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HPTLC Application LabHPTLC Application Lab

HPTLC Application Lab

Applications of HPTLC

Forensic Science

Opium is both a blessing and a curse, depending on who processes it. HPTLC is the only technique capable of quantifying all the six important alkaloids in opium from hundreds of samples when the seasonal crop is ready. These alkaloids can be sensitively and specifically detected by post chromatogram derivatisation using dragendorff reagent.

Food Science

Aflatoxins are a bane for our country. Their presence in grains has hurt our exports and also health of our population, since they are carcinogenic.

The aflatoxins B1, B2, G1, G2 being fluorescent, they are most amenable for HPTLC analysis in the PPT / PPB range. Here again, being commodity products, numerous samples have to be tested at a low cost and HPTLC is the obvious method of choice.

Pharmacology Science

Recently antibiotics and many other poultry feed additives have been in the news, for wrong reasons. Our lab has analysied monensin (an antibiotic) from chicken liver for co – relation with its content in the poultry feed.

Handling complex and “dirty” matrix is not a big issue in HPTLC, as the layer is disposable.

Herbal Fingerprint

One of the major applications is “HPTLC fingerprint” of herbal extracts and formulations. Govt. of India’s policy is that quality control of all AYUSH products is to be done by HPTLC fingerprint as a primary requirement .The Govt. of India has already published numerous quality guidelines which include such fingerprints. Now all pharma copias of the world include herbal raw materials and all them will have to be “identified” by HPTLC, in the near future.

Herbal Analysis – Quantification

In addition to the chemical “HPTLC fingerprint”, it is also possible to quantify desired fractions from herbal extracts and occasionally their formulations. The difficulty is that the reference compounds are either not available or too expensive. Only in HPTLC, samples cannot contaminate the system because they remain on the plate. Hence complex, “not so - clean” mixtures can still be analysed, without much clean – up.

Pharmaceutical Science

By far, the best application for HPTLC in pharmaceutical analysis is the “Uniformity of Content” test, which is prescribed for drug formulations with a small quantity of the drug. For this application, HPTLC is at least 10 times faster and 10 times cheaper than HPTLC!

Sample preparation is very simple as the excipients usually remain at the point of origin.

Synthetic Chemistry

Synthetic organic chemists are always in a hurry to know which compounds of interest have been synthesized in their reactions and which unwanted products have been formed. Mass spectrometry is an excellent technique for identification of expected products and associated reaction products. The new Camag HPTLC – MS hyphenation interface is the organic chemist’s dream come true. Here reaction mixtures are monitored using TLC and fractions of interest “only” are selectively fed into MS. This device’s high Throughput makes it a desirable accessory for LC – MS labs.

Pharmacokinetics

Blood withdraw period 15min, 1hr, 2hr

HPTLC is quite suitable for pharmacokinetic studies, as can be seen from the estimation of a natural product (!) curcumin from rat plasma.

Post Chromatographic Derivatization

Unlike in HPLC, post – chromatogram derivatisation is routinely used in HPTLC to obtain more info about the sample. In HPTLC, the chromatographed sample is retained on the plate. After normal scanning in UV and visible and fluorescence, one of 1100 known derivatisation reagents can be used for specific detection by functional groups and ultra sensitive detection using fluorophores. Moreover since the chromatogram is still on the plate, bioautography is also possible to study anti oxidants or toxic substances or certain other desirable properties.