A trade off between separation, detection and sustainability in liquid chromatographic fingerprinting — ASN Events
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  3. A trade off between separation, detection and sustainability in liquid chromatographic fingerprinting

A trade off between separation, detection and sustainability in liquid chromatographic fingerprinting (#239)

Cristiano S Funari 1 , Manish Khandagale 1 , Renato L Carneiro 2 , Alberto J Cavalheiro 3 , Emily F Hilder 1
  1. University of Tasmania, Sandy Bay, TAS, Australia
  2. Department of Chemistry, Federal University of São Carlos, São Carlos, São Paulo, Brazil
  3. Organic Chemistry, Institute of Chemistry, São Paulo State University, Araraquara, São Paulo, Brazil

It is estimated that HPLC analyses generate 34 million litres of solvent waste per year and acetonitrile is the most used organic solvent in them.1 2  Although this solvent presents propitious physicochemical properties for separation and detection in HPLC, it is potentially toxic to analysts and students involved in the research, has a half-life in water of 2-20 days and exhibits acute and chronic toxicity to aquatic life.3  Such characteristics classify it as an undesirable solvent from an environmental point of view.4  Acetone might be a greener alternative to replace of acetonitrile in reversed phase HPLC, since both are hydrogen bond acceptor solvents and present some similar solvatochromic properties and similar viscosities.5  However, as acetone’s UV cut-off extends out to 330 nm, it limits its use with UV absorbance based detectors. In this work, a benchmark method using acetonitrile and HPLC coupled to a UV photodiode array detector coupled to a corona CAD detector system (HPLC-PDA-Corona CAD) was developed to fingerprint an extract of Lippia alba (Verbenaceae). The possibility of replacing acetonitrile with acetone was investigated. Quality by design was adopted to maximize the number of peaks acquired (n) in gradient times of 60 minutes or less in both fingerprint developments. Up to six variables were investigated in a screening step (by means of fractional factorial designs) in each method, followed by a central composite design performed with the three most relevant variables. The methods with acetonitrile or acetone were successfully optimized and proved to be statistically similar when only n was taken into consideration (220±6 and 219±5 peaks, respectively). However, the superiority of the latter was evidenced when this response, n, was combined with the environmental assessment tool (HPLC-EAT)2  as the ratio n/HPLC-EAT (with values of 1.7 and 2.2 for acetonitrile and acetone, respectively). That is, because the acetone-based method consumed less environmental resources while providing the same amount of information. This ratio provides a more comprehensive view of the process of separation by taking into consideration the amount of information acquired, n, and also information about safety, health and environment impacts of the method given by HPLC-EAT. A time and resource saving approach was presented here, which is generic and applicable to other complex matrices. Furthermore, it is in line with environmental legislation and analytical trends.

Acknowledgements: Grant #012/15877-7, São Paulo Research Foundation (FAPESP) and Australian Research Council (ARC)

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