CN101256175A - Method for verification of color spectrum fingerprint pattern peak purity - Google Patents

Method for verification of color spectrum fingerprint pattern peak purity Download PDF

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Publication number
CN101256175A
CN101256175A CNA2007101900873A CN200710190087A CN101256175A CN 101256175 A CN101256175 A CN 101256175A CN A2007101900873 A CNA2007101900873 A CN A2007101900873A CN 200710190087 A CN200710190087 A CN 200710190087A CN 101256175 A CN101256175 A CN 101256175A
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peak
identified
chromatographic
spectrum
chromatographic peak
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邹纯才
鄢海燕
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Wannan Medical College
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Wannan Medical College
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/86—Signal analysis
    • G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
    • G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/86—Signal analysis
    • G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
    • G01N30/8689—Peak purity of co-eluting compounds

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Abstract

The invention discloses a method for testing purity of chromatographic fingerprint peak, which includes the following steps: processing the chromatographic fingerprint to the Chinese medicine by HPLC with DAD tester, obtaining whole information of the fingerprints; extracting chromatographic peaks to be identified in the chromatogram; deducting noise signals from the chromatographic peaks to be extracted and identified; setting standard windows; moving windows and establishing an orthogonal projection matrix; projecting chromatographic peaks to be identified to the orthogonal projection matrix, obtaining the residual spectrum; calculating the included angle between the chromatographic peak to be identified and the residual spectrum; determining whether the cosine of the included angle is less than the noise signal; if the cosine of the included angle is less than the noise signal, the chromatographic peak to be identified is a pure peak; if the cosine of the included angle is more than the noise signal, the chromatographic peak to be identified is a mixed peak. Compared with the prior art, the purity of the chromatographic peak is directly judged without standard or reference substance, the process is fast and easy, and is programmed.

Description

A kind of method of inspection of color spectrum fingerprint pattern peak purity
Technical field
The invention belongs to the method for inspection of chromatographic peak purity, belong to a kind of method of inspection of chromatographic fingerprints of Chinese materia medica peak purity especially, especially belong to the method for carrying out the check of chromatographic fingerprints of Chinese materia medica peak purity with self orthographic projection.
Background technology
Accurately the purity of identification chromatographic peak is the prerequisite of the qualitative and quantitative analysis of chromatogram.Because the chemical constitution of Chinese medicine mostly is secondary metabolite, chemical constitution difference to each other is very little, physicochemical property approximate, and its chromatographic peak unavoidably has overlapping on chromatographic fingerprinting, accurately differentiates chromatographic peak purity and becomes more important.Differentiate purity, the affirmation chromatographic peak of chromatographic fingerprinting chromatographic peak, not only increase the quantity of information of finger-print, and help further investigation Chinese medicine.The qualitative instrumental method and the chemometrics method of being summed up as of chromatographic peak purity, wherein back one method is again the basis of last method.Now Bao Dao chemometrics method has: 1. the progressive figure method of convolution-singular value ( Shen Weiyang, Hu Yuzhu. the application of the progressive figure method of convolution-singular value in chromatographic peak purity is differentiated, computing machine and applied chemistry, 2002,19 (3): 283-284.); 2. the two-dimensional convolution method (Shen Weiyang, Hu Yuzhu. two-dimensional convolution and the application in chromatographic peak purity is identified thereof. SCI, 2003,23 (3): 388-390.); 3. derivative chromatography (lucky peaceful, high Guoqiang, Chen Yu establishes. derivative chromatography is identified the high performance liquid chromatography peak purity. China Medicine University's journal, 1991,22 (5): 295-298.); 4. principal component analysis (PCA) (Prtnclpal component annlysis, be called for short PCA) (strongly fragrant building is etc. the quilitative method of chromatographic peak purity for Huang Fang, Kang Jihong. chromatogram, 1995,13 (1): 33-37.); 5. evolving factor analysis method (Evolving factor analysis, be called for short EFA) (Lorber A.Error propagation andfigures of merit for quantification by solving matrix equations.AnalChem, 1988,58,1167-1168.).
Above method all fails to make full use of the spectral information that respectively flows out component that the coupling chromatographic apparatus is write down in chromatographic separation process.
Summary of the invention
Technical matters to be solved by this invention provides a kind of need and can verify the purity at peak self and make it sequencing the rectangular projection analysis that chromatographic peak carries out self, need not the chromatographic peak purity check method of other reference substance or reference material.
The technical scheme of technical solution problem of the present invention is: a kind of method of inspection of color spectrum fingerprint pattern peak purity, comprise that the high performance liquid chromatograph that contains the DAD detecting device detects material, stored program digital machine, this stored program digital machine is carried out following steps:
A) in the full detail of collection of illustrative plates, extract chromatographic peak to be identified, and the deduction noise signal;
B) window that sets up standard in the chromatographic peak to be identified of deduction noise signal, moving window obtains spectroscopic data, as standard spectrum, and set up the rectangular projection battle array with it, with chromatographic peak to be identified each spectrum vector the rectangular projection battle array is carried out projection, deducted the spectroscopic data at the window place of getting again, get remaining spectrum along the chromatogram direction;
C) whether the angle of calculating chromatographic peak to be identified and remaining spectrum judges included angle cosine less than noise signal, and less than noise signal, chromatographic peak then to be identified is pure peak as if included angle cosine; If included angle cosine is greater than noise signal, chromatographic peak then to be identified is for mixing the peak.
Described noise signal is 0.05.
The application of the method for inspection of the color spectrum fingerprint pattern peak purity of stating in the Big Semen Cassiae finger-print.
The application of the method for inspection of described color spectrum fingerprint pattern peak purity in RADIX CURCUMAE volatile oil finger-print.
Ultimate principle of the present invention is: establish the N * M rank two-dimensional data matrix of X for the chromatographic fingerprinting that adopts the coupling chromatogram and record, their line direction has characterized the outflow information (M delivery time point) of chromatogram, and column direction has characterized spectral information (N absorbing wavelength).Use X aN * m rank the two-dimensional data matrix of the to be identified target components chromatographic peak of expression among the X is considered the background of instrument, can be matrix X aBe divided into two parts, promptly
X a＝X b+X c (1)
X cExpression is from the main information of target components, and X bThen represent background from instrument.In the zero-dose district before and after the target chromatographic peak, respectively choose 1 s (before the target chromatographic peak) and e (behind the target chromatographic peak) respectively, with this spectrum vector B of 2 sAnd B eAnd the spectrum vector B of contrast component c, form the spectrum matrix A.
A＝[B s，B e，B c] (2)
W is the moving window size, at first ends mobile window w from the s place to the e place in A and obtains spectroscopic data, as standard spectrum.Standard spectrum is whenever got a window, sets up the rectangular projection battle array with it:
M＝I-AA + (3)
Wherein, I is a unit matrix, A +Generalized Inverse Matrix for A.Each spectrum vector x with chromatographic peak A to be identified along the chromatogram direction CjM is carried out project (spectroscopic data at the deduction window place of getting)
x cj ＊＝Mx cj (4)
x CjExpression X cJ bar spectrum, x Cj *Expression x CjRemaining spectrum after the projection.Be to eliminate the inequality The noise, make the remaining spectrum x of gained after the projection Cj *With the original spectrum x before the projection CjIncluded angle cosine be r j, promptly
x j = x cj * x cj T / ( | | x cj * | | | | x cj T | | ) - - - ( 5 )
And with r jAs the criterion of differentiating the chromatogram peak purity.
Because M has deducted x CjThe spectral information relevant with A, x Cj *Kept x CjWith the incoherent spectral information of A, so r jBe worth more little, i.e. remaining spectrum x after the projection Cj *With the original spectrum x before the projection CjAngle big more, x is described Cj *Keep x CjSpectral information few more, x CjBig more with the A correlativity; Otherwise r jBe worth greatly more, x is described Cj *Keep x CjSpectral information many more, x CjMore little with the spectral correlation of A.Work as r jValue equals at 0 o'clock, x Cj *With x CjComplete quadrature, M has deducted x fully CjThe spectral information relevant with A, x Cj *Fully only remaining random noise.With projection matrix M to the target chromatographic peak X among the finger-print X cEach row spectrum carry out projection, can be along the spectrum residual error included angle cosine curve of chromatogram direction, if the r of this curve target chromatogram peak time scope jValue is zero or approaches zero, illustrate that then its pairing target chromatographic peak is one to divide identical one matter peak with control group, otherwise the target chromatographic peak is an overlap peak.
Because the complicacy of various material compositions and the delivery time of chromatogram are longer, in the mensuration of chromatographic fingerprinting, exist the background of instrument inevitably, therefore to carry out the background deduction of 2-D data, in order to avoid the chromatogram peak purity is produced erroneous judgement.
The present invention utilizes orthographic projection, by the 2-D data of target chromatographic peak (treat purity differentiate chromatographic peak), gets spectroscopic data with moving window, as standard spectrum, and sets up rectangular projection battle array M with it.Again with the target chromatographic peak along each spectrum vector of chromatogram direction M is carried out projection, computing (spectroscopic data at the deduction window place of getting) must remaining spectrum, included angle cosine with the original spectrum before remaining spectrum and the projection is the purity that identification beacon is estimated chromatographic peak again, can verify self peak purity quickly and easily, owing to adopted orthographic projection, the present invention does not need standard items or reference substance just can directly judge the purity of chromatographic peak, handle quick, easy, sequencing.
Description of drawings
Fig. 1 is a process flow diagram of the present invention.
Fig. 2 gives birth to the HPLC finger-print of product for Big Semen Cassiae.
Fig. 3 is the HPLC finger-print of aloe-emodin, Rhein, archen, Chrysophanol, Physcion standard solution.
Fig. 4 is the remaining spectrum and the original spectrum included angle cosine common pattern figure of Chrysophanol 15.
Fig. 5 is the remaining spectrum and the original spectrum included angle cosine common pattern figure of No. 1 chromatographic peak of need testing solution.
Fig. 6 is the HPLC finger-print of RADIX CURCUMAE volatile oil.
Fig. 7 is the HPLC collection of illustrative plates of germacrone standard items.
Fig. 8 is No. 67 remaining spectrum of peak germacrone and original spectrum included angle cosine mode chart
Fig. 9 is No. 61 not remaining spectrum and the original spectrum included angle cosine mode charts of principal component chromatographic peak.
Figure 10 is No. 64 not remaining spectrum and the original spectrum included angle cosine mode charts of principal component chromatographic peak.
Figure 11 is No. 65 not remaining spectrum and the original spectrum included angle cosine mode charts of principal component chromatographic peak.
In Fig. 1-11,1-16 is the chromatographic peak of Big Semen Cassiae purity to be measured, and 21 is aloe-emodin, and 22 is Rhein, and 23 is archen, and 24 is Chrysophanol, and 25 is Physcion.61-69 is the chromatographic peak of RADIX CURCUMAE volatile oil purity to be measured.
Embodiment
As shown in Figure 1, step 101 is: adopt the high performance liquid chromatograph that contains the DAD detecting device that material is carried out atlas analysis, utilize the MATLAB language to handle, obtain the full detail of collection of illustrative plates, described information comprises the peak area of respective peaks, peak height, retention time.
Step 102 is: extract chromatographic peak to be identified in the full detail of finger-print.
Step 103 is: will deduct the chromatographic peak to be identified that noise signal obtains not having the instrument interference in the chromatographic peak of identification to be extracted; Described noise signal is not more than 0.05.
Step 104 is: the window that sets up standard in the chromatographic peak of deduction noise signal, mobile normal window is set up the orthogonal projection matrix.
Step 105 is: chromatographic peak to be identified is aligned projection matrix carry out projection.
Step 106 is: the spectroscopic data of the projection result of step 105 being deducted the window place of getting obtains remaining spectrum.
Step 107: the angle that calculates chromatographic peak to be identified and remaining spectrum.
Step 108: whether judge included angle cosine less than noise signal, if included angle cosine less than noise signal, then is a step 110, chromatographic peak to be identified is pure peak; If included angle cosine greater than noise signal, then is a step 109, chromatographic peak to be identified is not pure peak.
The present invention is used to differentiate the cassia seed color spectrum fingerprint pattern peak purity.
1 instrument and reagent
Agilent 1100 high performance liquid chromatographs (comprising the Agilent1100 quaternary pump, Agilent 1100 diode array detector, Agilent 1100 chromatogram chem workstations); Cassia seed is given birth to the product medicinal material available from Kiamusze City's medicinal material company, originates in Anhui.Be accredited as the seed of Cassia tora (Cassia obtusifolia L.) through professor Liu Juan of pharmacognosy teaching and research room of Jiamusi University, meet 2005 editions regulations of the Pharmacopoeia of the People's Republic of China.Aloe-emodin reference substance (lot number: 766-200210), archen reference substance (lot number: 756-200110), Rhein reference substance (lot number: 774-200311), Chrysophanol reference substance (lot number: 796-200204), Physcion reference substance (lot number: 758-2004080) all available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute.Acetonitrile, methyl alcohol are chromatographically pure, and hydrochloric acid, phosphoric acid and chloroform are pure for analyzing.
2 specimen preparations
2.1 it is an amount of that reference substance solution takes by weighing aloe-emodin, Rhein, archen, Chrysophanol and Physcion reference substance, is solvent with methyl alcohol, is mixed with 200 μ gmL -1The mixing reference substance solution of aloe-emodin, Rhein, archen, Chrysophanol and five kinds of compositions of Physcion.
2.2 need testing solution depends on the pine torch sample, pulverizes, and crosses 80 mesh sieves, takes by weighing 2.0g, adds 1.5molL -1Hydrochloric acid solution 30mL, reflux 1.5h is put coldly, adds chloroform 25mL, backflow 20min takes out, and divides and gets chloroform layer, and the sour water layer adds chloroform 25mL again, backflow 20min divides and gets chloroform layer, and the same method operation adds chloroform refluxing extraction secondary again, merge four times chloroform extracted solution, add water 20mL jolting wash-out, discard water layer, the chloroform solution evaporate to dryness, residue adds methyl alcohol 15mL makes dissolving, filters, and quantitatively is transferred in the 25mL volumetric flask, adds methyl alcohol to scale, shake up, through 0.45 μ m filtering with microporous membrane, promptly.
3 chromatographic conditions
Chromatographic column: Zorbax Eclipse XDB-C 18(4.6mm * 250mm, 5 μ m), guard column: Zorbax SB-C 18Post (4.6mm * 12.5mm, 5 μ m); Moving phase: acetonitrile (A)-0.1% phosphate aqueous solution (B) is made gradient elution, 0min, 10%A-90%B; 5min, 15%A-85%B; 10min, 40%A-60%B; 40min, 70%A-30%B; 100%A behind the 41min.Flow velocity: 1.0mLmin -1Column temperature: room temperature; Detect wavelength: 278nm, reference wavelength: 550nm; Spectroscopic data recording interval 220～500nm, wavelength interval 2nm; Chromatogram 0～50min writing time; Sample size 10 μ L.
4 target peaks are determined
Depend on that the pine torch need testing solution measures by selected chromatographic condition, write down its chromatogram, the results are shown in Figure 2.Other gets aloe-emodin, Rhein, archen, Chrysophanol and Physcion reference substance mixed solution and measures with method, the results are shown in Figure 3.Comparative control product and cassia seed collection of illustrative plates, and utilize chromatographic work station peak decision-making function, determine that tentatively 12,15, No. 16 peaks among Fig. 2 are respectively archen, Chrysophanol, Physcion target peak.
Chromatogram spectrum 2-D data is gathered in zero component district before and after 12,15, No. 16 peaks of need testing solution chromatogram, is standard spectrum with got window, sets up rectangular projection battle array M with it, with each spectrum vector x along the chromatogram direction of chromatographic peak to be identified CjM is carried out project (spectroscopic data at the deduction window place of getting), calculate the included angle cosine of the spectrum rectangular projection front and back spectrum vector of chromatographic peak to be identified.Be respectively the chromatographically pure peak of archen, Chrysophanol, Physcion in 12,15, No. 16 peaks among Fig. 2 as can be known by result of calculation.Remaining spectrum and original spectrum included angle cosine common pattern figure such as Fig. 4 as No. 15 Chrysophanol.
In like manner, can determine other each purity of principal component chromatographic peak not, be principal component peak not as No. 1, need testing solution chromatographic peak, its remaining spectrum and original spectrum included angle cosine common pattern figure such as Fig. 5 thus, can determine that No. 1 peak is an impure peak.
Differentiate RADIX CURCUMAE volatile oil color spectrum fingerprint pattern peak purity with the present invention.
1 instrument and reagent
Agilent 1100 high performance liquid chromatographs (comprising the Agilent1100 quaternary pump, Agilent 1100 diode array detector, Agilent 1100 chromatogram chem workstations); RADIX CURCUMAE is purchased prepared slices of Chinese crude drugs factory in Beijing is emerging, and identify through professor Shi Quan of crude drug teaching and research room of Jiamusi University in Zhejiang, the place of production, meets the regulation under item of Chinese Pharmacopoeia (2005 editions).Germacrone reference substance (lot number: 111665-200401) available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute.Methyl alcohol is chromatographically pure, and anhydrous sodium sulfate, phosphoric acid are that analysis is pure.
The preparation of 2 test samples
2.1 the RADIX CURCUMAE meal 100g that the volatile oil preparation is learnt from else's experience and pulverized 30 mesh sieves puts in the 2000mL round-bottomed flask, the water logging bubble that adds 8 times of amounts spends the night, and presses the first method of determination of volatile oil among appendix X D of Chinese Pharmacopoeia (2005 editions), and steam distillation is extracted 8h.Collect the volatile oil part, behind the adding anhydrous sodium sulfate dehydration, sealing, dark place refrigeration.
2.2 RADIX CURCUMAE volatile oil 20 μ L are got in the test liquid preparation, put in the 10mL measuring bottle, add methyl alcohol to scale, shake up.Through 0.45 μ m filtering with microporous membrane, make the test liquid of RADIX CURCUMAE volatile oil.Get the about 10mg of germacrone reference substance, the accurate title, decide, and puts in the 100mL volumetric flask, adds methyl alcohol to scale, shakes up, and through 0.45 μ m filtering with microporous membrane, draws filtrate 1mL and put in the 10mL volumetric flask, adds methyl alcohol to scale, preparation germacrone reference substance solution.
3 chromatographic conditions
Chromatographic column: Zorbax Eclipse XDB-C 18(4.6mm * 250mm, 5 μ m), guard column: ZorbaxSB-C 18Post (4.6mm * 12.5mm, 5 μ m); Moving phase: methyl alcohol (A)-0.2% phosphate aqueous solution (B) is made gradient elution, 0min, 55%A-45%B; 5min, 55%A-45%B; 25min, 75%A-25%B; 40min, 75%A-25%B; 50min, 100%A.Flow velocity: 1.0mlmin -1Column temperature: room temperature; Detect wavelength: 210nm; Spectroscopic data recording interval 190～350nm, wavelength interval 2nm; Chromatogram 0～50min writing time; Sampling volume 20 μ L.
4 target peaks are determined
Get RADIX CURCUMAE volatile oil test liquid and measure, write down its chromatogram, the results are shown in Figure 6 by selected chromatographic condition.Other gets the germacrone reference substance and measures with method, the results are shown in Figure 7.Carry out the coupling of color spectrum fingerprint pattern peak through progressive window rectangular projection, No. 67 chromatographic peak is germacrone among Fig. 6 as can be known.
5 chromatographic peak purity are differentiated
Chromatogram spectrum 2-D data is gathered in zero component district before and after No. 67 chromatographic peak of RADIX CURCUMAE volatile oil finger-print, is standard spectrum with got window, sets up rectangular projection battle array M with it, with each spectrum vector x along the chromatogram direction of target chromatographic peak CjM is carried out project (spectroscopic data at the deduction window place of getting), and calculate the included angle cosine of the spectrum rectangular projection front and back spectrum vector of target chromatographic peak.By the result of calculation chromatographically pure peak that is germacrone, No. 67 peaks among Fig. 6 as can be known, its remaining spectrum and original spectrum included angle cosine mode chart are as shown in Figure 8.In like manner, can determine other each purity of principal component chromatographic peak not, as the 61st, 64, No. 65 in the test liquid the chromatographic fingerprinting not remaining spectrum and the original spectrum included angle cosine mode chart of principal component chromatographic peak, as Fig. 9, Figure 10, shown in Figure 11, thus, can determine that No. 61 peaks are a pure peak, 64, No. 65 chromatographic peaks respectively are a mixing peak.

Claims (4)

1, a kind of method of inspection of color spectrum fingerprint pattern peak purity, comprise that the high performance liquid chromatograph that contains the DAD detecting device detects material, testing result is imported stored program digital machine, it is characterized in that: this stored program digital machine is carried out following steps:
A) in the full detail of collection of illustrative plates, extract chromatographic peak to be identified, and the deduction noise signal;
B) window that sets up standard in the chromatographic peak to be identified of deduction noise signal, moving window obtains spectroscopic data, as standard spectrum, and set up the rectangular projection battle array with it, with chromatographic peak to be identified each spectrum vector the rectangular projection battle array is carried out projection, deducted the spectroscopic data at the window place of getting again, get remaining spectrum along the chromatogram direction;
C) whether the angle of calculating chromatographic peak to be identified and remaining spectrum judges included angle cosine less than noise signal, and less than noise signal, chromatographic peak then to be identified is pure peak as if included angle cosine; If included angle cosine is greater than noise signal, chromatographic peak then to be identified is for mixing the peak.
2, the method for inspection of a kind of color spectrum fingerprint pattern peak purity according to claim 1 is characterized in that: described noise signal is 0.05.
3, the application of the method for inspection in the Big Semen Cassiae finger-print of the described color spectrum fingerprint pattern peak purity of a kind of claim 1.
4, the application of the method for inspection of the described color spectrum fingerprint pattern peak purity of a kind of claim 1 in RADIX CURCUMAE volatile oil finger-print.
CNA2007101900873A 2007-11-21 2007-11-21 Method for verification of color spectrum fingerprint pattern peak purity Pending CN101256175A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103328966A (en) * 2011-01-21 2013-09-25 麦丝地科技有限公司 Background subtraction-mediated data-dependent acquisition
CN105518456A (en) * 2013-09-02 2016-04-20 株式会社岛津制作所 Chromatogram data processing device
CN105891397A (en) * 2015-01-26 2016-08-24 大连达硕信息技术有限公司 Comprehensive-two-dimensional-gas-chromatography peak detecting method
CN113624881A (en) * 2021-08-14 2021-11-09 昆明理工大学 Method for simultaneously detecting contents of six components in Sanwu capsule

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103328966A (en) * 2011-01-21 2013-09-25 麦丝地科技有限公司 Background subtraction-mediated data-dependent acquisition
CN103328966B (en) * 2011-01-21 2016-03-16 麦丝地科技有限公司 The data-dependent acquisition of background deduction mediation
US10984996B2 (en) 2011-01-21 2021-04-20 Massdefect Technologies, Llc Background subtraction-mediated data-dependent acquistion
CN105518456A (en) * 2013-09-02 2016-04-20 株式会社岛津制作所 Chromatogram data processing device
CN108918744A (en) * 2013-09-02 2018-11-30 株式会社岛津制作所 Chromatographic data processing method
CN105518456B (en) * 2013-09-02 2019-11-05 株式会社岛津制作所 Chromatography data system
CN105891397A (en) * 2015-01-26 2016-08-24 大连达硕信息技术有限公司 Comprehensive-two-dimensional-gas-chromatography peak detecting method
CN113624881A (en) * 2021-08-14 2021-11-09 昆明理工大学 Method for simultaneously detecting contents of six components in Sanwu capsule

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