CN107643341A - A kind of method for determining the active component in Cistanchis glycosides capsule - Google Patents
A kind of method for determining the active component in Cistanchis glycosides capsule Download PDFInfo
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Abstract
The present invention provides a kind of method for determining the active component in Cistanchis glycosides capsule, and control sample is detected by using ultra performance liquid chromatography, obtains the correction factor f of acteoside and tubulosideAActeoside/TubulosideA, acteoside and different acteoside correction factor fActeoside/Different acteoside, acteoside and echinacoside correction factor fActeoside/Echinacoside, and then by obtained correction factor and treat that the chromatogram of test sample obtains the content of other three kinds of compositions, wherein, when ultra performance liquid chromatography detects by making mobile phase be acetonitrile and 0.1vol% aqueous formic acids；And make acteoside as internal reference thing so that the content of tubulosideA, different acteoside and echinacoside that obtained correction factor is calculated is accurate, and a kind of reference substance is only used only and can be achieved to determine while four kinds of samples, greatlys save cost.
Description
Technical field
Analytical chemistry field of the present invention, more particularly to a kind of method for determining the active component in Cistanchis glycosides capsule.
Background technology
Cistanchis glycosides capsule is the folk prescription being prepared using extracting the Glycosides of Cistanche of gained in Cistanche tubulosa as raw material
Preparation, there is kidney tonifying marrow facilitating, the effect of brain tonic and intelligence development, be clinically used for the light moderate vascular dementia for treating marrow sea deficiency card.
According to Cistanchis glycosides capsule chemical composition result of study, clearly organic phenolic acid class and phenylethanoid glycoside are respectively green
Ortho acid, caffeic acid, echinacoside, acteoside, tubulosideA and different acteoside, and established above-mentioned benzyl carbinol glycoside
The quantitative approach that multicomponent determines simultaneously.But it is expensive because reference substance is not easy to obtain, it directly limit with conventional external standard method
To realize the purpose of its multi objective quality control, cause its method can not be applied in production practices.
Therefore it provides a kind of cost for reducing active ingredient in test Cistanchis glycosides capsule is the technology for needing to solve at present
Problem.
The content of the invention
In view of this, the technical problems to be solved by the invention are in the activity in a kind of measure Cistanchis glycosides capsule is provided
The method of composition, detection method detection testing cost provided by the invention are low.
The invention provides a kind of method for determining active component in Cistanchis glycosides capsule, including：
1) control sample is detected by using ultra performance liquid chromatography, obtains the school of acteoside and tubulosideA
Positive divisor fActeoside/tubulosideA, acteoside and different acteoside correction factor fActeoside/different acteoside, acteoside with
The correction factor f of echinacosideActeoside/echinacoside,
The ultra performance liquid chromatography detection is acetonitrile and 0.1vol% aqueous formic acids with mobile phase；
2) treated using with step 1) identical ultra performance liquid chromatography testing conditions detection Cistanchis glycosides capsule in test sample
The content of acteoside, obtain treating the content of acteoside and tubulosideA in test sample, different acteoside and Echinacea purpurea
The peak area of glycosides；
The content and tubulosideA of the acteoside that the correction factor and step 2 3) obtained by step 1) obtains,
The calculated by peak area of different acteoside and echinacoside obtains the content of tubulosideA, different acteoside and echinacoside.
Preferably, in the detection of the step 1), the elution of mobile phase is gradient elution.
Preferably, in the gradient elution, using acetonitrile as A phases, 0.1vol% aqueous formic acids elute journey as B phases
Sequence is：0~5min：9~13%A, 5~15min：13~15%A, 15~22min：15~17%A, 22~27min：17~
23%A, 27~35min：23~35%A.
Preferably, the flow velocity of mobile phase is 0.15~0.25mL/min in the step 1).
Preferably, the column temperature of detection is 28~32 DEG C in the step 1).
Preferably, the wavelength of detection is 325~335nm in the step 1).
Preferably, the Cistanchis glycosides capsule treats that test sample is prepared in accordance with the following methods：
Take content under Cistanchis glycosides capsule content uniformity item to be mixed with methanol, ultrasound, take supernatant, produce Cistanchis glycosides
Capsule treats test sample.
Preferably, the amount ratio of content and the methanol is 0.1g under the Cistanchis glycosides capsule content uniformity item：
(300~400) mL.
Preferably, the obtained fActeoside/tubulosideAValue be preferably 1.250~1.350, more preferably 1.280~
1.346 most preferably 1.346；The obtained fActeoside/different acteosideValue be preferably 1.250~1.330, more preferably 1.266
~1.270, most preferably 1.266；The obtained fActeoside/echinacosideValue be preferably 1.050~1.200, more preferably
1.135~1.145, most preferably 1.135；
The ultrasonic time is 20~40min.
Compared with prior art, the present invention provides a kind of method for determining the active component in Cistanchis glycosides capsule, passes through
Control sample is detected using ultra performance liquid chromatography, obtains the correction factor of acteoside and tubulosideA
fActeoside/tubulosideA, acteoside and different acteoside correction factor fActeoside/different acteoside, acteoside and Echinacea purpurea
The correction factor f of glycosidesActeoside/echinacoside, and detect desert using with detection control sample identical ultra performance liquid chromatography testing conditions
The total glycosides capsule of Rong treats the content of the acteoside in test sample, obtain treating in test sample the content of acteoside and tubulosideA,
The peak area of different acteoside and echinacoside；Then by obtained correction factor and treat that the chromatogram of test sample obtains it
The content of its three kinds of composition, wherein, when ultra performance liquid chromatography detects by making mobile phase be acetonitrile and 0.1vol% formic acid
The aqueous solution；And make acteoside as internal reference thing so that tubulosideA that obtained correction factor is calculated, different feltwort
The content of glucosides and echinacoside is accurate, and a kind of reference substance is only used only and can be achieved to determine while four kinds of samples, significantly
Cost is saved；Test result indicates that the content and standard for the treatment of each component in test sample that detection method provided by the invention obtains
There was no significant difference for the content for each component that curve method obtains；In addition, testing conditions provided by the invention can also be by chlorogenic acid
Separated well with caffeic acid.
Brief description of the drawings
Fig. 1 is the UPLC figures of mixed reference substance solution and need testing solution at 330nm.
Embodiment
The invention provides a kind of method for determining active component in Cistanchis glycosides capsule, including：
1) control sample is detected by using ultra performance liquid chromatography, obtains the school of acteoside and tubulosideA
Positive divisor fActeoside/tubulosideA, acteoside and different acteoside correction factor fActeoside/different acteoside, acteoside with
The correction factor f of echinacosideActeoside/echinacoside,
The ultra performance liquid chromatography detection is acetonitrile and 0.1vol% aqueous formic acids with mobile phase；
2) treated using with step 1) identical ultra performance liquid chromatography testing conditions detection Cistanchis glycosides capsule in test sample
The content of acteoside, obtain treating the content of acteoside and tubulosideA in test sample, different acteoside and Echinacea purpurea
The peak area of glycosides；
The content and tubulosideA of the acteoside that the correction factor and step 2 3) obtained by step 1) obtains,
The calculated by peak area of different acteoside and echinacoside obtains the content of tubulosideA, different acteoside and echinacoside.
According to the present invention, the present invention detects by using ultra performance liquid chromatography to control sample, obtains verbascose
The correction factor f of glycosides and tubulosideAActeoside/tubulosideA, acteoside and different acteoside correction factor
fActeoside/different acteoside, acteoside and echinacoside correction factor fActeoside/echinacoside；Wherein, the detection chromatographic column
To be preferably Agilent SB-C18RRHD；The detection is acetonitrile and 0.1vol% aqueous formic acids with mobile phase；The flowing
The elution of phase is preferably gradient elution, and in the gradient elution, using acetonitrile as A phases, 0.1vol% aqueous formic acids are as B
Phase, elution program are：0~5min：9~13%A, 5~15min：13~15%, 15~22min：15~17%A, 22~
27min：17~23%A, 27~35min：23~35%A；The flow velocity of the mobile phase is preferably 0.15~0.25mL/min, more
Preferably 0.19~0.21mL/min；The column temperature of the detection is preferably 28~32 DEG C, more preferably 30~31 DEG C；The detection
Wavelength be preferably 325~335nm, more preferably 330nm；The sample size is preferably 2 μ L.
The obtained fActeoside/tubulosideAValue be preferably 1.250~1.350, more preferably 1.280~1.346, it is optimal
Select 1.346；The obtained fActeoside/different acteosideValue be preferably 1.250~1.330, more preferably 1.266~1.270, most
Preferably 1.266；The obtained fActeoside/echinacosideValue be preferably 1.050~1.200, more preferably 1.135~1.145,
Most preferably 1.135；
In addition, in gradient elution, %A refers to volume ratio, such as 9~13%A refers in every 100mL eluant, eluents containing 9~
13mL A；Vol% refers to volumn concentration.
According to the present invention, the present invention also uses total with step 1) identical ultra performance liquid chromatography testing conditions detection desert cistanche
Glycosides capsule treats the content of the acteoside in test sample, obtains treating the peak of the content of acteoside and tubulosideA in test sample
The peak area of area, the peak area of different acteoside and echinacoside；Wherein, in the step, detected with step 1) identical
Condition is pointed out beyond sample difference to be detected, other test condition all sames, such as the mobile phase of detection, type of elution, detection
Column temperature, flow rate of mobile phase etc. it is all identical.
The Cistanchis glycosides capsule treats that test sample is preferably prepared in accordance with the following methods：Take under Cistanchis glycosides capsule content uniformity item
Content mixes with methanol, ultrasound, takes supernatant, produces Cistanchis glycosides capsule and treat test sample.The Cistanchis glycosides capsule loading amount is poor
The amount ratio of content and the methanol is preferably 0.1g under different item：(300~400) mL, more preferably 0.1g：(330~350)
mL；The ultrasonic time is 20~40min, more preferably 30~35min, and the ultrasonic power is preferably 200~300W,
More preferably 250~280W.
The content of the acteoside obtained according to the present invention, the correction factor and step 2 obtained by step 1) with
And the calculated by peak area of tubulosideA, different acteoside and echinacoside obtains tubulosideA, different acteoside and Echinacea purpurea
The content of glycosides, the present invention do not have particular/special requirement to the method for calculating, and survey the calculation formula for commenting method to provide according to one is calculated more
.
The present invention provides a kind of method for determining the active component in Cistanchis glycosides capsule, by using ultra high efficiency liquid phase color
Spectrum detects to control sample, obtains the correction factor f of acteoside and tubulosideAActeoside/tubulosideA, acteoside with it is different
The correction factor f of acteosideActeoside/different acteoside, acteoside and echinacoside correction factor fActeoside/echinacoside, and
Using the feltwort treated with detection control sample identical ultra performance liquid chromatography testing conditions detection Cistanchis glycosides capsule in test sample
The content of glucosides, obtain treating the peak of the content of acteoside and tubulosideA, different acteoside and echinacoside in test sample
Area；Then by obtained correction factor and treat that the chromatogram of test sample obtains the content of other three kinds of compositions, wherein, super
By making mobile phase be acetonitrile and 0.1vol% aqueous formic acids during high performance liquid chromatography detection；And make in acteoside conduct
Join thing so that the content of tubulosideA, different acteoside and echinacoside that obtained correction factor is calculated is accurate, and
A kind of reference substance is only used only to can be achieved to determine while four kinds of samples, greatlys save cost；And the detection method obtains
The content for treating each component that the content of each component and calibration curve method obtain in test sample there was no significant difference, in addition, of the invention
The testing conditions of offer can also separate chlorogenic acid well with caffeic acid.
It is clearly and completely described below in conjunction with the technical scheme of the embodiment of the present invention, it is clear that described implementation
Example only part of the embodiment of the present invention, rather than whole embodiments.It is common based on the embodiment in the present invention, this area
The every other embodiment that technical staff is obtained under the premise of creative work is not made, belong to the model that the present invention protects
Enclose.
Embodiment 1
1st, instrument and material
The Ultra Performance Liquid Chromatography instruments of Agilent 1290 (DAD UV-detectors)；Chromatographic column：Agilent SB-C18RRHD
(3.0 × 100mm, 1.8 μm), Agilent SB-C18RRHT (3.0 × 100mm, 1.8 μm), Agilent XDB-C18RRHT
(3.0 × 100mm, 1.8 μm).Acetonitrile, formic acid are chromatographically pure (import packing), and water is ultra-pure water, and remaining reagent is that analysis is pure.
Chlorogenic acid reference substance (lot number：110753-201415), caffeic acid reference substance (lot number：110885-200102), it is loose
Fruit chrysanthemum glycosides reference substance (lot number：111670-201505), acteoside reference substance (lot number：111530-201310) by China
Food and medicine calibrating research institute provides；TubulosideA reference substance (lot number：150629), different acteoside reference substance (lot number：
15111802) provided by Chengdu Puffy moral Bioisystech Co., Ltd.Cistanchis glycosides capsule (lot number：1310011,
1310021,1310031), provided by Kangyuan Pharmaceutical Co., Ltd., Jiangsu Prov.
2nd, method and result
The selection of 2.1 chromatographic conditions
2.1.1 the selection of Detection wavelength
This experiment composition to be measured in 190~400nm is to need testing solution has carried out full wavelength scanner, as a result chlorogenic acid,
Caffeic acid, echinacoside, acteoside, tubulosideA and different acteoside have maximum suction at 325~335nm wavelength
Peak is received, therefore selects 330nm as Detection wavelength, the results showed that under the conditions of this Detection wavelength, each composition separating degree to be measured is good
It is good.
2.1.2 the selection of column temperature
Investigated for the f durabilities of different column temperatures, using Aglient1290 ultra performance liquid chromatographies system and
Agilent C18Chromatographic column, influence of the different column temperatures (25,28,30,32,35 DEG C) to each relative correction factor has been investigated respectively,
As a result acteoside is not completely separated with tubulosideA under the conditions of 25 DEG C, under the conditions of 35 DEG C caffeic acid with adjacent chromatographic peak not
It can be kept completely separate, show the method to there is certain durable scope to be limited to column temperature, the good tolerance in the range of 28~32 DEG C.
2.1.3 specific detection method
Chromatographic column is Agilent SB-C18RRHD (3.0 × 100mm, 1.8 μm)；With the aqueous formic acid of acetonitrile (A) -0.1%
(B) it is mobile phase；Gradient elution：0~5min：9~13%A, 5~15min：13~15%, 15~22min：15~17%A,
22~27min：17~23%A, 27~35min：23~35%；Volume flow：0.2mL·min-1；Column temperature：30℃；Detect ripple
It is long：330nm；Sample size：2μL.Each composition separating degree to be measured is good under the conditions of this, and chromatogram is shown in Fig. 1, and Fig. 1 mixing reference substances are molten
The UPLC figures of liquid and need testing solution at 330nm, wherein, A mixing reference substance B. Cistanchis glycosides capsule test samples, 1. chlorogenic acids
2. 3. echinacoside of caffeic acid, 4. acteoside, 5. tubulosideA, 6. different acteoside
The preparation of 2.2 reference substance solutions and need testing solution
2.2.1 the selection of need testing solution (Cistanchis glycosides capsule treats test sample) preparation method
This experiment has carried out different solvents, and different extracting modes, different extraction times, different solvents dosage supplies examination
The investigation of product solution manufacturing method.By investigating 30% methanol, 50% methanol, 70% methanol, 100% methanol, 30% ethanol,
50% ethanol, 70% ethanol, 100% ethanol find, the peak shape obtained when 100% methanol is as Extraction solvent it is preferable and it is to be measured into
Divide content higher；Investigate different extracting modes (ultrasound, flow back) different extraction times (5min, 10min, 15min, 30min,
45min, 60min) find, ultrasound and reflux type are little on content influence, because ultrasound is more convenient, therefore select ultrasound to be used as extraction
Mode, and component content to be measured is in increasing trend with ultrasonic time, but changes of contents is little after 30min, therefore final choice is ultrasonic
30min。
2.2.2 the preparation of reference substance solution (control sample)
Precision weighs chlorogenic acid, caffeic acid, acteoside, tubulosideA, different acteoside and echinacoside reference substance
In right amount, it is separately added into methanol and dissolves and dilute to be made in every 1mL and contains 7.43 μ g, 10.20 μ g, 109.46 μ g, 123.97 μ g,
210.40 μ g and 1214.24 μ g mixed reference substance solution, shake up, and produce control sample (reference substance).
2.2.3 the preparation of need testing solution
Content about 0.15g under Cistanchis glycosides capsule content uniformity item is taken, it is accurately weighed, put in conical flask, precision adds
50mL methanol, ultrasonic 30min (250W, 40Hz), lets cool, then precision measures 1mL to 10mL measuring bottles, adds methanol dilution to scale,
Shake up, centrifuge (12000rmin-1, 5min), supernatant is taken, Cistanchis glycosides capsule is produced and treats test sample.
2.3 linear relationships are investigated
Accurate mixed reference substance solution 0.25,0.5,1.0,2.0,4.0,10.0mL of drawing is put in 10mL measuring bottles respectively, is added
Methanol dilution is shaken up to scale, accurate respectively to draw above-mentioned each 2 μ L of solution, is injected super Ultra Performance Liquid Chromatography instrument, is determined, with
Reference substance mass concentration is abscissa (X), using integrating peak areas value as ordinate (Y), carries out linear regression, obtains regression equation：
Chlorogenic acid：Y=26.11X+0.07223, r2=0.99998, the range of linearity：0.1858~7.4315 μ gmL-1；Caffeic acid：Y
=46.899X+2.2435, r2=0.99998, the range of linearity：0.2550~10.2000 μ gmL-1；Acteoside：Y=
16.21X+0.99338 r2=0.99998, the range of linearity：2.7364~109.4550 μ gmL-1；TubulosideA：Y=
12.104X-0.54241 r2=0.99995, the range of linearity：3.0993~123.9702 μ gmL-1；Different acteoside：Y=
12.825X+1.8102 r2=0.99999, the range of linearity：5.2602~210.4064 μ gmL-1；Echinacoside：Y=
14.792X-82.167 r2=0.9997, the range of linearity：30.3559~1214.2355 μ gmL-1。
2.4 precision test
Precision draws height, in, low various concentrations mixed reference substance solution, continuous sample introduction 6 times, peak area is recorded, it is as a result green
Ortho acid, caffeic acid, acteoside, tubulosideA, the RSD values of different acteoside and echinacoside peak area are respectively
0.44%, 0.19%, 0.16%, 0.15%, 0.20%, 0.95% (height)；1.37%, 1.17%, 1.05%, 1.04%,
1.06%, 1.66% (in)；0.87%, 0.60%, 0.58%, 0.60%, 0.74%, 0.75% (low), show in this condition
Lower sample introduction precision is good.
2.5 stability test
Take mixed reference substance solution and need testing solution (lot number：1310011), respectively at 0h, 2h, 4h, 8h, 12h, 16h,
20h, 24h sample introduction, peak area is recorded, calculate the RSD values of peak area.As a result mixed reference substance solution Content of Chlorogenic Acid, caffeic acid, hair
Stamen spends glucosides, and tubulosideA, the RSD values of different acteoside and echinacoside peak area are respectively 1.01%, 0.70%,
0.71%, 0.56%, 0.49%, 0.49%；Need testing solution Content of Chlorogenic Acid, caffeic acid, acteoside, tubulosideA, different hair
It is respectively 1.01%, 1.96%, 1.82%, 1.58%, 0.89% that stamen, which spends the RSD values of glucosides and echinacoside peak area,
1.16%, show that mixed reference substance solution and need testing solution are preferable in 24h internal stabilities.
2.6 replica test
Take need testing solution (lot number：1310011) 6 parts are prepared by need testing solution preparation method is parallel, by above-mentioned
Chromatographic condition surveys each composition quality fraction, calculates RSD values, as a result need testing solution Content of Chlorogenic Acid, caffeic acid, acteoside,
TubulosideA, the RSD values of different acteoside and echinacoside content are respectively 2.24%, 1.44%, 1.82%, 2.04%,
0.60%, 0.44%, show the repeatability of this method preferably.
2.7 sample-adding recovery tests
Precision weighs content (lot number under Cistanchis glycosides capsule content uniformity item：1310011), parallel 9 parts, 3 groups are divided, point
Do not sequentially add it is low, in, high 3 kinds of various concentrations (it is low, in, high concentration reference substance solution addition is is taken need testing solution to treat
Survey the 50% of composition amount, 100%, 150%) chlorogenic acid, caffeic acid, acteoside, tubulosideA, different verbascose
Glycosides and echinacoside reference substance, each composition quality fraction is determined by above-mentioned chromatographic condition, calculate average recovery and RSD values, knot
Light green ortho acid, caffeic acid, acteoside, tubulosideA, the mean sample recovery rate difference of different acteoside and echinacoside
For 97.34%, 100.96%, 99.29%, 103.27%, 102.41%, 98.72%；RSD values are respectively 1.05%,
1.97%, 1.61%, 1.30%, 1.50%, 1.75%, show that the degree of accuracy of the method is good.
Embodiment 2
The determination of relative correction factor
The measure of 1 relative correction factor
Mixed reference substance solution under Example 1 " 2.3 ", according to the detection method of 2.1.3 parts in embodiment 1, often
Individual sample feeding 2 times, the peak area of each composition is determined, the average peak area of each composition is calculated, according to the meter of relative correction factor
Formula is calculated, caffeinic f is calculated using chlorogenic acid as internal reference thing, tubulosideA, different hair are calculated using acteoside as internal reference thing
Stamen spends the f of glucosides and echinacoside, as a result f chlorogenic acids/caffeic acid, f acteosides/tubulosideA, f acteosides/different hair
Stamen spends glucosides, and f acteosides/echinacoside is respectively 0.538,1.346,1.266,1.135；RSD values are respectively 2.11%,
0.28%, 0.37%, 1.65%, it the results are shown in Table 1.
The relative correction factor of 1 each composition of table
Influence of the 2 different volumes flows to f
Different volumes flow is respectively 0.19mLmin-1, 0.20mLmin-1, 0.21mLmin-1When, it is right to investigate its
The influence of each ingredient f, as a result RSD values are respectively 0.39%, 1.90%, 1.48%, 1.95%, show to finely tune volume flow to each
Ingredient f does not make significant difference.
Influence of the 3 different column temperatures to f
When different column temperatures are respectively 28 DEG C, 30 DEG C, 32 DEG C, its influence to each ingredient f is investigated, as a result RSD values are respectively
0.50%, 2.65%, 1.90%, 2.47%, show that finely tune column temperature does not make significant difference to each ingredient f.
Influence of the 4 different Detection wavelengths to f
Different Detection wavelengths are respectively 328nm, when 330nm, 332nm, investigate its influence to each ingredient f, as a result RSD values
Respectively 2.06%, 0.71%, 1.77%, 0.00%, show that finely tune Detection wavelength does not make significant difference to each ingredient f.
Influence of the 5 different chromatographic columns to f
Investigate different chromatographic columns (the Agilent SB-C with brand same specification18RRHD (3.0 × 100mm, 1.8 μm),
Agilent SB-C18RRHT (3.0 × 100mm, 1.8 μm), Agilent XDB-C18RRHT (3.0 × 100mm, 1.8 μm)) it is right
The influence of each ingredient f, as a result RSD values are respectively 0.48%, 1.01%, 1.48%, 0.40%, show this 3 with brand chromatogram
Post does not make significant difference to each ingredient f.
Influence of the 6 different experiments personnel to f
Influence of the different experiments personnel (experimenter A, experimenter B, experimenter C) to each ingredient f is investigated, as a result
RSD values are respectively 1.69%, 1.79%, 0.64%, 1.09%, show that each ingredient f is without notable obtained by different experiments human users
Influence.
Combined influence of the 7 each durable sexual factors to f
In summary various factors, f chlorogenic acids/caffeic acid, f acteosides/tubulosideA, f acteosides/different hair
Stamen spends glucosides, and f acteosides/echinacoside RSD values are respectively 1.09%, 1.69%, 1.58%, 1.88%, be the results are shown in Table
2。
Combined influence of the various factors of table 2 to relative correction factor
Embodiment 3
The positioning of component chromatographic peak to be measured
Detection method according to 2.1.3 parts in embodiment 1 detects to determinand
The positioning of composition chromatographic peak to be measured, caffeic acid, tubulosideA, different feltwort are carried out by using relative retention time
The average relative retention time of glucosides and echinacoside is respectively that 0.790,0.987,0.928,1.603, RSD value is respectively
0.96%, 0.21%, 0.38%, 0.45%.
Embodiment 4
The comparison of QAMS methods and calibration curve method measurement result
Detection method according to 2.1.3 parts in embodiment 1 detects to determinand, using mark song method to Cistanchis glycosides
Capsule carries out Multiple components and determined simultaneously, then carries out quantitative calculating with the QAMS methods (survey comments method) established, to verify QAMS
Method is used for the accuracy of multi objective quality evaluation in Cistanchis glycosides capsule, the results are shown in Table 3, the benzyl carbinol glycoside that two methods measure
There was no significant difference for the amount of each composition, relative deviation (RD) ＜ 5%, and the method for showing to establish has reliability.
The mark song method (a) of table 3 and QAMS methods (b) measure tubulosideA, the result of different acteoside and echinacoside
When can be seen that by the result of table 3 to mark song method compared with the result that QAMS methods measure, benzyl carbinol glycoside
Each composition tubulosideA, the content deviation of different acteoside and echinacoside is respectively less than 5%, due to acteoside and pine nut
The content difference of chrysanthemum glycosides is larger so that the result error that two methods measure is slightly larger compared with other two kinds of compositions, but permits in deviation
Perhaps in the range of, show that the QAMS methods that this experiment is formulated can to the assay of phenylethanoid glycoside in Cistanchis glycosides capsule
Lean on.
And for two methods of the comparison of organic liposoluble ingredient measurement result
For organic liposoluble ingredient caffeic acid and chlorogenic acid, although other feasibility studies of QAMS methods are satisfied by requiring,
But the content results deviation that two methods measure is larger.The reason for its is possible is analyzed, first, caffeic acid is total in desert cistanche with chlorogenic acid
Content in glycosides capsule is relatively low, second, architectural difference, because both structures differ the structure of a quininic acid, UV absorption has
Certain difference, so as to cause the content results of QAMS methods and regression curve method difference.Therefore, from assay accuracy
Angle set out, it is proposed that chlorogenic acid and caffeic acid in Cistanchis glycosides capsule are contained using external standard method or equation of linear regression method
It is fixed to measure.
This research carries out multicomponent content to Cistanchis glycosides capsule using UPLC ultra-performance liquid chromatographies and determined simultaneously, pole
The earth has saved analysis time and mobile phase solvent dosage；And QAMS methods are applied to the multi objective matter of Cistanchis glycosides capsule first
Measure in evaluation model, and to the QAMS methods established while determine Multiple components and carried out feasibility study, the results showed that QAMS
Method reference substance is in short supply and multi-target ingredient testing cost costliness in the case of show big advantage, and then prove QAMS methods
There is good application prospect in the multi objective quality control of Chinese medicine and evaluation model.
The explanation of above example is only intended to help the method and its core concept for understanding the present invention.It should be pointed out that pair
For those skilled in the art, under the premise without departing from the principles of the invention, the present invention can also be carried out
Some improvement and modification, these are improved and modification is also fallen into the protection domain of the claims in the present invention.
Claims (10)
1. a kind of method for determining active component in Cistanchis glycosides capsule, including：
1) control sample is detected by using ultra performance liquid chromatography, obtain the correction of acteoside and tubulosideA because
Sub- fActeoside/tubulosideA, acteoside and different acteoside correction factor fActeoside/different acteoside, acteoside and pine nut
The correction factor f of chrysanthemum glycosidesActeoside/echinacoside,
The ultra performance liquid chromatography detection is acetonitrile and 0.1vol% aqueous formic acids with mobile phase；
2) using the hair stamen treated with step 1) identical ultra performance liquid chromatography testing conditions detection Cistanchis glycosides capsule in test sample
The content of flower glucosides, obtains treating the content of acteoside and tubulosideA in test sample, different acteoside and echinacoside
Peak area；
3) content and tubulosideA of the acteoside that the correction factor and step 2 obtained by step 1) obtains, different hair
Stamen spends the calculated by peak area of glucosides and echinacoside to obtain the content of tubulosideA, different acteoside and echinacoside.
2. the method for active component in measure Cistanchis glycosides capsule according to claim 1, it is characterised in that the step
1) in detection, the elution of mobile phase is gradient elution.
3. the method for active component in measure Cistanchis glycosides capsule according to claim 2, it is characterised in that the gradient
In elution, using acetonitrile as A phases, 0.1vol% aqueous formic acids are as B phases, elution program：0~5min：9~13%A, 5
~15min：13~15%A, 15~22min：15~17%A, 22~27min：17~23%A, 27~35min：23~35%
A。
4. the method for active component in measure Cistanchis glycosides capsule according to claim 1, it is characterised in that the step
1) flow velocity of mobile phase is 0.15~0.25mL/min in.
5. the method for active component in measure Cistanchis glycosides capsule according to claim 1, it is characterised in that the step
1) column temperature of detection is 28~32 DEG C in.
6. the method for active component in measure Cistanchis glycosides capsule according to claim 1, it is characterised in that the step
1) wavelength of detection is 325~335nm in.
7. the method for active component in measure Cistanchis glycosides capsule according to claim 1, it is characterised in that the desert cistanche
Total glycosides capsule treats that test sample is prepared in accordance with the following methods：
Take Cistanchis glycosides capsule 's content to be mixed with methanol, ultrasound, take supernatant, produce Cistanchis glycosides capsule and treat test sample.
8. the method for active component in measure Cistanchis glycosides capsule according to claim 7, it is characterised in that the desert cistanche
The amount ratio of total glycosides capsule content and the methanol is 0.1g：(300~400) mL.
9. the method for active component in measure Cistanchis glycosides capsule according to claim 7, it is characterised in that the ultrasound
Time be 20~40min.
10. the method for active component in measure Cistanchis glycosides capsule according to claim 1, it is characterised in that described
fActeoside/tubulosideAValue be preferably 1.250~1.350, more preferably 1.280~1.346, most preferably 1.346；It is described
fActeoside/different acteosideValue be preferably 1.250~1.330, more preferably 1.266~1.270, most preferably 1.266；It is described
fActeoside/echinacosideValue be preferably 1.050~1.200, more preferably 1.135~1.145, most preferably 1.135.
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|---|---|---|---|---|
| CN105628798A (en) * | 2014-10-29 | 2016-06-01 | 江苏康缘药业股份有限公司 | Cistanchis glycoside capsule fingerprint detection method |
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| CN112630327A (en) * | 2020-12-03 | 2021-04-09 | 武汉职业技术学院 | Quick detection method for active ingredients of cistanche |
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