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HP-TLC advantages include visibility of chromatogram & high throughput
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Tuesday, 03 September, 2019, 08 : 00 AM [IST]
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Purnima Parkhi
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The safety of consumers must be guaranteed by quality control ‘for all’ at the production site and ‘for some’ after distribution in the market. In the area of food and protein supplements, the analytical approach for quality requires special expertise, since the samples are very complex.
Adulteration is a common problem in protein supplements (PS). Among the illicit compounds present in PS, steroid hormones (e.g., androstenedione, nandrolone, stanozolol, testosterone and testosterone enanthate) are detected to enhance physical performances of body builders.
Chromatography is a group of scientific technique of separating a mixture into its components for individual identification and quantification. Planar Chromatography is one of them and utilises a flat (planar) stationary phase for separation. Sophisticated High Performance - Thin Layer Chromatography (HP-TLC) technique uses sophisticated instruments, controlled by software and a well defined methodology for analysis.
HP-TLC, having become an official technique in US and European statutes, well recognised as a distinct new technique. The main advantages of HP-TLC are ‘visibility of chromatogram,’ ‘high throughput,’ ‘low cost analysis,’ ‘nominal maintenance,’ ‘little or no sample clean up,’ ‘high sensitivity and accuracy,’ ‘hyphenated with MS and bio-assays in situ etc.’ The possibility of image acquisition of separated samples on the plate is an aspect of HP-TLC.
HP-TLC is particularly useful, when a laboratory has to analyse a very high number of similar samples, for detection of various adulterants. “HP-TLC screening” approach is very high throughput and economical to analyse the presence/absence of certain analytes in a series of samples. After screening, positive (or negative samples as the case may be) are considered in further analyses, if necessary, reducing significantly both costs and time spent for reaching the final results.
The presence of illegal Stanozolol steroid in the photographic image, two samples separated on this ‘plate’ is clearly visible, with both method of visualisation, at the nanogram level. A final confirmation is done using HP-TLC combined with mass spectrometry (MS). HP-TLC-MS instrumentation is indicated in
This simple HP-TLC method shows a good performance suited for fast detection of steroid hormones as adulterants in PS. Figure 2 illustrates the results obtained after chromatographic separation in HP-TLC of standard standards and protein supplements.
Stanozolol steroid observed in protein supplement at visible light and ultraviolet light 366 nm wavelength
Figure 3: Mass Spectrum of standard and corresponding adulteration fraction in the FS1 and FS2 samples are identical
(The author is marketing manager at Anchrom Enterprises (I) Pvt. Ltd. She can be contacted at purnima.parkhi@anchrom.in)
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