Validation of Optimized Method for Quantitative Estimation of Extracellular Ethanol from Micro-algae
Nishant Saxena1, Preeti Singh1, Debanjan Sanyal1* and Santanu Dasgupta2
1Research and Development, Reliance Industries Ltd, Jamnagar, India
2Research and Development, Reliance Corporate Park, Ghansoli, Navi-Mumbai, India
*Corresponding Author: Debanjan Sanyal, Research and Development, Reliance Industries Ltd, Jamnagar, India.
Received: April 01, 2021 ; Published: May 17, 2021
Identification and quantification of ethanol in biological samples such as algae, is of paramount importance for research and quality laboratories. An accurate quantification with minimum standard deviation is still a challenging task due to high interference of salts, ions, metals and biological contaminations. A rapid method for the determination of ethanol content in algae samples was validated for seven parameters using gas chromatography. The study was conducted here, to confirm it’s specificity, linearity, accuracy, precision, robustness, sensitivity, and suitability for ethanol estimation in micro-algae samples. During specificity analysis, retention time of ethanol were found 3.27 minutes for standard and 3.25 minutes for sample, which were statistically similar. The linearity of this method for ethanol estimation was found till 5000 ppm for both standard and sample and R2 were 1 and 0.9953 respectively. In the recovery study, the 100% ethanol were recovered from standard and sample with a standard deviation of less than 2%. Repeatability study demonstrated that, the method was found more precise and observed < 1% standard deviation for peak retention time and peak area, similarly < 2% standard deviation was observed among the 10 replicates of standard. In the ruggedness analysis, significant differences were not observed on ethanol content by different analysts on different days of analysis. In the robustness analysis the variation of flow rate, oven temperature and column did not affect the final results of ethanol content significantly. It was found from sensitivity analysis that, this analytical method can detect > 1.36 ppm and can accurately quantify > 4.13 ppm ethanol of algae sample. Validated data indicated that, GC-FID-HSS method is more specific, linear, accurate, repeatable, rugged, robust and sensitive for ethanol estimation. Hence this method can be used effectively to estimate ethanol content in algae samples without any interference from ions, salts, metals and biological contamination.
Keywords: Accuracy; Algae Sample; DB Wax; Ethanol Content; GC-FID-HS
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