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Related Concept Videos

Precipitation Titration: Endpoint Detection Methods01:19

Precipitation Titration: Endpoint Detection Methods

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In argentometric precipitation titrations, endpoints can be detected visually by the Mohr, Volhard, and Fajans methods. In the Mohr method, adding a soluble chromate indicator gives an initial yellow color to the analyte solution. As the titrant is added, the first excess of silver ions forms a red silver chromate precipitate, marking the endpoint. The solution pH should be maintained at about 8 by adding solid CaCO3.
In the Volhard method, a standard excess of AgNO3 is first added to the...
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Precipitation Gravimetry01:03

Precipitation Gravimetry

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Precipitation gravimetry is based on converting an analyte into a sparingly soluble precipitate, which is separated by filtration and weighed. An ideal precipitate should be pure, insoluble, of known composition, and easily filtered from the reaction mixture.
In determining nickel by gravimetric analysis, a precipitant of ethanolic dimethylglyoxime is added to a hot nickel salt solution. This is quickly followed by the dropwise addition of dilute ammonia solution until precipitation occurs. A...
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Precipitation Processes01:12

Precipitation Processes

5.1K
The experimental conditions in a gravimetric analysis should be optimized to maximize the particle size and purity of the obtained precipitate. Ideally, the concentration of the precipitating reagent should be low with effective stirring to maintain low relative supersaturation for the growth of large crystals. In homogeneous precipitation, the precipitant is slowly generated by a chemical reaction in the solution to avoid local reagent excesses. For example, urea decomposes gradually to...
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Types of Coprecipitation01:10

Types of Coprecipitation

5.6K
Coprecipitation is the contamination of a precipitate by otherwise soluble species and occurs via different processes. In colloidal precipitates, coprecipitation occurs via surface adsorption. For instance, barium sulfate has a primary layer of adsorbed barium ions and a secondary layer of nitrate counterions. This results in contamination of the precipitate by barium nitrate.
Sometimes, ions in a crystal lattice can undergo isomorphous replacement by inclusions of similar charge and size. For...
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Precipitation and Co-precipitation01:17

Precipitation and Co-precipitation

4.9K
Precipitation and coprecipitation methods can be used to separate a mixture of ions in a solution. In qualitative inorganic analysis, ions that form sparingly soluble precipitates with the same reagent are separated based on the differences in solubility products. For example, consider the separation of Cu(II) and Fe(II) ions by precipitation as insoluble sulfides. First, copper(II) sulfide is precipitated by the addition of acidic H2S, where the dissociation of H2S is suppressed. Adding H2S...
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Raman Spectroscopy Instrumentation: Overview01:26

Raman Spectroscopy Instrumentation: Overview

1.8K
A conventional Raman spectrophotometer includes a laser source, a sample holding system, a wavelength selector, and a detector.
The monochromatic laser source, typically using visible or near-infrared radiation, generates a highly focused beam of light. This light interacts with the molecules of the sample, scattering some of the light. Liquid and gaseous samples are usually tested in ordinary glass capillaries, while solids can be analyzed as powders packed in capillaries or as potassium...
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Related Experiment Video

Updated: May 6, 2026

Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
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Raman-based PAT for VLP precipitation: systematic data diversification and preprocessing pipeline identification.

Annabelle Dietrich1, Robin Schiemer1, Jasper Kurmann1

  • 1Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.

Frontiers in Bioengineering and Biotechnology
|June 17, 2024
PubMed
Summary

Raman spectroscopy enables real-time monitoring of virus-like particle (VLP) precipitation, a crucial step in biopharmaceutical manufacturing. This Process Analytical Technology (PAT) tool quantifies both VLPs and precipitants, improving process understanding and control.

Keywords:
Raman spectroscopychemometricspartial least squares regressionpipeline optimizationpreprocessingprocess monitoringprotein precipitationvirus-like particles

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Area of Science:

  • Biopharmaceutical Manufacturing
  • Process Analytical Technology (PAT)
  • Spectroscopy

Background:

  • Virus-like particles (VLPs) are key biopharmaceuticals for vaccines and drug delivery.
  • VLP capture typically involves selective precipitation from clarified lysates.
  • Current monitoring methods for VLP precipitation are time-consuming and lack direct product quantification.

Purpose of the Study:

  • To develop and validate a Raman spectroscopy-based Process Analytical Technology (PAT) tool for monitoring VLP precipitation.
  • To enable simultaneous quantification of precipitated VLPs and precipitant in real-time.
  • To overcome limitations of traditional, labor-intensive analytical methods.

Main Methods:

  • Application of Raman spectroscopy combined with chemometric methods (partial least squares models).
  • Development of a PAT tool using batch and fed-batch precipitation experiments of Hepatitis B core Antigen VLPs.
  • Systematic identification of a data preprocessing pipeline to handle complex mixtures and interferences.

Main Results:

  • Raman spectroscopy successfully predicted precipitant concentration with R² values of 0.98 (batch) and 0.97 (fed-batch).
  • The tool captured precipitation trends with R² values of 0.74 (batch) and 0.64 (fed-batch).
  • Effective elimination of lysate composition variations and interferences was achieved through optimized data preprocessing.

Conclusions:

  • Raman spectroscopy serves as a foundational PAT sensor for monitoring VLP precipitation processes.
  • This approach offers real-time insights into VLP precipitation dynamics, improving process control.
  • The methodology holds potential for application in other phase-behavior dependent processes and molecule monitoring.