There are very few unique plant metabolites for creating differentiation in chemical composition among extracts. In practice, many herbal formulations containing more than 3 herbal extracts are flooding in the market. So, how could we create method of anylysis to process such formulation for standardization and how could we finalise the marker for such analysis?
I'm working on herbal products since 1984 (import & distribution in Italy from Nepal different herbal preparations as food supplements with one herbal ingredient or with more herbal ingredients, but we don't use standardized extracts only full plant: leaves, rhizome, steam, cortex etc.). Es. by High Performance Liquid Chromatography (H P L C) defining and selecting a specific chromatographic technique: biomarker or marker of a plant it is important to be sure to avoid adulteration. It's a big question if we follow "herbal traditional preparations"!
It is the toughest task for all the researchers involved in herbal formulations. Standardisation can be either done in terms of their total metabolites or even as any unique and common marker present in all the herbs. But that is very difficult. Some of them develop fingerprints of their polyherbal formulations in HPLC or HPTLC. Still it wont be consistent enough batch to batch. You have to fix various factors like growth maintenance of the plant, location of plant collection, season, drying time, grinding method, extraction time, temperature, solvent etc, etc.
Standardisation of herbal formulations remains the biggest task among researchers. However, may not be accurate but there are certain standardisation techniques adopted in Industries for polyherbal formulations like
i. determination of total secondary metabolite contents like polyphenols, flavonoids, alkaloids, saponins etc.
ii. if any common marker exist among the various extracts used then the polyherbal formulation can be standardised with respect to the specific marker
iii. at the last stage a fingerprint can be obtained by HPTLC or HPLC methods based on their rich metabolites.
All the above mentioned methods suffer from consistency again. So many factors influence the chemical content of herbal extracts like location of geographical location, soil fertility, seasonal variation,time of collection, drying time etc etc. Thus stringent procedures has to be followed from the growth of herbs itself to obtain consistency in every batch.
Dear All
TLC fingerprinting using HPTLC is really questionable. Even same extract of plant show variation in fingerprinting. One extract itself contain thousands of metabolites. Many time we get good resolution in TLC and more then 20-30 band is separated. It means if one extract is capable of separation of such a large number of band in TLC. Then what about mixture. Fingerprinting of polyherbal mixture is always doubtful. After all our purpose is standardisation. Many allied species of plant or adulterated species gives such bands on TLC.
I am strictly not in favor of fingerprinting as a tool of standarization by any means (either HPTLC, HPLC, IR, NMR etc) for polyherbal formulation.
Their is only one possibility if we work on identification of differentiation markers for particular formulation. Which is very limited.
Yes. Fingerprinting cannot be consistent within batches as various factors influences it. But identification of a standard marker for a mixture of herbal extracts is very tough. Atleast with a single herbal extract it can be done. But when you are going for a mixture of extracts, then it is tough to detect specific markers of individual extracts as the quantity in polyherbal formulation is very very meagre - will not be under detection limit.
Today’s sophisticated software offers the user such choices of data treatments as multiple linear regression (MLR), partial least squares (PLS), principal components regression (PCR), factor analysis (FA), neural networks (NN), and Fourier transform (FT), among others. The trade-off is a good one: even after several hours (or days) of
calibrating, the multiple advantages of analysis by NIR far outweigh the time required for method development. and MCR can also be used these way you can minimize noise and able to calibrate some components in multivariate environment when traditional systems do not accept isolation techniques in regular dosage form.
Dear Dr Sethiya
You will go for LC-MS analysis for Standardization of multicomponent herbal formulation.
If you really interested in standardization of poly herbal formulation in multivariate norm, you can developed multivariate calibration method with hyphenated technique(HPLC-PDA, LC-MS), non-destructive technique (UV-VIS NIR, 2D-NMR) target class of moiety.
Various multi-way modelling like PARAFAC, Tucker, N-PLS on various open source software like R, Weka and commercial Matlab, unscrambler software you can decomposed mathematically component. You can developed the database.
On the other hand, In controlling the formulation batch to batch , you can visualize the data control chart.
Finally say any one not to control nature.
Select at least one marker of each ingredient and analyse quantitatively by Chromatography. For non volatile markers you may go for HPLC while for volatile Gas chromatography is recommended. However as per recent guidelines quantification of four markers are necessary in each and every drug either single plant base medicine or compond formulation prepared by adding different plants.
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