.But the range that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to example below.It is all to be based on above content of the present invention
The technology realized all belongs to the scope of the present invention.
Detailed description of the invention
Fig. 1 is dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae standard colour chart feature spectrogram；
Fig. 2 is that the specificity of negative control tests HPLC spectrogram；
Fig. 3 is that the specificity of aurantiamarin tests HPLC spectrogram；
Fig. 4 is that the specificity of synephrine tests HPLC spectrogram；
The specificity that Fig. 5 is atractylodes lactone III tests HPLC spectrogram；
The specificity that Fig. 6 is atractylodes lactone I tests HPLC spectrogram；
Fig. 7 is that the specificity of atractylone tests HPLC spectrogram；
Fig. 8 is the specificity test HPLC spectrogram for frying Rhizoma Atractylodis Macrocephalae sample；
Fig. 9 is that the specificity of dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae+fry Rhizoma Atractylodis Macrocephalae sample tests HPLC spectrogram；
Figure 10 is that the specificity of dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae sample tests HPLC spectrogram；
Figure 11 is 10 batch dried immature fruit of citron orange atractylodes slice HPLC feature spectrograms (S- atractylodes lactone III)；
Figure 12 is the HPLC feature spectrogram obtained when organic phase is acetonitrile in mobile phase；
Figure 13 is the HPLC feature spectrogram obtained when organic phase is methanol in mobile phase；
Figure 14 is the HPLC feature spectrogram obtained when organic phase is acetonitrile-methanol (1:1) in mobile phase；
Figure 15 is the HPLC feature spectrogram obtained in experimental example 4 using condition of gradient elution 1；
Figure 16 is the HPLC feature spectrogram obtained in experimental example 4 using condition of gradient elution 2；
Figure 17 is the HPLC feature spectrogram obtained in experimental example 4 using condition of gradient elution 3；
Figure 18 is the HPLC feature spectrogram obtained in experimental example 4 using condition of gradient elution 4；
Figure 19 is the HPLC feature spectrogram obtained in experimental example 4 using condition of gradient elution 5；
Figure 20 is the HPLC feature spectrogram obtained in experimental example 4 using condition of gradient elution 6.
Specific embodiment
Raw material, equipment used in the specific embodiment of the invention are known product, are obtained by purchase commercial product.
The HPLC characteristic spectrum of the dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae of the present invention of embodiment 1
1 instrument and material
Agilent1260 type high performance liquid chromatograph, Agilent company；1/,100,000 electronic analytical balance of BT25S type, north
Jing Saiduoli scientific instrument Co., Ltd；1/1000 electronic analytical balance of JA2003 type, upper current chart level instruments and meters Co., Ltd；
BT25S electronic balance, Sai Duolisi scientific instrument (Beijing) Co., Ltd.
Acetonitrile, methanol are chromatographic grade, Fisher Scientific company；Water is ultrapure water, other reagents are analysis
Pure (Chengdu section dragon chemical reagent).
Fructus Aurantii Immaturus, lot number 1603012 are purchased from Kelun Pharm Ind Co., Ltd., Sichuan, through in Chengdu University of Traditional Chinese Medicine
Medicine identification teaching and research room Lu elder generation penetrating judgment award be accredited as rutaceae bitter orange (Citrus aurantiumL.) and its variety or
The drying young fruit of sweet orange (C.sinensisOsbeck), Rhizoma Atractylodis Macrocephalae are compositae plant Rhizoma Atractylodis Macrocephalae
(AtractylodesmacroephalaKoidz.) dry rhizome.(the Chinese pharmaceutical biological product calibrating of synephrine reference substance
Institute, lot number 110727-201107)；Aurantiamarin reference substance (Nat'l Pharmaceutical & Biological Products Control Institute, lot number 110721-201316)；
Atractylodes lactone I reference substance (Nat'l Pharmaceutical & Biological Products Control Institute, lot number 111975-201501)；Atractylodes lactone III reference substance
(Nat'l Pharmaceutical & Biological Products Control Institute, lot number 111978-201501).
Dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae: self-control, preparation method: the extraction of dried immature fruit of citron orange juice: weighing dried immature fruit of citron orange 100g, adds 1430mL water logging
Steep 30min, boil 1.5h, filter, the dregs of a decoction add again 1200mL boiling boil 1.5h, filtration, merging filtrate, be concentrated into 500mL to get
Dried immature fruit of citron orange juice, wherein 1mL dried immature fruit of citron orange juice is equivalent to 0.2g Fructus Aurantii Immaturus；It processes: taking raw Rhizoma Atractylodis Macrocephalae, dried immature fruit of citron orange juice 50mL is added, it is corresponding
Fructus Aurantii Immaturus dosage be the 10% of Rhizoma Atractylodis Macrocephalae, impregnate for 24 hours, stir frequently, until use up medicine saturating for juice, be 110 DEG C in medicinal material temperature and fry
6min processed takes out, in 40 DEG C of baking ovens bake for 24 hours to get.
2 methods and result
2.1 chromatographic condition
Using octadecylsilane chemically bonded silica as filler (column length 25cm, internal diameter 4.6mm, partial size are 5 μm)；With first
Alcohol is mobile phase A, with water (lauryl sodium sulfate containing 0.1%w/v and 0.1%w/v phosphoric acid) for Mobile phase B, by the regulation of table 1
Carry out gradient elution；Flow velocity is 1.0ml per minute；Detection wavelength is 225nm；Column temperature: 30 DEG C.Number of theoretical plate presses atractylodes lactone
The peak III, which calculates, should be not less than 140000.
1 linear gradient elution mobile phase of table
The preparation of 2.2 reference solutions
The preparation method of reference substance solution: take aurantiamarin, synephrine, atractylodes lactone III and atractylodes lactone I reference substance suitable
Amount, it is accurately weighed, respectively plus methanol be made solution of every 1ml containing 0.06mg, 0.1mg, 0.1mg, 0.04mg to get.
The preparation of 2.3 test solutions
Sample powder (crossing No. five sieves) 2.0g is taken, accurately weighed, methanol 40ml is added in precision, and weighed weight sets 80 DEG C of water
It is kept in bath slightly boiled 2 hours, lets cool, then weighed weight, the weight of less loss is supplied with methanol, is shaken up, filtered, precision measures continuous filter
Liquid 30ml is concentrated and is transferred in 5ml measuring bottle, cross 0.45um miillpore filter to get.
2.4 HPLC analysis
It takes 10 μ L of sample solution, injects high performance liquid chromatograph, when recording the reservation of chromatogram (Fig. 1) and each chromatographic peak
Between, the relative peak area (AR) of each chromatographic peak is calculated with area normalization method, each maximum 5 peaks of middle peak of spectrogram area are the last 5
Peak.
2.5 methodological study
2.5.1 specificity and characteristic
Blank solvent, aurantiamarin reference substance solution, synephrine reference substance solution, atractylodes lactone III reference substance are prepared respectively
Solution, atractylodes lactone I reference substance solution, atractylone reference substance solution, Rhizoma Atractylodis Macrocephalae sample solution (press sample solution preparation method system
It is standby), Rhizoma Atractylodis Macrocephalae (dried immature fruit of citron orange system) sample solution (by sample solution preparation method prepare), Rhizoma Atractylodis Macrocephalae+Rhizoma Atractylodis Macrocephalae (dried immature fruit of citron orange system) sample solution
(by sample solution preparation method prepare, sampling amount be Rhizoma Atractylodis Macrocephalae, each 1g of Rhizoma Atractylodis Macrocephalae (dried immature fruit of citron orange system)), according to above-mentioned chromatographic condition into
Row test, spectrogram are shown in Fig. 2~10, and specificity and characteristic are strong.
2.5.2 precision test
By sample preparation methods, randomly selects and prepare test liquid with a collection of dried immature fruit of citron orange atractylodes slice, continuous sample introduction 5 times, survey
Determining HPLC chromatogram, 5 shared peaks is selected to be compared, the results showed that each shared peak relative retention time RSD is 0.07%~
1.68%, the RSD of each shared peak relative peak area is 0.14%~1.10%, shows that instrument precision is good.
2.5.3 reappearance test
It is taken at random with a collection of dried immature fruit of citron orange atractylodes slice, by sample preparation methods operation repetitive, prepares 5 portions of dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae drinks
Piece test solution measures its HPLC chromatogram.As a result each shared peak relative retention time RSD is 0.07%~0.16%, respectively
Shared peak relative peak area RSD is 1.07%~3.80%.
2.5.4 stability test
The test solution with a collection of dried immature fruit of citron orange atractylodes slice is taken, measures HPLC respectively at 0,5,10,15,20,25,48h
Chromatogram is compared 5 shared chromatographic peaks.As a result each shared peak relative retention time RSD0.79%~1.12%, it is each total
The relative peak area RSD for having peak is 0.33%~4.72%, shows that sample is good in 48h internal stability.
The building of 2.6 Chromatographic Fingerprints
2.6.1 the calculating of relative retention time α
Select appearance time it is relatively placed in the middle, in each sample existing component as reference peak (α=1).Find out sample institute
There is the relative retention time at peak are as follows: tRi/tRα, tR in formulaiFor the appearance time of each component, tRαFor the appearance time referring to peak.
2.6.2 referring to the selection at peak
Compared by chromatogram, choosing the 3rd strong peak (No. 3 peaks) in chromatogram is to calculate the opposite of each peak referring to peak
Retention time α.
2.6.3 the foundation and evaluation of characteristic spectrum
Take the dried immature fruit of citron orange of 10 batch different sources with Rhizoma Atractylodis Macrocephalae respectively by legal system available test sample solution below " 2.3 " item, then
By condition sample introduction under " 2.1 " item and record chromatogram.The characteristics of according to 10 batches of sample characteristic spectrums, can determine 5 peak conducts
Common characteristic peaks, and four peaks therein have been demarcated, No. 1 peak is aurantiamarin peak, and No. 2 peaks are synephrine peak, and No. 3 peaks are Rhizoma Atractylodis Macrocephalae
The peak lactone II I, No. 4 peaks are the peak atractylodes lactone I, these three peak separating degrees are preferable, and retention time is suitable for, therefore can be used as reference
Peak, chromatogram are shown in Fig. 1.The characteristic spectrum signal of each batch of Fructus Aurantii Immaturus is imported into " the Chinese medicine that exploitation is presided over by Chinese Pharmacopoeia Commission
Fingerprint similarity evaluation software (2009 editions) ", the common pattern of 10 batches of medicinal material characteristic spectrums is shown in Figure 11.Calculate comparative diagram
Similarity data between spectrum and 10 batches of sample maps, similarity calculation the results are shown in Table 4.Similarity thinks to meet 0.90 or more
It is required that.As a result, it has been found that the similarity of 10 batches of medicinal materials shows that this experimental method is reliably effective 0.90 or more.
The measurement of 2.7 test sample characteristic spectrums
The dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae for taking 10 batches to be prepared by the dried immature fruit of citron orange, the Rhizoma Atractylodis Macrocephalae of different sources, is prepared by test solution
Method operation, precision draw the measurement of 10 μ L sample introductions, obtain each sample HPLC characteristic spectrum.The dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae of 10 batch different sources
Medicine materical crude slice retention time and peak area and relative retention time the results are shown in Table 2, table 3, table 4.Chromatogram is shown in Figure 11.
2 10 batch different sources dried immature fruit of citron orange atractylodes slice characteristic spectrum retention time of table
3 10 batch different sources dried immature fruit of citron orange atractylodes slice characteristic spectrum peak area of table
4 10 batch dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae characteristic spectrum relative retention time of table
Test result shows that the shared chromatographic peak of the dried immature fruit of citron orange atractylodes slice of 10 batches of different sources is 5, wherein in the dried immature fruit of citron orange
There are 2 peaks, there are 3 peaks in Rhizoma Atractylodis Macrocephalae, efficient liquid phase detection is carried out to the dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae of 10 batches of different sources, is compared from chromatogram
Compared with, though the amount of each batch of sample composition is variant, chromatogram general picture is consistent, illustrate the medicinal material group consistence of composition (including
The consistency of ingredient type and mass fractions relative), and stablize, characteristic spectrum can be to the consistent of Rhizoma Atractylodis Macrocephalae component population characteristic
Property judges.Dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae characteristic spectrum has the feature of itself chemical bar code, is its intrinsic chemical ingredient type and quantity
Reflection.In the common characteristic peaks of dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae characteristic spectrum, overall separation degree is relatively good；Different batches medicinal material characteristic pattern
Spectrum general picture also has preferable consistency, can sufficiently reflect the feature of dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae chemical component.
Beneficial effects of the present invention are further illustrated below by way of experimental example.
The preparation method of 1 test solution of experimental example
In the preparation process of test solution, it is contemplated that effective component synephrine and orange peel methods of glycosides and Rhizoma Atractylodis Macrocephalae in the dried immature fruit of citron orange
Middle atractylodes lactone constituents and atractylone ingredient, final choice is using methanol as solvent extraction.
This experiment takes same amount of sample, prepares test sample, detection knot respectively with methanol eddy, ultrasonic two kinds of extracting methods
Fruit see the table below:
The peak area result of the different methanol extraction processes of table 5
Experimental result: with methanol ultrasonic extraction, causing synephrine and aurantiamarin in dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae low, therefore selects methanol
Reflux concentration, to improve in test solution the content of the dried immature fruit of citron orange and accurately carry out qualitative analysis to dried immature fruit of citron orange Rhizoma Atractylodis Macrocephalae.
Influence of 2 ion-pairing agent of experimental example to separating effect
The characteristics of due to composition of alkaloids, joined ion-pairing agent in the mobile phase of efficient liquid phase with realize its
Reservation in chromatographic column.
Preliminary experiment has been respectively compared addition lauryl sodium sulfate-phosphoric acid, dodecyl sodium sulfate-phosphorus in mobile phase
Acid, dodecyl sodium sulfate-potassium dihydrogen phosphate-glacial acetic acid, diethylamine and triethylamine, the results showed that with dodecyl sodium sulfate-
Phosphoric acid is ion-pairing agent, and separating effect is best.
Influence of 3 organic phase of experimental example to separating effect
This experiment with the organic phase that acetonitrile, methanol, acetonitrile-methanol (1:1) are in mobile phase, is examined processed product respectively
It surveys, the result is shown in Figure 1 2~14.
The result shows that the organic phase in using methanol as mobile phase, can reach good separating effect.
Influence of 4 elution requirement of experimental example to separating effect
Since ingredient is the entirely different element of the first species of two polarity, final choice gradient in the ingredient and Rhizoma Atractylodis Macrocephalae in the dried immature fruit of citron orange
Elution.
The condition of gradient elution of this experiment screening is as follows:
Condition 1, mobile phase A: methanol, B: water (containing 0.1%w/v lauryl sodium sulfate and 0.1%w/v phosphoric acid) gradient
Elution:
Condition 2, mobile phase A: methanol, B: water (containing 0.1%w/v lauryl sodium sulfate and 0.1%w/v phosphoric acid) gradient
Elution:
Condition 3, mobile phase A: methanol, B: water (containing 0.1%w/v lauryl sodium sulfate and 0.1%w/v phosphoric acid) gradient
Elution:
Condition 4, mobile phase A: methanol, B: water (containing 0.1%w/v lauryl sodium sulfate and 0.1%w/v phosphoric acid) gradient
Elution:
Condition 5, mobile phase A: methanol, B: water (containing 0.1%w/v lauryl sodium sulfate and 0.1%w/v phosphoric acid) gradient
Elution:
Condition 6, mobile phase A: methanol, B: water (containing 0.1%w/v lauryl sodium sulfate and 0.1%w/v phosphoric acid) gradient
Elution:
Separating resulting is successively shown in Figure 15~20.
Claims (7)
1. a kind of method for obtaining dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae efficient liquid phase characteristic spectrum, it is characterized in that: including the following steps:
A, it preparing test solution: taking dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae, methanol extracts, concentration,
It is spare；
B, test solution is detected using following chromatographic condition:
Stationary phase: using octadecylsilane chemically bonded silica as filler；
Mobile phase: methanol-water solution gradient elution, methanol percent by volume are 30~80%；Wherein, water phase contains 0.1%w/v
Lauryl sodium sulfate and 0.1%w/v phosphoric acid；The eluent gradient elution requirement is as follows:
2. the method as described in claim 1, it is characterized in that: the stationary phase is XAquaC18 chromatographic column；Chromatographic column specification:
Column length is 25cm, and internal diameter 4.6mm, partial size is 5 μm.
3. the method as described in claim 1, it is characterized in that: column temperature is 30 DEG C, flow velocity is 1.0ml per minute.
4. the method as described in claim 1, it is characterized in that: Detection wavelength is 225nm.
5. the method as described in claim 1, it is characterized in that: the every 1ml of the test solution corresponds to dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae
0.4g。
6. the method as described in claim 1, it is characterized in that: a step methanol extraction conditions are as follows: dried immature fruit of citron orange processed products of Rhizoma Atractylodis Macrocephalae is taken,
20 times of volume methanol are added, are extracted 2 hours in 80 DEG C.
7. the method as described in claim 1, it is characterized in that: the sample volume of the test solution is 10 μ L.
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