CN104390926A - Online rapid detection method of herba andrographis concentrated decolorization process - Google Patents
Online rapid detection method of herba andrographis concentrated decolorization process Download PDFInfo
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Abstract
The invention provides an online rapid detection method of a herba andrographis concentrated decolorization process. An online UV detection device is installed, a concentrated solution sample and a corresponding online UV spectrum are collected, the content of a quality control index in a sample solution is detected by using a high performance liquid chromatography and a partial least squares method is used for establishing a quantitative calibration model and the test is implemented. The model established by the invention has good calibration and prediction abilities, and has the advantages that the contents of andrographolide and dehydroandrographolide in the herba andrographis extract concentrated decolorization process can be detected more quickly and accurately compared with the traditional analysis method; the detection method is favorable for improving the quality control level of traditional Chinese medicine products in the production process, and the stability and the reliability of product quality are ensured.
Description
Technical field
The invention belongs to process analysis technique field, be specifically related to the online ultraviolet spectrum method for quick that a kind of Herba Andrographitis concentrates decolorization.
Background technology
The concentrated decolorization of Herba Andrographitis extract is one of main production run of Herba Andrographitis Chinese patent drug, and substantial connection effect and the quality stability of patent medicine.Obtain extract with 95% alcohol steep Herba Andrographitis medicinal material in commercial production, heating is concentrated to add medicinal carbon simultaneously and decolours.In this process, andrographolide and Dehydro and drographolide are principal ingredient.Because the active constituent content of Herba Andrographitis different parts is different, usually make each batch of initial concentration entering concentrated decolorization process inconsistent; On the other hand, the quality controling mode many employings off-line in production or empirical means, as high performance liquid chromatography, densimetry etc., be unfavorable for Real-Time Monitoring product quality, be both unfavorable for quality stability and the validity of final products, make again production not have economy and high efficiency.Therefore one in real time, fast and accurately quality determining method urgently research and develop, this contributes to ensureing Herba Andrographitis product quality, and progress and the development of centering medicine engineering are significant.
Online ultraviolet technology be a kind of fast, harmless analytical technology, have express-analysis, sample process simple, without the need to consuming the features such as reagent.Must set up quantitative calibration models when using the ultraviolet band of 200 nm to 400 nm to carry out fast quantitative analysis, the multivariate statistical method being most commonly used to Modling model is at present partial least square method (Partial Least Squares Regression, PLSR).The research that current quality control in Chinese medicine production critical process is applied to online ultraviolet technology has part document patent, as patent (number of patent application: 201320490161.4,201310350212.8) etc., what study is that online ultraviolet is for the quick detection in extraction of traditional Chinese medicine, document has reports such as " the red extract quality of the pellet based on ultraviolet spectrum express-analysis technology-Yan Binjun ", and online ultraviolet is used for Herba Andrographitis and concentrates the research of decolorization there is not been reported.
Summary of the invention
A kind of Herba Andrographitis is the object of the present invention is to provide to concentrate decolorization on-line quick detection method, application the method can measure the content of Herba Andrographitis extract andrographolide and Dehydro and drographolide in concentrated decolorization quickly and accurately, is corresponding production run and quality control supplying method.
The present invention is achieved through the following technical solutions:
(1) online ultraviolet detector is installed
Online ultraviolet detector is formed primarily of heating arrangement, reactor, peristaltic pump, filtrator, sample tap, flow cell, ultraviolet-visible radiant, online ultraviolet spectrometer (UVS), worktable, circulation line, cooling pump, distillate receiving bottle, reactor connects peristaltic pump, filtrator, sample tap, flow cell formation loop by circulation line, ultraviolet-visible radiant, flow cell, online ultraviolet spectrometer (UVS), worktable connect with cascade successively, one end coupled reaction still of cooling pump, the other end connects distillate receiving bottle.
Solution in reactor pumps into circulation line by wriggling, and circulation line adopts medical rubber hose.Light path is that the flow cell of 2 mm is connected with UV, visible light radiant and online ultraviolet spectrometer (UVS) respectively by optical fiber, carries out spectra collection.Spectra collection is dynamic acquisition, and with 95% ethanolic solution for background, spectral scan scope is 200 ~ 900 nm, and resolution is 0.8 nm.
In device, distillation fraction uses the condensation of ethanol coolant circulation pump to distillate solvent, and condensate temperature is set to-10 ~-5 DEG C, and distillate is received by conical flask.
(2) the concentrate sample of online acquisition Herba Andrographitis and online ultraviolet spectrum corresponding to sample
Add Herba Andrographitis extract and medicinal carbon in a kettle., the collection of sample solution and spectrum is carried out according to specific time point, after starting heating, open peristaltic pump simultaneously, with the rotating speed conductive solution of 48 rpm, by sample tap collecting sample liquid, control to gather corresponding online ultraviolet spectrum by line ultraviolet spectrometer (UVS) by worktable at flow cell, filtrator is for the activated charcoal in filtering solution and other solid impurities simultaneously.In online ultraviolet detector, solution is successively through peristaltic pump, filtrator, sample tap, flow cell, returns reactor along circulation line.Gather corresponding online ultraviolet spectrum by line ultraviolet spectrometer (UVS) while gathering concentrate sample, collect the experiment sample of some batches, divide the foundation that calibration set participates in quantitative calibration models, all the other carry out the inspection of model prediction ability as checking collection.
(3) the quality control index content in high effective liquid chromatography for measuring concentrate sample is adopted
Adopt andrographolide, the Determination of Dehydroandrographoliin in high effective liquid chromatography for measuring Herba Andrographitis sample liquid.Liquid-phase condition is: chromatographic column is C18 (5 μm of 250 × 4.6 mm); Mobile phase is methanol-water gradient elution, flow velocity 0.8 mL/min; Column temperature 35 DEG C; Initial wavelength 225 nm, 16 min are converted to 254 nm.Elution program is: 0 ~ 15 min, 55%B; 15 ~ 16 min, 55%B → 65%B; 16 ~ 26 min, 65%B.Extracting sample solution, the syringe filter through 0.45 μm of miillpore filter filters, and gets the analysis of subsequent filtrate sample introduction.
(4) foundation of quantitative calibration models and checking
Use partial least square method each index to be carried out to the foundation of quantitative modeling, the lactone content that the spectroscopic data collected and high performance liquid chromatography record is associated.First carry out exceptional spectrum before modeling and contain surveying discriminating data, rejecting abnormalities point, to improve model performance.With first order derivative, second derivative, Savitzky-Golay is level and smooth, Norris is level and smooth etc., and method carries out pre-service to spectrum, and selects suitable modeling wave band, by more each model index determination optimization model.Wherein, with model coefficient R, calibration set error mean square root RMSEC, checking collection error mean square root RMSEP, cross validation error root mean square RMSECV is model performance index.R is more less and more close close to 1, RMSEC and RMSEP, then model performance is better.RSEC and RSEP is the smaller the better, and General Requirements is less than 10%.
Through software computing, andrographolide model selection original spectrum does not make smoothing processing, and modeling wave band is 201 ~ 298 nm, has good modeling effect; The preprocess method of Dehydro and drographolide model selection is second derivative method and Norris smoothing processing, and modeling wave band is 263 ~ 311 nm and 326 ~ 341 nm, can build good quantitative calibration models; The coefficient R of the quantitative calibration models of result display andrographolide and Dehydro and drographolide is more than 0.9, RMSEP and RMSEC is less and close, and RSEP value is all below 5%, and visible model tuning effect is better, and predictive ability is strong.
Be taken at line ultraviolet spectrum, Stochastic choice is a collection of as checking collection, all the other participate in the foundation of calibration model, not make the original spectrum of smoothing processing, carry out the foundation of the calibration model of andrographolide at 201 ~ 298 nm wave bands, separately using second derivative and with Norris smoothly as preprocess method, choose 263 ~ 311 nm and 326 ~ 341 nm as modeling wave band, set up the calibration model of Dehydro and drographolide, to verify that the set pair analysis model is tested.
Online ultraviolet technology is incorporated into the concentrated decolorization of Chinese medicine by the present invention, adopts PLSR algorithm, achieves the detection fast and accurately of andrographolide and Dehydro and drographolide when concentrating decolouring to Herba Andrographitis extract.Model tuning and the predictive ability of the present invention's foundation are good, compared with traditional analysis, the content that Herba Andrographitis extract concentrates andrographolide and Dehydro and drographolide in decolorization can be measured quickly and accurately, be conducive to the quality control level improving tcm product, ensure the stable, reliable of product quality.The present invention is easy and simple to handle, and model prediction ability is good, and stability is strong.
Accompanying drawing explanation
Accompanying drawing 1 is that Herba Andrographitis extract concentrates the online ultraviolet detector figure of decolorization.
Accompanying drawing 2 is Determination of Andrographolide changing trend diagrams that 9 experimentai batches record.
Accompanying drawing 3 is that 9 experimentai batches record Determination of Dehydroandrographoliin changing trend diagram.
Accompanying drawing 4 is online ultraviolet primary light spectrograms of a typical batch.
Accompanying drawing 5 is andrographolide checking collection actual value and the figure that predicts the outcome.
Accompanying drawing 6 is Dehydro and drographolide checking collection actual value and the figure that predicts the outcome.
Embodiment
The present invention is further described in conjunction with the accompanying drawings and embodiments.
embodiment 1:
1. online ultraviolet detector is installed
See Fig. 1, online ultraviolet detector forms by with lower part: described online UV detect system mainly comprises heating arrangement 1, reactor 2, peristaltic pump 3, filtrator 4, sample tap 5, flow cell 6, ultraviolet-visible radiant 7, online ultraviolet spectrometer (UVS) 8, worktable 9, circulation line 10, cooling pump 11, distillate receiving bottle 12, reactor 2 connects peristaltic pump 3 by circulation line 10, filtrator 4, sample tap 5, flow cell 6 forms loop, ultraviolet-visible radiant 7, flow cell 6, online ultraviolet spectrometer (UVS) 8, worktable 9 cascade connects, one end coupled reaction still 2 of cooling pump 11, the other end connects distillate receiving bottle 12.
Solution in reactor 2 imports circulation line 10 by peristaltic pump 3, and circulation line 10 adopts medical rubber hose.Light path is that the flow cell 6 of 2 mm is connected with UV, visible light radiant 7 and online ultraviolet spectrometer (UVS) 8 respectively by optical fiber, carries out spectra collection.
In device, distillation fraction uses the condensation of ethanol coolant circulation pump 11 to distillate solvent, and condensate temperature is set to-10 ~-5 DEG C, and distillate is received by conical flask 12.
2. the concentrate sample of online acquisition Herba Andrographitis and the online ultraviolet spectrum of corresponding sample
3 L Herba Andrographitis extracts and 20.30 g(15% quality of medicinal materials are added in reactor 2) medicinal carbon, open peristaltic pump 3 simultaneously, with the rotating speed conductive solution of 48 rpm, by sample tap 5 collecting sample liquid, control to gather online ultraviolet spectrum by worktable 9 at flow cell 6.Filtrator 4 is for the activated charcoal in filtering solution and other solid impurities.Through preliminary experiment, 68 min are needed with the concentrated decolorization that 50% solvent distillates as terminal, after starting heating, the collection of sample solution and spectrum is carried out according to specific time point, specifically in table 1, gather corresponding online ultraviolet spectrum while gathering concentrate sample, each sample solution gathering 10 mL volumes, be stored in 15 mL EP pipes, each batch is collecting sample liquid and each 36 parts of spectrum altogether.Have collected the experiment sample of 9 batches altogether, divide the foundation that calibration set participates in quantitative calibration models, all the other carry out the inspection of model prediction ability as checking collection.
Spectra collection is dynamic acquisition, and with 95% ethanolic solution for background, spectral scan scope is 200 ~ 900 nm, and resolution is 0.8 nm.
3. adopt the quality control index content in high effective liquid chromatography for measuring concentrate sample
Adopt andrographolide, the Determination of Dehydroandrographoliin in high effective liquid chromatography for measuring Herba Andrographitis sample liquid.
Agilent liquid-phase condition: chromatographic column is Dikma Technologies Diamonsil C18 (5 μm of 250 × 4.6 mm); Mobile phase is methanol-water gradient elution, flow velocity 0.8 mL/min; Column temperature 35 DEG C; Sample size 5 μ L; Initial wavelength 225 nm, 16 min are converted to 254 nm.Elution program is: 0 ~ 15 min, 55%B; 15 ~ 16 min, 55%B → 65%B; 16 ~ 26 min, 65%B.Sample this liquid about 1 mL, the syringe filter through 0.45 μm of miillpore filter filters, and gets the analysis of subsequent filtrate sample introduction.
Sample introduction analysis after gradient dilution reference substance solution, andrographolide is good in 0.04 ~ 1.92 mg/mL concentration range internal linear, and linear equation is y=1158x-27.20, R
2=1.000, Dehydro and drographolide is good in 0.03 ~ 1.48 mg/mL concentration range internal linear, and linear equation is y=759.1x-9.55, R
2=1.000.
The situation of change of concentrated decolorization each batch of andrographolide and Determination of Dehydroandrographoliin is see Fig. 2 ~ Fig. 3.
4. the foundation of quantitative calibration models and checking
Use the partial least square method of TQ Analyst software Determination of Andrographolide to be carried out to the foundation of quantitative model, the lactone content that the spectroscopic data collected and high performance liquid chromatography record is associated.First carry out exceptional spectrum before modeling and contain surveying discriminating data, rejecting abnormalities point, to improve model performance.With first order derivative, second derivative, Savitzky-Golay is level and smooth, Norris is level and smooth etc., and method carries out pre-service to spectrum, to eliminate the impact such as baseline wander, noise, and selects suitable modeling wave band, by more each model index determination optimization model.
Get batch 5 conduct checking collection in the experimental data of 9 batches, all the other batches participate in the foundation of calibration models, and Fig. 4 is the original online ultraviolet spectrogram collected.Through the computing of TQ software, andrographolide, at the wave band of 201 ~ 298 nm, adopts original spectrum and does not do smoothing processing, have good modeling effect, calibration model coefficient R is 0.967, RMSEC is 0.547, RMSEP is 0.325, and modelling verification result RSEP is 3.27%.The model prediction result of andrographolide is see Fig. 5.
As can be seen from above modeling result, the coefficient R of the quantitative calibration models of andrographolide is more than 0.9, RMSEP and RMSEC is less and close, and RSEP value is all below 5%, and visible model tuning effect is better, and predictive ability is strong.
embodiment 2:
According to the method for embodiment 1, difference part is to set up quantitative calibration models to Dehydro and drographolide.
1. select the sample data identical with embodiment 1
In the experimental data of 9 batches, get batch 5 conduct checking collection, all the other batches participate in the foundation of calibration models.First carry out exceptional spectrum before modeling and contain surveying discriminating data, rejecting abnormalities point, to improve model performance.
2. the foundation of the quantitative calibration models of Dehydro and drographolide
Use the partial least square method of TQ Analyst software Determination of Dehydroandrographoliin to be carried out to the foundation of quantitative model, the lactone content that the spectroscopic data collected and high performance liquid chromatography record is associated.Through the computing of TQ software, Dehydro and drographolide carries out modeling at these two wave bands of 263 ~ 311 nm and 326 ~ 341 nm, and adopt second derivative method and Norris smoothing processing as preprocess method, good quantitative calibration models can be built, calibration model related coefficient is 0.938, RMSEC is 0.185, RMSEP is 0.123, and modelling verification result RSEP is 4.84%.The model prediction result of Dehydro and drographolide is see Fig. 6.
As can be seen from above modeling result, the coefficient R of the quantitative calibration models of Dehydro and drographolide is more than 0.9, RMSEP and RMSEC is less and close, and RSEP value is all below 5%, and visible model tuning effect is better, and predictive ability is strong.
Claims (5)
1. Herba Andrographitis concentrates a decolorization on-line quick detection method, it is characterized in that, is realized by following steps:
(1) online ultraviolet detector is installed: this device is primarily of heating arrangement (1), reactor (2), peristaltic pump (3), filtrator (4), sample tap (5), flow cell (6), ultraviolet-visible radiant (7), online ultraviolet spectrometer (UVS) (8), worktable (9), circulation line (10), cooling pump (11), distillate receiving bottle (12) is formed, reactor (2) connects peristaltic pump (3) by circulation line (10), filtrator (4), sample tap (5), flow cell (6) forms loop, ultraviolet-visible radiant (7), flow cell (6), online ultraviolet spectrometer (UVS) (8), worktable (9) connects with cascade successively, one end coupled reaction still (2) of cooling pump (11), the other end connects distillate receiving bottle (12),
(2) the online ultraviolet spectrum of online acquisition sample and correspondence: add Herba Andrographitis extract and medicinal carbon in reactor (2), start heating, open peristaltic pump (3) simultaneously, with the rotating speed conductive solution of 48 rpm, by sample tap (5) collecting sample liquid, control to gather corresponding online ultraviolet spectrum by line ultraviolet spectrometer (UVS) (8) by worktable (9) at flow cell (6), filtrator (4) is for the activated charcoal in filtering solution and other solid impurities simultaneously;
(3) each index content in high effective liquid chromatography for measuring sample solution is adopted: adopt andrographolide, the Determination of Dehydroandrographoliin in high effective liquid chromatography for measuring Herba Andrographitis sample liquid, liquid-phase condition is: chromatographic column is C18 (5 μm of 250 × 4.6 mm); Mobile phase is methanol-water gradient elution, flow velocity 0.8 mL/min; Column temperature 35 DEG C; Initial wavelength 225 nm, 16 min are converted to 254 nm; Elution program is: 0 ~ 15 min, 55%B; 15 ~ 16 min, 55%B → 65%B; 16 ~ 26 min, 65%B, sample this liquid, and the syringe filter through 0.45 μm of miillpore filter filters, and gets the analysis of subsequent filtrate sample introduction;
(4) foundation of quantitative calibration models and checking: be taken at line ultraviolet spectrum, Stochastic choice is a collection of as checking collection, all the other participate in the foundation of calibration model, not make the original spectrum of smoothing processing, carry out the foundation of the calibration model of andrographolide at 201 ~ 298 nm wave bands, separately using second derivative and with Norris smoothly as preprocess method, choose 263 ~ 311 nm and 326 ~ 341 nm as modeling wave band, set up the calibration model of Dehydro and drographolide, to verify that the set pair analysis model is tested.
2. Herba Andrographitis according to claim 1 concentrates decolorization on-line quick detection method, it is characterized in that, in online ultraviolet detector, solution is successively through peristaltic pump (3), filtrator (4), sample tap (5), flow cell (6), returns reactor (2) along circulation line (10).
3. Herba Andrographitis according to claim 1 concentrates decolorization on-line quick detection method, it is characterized in that, in online ultraviolet detector, flow cell (6) light path is 2 mm, spectra collection is dynamic acquisition, with 95% ethanolic solution for background, spectral scan scope is 200 ~ 900 nm, and resolution is 0.8 nm.
4. Herba Andrographitis according to claim 1 concentrates decolorization on-line quick detection method, it is characterized in that, andrographolide model selection original spectrum does not make smoothing processing, modeling wave band is 201 ~ 298 nm, the preprocess method of Dehydro and drographolide model selection is that second derivative adds Norris smoothing processing, and modeling wave band is 263 ~ 311 nm and 326 ~ 341 nm.
5. Herba Andrographitis according to claim 1 concentrates decolorization on-line quick detection method, it is characterized in that, circulation line (10) adopts medical rubber hose.
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| CN114627120A (en) * | 2022-05-16 | 2022-06-14 | 河南源宏高分子新材料有限公司 | Method for distinguishing decoloration of regenerated polyester material |
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