"Colloidal" Phosphorus Errors in Gel Chromatography

1983 ◽  
Vol 40 (10) ◽  
pp. 1614-1621 ◽  
Author(s):  
B. Kent Burnison

Gel chromatography has been used for the separation of 32PO4 and a high molecular weight "colloidal" 32P-labeled fraction from 32PO4-labeled lakewater. When the labeled filtrate is treated with reagents required for the molybdenum blue method for orthophosphate analysis, only a small fraction of the "colloidal" peak is hydrolyzed to orthophosphate. As the reduced molybdophosphoric acid is strongly adsorbed to the dextran gel, quantitative elution of 32PO4 can be achieved with 0.05 mol∙L−1 NaOH and 0.3% NaCl. In hardwater lakes, care must be taken to eliminate the possibility of orthophosphate precipitation at higher pH. In these lakes, it is unlikely that the discrepancy between 32PO4 bioassays and chemical analyses can be solely attributed to acid hydrolysis of "colloidal" phosphorus. Microparticulate apatite also has the potential to release soluble reactive phosphate when the acidic molybdenum blue method is used.

1981 ◽  
Vol 38 (10) ◽  
pp. 1215-1219 ◽  
Author(s):  
E. White ◽  
G. Payne ◽  
S. Pickmere ◽  
F. R. Pick

Molybdate reactive high molecular weight (MW > 5000) phosphorus (RHMW-P) in solution was separated from low molecular weight material (PO4-P) by Sephadex gel (G25–150). PO4-P is as close to orthophosphate as has been possible to achieve with molybdenum blue technology. Chromatograms of near-surface waters of 32 lakes showed that RHMW-P was rarely the dominant component of dissolved reactive phosphorus (DRP), and so cannot be the major influence causing discrepant estimates of orthophosphate concentrations derived from molybdenum blue and radiotracer technology. The low molecular weight material (PO4-P) may still contain chemically reactive organic phosphorus which could lead to overestimation of orthophosphate. Recent literature points to discrepancies in radiotracer estimates too, so further effort is required to provide reliable assessment of orthophosphate in lake water. Estimates of orthophosphate flux from solution to particulate material based on 32PO4-derived turnover times are likely to be in error irrespective of the source of orthophosphate estimate.Key words: molybdenum blue method, radiotracer, orthophosphate flux, reactive high molecular weight phosphorus, gel chromatography, radiobioassay


1978 ◽  
Vol 35 (12) ◽  
pp. 1636-1639 ◽  
Author(s):  
Malcolm T. Downes ◽  
Hans W. Paerl

A method for the separation of two dissolved reactive phosphorus (DRP) fractions in unconcentrated lakewater is described. A reactive high molecular weight phosphorus (RHMW-P) fraction has been found in all waters studied and a second fraction, corresponding to orthophosphate (PO4-P) is frequently detected. The hydrolysis of RHMW-P to PO4-P by acid in the analytical reagents is shown to be the most likely reason for its determination as DRP. Key words: freshwater analysis, reactive phosphorus, orthophosphate, molybdenum blue method, gel filtration


2020 ◽  
Vol 81 (2) ◽  
pp. 301-308 ◽  
Author(s):  
Wenzhe Song ◽  
Yu Zhang ◽  
Amir Hossein Hamidian ◽  
Min Yang

Abstract The biodegradation of polyacrylamide (PAM) includes the hydrolysis of amino groups and cleavage of the carbon chain; however, the effect of molecular weight on the biodegradation needs further investigations. In this study, biodegradation of low molecular weight PAM (1.6 × 106 Da) was evaluated in two aerobic (25 °C and 40 °C) and two anaerobic (35 °C and 55 °C) reactors over 100 days. The removal of the low molecular weight PAM (52.0–52.6%) through the hydrolysis of amino groups by anaerobic treatment (35 °C and 55 °C) was much higher than that of the high molecular weight (2.2 × 107 Da, 11.2–17.0%) observed under the same conditions. The molecular weight was reduced from 1.6 × 106 to 6.45–7.42 × 105 Da for the low molecular weight PAM, while the high molecular weight PAM declined from 2.2 × 107 to 3.76–5.87 × 106 Da. The results showed that the amino hydrolysis of low molecular weight PAM is easier than that of the high molecular weight one, while the cleavage of its carbon chain is still difficult. The molecular weights of PAM in the effluents from the two aerobic reactors (25 °C and 40 °C) were further reduced to 4.31 × 105 and 5.68 × 105 Da by the biofilm treatment, respectively. The results would be useful for the management of wastewater containing PAM.


1975 ◽  
Author(s):  
Y. Benabid ◽  
E. Concord ◽  
M. Suscillon

Purified fibrinogen solutions, incubated with thrombin. CNBr. Sepharose, were subjected to agarose gel chromatography and eluted at different pH (6.5; 7.5; 8.5). Among high molecular weight derivatives formed by thrombin, the major component was a dimer. Gel chromatography at pH 8.5 showed a complexes peak distinct of that from fibrinogen, whereas at pH 6.5, only the fibrinogen peak appeared: fibrin monomer was eluted with fibrinogen as demonstrated by polyacrylamid gel electrophoresis 3.75% pH 8.9. SDS urea electrophoresis after reduction indicated that complexes peak contained two α-chains (α and α′). When fibrinogen was incubated with thrombin in the presence of FSF and calcium, several derivatives with higher and higher molecular weights were formed besides the dimer, and elution profiles of chromatography were identical at pH 6.5 and 8.5, thus indicating stable complexes formation. If fibrinogen-fibrin monomer mixture was subjected to FSF action at different pH, no complexes were formed at pH 6.5. These results confirm that at pH 6.5, any association was prevented.


1977 ◽  
Author(s):  
Milica Jakábová ◽  
David R. Phillips

The effect of calcium on human platelet polypeptides was investigated. When lysed platelets were incubated with mM Ca++, two major intracellular polypeptides (Mr = 255,000 and 230,000) were found to rapidly disappear. A similar phenomenon was also observed when intact platelets were treated with the calcium ionophore A-23187 in the presence of mM Ca++. Determinations of lactic dehydrogenase activity in supernatant fractions demonstrated that these losses occurred before platelet lysis. Investigations into the identity of the high molecular weight polypeptides revealed that one (Mr = 255,000) had similar properties to actin binding protein. The loss of the high molecular weight polypeptides was accompanied by formation of lower molecular weight polypeptides (Mr = 135,000, 93,000 and 48,000), indicating that Ca++ activates a polypeptide cleavage mechanism. The Ca++-activated polypeptide cleavages were rapid, with significant changes being observed within the first 0.5 min of incubation. An obvious explanation for these effects is. that there is Ca++-activated proteolytic activity within platelets. The Ca++-activated proteolytic activity was determined by the hydrolysis of the artificial substrate azocasein. We found that more than 90% of the proteolytic activity in lysed platelets was due to Ca++-activated proteases. These studies show that Ca++-activated proteases may play an important role in platelet activation.


Marine Drugs ◽  
2019 ◽  
Vol 17 (1) ◽  
pp. 52 ◽  
Author(s):  
Weihua Jin ◽  
Wanli Wu ◽  
Hong Tang ◽  
Bin Wei ◽  
Hong Wang ◽  
...  

Sulfated galactofucan (ST-2) was obtained from Sargassum thunbergii. It was then desulfated to obtain ST-2-DS, and autohydrolyzed and precipitated by ethanol to obtain the supernatant (ST-2-S) and precipitate (ST-2-C). ST-2-C was further fractionated by gel chromatography into two fractions, ST-2-H (high molecular weight) and ST-2-L (low molecular weight). Mass spectrometry (MS) of ST-2-DS was performed to elucidate the backbone of ST-2. It was shown that ST-2-DS contained a backbone of alternating galactopyranose residues (Gal)n (n ≤ 3) and fucopyranose residues (Fuc)n. In addition, ST-2-S was also determined by MS to elucidate the branches of ST-2. It was suggested that sulfated fuco-oligomers might be the branches of ST-2. Compared to the NMR spectra of ST-2-H, the spectra of ST-2-L was more recognizable. It was shown that ST-2-L contain a backbone of (Gal)n and (Fuc)n, sulfated mainly at C4 of Fuc, and interspersed with galactose (the linkages were likely to be 1→2 and 1→6). Therefore, ST-2 might contain a backbone of (Gal)n (n ≤ 3) and (Fuc)n. The sulfation pattern was mainly at C4 of fucopyranose and partially at C4 of galactopyranose, and the branches were mainly sulfated fuco-oligomers. Finally, the anti-tumor and anti-angiogenic activities of ST-2 and its derivates were determined. It was shown that the low molecular-weight sulfated galactofucan, with higher fucose content, had better anti-angiogenic and anti-tumor activities.


2021 ◽  
Vol 101 (1) ◽  
pp. 91-98
Author(s):  
K.K. Pirniyazov ◽  
◽  
V.E. Tikhonov ◽  
S.Sh. Rashidova ◽  
◽  
...  

Oligochitosan samples were obtained by acid hydrolysis of high molecular weight chitosan isolated from Bombyx mori (B.M.). Carrying out acid hydrolysis for 6 hours, it was found that after 4 hours and further with increasing duration, the molecular weight of chitosan decreases to a value corresponding to the ranges of mo-lecular weights of oligochitosan 2–16 kDa. It has been seen that the optimal duration of hydrolysis, leading to the production of oligochitosan with a molecular weight of less than 16 kDa, should be considered 4–5 hours. Depolymerization of chitosan with a molecular weight of 177 kDa was carried out using sodium nitrite in so-lution to obtain oligochitosan with a molecular weight of 6 kDa. On the basis of oligochitosan samples ob-tained by two methods, their ascorbates were received. Under constant conditions with varying the ratio of the components (ChS:AA) and the pH of the solution, the reaction of chitosan ascorbate formation was car-ried out on the basis of the suspension method. The composition, structure, and molecular weight characteris-tics of oligochitosan ascorbate and oligochitosan Bombyx morisamples were confirmed by physicochemical methods. It has been seen that the obtained samples have antimicrobial properties against Fuzarium oxysporum.


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