Impurities molecular mass

Some substances under El conditions fragment so readily that either no molecular ions survive or so few survive that it is difficult to be sure that the ones observed do not represent some impurity. Therefore, there is either no molecular mass information or it is uncertain. Under Cl conditions, very little fragmentation occurs and, depending on the reagent gas, ions [M + X]+ (X = H, NH4, NO, etc.) or [M - H] or [M - H]" or [M -1- X] (X = F, Cl, OH, O, etc.) are the abundant quasi-molecular ions, which do give molecular mass information. 

A SSIMS spectrum, like any other mass spectrum, consists of a series of peaks of dif ferent intensity (i. e. ion current) occurring at certain mass numbers. The masses can be allocated on the basis of atomic or molecular mass-to-charge ratio. Many of the more prominent secondary ions from metal and semiconductor surfaces are singly charged atomic ions, which makes allocation of mass numbers slightly easier. Masses can be identified as arising either from the substrate material itself from deliberately introduced molecular or other species on the surface, or from contaminations and impurities on the surface. Complications in allocation often arise from isotopic effects. Although some elements have only one principal isotope, for many others the natural isotopic abundance can make identification difficult. 

The resolution required in any analytical SEC procedure, e.g., to detect sample impurities, is primarily based on the nature of the sample components with respect to their shape, the relative size differences of species contained in the sample, and the minimal size difference to be resolved. These sample attributes, in addition to the range of sizes to be examined, determine the required selectivity. Earlier work has shown that the limit of resolvability in SEC of molecules [i.e., the ability to completely resolve solutes of different sizes as a function of (1) plate number, (2) different solute shapes, and (3) media pore volumes] ranges from close to 20% for the molecular mass difference required to resolve spherical solutes down to near a 10% difference in molecular mass required for the separation of rod-shaped molecules (Hagel, 1993). To approach these limits, a SEC medium and a system with appropriate selectivity and efficiency must be employed. 

The selectivity of a gel, defined by the incremental increase in distribution coefficient for an incremental decrease in solute size, is related to the width of the pore size distribution of the gel. A narrow pore size distribution will typically have a separation range of one decade in solute size, which corresponds to roughly three decades in protein molecular mass (Hagel, 1988). However, the largest selectivity obtainable is the one where the solute of interest is either totally excluded (which is achieved when the solute size is of the same order as the pore size) or totally included (as for a very small solute) and the impurities differ more than a decade in size from the target solute. In this case, a gel of suitable pore size may be found and the separation carried out as a desalting step. This is very favorable from an operational point of view (see later). 

In the early days of polymer science, when polystyrene became a commercial product, insolubility was sometimes observed which was not expected from the functionality of this monomer. Staudinger and Heuer [2] could show that this insolubility was due to small amounts of tetrafunctional divinylbenzene present in styrene as an impurity from its synthesis. As little as 0.02 mass % is sufficient to make polystyrene of a molecular mass of 2001000 insoluble [3]. This knowledge and the limitations of the technical processing of insoluble and non-fusible polymers as compared with linear or branched polymers explains why, over many years, research on the polymerization of crosslinking monomers alone or the copolymerization of bifunctional monomers with large fractions of crosslinking monomers was scarcely studied. 

SDS polyacrylamide gel electrophoresis (SDS-PAGE) represents the most commonly used analytical technique in the assessment of final product purity (Figure 7.1). This technique is well established and easy to perform. It provides high-resolution separation of polypeptides on the basis of their molecular mass. Bands containing as little as 100 ng of protein can be visualized by staining the gel with dyes such as Coomassie blue. Subsequent gel analysis by scanning laser densitometry allows quantitative determination of the protein content of each band (thus allowing quantification of protein impurities in the product). 

Calibration with standards allows accurate determination of the molecular mass of the product itself, as well as any impurities. Batch-to-batch variation can also be assessed by comparison of chromatograms from different product runs. 

Figure 2. Resolution required to resolve molecular impurities around mass 88 on...

The molecular mass would be too high because the nonvolatile impurities would contribute additional mass. The contribution to volume would be negligible. 

Various liquid chromatographic techniques have been frequently employed for the purification of commercial dyes for theoretical studies or for the exact determination of their toxicity and environmental pollution capacity. Thus, several sulphonated azo dyes were purified by using reversed-phase preparative HPLC. The chemical strctures, colour index names and numbers, and molecular masses of the sulphonated azo dyes included in the experiments are listed in Fig. 3.114. In order to determine the non-sulphonated azo dyes impurities, commercial dye samples were extracted with hexane, chloroform and ethyl acetate. Colourization of the organic phase indicated impurities. TLC carried out on silica and ODS stationary phases was also applied to control impurities. Mobile phases were composed of methanol, chloroform, acetone, ACN, 2-propanol, water and 0.1 M sodium sulphate depending on the type of stationary phase. Two ODS columns were employed for the analytical separation of dyes. The parameters of the columns were 150 X 3.9 mm i.d. particle size 4 /jm and 250 X 4.6 mm i.d. particle size 5 //m. Mobile phases consisted of methanol and 0.05 M aqueous ammonium acetate in various volume ratios. The flow rate was 0.9 ml/min and dyes were detected at 254 nm. Preparative separations were carried out in an ODS column (250 X 21.2 mm i.d.) using a flow rate of 13.5 ml/min. The composition of the mobile phases employed for the analytical and preparative separation of dyes is compiled in Table 3.33. 

Webster s definition for database is a large collection of data in a computer, organized so that it can be expanded, updated, and retrieved rapidly for various uses. An LC/MS database established for drug impurities contains multi-dimensional information such as relative retention times, UV spectra, molecular mass and substructural information. In order for the information to be updated and expanded, the methods used for information collection need to be unified. A generic LC/MS method allows relevant information to be collected in a consistent... 

Rejection of a fraction of macromolecules, or of impurities, or both, from growing crystals. Note The rejected macromolecules are usually those of insufficient relative molecular mass, or differing in constitution or configuration (e.g., branching, tacticity, etc.). 

C) Examine the electropherograms obtained under reducing conditions in the test for impurities of molecular masses differing from that of interferon-a.2. The principal band in the electropherogram obtained with test solution (a) corresponds in position to the principal band in the electropherogram obtained with reference solution (a). 

Impurities of molecular mass differing from that of interferon-a.2... 

Although CuBS2 elutes at approximately the same position in the salt gradient as CdBSl, this peak is not exclusively PC or glutathione. The amino acid composition of essentially pure CuBS2 (Table 2) reveals a complex mixture of amino acids although high in Glx, Cys and Gly (50%), the amino acid composition is not characteristic of other impure or purified PCs which have 75-90% Glx, Cys and Gly (Table 1 Salt et al., 1989 Tomsett etal., 1989). Furthermore, it is estimated to have a molecular mass of 2700 Da and is remarkably similar to the class I MT from A. bisporus (Table 2). 

Mass spectrometry Purity and impurities, molecular weight, glycosylation... 

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