Why dna precipitates in ethanol

This method can improve the consistency of the active component contents in the supernatant [ 70 ]. The authors noticed that the refrigeration temperature for EP in the production of pharmaceutical companies is affected by the season. Therefore, it is proposed to set the refrigeration temperature as the noise parameter and optimize the range of other easily controlled parameters to reduce the impact of noise parameter fluctuation [ 74 ].

Operating process parameters with design space, varying process parameters according to the change of concentrate quality, or adjusting controllable parameters to lower the effects of noise parameters can all improve the batch-to-batch consistency of supernatant quality after EPP.

In the production of TCM, an ethanol meter is widely used to detect the apparent ethanol content of the supernatants on the spot. This method is simple and practical, but only the density information of the liquid can be obtained.

The monitoring technology and indicators of EPP in the literature are listed in Table 5. At present, near-infrared spectroscopy NIR is widely used due to its simple sample preprocessing, fast speed, losslessness, large amount of information collected, etc. Spectral preprocessing methods have a great influence on the modeling results. Common preprocessing methods include standard normal variate, multiplicative scatter correction, Savitzky-Golay smoothing, Norris-Williams smoothing, first derivative, second derivative, etc.

By establishing a multivariate statistical process control model, the control limit of the process operation statistics such as Hotelling T 2 , squared prediction error, and principal component score is set up, and the process trajectory diagram is drawn. The multivariate statistical process control model can monitor the EPP in real-time and sensitively judge the normal operation state of the process. The establishment of a multivariate statistical process control model is helpful further to implement the feedback control of the EPP.

In general, spectrum of EP system is rich in information. The process monitoring method based on spectrum can not only judge the process state, but also quantify the concentrations of specific components in combination with chemometrics. Based on the extensive review, great progress has been made in the study of process parameters, optimization methods, and process monitoring methods of EP of TCM.

Problems still exist in industrial EP, including the loss of active components, the long time necessary for refrigeration, the quality difference between batches of EP supernatants, etc. In the future, EP technology research can be carried out from the following directions:. The difference in concentrates between batches is mainly reflected in the fluctuation of the content of the components.

At present, there have been reports about the influence of ethanol content in the supernatant on the solubility of Chinese herbal medicinal components. Nevertheless, there is no study on the influence of the content of Chinese herbal medicinal components on the solubility of other components. Therefore, it is not yet possible to describe the effect of the composition change of concentrate on the effect of EP. It is also difficult to accurately predict the material transfer and drug delivery rule of EPP.

The relationship between TCM substances and its quality is generally nonlinear. Therefore, some newly developed artificial intelligence technology can probably be used for the investigation of EPP and TCM quality. For example, as a typical algorithm of deep learning, convolutional neural network CNN can be a useful tool to deal with nonlinear quantitative problems [ , ].

At present, the concentrate quality in the industry is mostly controlled by density or volume. However, less attention has been paid to the chemical composition of the concentrate. It is recommended that the concentrate be used as one of the critical intermediates, and the quality standard of its composition should be set.

This work provides a scientifically based method to set the quality standard of the concentrates. Where permitted by regulations, it can be considered that EP can be carried out after a mixed concentrate is prepared with different batch concentrates, which will help to improve the consistency of the components of the supernatant.

NIR has many advantages, but the equipment cost is high, and the renewal and maintenance of the multivariate statistical model require professionals.

In addition, there is still no means to detect the amount of encapsulation loss. Therefore, it is still necessary to develop simpler and easier-to-use detection technology. At present, the structure of EP equipment is simple, and process control relies heavily on manual work. The energy and material consumption are still high.

Therefore, a complete set of intelligent EP equipment should be developed based on multidisciplinary technology. This equipment should be able to improve the efficiency of heat and mass transfer, quickly collect and analyze process data, and realize the automatic control of EPP.

Comparison of two separation technologies applied in the manufacture of botanical injections: second ethanol precipitation and solvent extraction.

Ind Eng Chem Res. CAS Google Scholar. Removing tannins from medicinal plant extracts using an alkaline ethanol precipitation process: a case study of Danshen injection. Establishment of determination of fractal dimension of ethanol-precipitated flocs of two root medicinal herbs. Chin J Exp Med Formul. Google Scholar.

State Pharmacopoeia Commission. Beijing: China Medical Science Press; Properties of sugar, polyol, and polysaccharide water—ethanol solutions. J Chem Eng Data. Solid-liquid equilibria of d -glucose, d -fructose and sucrose in the mixture of ethanol and water from Chin J Chem Eng. Solubility of xylose, mannose, maltose monohydrate, and trehalose dihydrate in ethanol-water solutions.

Solubilities of protocatechuic aldehyde, caffeic acid, d -galactose, and d -raffinose pentahydrate in ethanol-water solutions. Precipitation of inulins and oligoglucoses by ethanol and other solvents. Food Chem. Dispersion of food proteins in water-alcohol mixed dispersants.

J Mol Liq. Solubility and solid phase studies of isomeric phenolic acids in pure solvents. Thermodynamic solubility modeling, solvent effect and preferential solvation of curcumin in aqueous co-solvent mixtures of ethanol, n -propanol, isopropanol and propylene glycol. J Chem Thermodyn. Measurement and correlation of solubilities of apigenin and apigenin 7-O-rhamnosylglucoside in seven solvents at different temperatures.

Solubility and thermodynamic properties of baicalein in water and ethanol mixtures from Chem Eng Commun. Article Google Scholar. Solubility of chrysin in ethanol and water mixtures. Solubility of Genistein in water, methanol, ethanol, propanol, 1-butanol, and ethyl acetate from to K. Solubility, thermodynamic properties and solute-solvent molecular interactions of luteolin in various pure solvents.

Liu LX, Chen J. Solubility of hesperetin in various solvents from Solubility of naringenin in ethanol and water mixtures. Solubility of naringin in ethanol and water mixtures from Solubility of daidzein in the binary system of ethanol and water. Solubility, molecular interactions and mixing thermodynamic properties of piperine in various pure solvents at different temperatures.

Solubility of osthole in a binary system of ethanol and water. Fluid Phase Equilib. J Solution Chem. Purification of a water extract of Chinese sweet tea plant Rubus suavissimus S.

Lee by alcohol precipitation. J Agric Food Chem. Ethanol precipitation of Codonopsis Radix concentrate with a membrane dispersion micromixer. J Clean Prod. Separation characteristics of ethanol precipitation for the purification of the water extract of medicinal plants. Sep Purif Technol. The determination of dissociation constants for active ingredients from herbal extracts using a liquid—liquid equilibrium method. Study on the separation and purification process of emodin from Polygonum cuspidatum.

Chin Tradit Patent Med. Polysaccharides from Panax japonicus C. Meyer and their antioxidant activities. Carbohydr Polym. Purification, structural characterization, and antioxidant activity of the COP-W1 polysaccharide from Codonopsis tangshen Oliv.

Characterization and anti-tumor activity of a polysaccharide isolated from Dendrobium officinale grown in the Huoshan County. Chin Med. Immunomodulatory effects of polysaccharide extract and its uptake behaviors at the cellular level. The chemical character of polysaccharides from processed Morindae officinalis and their effects on anti-liver damage. Int J Biol Macromol. Effect of step alcohol precipitation on compositions and antioxidant activity of maca polysaccharide. Comparison of the abilities of universal, super, and specific DNA barcodes to discriminate among the original species of Fritillariae cirrhosae bulbus and its adulterants.

Optimization of alcohol precipitation technology of Wubie granules intermediates based on pharmacodynamic screening. Study on optimizing of the alcohol precipitation refining technology for Yanshuning compound water extract. Fujian Med J. Optimization of the water extraction and ethanol precipitation technology of Zhuang Medicine Baijin granules by pharmacodynamics combined with orthogonal test. China Pharm. Zhang L, Liu JT. Optimization of alcohol precipitation technology for dingtongning granule based on index components and pharmacodynamics.

China Pharmacist. Optimization of alcohol precipitation technology of changkang granules based on index components and pharmacodynamics. Optimization of extraction technology and precipitation process for Eryan Huguo decoction by orthogonal test. Optimization of water extraction and alcohol precipitation of Xikebao decoction and comparison of anti-inflammatory and analgesic effects. Optimization of purification technology for Shenqi Sherong pills by chemical and pharmacodynamic indicators.

A comparative study on efficacy of Gualou-Xiebai extracts in different alcohol precipitation oncentration. Chin Hosp Pharm J. Optimization of alcohol precipitation for Lidan Paidu prescription based on index compotents and pharmacodymamics. Cent South Pharm. Anti-hypertensive effect of aqueous extract and supernatants of alcohol extracting-water precipitation of Prunella vulgari.

Research on changes of composition and efficacy of Scutellariae radix extract in process of preparation. Polysaccharides obtained from bamboo shoots Chimonobambusa quadrangularis processing by-products: new insight into ethanol precipitation and characterization. Optimization of extraction technology of Guizhi Zhumian Capsule by orthogonal design and pharmacodynamics test. China Tradit Herb Drugs. Zheng X. Chinese Medicine Pharmaceutical Engineering and Technology.

Wang Q. Talking about ethanol precipitation technology and equipment of Chinese medicine. Mechanical and Electrical Information.

Application of continuous mixing technology in ethanol precipitation process of Salvia miltiorrhiza by using micromixer. China J Chin Materia Med. Device and method of alcohol precipitation in pressure type mechanical atomization combination continual alcohol precipitation patent. Ethanol Precipitation Tank patent. Zhao ZY. Ethanol settling tank with efficient stirring device patent. Efficient Ethanol Precipitation Tank patent.

Optimizing process of radix salvia miltiorrhiza ethanol precipitation by response surface methodology.

Chin JMAP. Optimization of alcohol precipitation process of Salviae miltiorrhizae water-extraction solution by orthogonal design with multi-index comprehensive evaluation. Chin J Pharms. Correlations of three important technological parameters in first ethanol precipitation of Danshen. Optimization of alcohol precipitation process of Cinobufacini extract by multi-index comprehensive evaluation method.

Lishizhen Med Mater Med Res. Orthogonal test for optimizing ethanol precipitated technique of Kuanxin oral liquor. Optimizing the alcohol precipitation of Danshen by response surface methodology. Sep Sci Technol. An approach to determine critical process parameters for ethanol precipitation process of Danhong injection.

Feedforward control strategy and its application in quality improvement of ethanol precipitation process of Danhong injection. Application of quality by design to the process development of botanical drug products: a case study. Network analysis of ethanol precipitation process for Schisandrae Chinensis Fructus. Optimization for the ethanol precipitation process of botanical injection: indicator selection and factor influences.

Control the effects caused by noise parameter fluctuations to improve pharmaceutical process robustness: a case study of design space development for an ethanol precipitation process. Optimization of alcohol precipitation technology for Qingmai granules by orthogonal test.

Optimization of water extraction and alcohol precipitation process for Gardeniae Fructus by orthogonal design. Optimization of water extraction and alcohol precipitation technology for hemorrhoid fumigants by orthogonal test. Optimization of alcohol precipitation process for extra-cellular polysaccharides from fermentative fluid of cordyceps.

Optimization of alcohol precipitation technology for safflower injection with orthogonal test method. Optimization of extraction technology of ephedran. China Pharmaceuticals. Optimization of water extraction and alcohol precipitation process of Lanshen Lipidlowering prescription by orthogonal test.

If the DNA concentration in the sample is low, isopropanol may work better than ethanol to precipitate the available proteins. In addition, isopropanol is often used for precipitating DNA from large volumes as less alcohol is used see protocols below. The ethanol and isopropanol can also wash away the remaining salt residue. After being washed in alcohol and subjected to a centrifuge, the precipitated DNA protein will form a pellet, which can be washed in alcohol again, dried, and re-suspended in a Tris or TE buffer.

Be careful not to overdry the sample, since this can denature the DNA; just leave the washed pellet on the lab table for a few minutes. If isopropanol has been used during the extraction instead of ethanol, the sample may not adhere as tightly to the tube and may require a longer drying time. Incubate on ice for 15 minutes. In case of small DNA fragments or high dilutions overnight incubation gives best results.

Discard supernatant by decanting or pipetting, being careful not to throw out DNA pellet which may or may not be visible. Add 0.

Calculate after addition of sodium acetate. Add comment Cancel. Submit a comment. Comment title. You have entered an invalid code. Submit Cancel. Thank you for submitting a comment on this article.

Your comment will be reviewed and published at the journal's discretion. Please check for further notifications by email. Sign in Don't already have an Oxford Academic account? You could not be signed in. Sign In Forgot password? Don't have an account? Sign in via your Institution Sign in.

This article is also available for rental through DeepDyve. View Metrics. Email alerts Article activity alert. Advance article alerts. New issue alert.