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Its technique, tools and first applications have been e-published as a free book :
www.planar-chromatography-by-kaiser.com
Sample size: below 1 nl to over 1 ml. 2 to 6 samples partially overlapped on 10x10 cm plates circular positioned. Strictly focussed in the plate centre as -circle. Completely dried (air / N2 , 2 L/min, 20 degr C). Constant flow mobile phase . Gas phase reactions, if needed. Digital photo multi integration, if needed.
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Phase inlet position + - 0.2 mm central. Plate positioning strictly vertical. Phase flow, drying and gas phase treatment at constant temperature and strictly symmetrical .
Only this new technique offers 100% safety of results, see the simple facts in the green triangles :
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To figure 10:
In the green located plate triangle there is any detail of physics, physical chemistry and chemistry strictly equal for the substance bows from two samples which partially overlap. In addition: the TIME is just the same for all visible details. Thus if the bows of the two overlapping samples DIFFER, the two samples differ for 100% sureness. Only if NO difference is seen there may exist all the standard uncertainty two differing samples may have although they look equal. Than there is NO sureness that two are really equal.
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mobile phase
plate
virtual -PLC chamber by 1 mm distance of the cover glass at all four corners, see
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To figure 13:
This is the main part of a -PLC instrument: It is the thick glass cover over the PLC plate enclosing a 1 mm thick gas volume between the cover glass and the PLC stationary phase. In the cover glass centre is the 5 mm i.d. hole which holds a PTFE tube surrounding a wick. This wick transfers the mobile phase from the 1.5 ml glass bottle into the PLC layer centre. The mobile phase flow is constant from the moment the wick touches the PLC layer surface till the manual removal of the mobile phase glass. The plate position remains fixed (by three aluminium rods) and must have a perfect vertical position. Otherwise the mobile phase vapor would flow out of the 1 x 100 x 100 mm gas phase.
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To figure 15:
The UV lamp illuminates the dried -PLC plate in 100 mm distance. The picture is photographed by a digital photo camera protected from visible light by a thick black mini curtain. One uses UV 254 nm and 366 nm light.
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To figure 17:
With micro brushes no. 6 to 10, wetted but not overloaded by methylene chlorid, we touch and extract by slight brush movements about six to 12 1 cm square fruit surface positions. The visible blue lines illustrate brush extracted spots only for this picture no. 17. Brushing is done per spot within a few seconds. The still wet brush than contacts for 2..3 seconds a selected plate position next to the centre but for each fruit at the same position see the next figures for more details.
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The fruit surface is covered by natural products solvable in CH2Cl2. They chemisorb on silicagel. Impregnated fruit surfaces add further material, which may be visible by planar chromatography, see the BIO-orange material below:
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Under normal conditions in overlapped sampling focussing and separation works fine under -PLC
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This way it is easy to identity falsified medical products example VIAGRA : only A is authentic. Q1, Q2, Q3 are faiked. Quantitation if necessary with N up to 16 repetitions
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To figure 25:
The red lined plate portions show where sections of the plate surface photo scan the plate surface producing single sets of integration data. Each scan track differs by angle position, which is selectable by software and in length and width. At least four but up to 16 scan track data are added and the totals divided by four up to 16 thus keeping the substance bow integral signal at a constant if the bow is equally strong substance loaded along its length. But all plate structure noise signals are reduced by a factor four up to 16 thus drastically improving the signal-to-plate-structure relation. This reduces significantly the well known bad relation of signal-to-noise in TLC quantitation. NOTE: in fact the plate structure is no noise but a systematical quantitation error, as plate structure has nothing to do with substance or detection. It is just the local inhomogeneity of the stationary phase. But at the final result this trick of data improvement is based on measurement, not on mathematical noise reducing statistics. This ends up with an otherwise impossible PLC data improvement towards repeatability (or better: comparability-) standard deviations of +- 0.5 to +- 0.05 % for main compounds.
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Three of the clearly falsified VIAGRAs have been ordered through the Internet.
I got these samples from my Apotheker who asked me for a critical compare analysis because of problems his customers had.
A few days ago we got from a pharma company one original drug and its remake as important anti cancer medicine. Remake and Original DIFFER strongly!
Remake
7 integr.chrom. each
Original
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To figure 30:
Why illegally ? Well: the accepted fruit transportation and storage protection chemicals are limited by law to three, which in chromatograms would result in three sharp lines (or bows). But the BIO lemon surface extract chromatogram in figure 30 does not show 1 to 3 sharp lines, instead we see a huge number and amount of UV absorbing substances. These materials change the smell, the taste and who knows the health value of a fruit. Increasing effects of allergy, stomach pain, headache and other side effects exist next to a real poisoning. See in the next figure the lemon skin problem: washing does not help, so you get the stuff in your tea when you add a lemon disk.
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1 = IM-mandarin1 [ shop A (minimum sampling) ] OK 2 = mandarin2 [ shop B same town, same road,] NOK 3 = OP (ortho phenylphenol) test substance 4 = Bio-lemon [ shop B (minimum sampling) ] NOK 5 = TH (Thiabendazol) 6 = Bio-orange [ shop B (minimum sampling) ] NOK
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20 diff. plant surface analyses would to day show >10 not acceptable results: (as a mean)
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Thiabendazol (TH)
o-Phenylphenol (PH)
Imazalil (IM)
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TH, IM and PH also used on potatoes, bananas, grain. Information about treated goods is often regulated by law. Critical concentrations. Lots of health warnings / strict regulation / time of future usage is by part already limited. http://de.wikipedia.org/wiki/Imazalil http://www.sciencelab.com/xMSDS2_Phenylphenol-9926513 http://www.private-health-organisation.de/ About the chemistry and toxicity of falsified materials is by now NOTHING KNOWN.
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purity of test substance a = 97.8 +- 0.09 % purity of test substance b = 97.3 +- 0.11 %
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Total areaY in % =
RpY x areaY x 100 / sum (RpYn x areaYn) Reason: circular PLC with multiple runs; N always > 4 Rp is the relative bow position from centre to integration border. Here no Rf value acceptable. Quantitation in PLC: by non linear calibration lines only !
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The fundamental law of NON linearity in PLC calibration lines needs graphical statistics at best using Polynomial Interpolation You have it ? (It exists as PI-rek-E11 program)
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Sorry: this was a polynom 2nd degree. You better use a 3rd degree Polynom:
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A is NOT Q
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To figure 45:
We just started to make a small video about how -PLC is prepared and running. We took from two medicine containers the same drug and planned to show how equal products look how sharp substance bows overlap or if a remake really shows visibly differences to a standard product now out of patent regulation. Unexpectedly: the bows did not line up. A next example of medicine falsification was found. The authentic pill A differs by 100 % safety from Q. Up to over 90% falsification is found in internet sales, up to 50% in official drugstores. The drug falsification is critical in case of cancer, HIV, blood pressure and far over thousand other health problems. For a save detection of non conformity we never need forensic analytical tecniques nor a big lab instrumentation. Micro PLC does it. Quantitation more in details: Multi integration is a software concept: the program resulted from a close cooperation of IfC with the Sorbfil programmers. See www.sorbfil.com (News; select version 2 or higher)
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Integration can be checked in single steps. This costs time but brings quality.
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VIS
Light optmization is possible by multi integration software.
FLU UV
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Thank You !