Sec Analyses

FIGURE 2.1 SEC analysis of a polymeric dextran sample (Mw = 1000) on Superdex peptide HR 10/50. The very high resolution between individual components of the sample is obtained by using two columns in series. Courtesy of T. Andersson. (Reproduced with permission from Amersham Pharmacia Biotech.)... 

For organic SEC separations the use of polystyrene/divinylbenzene (PS/ DVB) particles is almost universal throughout the industry. Polymer Laboratories PS/DVB material, PLgel, which is produced in a series of individual pore sizes, formed the basis for the original product line of SEC columns. Developments in the refinement of particle sizing introduced the benefits of smaller particle size and more efficient columns, which significantly reduced SEC analysis time through a reduction in the number of columns required for... 

Several other examples of modified mobile phases are given in Figs. 13.58 and 13.59 using 90/5/5 TEIF/MeOH/ACN and 95/5 chloroform/w-butylamine for the SEC analysis of poloxamer and nitrile-butadiene rubber samples, respectively. 

Even with mobile-phase modifiers, however, certain polymer types cannot be run due to their lack of solubility in organic solvents. In order to run aqueous or mixed aqueous/organic mobile phases, Jordi Associates has developed several polar-bonded phase versions of the PDVB gels as discussed earlier. Figures 13.60 thru 13.99 detail examples of some polar and ionic polymers that we have been able to run SEC analysis of using the newer bonded PDVB resins. 

A large excess of 1,2-ethanediol (threefold) is reacted with adipic acid until carboxyl groups have reacted. The excess of 1,2-ethanediol is distilled off under vacuum in the presence of Ti(OBu)4 as hydroxy-ester interchange catalyst. The reaction can be followed by SEC analysis of samples withdrawn from reaction medium and stopped when the desired molar mass has been reached (Scheme 2.48). 

Under argon, a mixture of 145 (0.046 mmol), 132 (0.046 mmol), Pd(PPli3)4 (0.2 (junol), Cul (0.2 [miol), and diisopropylamine (0.015 mmol) in THF (4 mL) was stirred in the dark at 50°C for 2 days. Ethynylbenzene (0.92 mmol) was then added and stirred at 50°C overnight. After concentration, die residue was dissolved in CHCI3 and filtered. The filtrate was subjected to preparative SEC with CHCI3 as eluent in order to remove catalyst residues and unreacted starting materials. Polymer 42 was obtained as a yellow solid in 85% yield. SEC analysis (THF, polystyrene standards) Mw = 280,000 (PDI = 6.5). 

Size Exclusion Chromatography (SEC) Analysis of Various Polyisobutylene PIB Architectures... 

The discussion and experimental approach presented here rely on the principles and use of universal calibration for SEC analysis. For a review of this method the reader is referred to several useful articles (1, 3,8,9 ). 

Data V(ml) Table II from SEC/LALLS and conventional SEC analysis of NBS SRM706 polystyrene /M (MJvXlO" (M )vX10-5 g ... 

Data from SEC/LALLS and conventional SEC analysis of two PVA Samples... 

SEC analysis shows that some samples have a blmodal MWD. At this time it is not possible to tell whether the bimodallty is an artifact of the polymerization mechanism or, perhaps, a consequence of partial hydrolysis of the polymer i.e., the high MW shoulder in Figure 5 may be due to the formation of aggregates through Intermolecular dipolar interactions of P-OH side groups or to polymer molecules crosslinked by P-O-P bonds. 

The SEC mechanism demands only an isocratic (constant composition) solvent system with normally a single solvent. The most frequently used organic solvents are THF, chloroform, toluene, esters, ketones, DMF, etc. The key solvent parameters of interest in SEC are (i) solubility parameter (ii) refractive index (iii) UV/IR absorbance (iv) viscosity and (v) boiling point. Sample solutions are typically prepared at concentrations in the region of 0.5-5 mg mL-1. In general an injection volume of 25-100p,L per 300 x 7.5 mm column should be employed. For SEC operation with polyolefins chlorinated solvents (for detector sensitivity and increased boiling point) and elevated temperatures (110 to 150 °C) are required to dissolve olefin polymer. HFIP is the preferred solvent for SEC analysis of polyesters and polyamides. 

In SEC analysis of additive extracts from polymers, the effect of the extraction solvent on the mobile phase is less critical than in HPLC analysis. The extraction solvents typically employed generally do not interfere with the SEC mobile phases. Moreover, the same solvents are often used both as extraction solvent and as mobile phase. Therefore, there is no need to evaporate the extract to dryness prior to analysis and then to redissolve it in a suitable solvent. Typical extraction procedures often produce extracts that generally contain a small amount of wax. Frequently, removal of such oligomers from an extract is necessary, e.g. by means of precipitation, centrifuging, precolumn filtration or protection (use of a reversed-phase guard column). In SEC separations the presence of polyolefin wax does not usually disturb provided that the MW of the wax is higher than that of the analysed compounds. 

SEC-RI/UV has also been used to analyse some 26 thioorganotin compounds, organotin carboxylates and chlorides, essentially PVC stabilisers, and some of their main by-products and related compounds (thioesters and dithioesters, n-alkanes) [803]. Not all organotin chlorides were stable in the adopted analysis conditions. N, Ai -ethylene-bis-stearamide and -oleamide in common plastics (ABS, SAN, PUR, LDPE, PA6.6) can be analysed by SEC after derivatisation with trifluo-roacetic anhydride. SEC analysis of fatty alcohol ethoxy-lates (FAE), used as nonionic surfactants, has also been described [759]. 

For samples that meet the solubility requirements of the SEC approach, analyses were also reported for additives in polymers such as PVC and PS [28,29]. Direct SEC analysis of PVC additives such as plasticisers and thermal stabilisers in dissolution mode has been described [28,30,31 ]. In the analysis of a dissolved PS sample using a SEC column of narrow pore size, the group of additives was separated on a normal-phase column after elution of the polymer peak [21]. Column-loading capacity of HPSEC for the analysis of additives, their degradation products and any other low-MW compounds present in plastics has been evaluated for PS/HMBT, PVC/TNPP and PVC/TETO (glyceryl tri[l-14C] epoxyoleate) [31]. It was shown that HPSEC can be used to separate low-MW compounds from relatively large amounts of polymers without serious loss of resolution of the additives the technique has also been used for the group analysis of chlorohydrin transformation products of the TETO model compound [32]. 

FIGURE 17.25 LCCC separation of a poly(l,6-hexanediol adipate) (a) and SEC analysis of the functionality fractions (b) from Kriiger et al., 1994 (Copyright 1994 from Journal of Liquid Chromatography, 1994, p. 17 by Kruger et al. Reproduced by permission of Taylor Francis Group, LLC., http //www.taylorandfrancis.com). 

GTP was employed for the synthesis of block copolymers with the first block PDMAEMA and the second PDEAEMA, poly[2-(diisopropylamino)e-thyl methacrylate], PDIPAEMA or poly[2-(N-morpholino)ethyl methacrylate], PM EM A (Scheme 33) [87]. The reactions took place under an inert atmosphere in THF at room temperature with l-methoxy-l-trimethylsiloxy-2-methyl-1-propane, MTS, as the initiator and tetra-n-butyl ammonium bibenzoate, TBABB, as the catalyst. Little or no homopolymer contamination was evidenced by SEC analysis. Copolymers in high yields with controlled molecular weights and narrow molecular weight distributions were obtained in all cases. The micellar properties of these materials were studied in aqueous solutions. 

Employing similar procedures, PPO-fc-POEGMA block copolymers and POEGMA-fc-PPO-fc-POEGMA triblock copolymers were prepared from the corresponding PPO macroinitiators [129]. The polymerizations were performed in a isopropanol/water (70/30) mixture at 20 °C using CuCl and bpy. The methacrylate monomer was almost quantitatively polymerized, and the polydispersities were lower than 1.25 in most cases. Less than 5% PPO homopolymer contamination was detected by SEC analysis. 

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