CN104013673B - A kind of Salvia root P.E and preparation method thereof - Google Patents
A kind of Salvia root P.E and preparation method thereof Download PDFInfo
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Abstract
The invention provides the extracting method of the red sage root, it includes following operating procedure：Salvia piece is taken, is added water, under reduced pressure, 70 DEG C of boilings are extracted, and merge extract solution, you can.Present invention also offers Salvia root P.E and preparation method thereof.Present invention also offers the detection method of Salvia root P.E.Red sage root extracting method of the invention, extract obtained middle content of danshinolic acid B is far above high temperature reflux extract, and other compositions and impurity content higher temperatures refluxing extraction are low, and Salvia root P.E tanshin polyphenolic acid B purity obtained in low-temperature boiling dynamic method is more than high temperature reflux extract.Detection method, effectively can efficiently separate the multiple compositions in Salvia root P.E, and peak shape is good, and baseline is steady, for the quality control of Salvia root P.E provides reliable basis.
Description
Technical field
The present invention relates to a kind of Salvia root P.E and preparation method thereof.The invention further relates to the detection side of Salvia root P.E
Method.
Background technology
The red sage root is the dry root and rhizome of labiate red sage root Salvia miltiorrhiza Bge., first recorded in《God
Agriculture book on Chinese herbal medicine warp》, top grade is classified as, it is clinical parts of generic medicinal plants.With promoting blood circulation, relieving pain of dysmenorrhea, relieving restlessness that clears away heart-fire, cool blood to disappear carbuncle etc.
Effect, clinic is widely used in the card such as chest impediment and cardialgia, gastral cavity abdomen hypochondriac pain, lumps in the chest and abdomen, hot numbness pain, dysphoria and insomnia, irregular menstruation.
The red sage root contains fat-soluble diterpene quinone and water miscible phenolic acid compound.Soluble salvianolic acid is the red sage root
Main active component, including danshensu, tanshin polyphenolic acid B, salvianolic acid C and composition etc. protocatechualdehyde, with content of danshinolic acid B highest.
Tanshin polyphenolic acid B all has protective effect to cardiovascular system, brain, liver, kidney etc., is effective water soluble ingredient of the red sage root.
The red sage root is extracted and uses refluxing extraction more, and Extracting temperature is solvent boiling point under atmospheric pressure, and temperature is higher, danshinolic acid
To thermally labile, long-time heating, tanshin polyphenolic acid B is easily decomposed destruction B, makes the reduction of its content.It would therefore be highly desirable to one kind can be carried effectively
Tanshin polyphenolic acid B is taken, and less red sage root extracting method is extracted to impurity.
The content of the invention
The technical scheme is that there is provided a kind of content of danshinolic acid B Salvia root P.E higher, it is of the invention another
Technical scheme there is provided the preparation method of Salvia root P.E.
The invention provides the extracting method of the red sage root, it includes following operating procedure：Salvia piece is taken, is added water, depressurize bar
Under part, 70 ± 5 DEG C of boilings are extracted, and merge extract solution, you can.
Said extracted temperature, preferably uses 70 DEG C.
Heretofore described boiling is extracted, i.e., at reduced pressure conditions so that water is issued to boiling in corresponding Extracting temperature
State is risen, is extracted under the conditions of this temperature and pressure, wherein, vacuum pressure ensures water to meet under a certain temperature conditionss
On the basis of boiling.
Wherein, amount of water is 5~15 times, preferably 12 times of red sage root dry weight.
Wherein, extract 1~3 time, 0.5~3h every time；It is preferred that extracting 3 times, each 2h.
Present invention also offers a kind of preparation method of Salvia root P.E, it includes following operating procedure：
（1）Extract as stated above；
（2）Take step（1）Extract solution, after solvent, freeze-drying is recovered under reduced pressure, obtain final product Salvia root P.E.
Wherein, it is described solvent is recovered under reduced pressure condition be：- 0.05~-0.095MPa, 40~90 DEG C；It is preferred that 70 ± 5 DEG C.
Present invention also offers a kind of Salvia root P.E, the Salvia root P.E contains 5~15% tanshin polyphenolic acid B.
Further, the preparation method of the Salvia root P.E is as described above.
Present invention also offers the detection method of Salvia root P.E, it is measured with high performance liquid chromatography, tool
Body operating procedure is as follows：
（1）Salvia root P.E to be measured is taken, with methyl alcohol as solvent or diluent, need testing solution is prepared；
（2）Need testing solution is injected in high performance liquid chromatograph, detection is obtained final product, and its chromatographic condition is as follows：
Chromatographic column：Octadecyl silane is fixing phase；
Mobile phase：Methyl alcohol A-1.0% glacial acetic acid aqueous solution B, gradient elution, elution program A：0min7%, 10min7%,
12min9%, 31min33%, 47min39%, 60min58%, 70min62%；
Detection wavelength：286±5nm.
Further, in the high performance liquid chromatography, with the one kind in tanshin polyphenolic acid B, protocatechualdehyde or Sodium Danshensu or
Two or more is reference substance, prepares reference substance solution.
Further, detection column temperature is 30 ± 5 DEG C.
Using extracting method of the present invention, content of danshinolic acid B is far above high temperature reflux extract in obtained Salvia root P.E,
Other compositions and impurity content higher temperatures refluxing extraction are low, Salvia root P.E tanshin polyphenolic acid B purity obtained in low-temperature boiling dynamic method
More than high temperature reflux extract.
For higher temperatures refluxing extraction, suitable low temperature extracts the maximum dissolution that both ensure that liposoluble ingredient, also prevents it
Be decomposed destruction at high temperature, and the liposoluble ingredient in the red sage root has been extracted to greatest extent；Simultaneously as low temperature, impurity and
Other compositions stripping quantity is reduced, while liposoluble ingredient is efficiently separated from the red sage root, impurity in extract solution
Content with other compositions is also reduced, and is conducive to follow-up separation purifying technique, improves product quality and added value.
Detection method, effectively can efficiently separate the multiple compositions in Salvia root P.E, and peak shape is good, baseline
Steadily, for the quality control of Salvia root P.E provides reliable basis.
Brief description of the drawings
Fig. 1：Different Extracting temperature content of danshinolic acid B figures
Fig. 2：Red sage root extract surface tension
Fig. 3：Red sage root extract Zeta potential
Fig. 4：Red sage root extract particle diameter distribution
Fig. 5：Red sage root extract finger-print chromatogram
Fig. 6：10 crowdes of red sage root extract finger-print stacking charts
Fig. 7：Salvia root P.E grain size distribution
Fig. 8：Salvia root P.E Moisture percentage curve
Fig. 9：10 crowdes of Salvia root P.E finger-print stacking charts
Specific embodiment
The red sage root extracting method of embodiment 1
Take salvia piece, plus red sage root dry weight 12 times of amount water, under reduced pressure, 70 DEG C of boilings are extracted 3 times, and each 2h is closed
And extract solution, you can.
The research of the red sage root extracting method of embodiment 2
1 instrument and reagent
Shimadzu LC-10A high performance liquid chromatographs, SCL-6A system controllers, N2000 Data Processing in Chromatography Workstation, TC-100
Insulating box（Japanese Shimadzu Corporation）；Agilent1200 high performance liquid chromatographs series（Agilent companies of the U.S.）；Mettler
The a ten thousandth electronic balances of AE240 ten（German Mettler companies）；FA1104 a ten thousandth electronic balances（Shanghai balance equipment
Factory）；KQ3200 type ultrasonic cleaning machines（Kunshan Ultrasonic Instruments Co., Ltd.）；SHZ-D（Ⅲ）Circulating water type vavuum pump（Gongyi City
Yu Hua instruments Co., Ltd）；Thermostat water bath（Beijing Zhong Xing great achievements Instrument Ltd.）；RE-2000 type rotary evaporations
Instrument（Shanghai Yarong Biochemical Instrument Plant）, ALPHA1-4LSC freeze-dryers（CHRIST companies）, PHS-3E types PH meters（Upper Nereid
Close scientific instrument Co., Ltd）；OCAT21 surface/interface tensiometers（German Dataphysics companies）；Conductivity meter（Lei Ci
DDS-307A, upper Nereid section）Malvern nano particle size instrument（Model Nano-ZS）.
Red rooted salvia（The natural pharmaceutcal corporation, Ltd in Sichuan Cologne）；Tanshin polyphenolic acid B reference substance（Lot number：MUST-12020104,
Chengdu Man Site bio tech ltd）, Sodium Danshensu reference substance（Lot number：MUST-12020803, Chengdu Man Site are biological
Science and Technology Ltd.）, protocatechualdehyde reference substance（Lot number：110810-200205, Nat'l Pharmaceutical ＆ Biological Products Control Institute）, methyl alcohol
It is chromatographically pure, water is redistilled water（Self-control）, it is pure that other reagents are analysis.
The foundation of 2 content of danshinolic acid B mensuration methodologies
Chromatographic condition chromatographic column：Octadecylsilane chemically bonded silica post；Agilent TC-C18（250mm×4.6mm）Color
Spectrum post, 5 μm；The phosphoric acid water of acetonitrile -0.5%（28:72）It is mobile phase, Detection wavelength 286nm, flow velocity 1.0mL/min；Column temperature：30
℃。
The preparation precision of the preparation reference substance solution of reference substance solution weighs tanshin polyphenolic acid B reference substance 14.08mg and is measured in 10mL
In bottle, 75% methanol solution constant volume shakes up, as stock solution；Precision measures 1mL in 10mL measuring bottles, 75% methanol solution constant volume,
Shake up, obtain final product 0.1408mg/mL tanshin polyphenolic acid B reference substance solutions.
The preparation of need testing solution is accurate to draw extract solution in right amount in 25mL measuring bottles, and 75% methanol solution constant volume shakes up,
0.45 μm of miillpore filter filtration, takes subsequent filtrate as need testing solution.
Range of linearity experiment takes reference substance solution, and the accurate μ L of sample introduction 2,4,8,16,20, determine according to above-mentioned chromatographic condition respectively
Integrating peak areas value, is returned with peak area to sample size, obtains regression equation：Y=131290X+46157, R=0.9991.Knot
Fruit shows, is that linear relationship is good in the range of 0.2816mg~2.8160mg in content of danshinolic acid B.
Precision test is accurate to draw reference substance solution 10uL injection liquid chromatographs, and continuous sample introduction 6 times records peak face
Product, it is 1.16% to calculate RSD, shows that instrument precision is good.
Stability test is accurate to draw need testing solution, respectively at 0h, 2h, 4h, 6h, 8h sample introduction 5uL, records peak area,
RSD is 1.08%, shows that sample is good in 8h internal stabilities.
Replica test prepares 6 parts of need testing solutions by " preparation of need testing solution ", accurate respectively to draw 5uL injection liquid
Chromatography, records peak area, and RSD is 2.14%, shows that method repeatability is good
Recovery test precision weighs the Danshen Root 0.1g of known content, and precision adds tanshin polyphenolic acid B reference substance
3.8016mg, to 50mL, weighed weight, ultrasonic 1h takes out 75% methanol constant volume, lets cool, then weighed weight, is mended with 75% methyl alcohol
The weight of sufficient less loss, shakes up, filtration, takes subsequent filtrate, and the μ L of sample introduction 5 determine content, calculate the rate of recovery, and average recovery rate is
101.77%, RSD are 2.89% (n=6).
Content of danshinolic acid B is determined and takes Danshen Root (crossing No. three sieves) about 0.2g in red rooted salvia, accurately weighed, puts tool plug cone
In shape bottle, precision adds 75% methyl alcohol 50mL, and weighed weight, ultrasonic 1h takes out, lets cool, then weighed weight, is mended with 75% methyl alcohol
The weight of sufficient less loss, shakes up, filtration, takes subsequent filtrate, and the μ L of sample introduction 5 determine content, and content of danshinolic acid B is in measuring red rooted salvia
3.81%。
3 Orthogonal Experiment and Designs
Weigh salvia piece 30g, with water as solvent, under the Extracting temperature that seethed with excitement at vacuumizing 70 DEG C to extraction time, molten
Agent consumption, three factors of extraction time carry out L9(34) orthogonal experiment, sample solution is prepared, content of danshinolic acid B is determined, as a result such as
Under：
The Orthogonal Experiment and Design of table 1 and result
The variance analysis of table 2
Note：P ＜ 0.05 indicate conspicuousness, and P ＜ 0.01 indicate pole conspicuousness
Result shows that each factor effect primary and secondary is A>C>B, A extract influence maximum on root of red-rooted salvia phenolic acid, are secondly C factors, B
Factor influence is minimum.The results of analysis of variance is：A, B, C factor all have a significant impact to extraction effect.Comprehensive analysis directly perceived and side
The result of difference analysis, determines that optimize technique is A3B3C3Plus 12 times of amount water, i.e., extract 3 times in 70 DEG C of boiling refluxs of low temperature, every time
2h。
The checking of orthogonal test according to the result of above orthogonal test, to optimum process condition A preferably out3B3C3Enter
Row checking test, in triplicate, tanshin polyphenolic acid B and dried cream yield are respectively 3.11%, 40.36%.
The difference of different Extracting temperatures weighs salvia piece 30g, using the process conditions of optimization of orthogonal test, in 50,60,
70th, red sage root extract is prepared at 80,90,100 DEG C, content of danshinolic acid B and dried cream yield is determined.Result such as table 3, Fig. 1：
The influence of the different Extracting temperatures of table 3（n=3）
Data above shows, with the rising of Extracting temperature, content of danshinolic acid B first increases and then decreases, 70 DEG C of content highests
It is 3.14%, 100 DEG C of contents minimum 2.46%；Dried cream yield is raised with temperature and gradually increased, 50 DEG C minimum 38.95%, 100
DEG C up to 48.50%.When Extracting temperature is less than 70 DEG C, tanshin polyphenolic acid B can not effectively dissolution；During higher than 70 DEG C, with the heat time
Extension, tanshin polyphenolic acid B is decomposed destruction；Therefore 70 DEG C is the suitable Extracting temperature of tanshin polyphenolic acid B.Due in material solvent solubility it is general with
Temperature to raise and increase, thus, material dissolution is few during low temperature, and dried cream yield is low, and as temperature is raised, substance dissolved quantity increases
Greatly, dried cream yield increases.At 70 DEG C, tanshin polyphenolic acid B can effectively dissolution, be not destroyed, other compositions and impurity dissolve in solution
Also higher temperatures are low for amount, are conducive to later separation to purify.
4 extract solution physical parameter measurements
（1）The measure of red sage root extract surface tension
The surface tension of red sage root high temperature and low temperature extract solution is determined using OCAT21 surface/interfaces tensiometer, is used
SPSS17.0 statistical softwares carry out independent samples t test comparative analysis, measurement result such as table 4, Fig. 2 to test data.
The surface tension test of the red sage root high temperature extract solution of table 4 and low temperature extract solution（Unit：mN）
Compare with high temperature extract solution, * P ＜ 0.05
Statistic analysis result shows that two groups of data variances are uneven, two-sided test P=0.027 ＜ 0.05, high temperature extract solution with
The surface tension of low temperature extract solution has significant difference, and low temperature extract solution surface tension is noticeably greater than high temperature extract solution surface
Power, this may be relevant with extraction process.
（2）The measure of red sage root extract electrical conductivity
The electrical conductivity of red sage root high temperature and low temperature extract solution is determined using conductivity meter, with SPSS17.0 statistical softwares to examination
Testing data carries out independent samples t test comparative analysis, and measurement result is shown in Table 5.
The conductance measurement of the red sage root high temperature extract solution of table 5 and low temperature extract solution（Unit：μs/cm）
Statistical analysis, two groups of data variances are neat, and two-sided test P=0.190 ＞ 0.05 show high temperature extract solution and low temperature
Extract solution electrical conductivity is not significantly different from.
（3）Red sage root extract pH value is determined
The pH of red sage root high temperature and low temperature extract solution is determined using pHS-3C Accurate pHs meter, with SPSS17.0 statistical softwares pair
Test data carries out independent samples t test comparative analysis, and measurement result is shown in Table 6.
The red sage root high temperature extract solution of table 6 is determined with the pH of low temperature extract solution
Compare with high temperature extract solution, * P ＜ 0.05
Statistical result showed, two groups of data variances are neat, two-sided test P=0.000 ＜ 0.05, and both have notable statistics poor
Different, low temperature extracting liquid pH value is substantially less than high temperature extract solution；Because both Extracting temperatures are different, dissolution of the material in both is not
Together, classes of compounds contained by extract solution is variant with quantity, therefore the pH value of extract solution is different.
（4）The measure of red sage root extract Zeta potential
Red sage root high temperature and low temperature extract solution Zeta potential are determined using zeta potential instrument, with SPSS17.0 statistical softwares pair
Test data carries out independent samples t test comparative analysis, and measurement result is shown in Table 7, Fig. 3.
The conductance measurement of the red sage root high temperature extract solution of table 7 and low temperature extract solution（Unit：vm）
Compare with high temperature extract solution, * P ＜ 0.05
Statistical analysis shows that two groups of data variances are neat, two-sided test P=0.008 ＜ 0.05, and both have significant statistics
Difference is learned, low temperature extract solution Zeta potential is substantially less than high temperature extract solution Zeta potential, shows high temperature extract solution stability than low
Warm extract solution is good.
（5）The measure of red sage root extract nanometer particle size
Red sage root high temperature and low temperature extract solution nanometer particle size are determined using Malvern nano particle sizes instrument, is united with SPSS17.0
Meter software carries out independent samples t test comparative analysis to test data, and measurement result is shown in Table 8, Fig. 4.
The conductance measurement of the red sage root high temperature extract solution of table 8 and low temperature extract solution（Unit：d.nm）
Compare with high temperature extract solution, * P ＜ 0.05
Statistical result showed, two groups of data variances are neat, two-sided test P=0.000 ＜ 0.05, and both have notable statistics poor
Different, low temperature extract solution nanometer particle size average value is noticeably greater than high temperature extract solution nanometer particle size average value.From the surface of intensity distribution,
High temperature extract solution particle diameter distribution is coarse dispersion in 400~800nm, low temperature extract solution particle diameter distribution in 2000~6000nm
System, low temperature extract solution nanometer particle size is much larger than high temperature extract solution.
The finger-print research of the extract solution of embodiment 3
The preparation of need testing solution weighs red rooted salvia 30g, plus 12 times of water, in 70 DEG C of refluxing extractions 3 times, each 2h, filter
Cross, merging filtrate, be settled to 1000mL, precision draws 5mL in measuring bottle, and methanol constant volume shakes up to 25ml, obtains final product red sage root low temperature
Extract solution need testing solution.
Red rooted salvia 30g, plus 12 times of water are weighed, refluxing extraction 3 times, each 2h, filtration, merging filtrate is settled to
1000mL, precision draws 5mL in measuring bottle, and methanol constant volume shakes up to 25ml, obtains final product red sage root high temperature extract solution need testing solution.
Reference substance solution to prepare accurately weighed tanshin polyphenolic acid B, protocatechualdehyde, Sodium Danshensu reference substance appropriate in 10mL measuring bottles
In, methanol constant volume obtains final product 0.1466mgmL containing tanshin polyphenolic acid B-1, the μ gmL of protocatechualdehyde 8.533-1, the μ g of Sodium Danshensu 33.4
mL-1Mixed reference substance solution.
（1）The selection of chromatographic condition
The selection of Detection wavelength quantifies to ensure the maximum information of finger-print, and sample overall picture is reflected as much as possible,
Using the glacial acetic acid aqueous solution binary gradient elutes of methyl alcohol -1.0%, under 30 DEG C of column temperatures, record 254,270,280,286,310nm ripples
Lower red sage root extract chromatogram long, and determine Detection wavelength with reference to 3D-plot figures.Result shows, chromatographic peak under 286nm wavelength
Preferably, all chromatographic peaks can embody separating effect, can fully reflect the situation of red sage root sample, therefore, selection 286nm is inspection
Survey wavelength.
The selection of mobile phase species under 30 DEG C of column temperatures, with the phosphate aqueous solution of methyl alcohol -0.5%, the glacial acetic acid water of methyl alcohol -1.0%
Solution, the aqueous formic acid of methyl alcohol -1.0%, the phosphate aqueous solution of acetonitrile -0.5% are mobile phase, carry out gradient elution, record chromatogram.
Result shows, steady with the glacial acetic acid aqueous solution gradient elution of methyl alcohol -1.0% baseline, separates preferably, therefore with the glacial acetic acid of methyl alcohol -1.0%
The aqueous solution is mobile phase.
The selection of gradient elution program with the glacial acetic acid aqueous solution of methyl alcohol -1.0% as mobile phase, under 30 DEG C of column temperatures, using not
Same gradient elution program, determines finger-print, as a result shows in gradient elution program（A：0min7%, 10min7%, 12min9%,
31min33%, 47min39%, 60min58%, 70min62%）Preferably, characteristic peak is obvious for lower separating degree, therefore with the gradient elution journey
Sequence is eluted.
The selection of chromatographic column, as mobile phase, under 286nm, is respectively adopted with the glacial acetic acid aqueous solution (B) of methyl alcohol (A) -1.0%
Agilent TC-C18, Dikma Kromasil100A C18, Scienhome Kromasil C18 chromatographic columns determine fingerprint image
Spectrum, as a result with Agilent TC-C18 chromatographic columns separating effect preferably, therefore selection Agilent TC-C18 chromatographic columns are measured.
Result shows that red sage root finger-print chromatographic condition is：Chromatographic column is Agilent TC-C18 chromatographic columns；Mobile phase is
The glacial acetic acid aqueous solution (B) of methyl alcohol (A) -1.0%, binary gradient elutes, elution program A：0min7%, 10min7%, 12min9%,
31min33%, 47min39%, 60min58%, 70min62%；Column temperature is 30 DEG C,；Volume flow is 1.0mLmin-1, detect ripple
A length of 286nm, analysis time 70min, test sample, reference substance solution sample size are 15 μ L.
（2）Methodological study
Precision test takes No. 1 red sage root normal pressure extract solution need testing solution continuous sample introduction 5 times, records finger-print, calculates
Total peak relative retention time, relative peak area and RSD values, respectively less than ＜ 3% show that instrument precision is good.
Stability test takes precision test need testing solution, and fingerprint image is determined respectively at 0h, 2h, 4h, 8h, 16h, 36h
Spectrum, calculates total peak relative retention time, relative peak area and RSD values, and respectively less than ＜ 3% shows that sample 36h internal stabilities are good
It is good.
Replica test takes 6 parts of red rooted salvias of same batch, accurately weighed, is prepared by " preparation of need testing solution " method
Need testing solution, determines finger-print, calculates total peak relative retention time, relative peak area and RSD values.Result shows, common
There are fingerprint peakses relative retention time, relative peak area RSD ＜ 3%, assay method repeatability is good.
（3）The foundation of red sage root finger-print
The detection of red sage root finger-print takes 10 batch red rooted salvias respectively, accurately weighed, by " the system of need testing solution
It is standby " method prepares need testing solution, and detection, chromatogram in record 70min, by relatively more each peak are prepared by experiment chromatographic condition
Retention time selects the total peak that each collection of illustrative plates is produced, the total peak-to-peak area of record to be shown in Table 9, table 10, Fig. 5, Fig. 6.
The red sage root low temperature extract solution peak area of table 9
The red sage root high temperature extract solution peak area of table 10
Fingerprint similarity calculates the chromatographic fingerprints of Chinese materia medica similarity evaluation published using the Chinese Pharmacopoeia committee
Software (2004A editions) carries out Similarity Measure, is shown in Table 11.
The red sage root extract Similarity Measure result of table 11
From the point of view of extract solution finger-print, both differ greatly；Red sage root high temperature extract solution finger-print chromatographic peak quantity compared with
Many, red sage root low temperature extract solution finger-print chromatographic peak negligible amounts show that classes of compounds contained by red sage root high temperature extract solution is relatively low
Warm extract solution is more, and the dissolution when low temperature is extracted of some compositions is reduced.High temperature extract solution has 6 total peaks with low temperature extract solution,
13min or so chromatographic peaks are Sodium Danshensu, and 21min or so chromatographic peak is protocatechualdehyde, and 56min or so chromatographic peaks are tanshin polyphenolic acid B.
From the point of view of total peak-to-peak area, both are also variant；Low temperature extract solution Sodium Danshensu, protocatechualdehyde peak area are carried much smaller than high temperature
Liquid is taken, tanshin polyphenolic acid B chromatographic peak peak area is much larger than high temperature extract solution.Test above, red sage root low temperature extract solution content of danshinolic acid B
Height, impurity and other compositions are few, and tanshin polyphenolic acid B purity is higher in extract solution.
The parametric measurement of the Salvia root P.E of embodiment 4
Take low temperature（Embodiment 1）, high temperature extract solution in 200mL is concentrated at 70 DEG C of Rotary Evaporators, concentrate is in -5 DEG C
Lower refrigeration 12 hours, forms ice cube, and 24h is dried in freeze-dryer, takes out, and weighs, and in being kept in dark place in drier, determines red
The content of phenolic acid B, low temperature extract is 7.38%, and high temperature extract is 5.62%, and low temperature extract content of danshinolic acid B is significantly higher than
High temperature extract.
The measure of 1 extract physical parameter
（1）The measure of Salvia root P.E specific surface area
Take extract powder in right amount, the specific surface of red sage root high temperature and low temperature extract is determined using specific surface area measuring instrument
Product, independent samples t test comparative analysis is carried out with SPSS17.0 statistical softwares to test data, and measurement result is shown in Table 12
The specific area measuring of the red sage root high temperature extract of table 12 and low temperature extract（Unit：m2/g）
Compare with high temperature extract, * P ＜ 0.05
Statistical analysis, two groups of data variances are neat, the P ＜ 0.05 of two-sided test, it is impossible to think two kinds of Salvia root P.Es
Specific surface area is identical, because the specific surface area average of low temperature extract is more than high temperature extract, therefore the specific surface area of low temperature extract
Noticeably greater than high temperature extract specific surface area, points out the adsorption ability of low temperature extract may be stronger than high temperature extract.
（2）Salvia root P.E porosity measurement
Take extract powder in right amount, the specific surface of red sage root high temperature and low temperature extract is determined using specific surface area measuring instrument
Product, independent samples t test comparative analysis is carried out with SPSS17.0 statistical softwares to test data, and measurement result is shown in Table 13.
The porosity measurement of the red sage root high temperature extract of table 13 and low temperature extract（Unit：cm2/g）
Compare with high temperature extract, * P ＜ 0.05
Statistical analysis, two groups of data variances are neat, the P ＜ 0.05 of two-sided test, and both have notable significant difference, low temperature
The porosity of extract is noticeably greater than high temperature extract porosity.
（3）The particle size determination of Salvia root P.E
Salvia root P.E particle diameter is determined using powder dry method, measurement result is shown in Table 14, Fig. 7.
The particle size determination of the red sage root high temperature extract of table 14 and low temperature extract（Unit：μm）
Result shows that low temperature extract d (0.1), d (0.9) particle diameter are less than high temperature extract, low temperature extract d (0.5) grain
Footpath is more than high temperature extract, and red sage root high temperature extract particle diameter distribution exists in 0.8~2000nm, red sage root low temperature extract particle diameter distribution
0.8~700nm, low temperature extract higher temperatures extract particle diameter distribution is more concentrated.
（4）The measure at Salvia root P.E angle of repose
Salvia root P.E angle of repose is determined using fixed funnel method, funnel is fixed on the top of the drawing paper of horizontal positioned
Certain distance, the extraction powder that will carefully prepare is poured into funnel, and cone is piled on graph paper, measures circular cone
The diameter 2R and height H of body, according to tan θ=H/R, calculate angle of repose θ, are shown in Table 15.
The red sage root high temperature extract of table 15 is determined with the angle of repose of low temperature extract（Unit：Degree）
Independent samples t test comparative analysis, two master data variances are carried out to data above with SPSS17.0 statistical softwares
Together, two-sided test P=0.067 ＞ 0.05, both are less than 40 ° without notable significant difference, high temperature with low temperature extract angle of repose,
Mobility preferably, can meet production needs, and low temperature extract good fluidity is in high temperature extract.
（5）Salvia root P.E heap density and tap density are determined
Salvia root P.E powder 10g is weighed, is placed in 25mL graduated cylinders, volume V is read after gently striking off0, vibration graduated cylinder is extremely
Powder volume reads volume V not in changef, heap density is calculated, tap density the results are shown in Table 16.
The red sage root high temperature extract of table 16 is determined with the heap density and tap density of low temperature extract（Unit：g/mL）
Compare with high temperature extract, * P ＜ 0.05
Independent samples t test comparative analysis is carried out to data above with SPSS17.0 statistical softwares, variance is neat, bilateral inspection
Test heap density and be 0.000 ＜ 0.05 with tap density P values, there is notable significant difference, low temperature extract heap density and jolt ramming
Density is significantly less than high temperature extract.
（6）Salvia root P.E Moisture percentage is determined
Take clean measuring cup constant weight to weigh, take Salvia root P.E tiling（1~2mm of thickness）In measuring cup, in 60 DEG C
Dry 6h, take out, be put in fill discoloration silica gel drier in be dehydrated 12h, weigh, measuring cup is put in and fills supersaturated chlorination
Sodium solution（Relative humidity is 75%）Drier in, close drying device is placed at room temperature, respectively at 0.5,1,2,4,8,12,
24th, 48,72,96,120,144,168h takes out and weighs, by hydroscopicity（%）=（Extract weight before extract weight-moisture absorption after moisture absorption
Amount）Extract weight × 100% formula before/moisture absorption, calculates Moisture percentage, is plotted against time with Moisture percentage, obtains moisture absorption and puts down
Weighing apparatus curve, is shown in Fig. 8：
Data display, during 0~120h, as time went on, Salvia root P.E Moisture percentage is significantly increased, super when the time
After crossing 120h, as time went on, Moisture percentage slowly increases, and shows that Salvia root P.E reaches moisture equilibrium at dry side in 120h.It is red
Darkened after conopsea extraction moisture absorption, and liquefied.Extract balance Moisture absorption percentage is all higher than 15%, illustrates Salvia root P.E pole
Tool draws moist.Low temperature extract balance Moisture absorption percentage is more than high temperature extract balance Moisture absorption percentage, and low temperature extract is higher
Warm extract is more easy to moisture absorption, it may be possible to two kinds of differences of extract ingredient, causes its hygroscopicity to change.
The Salvia root P.E finger-print of embodiment 5 is studied
The preparation precision of need testing solution weighs Salvia root P.E equivalent to crude drug amount 0.15g in 25mL measuring bottles, first
Alcohol constant volume, ultrasonic 30min dissolvings, filtration is removed subsequent filtrate, obtains final product Salvia root P.E need testing solution.
Reference substance solution is prepared by the preparation method of reference substance solution under " research of extract solution finger-print ".
（1）The foundation of finger-print
Need testing solution and reference substance solution are taken, detection, record are prepared by extract solution finger-print experiment chromatographic condition
Chromatogram in 70min, the total peak for selecting each collection of illustrative plates to produce by the retention time at relatively more each peak calculates total peak
Peak area, is shown in Table 17, table 18, Fig. 9.
The red sage root low temperature extract peak area of table 17
The red sage root high temperature extract peak area of table 18
（2）Fingerprint similarity is calculated
The chromatographic fingerprints of Chinese materia medica similarity evaluation software (2004A editions) published using the Chinese Pharmacopoeia committee carries out phase
Calculated like degree, be shown in Table 19.
The Salvia root P.E Similarity Measure result of table 19
From the foregoing, it will be observed that Salvia root P.E finger-print differs greatly.High temperature extract finger-print chromatographic peak is more, low temperature
Extract chromatographic peak is less, and high temperature extract ingredient species is more compared with low temperature extract；High temperature extract danshensu and former youngster
Tea aldehyde chromatographic peak peak area is more than low temperature extract, and tanshin polyphenolic acid B peak area is less than low temperature extract.Show, red sage root low temperature is extracted
Content of danshinolic acid B is more than high temperature extract in thing, and impurity and other compositions content higher temperatures extract are few, low temperature extract pellet phenol
Sour B purity is higher than high temperature extract.
9 conclusions
Tested more than, red sage root low-temperature boiling Dynamic Extraction, tanshin polyphenolic acid B can dissolution to greatest extent, prevent by high temperature point
Solution destruction, impurity and other compositions dissolution are reduced；Higher temperatures are extracted, and low temperature extraction content of danshinolic acid B is high, and purity is big.Due to carrying
Take temperature different, classes of compounds is different from quantity, and extract solution physical parameter and extract powder property are also variant.
Detection method, effectively can efficiently separate the multiple compositions in Salvia root P.E, and peak shape is good, baseline
Steadily, for the quality control of Salvia root P.E provides reliable basis.
Claims (9)
1. the extracting method of the red sage root, it is characterised in that：It includes following operating procedure：Salvia piece is taken, is added water, reduced pressure
Under, 70 ± 5 DEG C of boilings are extracted, and merge extract solution, you can.
2. extracting method according to claim 1, it is characterised in that：Amount of water is 5~15 times of red sage root dry weight.
3. extracting method according to claim 2, it is characterised in that：Amount of water is 12 times of red sage root dry weight.
4. extracting method according to claim 1, it is characterised in that：Extract 1~3 time, 0.5~3h every time.
5. extracting method according to claim 4, it is characterised in that：Extract 3 times, each 2h.
6. a kind of preparation method of Salvia root P.E, it is characterised in that：It includes following operating procedure：
(1) extracted by claim 1-5 any one methods described；
(2) extract solution of step (1) is taken, after solvent, freeze-drying is recovered under reduced pressure, Salvia root P.E is obtained final product.
7. the preparation method of Salvia root P.E according to claim 6, it is characterised in that：The condition that solvent is recovered under reduced pressure
For：- 0.05~-0.095MPa, 40~90 DEG C.
8. the preparation method of Salvia root P.E according to claim 7, it is characterised in that：The condition that solvent is recovered under reduced pressure
For：70±5℃.
9. the Salvia root P.E that prepared by claim 6 to 8 any claim methods described, it is characterised in that：The red sage root is carried
Take the tanshin polyphenolic acid B that thing contains 5~15%.
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