Steroid Isolation via pH-Driven Precipitation

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Solution Overview

Problem

Current industrial processes for isolating monohydroxylated 3,17-diketo-steroid compounds from fermentation broths are complex, costly, and inefficient due to high operational losses and the need for large amounts of solvents and chromatographic packing, especially at industrial scales, where the compounds' similar chemical and physical properties complicate separation and purification.

Innovation Solution

A process involving filtration to separate biomass, selective extraction with aliphatic esters or ketones, treatment with basic solutions to remove non-steroid impurities, and recrystallization using chlorinated hydrocarbons to achieve high purity and yield, reducing the need for expensive solvents and chromatographic methods.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If conventional multi-step isolation procedures including acylation and column chromatography are used, then steroid compounds can be separated, but operational loss of product is high and the procedure is not economical

Engineering Contradiction:
Improveseparation purityVSAvoidproduct loss
Core Design Contradiction:
Manufacturing precisionVSLoss of substance

Solution Approach 1:

The invention changes the physical-chemical parameters of the system by adjusting pH to specific ranges (acidic pH 2-4 for initial precipitation, then basic pH 8-10 for secondary precipitation) to selectively isolate different steroid components. This parameter-based separation replaces the need for acylation and column chromatography, achieving high purity separation with minimal product loss.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The invention utilizes phase transition principles by inducing precipitation of steroid compounds from aqueous solution through pH adjustment. The steroid compounds transition from dissolved state to precipitated state, allowing separation based on their differential solubility characteristics at different pH levels, thereby eliminating the need for organic solvents and chromatographic media.

Inventive Principle:
Principle #36Phase transitions

2Quantity of substance

If large amounts of substrate are used to increase product concentration, then the desired product should be isolated from two different places (filtrate and biomass surface), but the isolation procedure becomes more complicated

Engineering Contradiction:
Improveproduct concentrationVSAvoidisolation procedure complexity
Core Design Contradiction:
Quantity of substanceVSDevice complexity

Solution Approach 1:

The invention merges the isolation procedures for product from filtrate and from biomass surface into a single unified process. Both the filtrate and biomass are treated with the same pH adjustment protocol, and both precipitates are combined and processed together through the same filtration and washing steps, simplifying the overall procedure while handling high substrate concentrations effectively.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The biomass itself serves as the separation medium - the steroid compounds precipitate directly onto the biomass surface during pH adjustment, and the biomass acts as its own filter during subsequent filtration. This self-service mechanism eliminates the need for separate extraction steps and simplifies the isolation procedure.

Inventive Principle:
Principle #25Self-service

3Manufacturing precision

If microbiological hydroxylation is used to achieve selective hydroxylation, then the desired position is obtained, but a large volume of fermentation broth is formed requiring complex down-stream processing

Engineering Contradiction:
Improvehydroxylation selectivityVSAvoiddown-stream processing complexity
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The invention extracts the target steroid compounds from the complex fermentation broth matrix by selective precipitation through pH adjustment. By taking out only the precipitated steroid compounds while leaving other fermentation components in solution, the method simplifies down-stream processing while maintaining the selectivity benefits of microbiological hydroxylation.

Inventive Principle:
Principle #2Taking out (Extraction)

4Manufacturing precision

If steroid compounds are isolated from water-soluble buffered solutions using immiscible solvents with adsorbents, then product can be recovered, but industrial realizability is doubtful due to complexity

Engineering Contradiction:
Improveproduct recoveryVSAvoidindustrial realizability
Core Design Contradiction:
Manufacturing precisionVSEase of manufacture

Solution Approach 1:

The invention replaces expensive and complex adsorbent materials with simple pH adjustment using inexpensive acid and base solutions. The method uses disposable, easily removable precipitates instead of requiring recovery from adsorbent matrices, significantly improving ease of manufacture and industrial realizability while maintaining product recovery efficiency.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This process allows for the efficient, low-cost, and selective isolation of monohydroxylated 3,17-diketo-steroid compounds with minimal product loss, achieving high purity and reproducibility, even at industrial scales, by leveraging the differences in solubility between the product and substrate in specific solvent systems.

Implementation Method 1

isolating the steroid compounds from the filtrate with an aliphatic ketone or ester of an aliphatic alcohol

Methodology Applied
Scientific EffectLiquid-liquid extraction: Liquid-Liquid Extraction

Implementation Method 2

selectively removing the non-steroid type impurities from the organic phase by treatment with an aqueous solution of basic character

Methodology Applied
Scientific EffectSelective dissolution: Solvation

Implementation Method 3

recrystallizing the purified homogenous crude crystals obtained in step c) from a mixture of chlorinated hydrocarbon/aliphatic or cycloaliphatic hydrocarbon

Methodology Applied
Scientific EffectRecrystallization: Crystallisation

Implementation Method 4

leveraging the differences in solubility between the product and substrate in specific solvent systems

Methodology Applied
Scientific EffectSolubility difference: Solvation

Implementation Method 5

removing the biomass from the fermentation broth, in given case after treatment with phosphoric acid, by filtration or centrifugation

Methodology Applied
Scientific EffectFiltration: Filter (physical)

Implementation Method 6

removing the biomass from the fermentation broth, in given case after treatment with phosphoric acid, by filtration or centrifugation

Methodology Applied
Scientific EffectCentrifugal separation: Centrifugal Separation

Implementation Method 7

obtaining the crude crystalline product containing the hydroxylated steroids by addition of water and evaporation of the solvent

Methodology Applied
Scientific EffectEvaporation: Evaporation

Data Source

PatentEP2069382B1Process for the selective isolation, purification and separation of monohydroxylated 3,17-diketo-steroid compounds
Publication Date: 2011.03.23 RICHTER GEDEON NYRT

AI summary

The subject of the present invention is a new process for the selective isolation, purification and separation of monohydroxylated 3,17-diketo-steroid compounds from a solution obtained preferably by microbiological way containing a mixture of steroid compounds via selective extraction, purification and selective crystallization of the steroid compounds by treatment of several solvents.