452 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS 1.0 0.9 0.8 0.7 0.6' 0.4' uclease A 13,700 0.3 %Chymotrypsinogen A 25,000 t 0 1 Ibumin 45,000 I Aldolase 158,000 lX103 lX104 lX10 s Molecular Weight Fiõure 2. Calibration curve, G-?5 lX10 s 20- 18- 14- 12- 8- m Enzyme Hydrolysate ß ,.m.• Acid Hydrolysate • Steam Hydrolysate 8 10 12 14 16 18 20 22 24 Molecular Weight X 103 Figure 3. Molecular weight distribution curves 26 28 30
COSMETIC PROTEIN HYDROLYSATES 453 the column with the low molecular weight material. When preparing the molecular weight distribution curve of the enzyme hydrolysate, the absorbance due to the pre- servative had to be subtracted. Figure 2 is the calibration curve, and shows the relationship of molecular weight to elu- tion volume. The elution volume is expressed as Kav, which is the relationship of the elution volume to the void volume and the total bed volume. Kav= Ve - Vo Ve - Vo As can be seen from the calibration curve, the elution volume has a straight line rela- tionship to the logarithm of the molecular weight over most of the range between the void volume and the total bed volume. From the elution diagrams and the calibration curve, molecular weight distribution curves were drawn. These graphs are shown in Fig. 3. These distribution curves are used to calculate both the weight average and the number average molecular weights. The weight average and the number average molecular weights were calculated as follows. Pi (Pi/Mi) where Mw = weight average molecular weight P = per cent of material at a constant elution volume interval Mi = molecular weight at this interval Mn = P•/(P•/M•) Mn = number average molecular weight P• = percentage of material at a constant elution volume interval M• = molecular weight at this interval In Table I the molecular weight averages calculated from the molecular weight dis- tribution curves are shown. ULTRACENTRIFUGATION In Table II, the results of the ultracentrifuge run are compared to the gel filtration results. Table I Protein Hydrolysate Weight Average Number Average Enzyme 4,000 + 10 per cent 2,100 + 10 per cent Acid 9,300 +'10 per cent 2,500 - 10 per cent Steam 7,800 - 10 per cent 1,800 - 10 per cent
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