Paper chromatography spotting

The diuretic and cholagog effect of Ononis arvensis motivated the development of a new two-dimensional paper chromatographic and TLC method for the measurement of onion in its roots and aerial parts. Samples were dried, ground and extracted with methanol (70 per cent, w/v) for 2 h. The supernatant was diluted and used for TLC separation on a cellulose stationary phase. The first eluent was 3 per cent formic acid, and the second was n-butanol-acetic acid-water (4 1 5, v/v) for both TLC and paper chromatography. Spots were scraped off, extracted with methanol and the absorption was measued at 260 nm. It was found that the average ononin content in roots and aerial parts was 0.153 0.0278 (per cent) and 0.498 0.045 (per cent), respectively. Because of the simplicity, the method was... 

Figure 1. Tracings of separation of total capsaicinoids of capsicum extracts and reference pure compounds by paper chromatography Spot (1) red capsicum Spot (2) natural capsaicinoids pure Spot (3) green capsicum fresh and Spot (4) synthetic capsaicin (C) color (F) fat, and (Cv), capsaicinoids/capsaicin ((27) and...

Paper chromatography spot urine and spots of amino acids it isthoughtto contain on the paperrun chromatogram usinga suitable solvent (water) develop colourless spots by spraying with ninhydrin. 

Purified by paper chromatography using /crt-butanol-water, cutting out the main spot and eluting with water. Also purified by adsorption onto an apo-flavodoxin column, followed by elution and freeze drying Crystd from acidic aqueous soln. [Mayhew and Strating Eur J Biochem 59 539 1976.]... 

Chromatography is one of the most powerful and widely used means for separating mixtures, because it is often inexpensive and it can be used to provide quantitative as well as qualitative information. The simplest method is paper chromatography. A drop of solution is placed near the bottom edge of the stationary phase, an absorbant support, such as a strip of paper. The mobile phase, a fluid solvent, is added below the spot and the solvent is absorbed on the support. As the mobile phase rises up the stationary phase by capillary... 

Jalal and Collin19 used paper chromatography and TLC to determine caffeine in both coffee and tea, and theobromine in tea. Their TLC method used cellulose plates that were developed with butanokhydrochloric acid water (I 00 1 1 28) for 4 h. The spots were eluted from the plates with ammonium hydroxide and measured spectrophotometrically against a blank at 272 nm for caffeine and 274 nm for theobromine. 

All of these methods fail to show the presence of the epimeric form of the tetracyclines and in most instances streaking of the spots is a problem. A basic improvement in the paper chromatography of these antibiotics was achieved by Selzer and Wright (47) and Kelly and Bryske (48) when they reported methods for the pretreatment of the paper with com-plexing agents to bind the metallic ions which may be present. 

The column chromatography technique using Dowex 50 ion-exchange resin, introduced in 1951 (M2) and improved in 1954 (M3) by Moore and Stein, first made possible the precise quantitative analysis of amino acids liberated in the course of acid hydrolysis of urine. Similar results were also obtained by Muting in 1954 (M4), who used paper chromatography methods. In this procedure amino acids were quantitatively determined after staining on the paper and elution of the resulting spots. 

High-voltage electrophoresis and subsequent paper chromatography of the fractions obtained made possible the isolation from the analyzed mixture of twenty-two components giving colored spots with ninhydrin and isatin. Among these, fourteen were identified as peptides and their amino acid composition established (Table 5). In the case of eight peptides, also N- and C-terminal amino acids were determined (Table 6). 

Quantitative analysis is also possible. The spot representing the component of interest can be cut (in the case of paper chromatography) or scraped from the surface (TLC), dissolved, and quantitated by some other technique, such as spectrophotometry. Alternatively, modern scanning densitometers, which utilize the measurement of the absorbance or reflectance of ultraviolet or visible light at the spot location, may be used to measure quantity. 

Figure 11.22 represents a paper electrophoresis apparatus. The soaked cellulose sheet is sandwiched between two horizontal glass plates with the ends dipped into vessels containing more electrolyte solution. The electrodes are also dipped into these vessels, as shown. The sample is spotted in the center of the sheet, and the oppositely charged ions then have room to migrate in opposite directions on the sheet. Qualitative analysis is performed much as with paper chromatography, by comparing the distances the... 

The transmethylation hypothesis depended on the psychosis of mescaline as an example of how methylated compounds similar in structure to the monoamine neurotransmitters could be psychotogenic, and demonstrated how methionine, the precursor of the methyl donor S-adenosylmethionine, could exacerbate the psychotic symptoms of schizophrenia in patients. This theory was fed by studies of the now notorious pink spot, an amine found in paper chromatography of urine extracts from schizophrenics and thought to be 3,4-dimethoxyphenylethylamine (i.e., O-methylated dopamine). Subsequent studies eventually identified this as another compound or compounds, primarily of dietary origin. Another methylated derivative erroneously proposed to be found in higher quantities in schizophrenia was dimethyltryptamine. This compound is similar in structure to LSD, the hallucinogenic nature of which was the key to the serotonin deficiency hypothesis, which proposed that the known antagonism of serotonin (5-HT) by LSD indicated that psychotic disorders such as schizophrenia may result from a hypofunction of 5-HT. 

The specific radioactivities of uracil were 14C, 0.6 mC/mAf T, 7.75 inC/mAf ratio T/14C = 12.9. The concentration of uracil was 10 3M. Photoproducts were isolated by paper chromatography with n-butanol/water, 86.14. Spots were eluted with water and radioactivity determined in a Tri Carb 314 EX (Packard Instruments). 

Paper chromatography was also used to separate the complex obtained by polymerization of acrylic acid in the presence of poly(ethylene imine). In this case, both the complex obtained by mixing of two polymers and the complex obtained in template polymerization gave two distinct spots. No trace was found of any graft copolymer. 

---