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Identification of unknown compounds by using a novel retention index system in liquid chromatography

A liquid chromatography, retention index technique, applied in the field of identification of unknown compounds by using a novel retention index system in liquid chromatography, which can solve the problems of poor affordability and chemical stability, hindrance, unreliable identification tools, etc.

Active Publication Date: 2019-07-09
CHROMALEONT SRL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0034] However, in WO2013 / 134862 there is no suggestion about identifying unknown compounds by means of other detectors, in particular by means of detectors which do not give information about the structure of the compound, such as for example ELSD (Evaporative Light Scattering Detector) detectors
The method described in WO2013 / 134862 is only valid when coupled to mass spectrometry and is therefore not reliable in itself as an identification tool in liquid chromatography
In addition, the compounds included in the same series have not been studied from a toxicological point of view and may be highly toxic due to the presence of the pyridine ring
Another limitation of these compounds concerns their relatively limited availability in the market
[0035] Limitations (such as low UV absorption, limited coverage of chromatographic space, high toxicity, poor affordability, and chemical stability) encountered when using each of the above synchronous series, as well as any constraints imposed by analytical parameters, instrumentation, or even column packing Low retention time reproducibility of varying experiments hinders widespread use of retention index databases in liquid chromatography at both the intra-laboratory and inter-laboratory levels

Method used

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  • Identification of unknown compounds by using a novel retention index system in liquid chromatography
  • Identification of unknown compounds by using a novel retention index system in liquid chromatography
  • Identification of unknown compounds by using a novel retention index system in liquid chromatography

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Embodiment approach

[0215] The embodiments and examples described herein are for illustrative purposes only, and they are not meant to limit the scope of the invention as claimed. Identification of other lipid species such as free fatty acids, mono- and di-acylglycerols, as well as sterols, carotenoids and waxes falls within the scope of the present invention. In fact, the inventors plan to adapt this method for bulk lipid analysis on different real samples in the near future. It is also clear that the retention index database of the present invention can be expanded by including retention indices of other lipid classes, such as, for example, free fatty acids, mono- and diglycerides, and sterols, carotenoids, and waxes, using to achieve the retention behavior of all lipid species (e.g., t R and RI) comprehensive database. The inventors also plan to apply the retention index system to a chromatography method that is also suitable for the determination of polar lipids such as phospholipids.

Embodiment 1

[0218] Embodiment 1: Vegetable sample preparation

[0219] Vegetable oil samples were prepared by dissolving 10 mg of the oil in 10 mL of 2-propanol, except for borage oil, where 10 mg was dissolved in 1 mL of the same solvent.

Embodiment 2

[0220] Example 2: Milk sample preparation (by Folch)

[0221] Treat 10 mL of milk with 40 mL of a chloroform:methanol mixture (1:2 v / v) and place in an ice bath with magnetic stirring for 30 min; transfer the contents to a separatory funnel and shake vigorously for 5 min; then , the mixture was collected into separate test tubes and centrifuged at 3000 rpm for 15 minutes. Once phase separation is obtained, pool the lower phase (including the lipid fraction) and add the upper phase (aqueous phase) to the separatory funnel again for further extraction with 20 mL of chloroform:methanol mixture (2:1v:v) . Repeat the last step again. The pooled organic phases were filtered over anhydrous sodium sulfate and evaporated using a rotary evaporator.

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Abstract

A method for the identification of unknown compounds based on a novel Retention Index System comprising a TAGs homologous series, wherein such identification is performed by means of liquid chromatography (LC), or liquid chromatography coupled with mass spectrometry (LC-MS) is disclosed.

Description

technical field [0001] The present invention relates to a method for the identification of unknown compounds based on a novel retention index system, wherein such identification is performed by means of liquid chromatography (LC) or liquid chromatography coupled to mass spectrometry (LC-MS). Background technique [0002] Before the introduction of retention index systems, qualitative chromatography relied on absolute retention times (t R ) and / or relative retention time (t R(Rel) ) as an identification parameter. [0003] The absolute retention time method consists in comparing the analyte retention time with that of a known compound (e.g., the t of a listed reference compound). R ) for comparison: if the peak's t R If it is the same as the reference substance, a positive identification can be carried out. [0004] However, such methods are only reliable under constant chromatographic conditions: even small differences can lead to shifts in retention times, making identi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/86
CPCG01N30/8668G01N30/8682G01N30/8686
Inventor 路易吉·蒙代洛弗兰切斯卡·里加诺
Owner CHROMALEONT SRL