Collagen hydrolysate

Retention time reproducibility is also enhanced with a new HPLC system, especially in the shallow region of the gradient profile. Analysis of collagen hydrolysates and cell culture supernatants are shown as representative applications. 

Figure 3. Separations using optimized conditions for Mixture I separation. Eluent B = eluent titrated to pH 6.80 with NaOH eluent C = acetonitrile. The gradient profile is given in Table 1. The column temperature was 34°C. All other conditions were identical to those in Figure 2B. (A) Mixture I standard, (B) collagen hydrolysate sample, prepared as described in the methods section. Total run time was 54 min.

Human urine, serum and rat brain tissue extracts were treated with sulfosalicylic acid to precipitate protein 20 pi of 35% sulfosalicylie acid was added to 200 pi of each sample. These solutions were vortexed and allowed to sit at room temperature at least 20 minutes before proceeding. The samples were then spun in a microfuge for 2 minutes and the supernatants were collected. Collagen samples were hydrolyzed in 6 N HCl, 110 C for 24 hours. The hydrolysates were then dried down and resuspended in 250 pg/ml K4EDTA. Ant hemolymph was not pretreated before analysis. Samples were loaded on to the analyzer as follows Urine-10 pi of a 1 2 dilution of the supernatant, serum -10 pi of undiluted supernatant, rat brain extract-10 pi of the undiluted supernatant, collagen hydrolysate-1.2 pg in 15 pi, fire ant hemolymph-4 pi of a 1 9 dilution. 

Figure 6. Chromatographic separation of amino acids after derivatization with phenylisothiocyanate (PITC)using physiological conditions A. Free amino acids found in rat brain tissue. B. Free amino acids in ant hemolymph. C. Bone collagen hydrolysate.

The eluents suitable for the separation of amino acids on latex cation exchangers do not comprise the classical citrate/borate buffers but mixtures of nitric acid and potassium oxalate. In comparison to buffers composed of sodium citrate and borate, these components may be obtained at much higher purity. The retention of the amino acids to be analyzed, however, is possibly affected by the sample pH due to the limited buffer capacity of the eluents that are based on nitric acid and potassium oxalate. Fig. 4-21 shows the separation of a calibration standard for collagen hydrolysates on an Amino Pac PA-1 latex cation exchanger at ambient temperature. The advantage is the short... 

Fig. 4-21. Separation of a calibration standard for collagen hydrolysates on a latex cation exchanger AminoPac PC-1. — Eluent (A) 0.00025 mol/L HN03 (B) 0.006 mol/L K2C2O4 + 0.0056 mol/L HNO3, (C) 0.009 mol/L K2C2O4, regenerent 0.005 mol/L HN03 flow rate 1.2 mL/min detection see Fig. 4-19.

Collagen hydrolysates (INCI/ CTFA Hydrolyzed Collagen) ... 

Nutrilan I/I-50/1 Powder Partial collagen hydrolysate pH 4.0-5.0 38-40, 50-52 and 94-97 Care additive in surfactant preparations, hair after-treatment aids and after-sun preparations... 

Z. Zhang, G. Li, B. Shi, Physicochemical properties of collagen, gelatin and collagen hydrolysate derived from bovine limed split wastes, J. Soc. Leather Technol. Chem. 90 (2012) 23-28. 

See Aluminum isostearates/laurates/stearates Aluminum trisulfate. See Aluminum sulfate Aluminum undecylenoyl collagen amino acids Synonyms Collagens, hydrolysates, reaction prods, with undecenoyl chloride, aluminum salts... 

Ammonium hydrolyzed animal protein. See Ammonium hydrolyzed collagen Ammonium hydrolyzed collagen CAS 68951-88-2 222400-34-2 Synonyms Ammonium hydrolyzed animal protein Collagen, hydrolysates, ammonium salts Proteins, hydrolysates, ammonium salts Definition Ammonium salt of hydrolyzed collagen Uses Antistat, hair conditioner, skin conditioner in cosmetics... 

Synonyms Collagen, hydrolysates, isostearoyl, compds. with 2-amino-2-methyl-1,3-propanediol... 

Synonyms Collagens, hydrolysates, prod, with rosin and 2-amino-2-methyl-1,3-propanediol Collagens, hydrolysates, resin acyl, compds. with 2-amino-2-metnyl-1,3-propanediol... 

See AMP isostearoyl hydrolyzed collagen AMP isostearoyl hydrolyzed collagen CAS 95032-84-1 222400-36-4 Synonyms AMP-isostearoyl hydrolyzed animal protein Collagen, hydrolysates, isostearoyl, compds. with 2-amino-2-methyl-1-propanol Definition Aminomethyl propanol salt of isostearoyl hydrolyzed collagen... 

Synonyms Benzyltrimonium hydrolyzed animal protein Collagens, hydrolysates, N,N,N-trimethylbenzenemethanaminium salts... 

Synonyms Capryloyl hydrolyzed animal protein Collagen, hydrolysates, reaction prods, with capryloyl chloride... 

Synonyms Cocamidopropyl dimethylamine hydrolyzed animal protein Collagen hydrolysates, salts with N-(3-dimethylaminopropyl) coco-amides... 

Collagen hydrolysate Ethyl acrylate Single Tg - Mindni et al. (1983)... 

FIGURE 35.9 Cation-exchange chromatogram from the preparative separation of collagen hydrolysate by Hare and coworkers in 1991 (figure with kind permission from Hare et al. [62]). 

The same authors studied the effects of application of collagen hydrolysates on the stress-strain characteristics of different hair 5 cm of virgin, bleached, and cold-wave-tteated hair were taken and stretched to 30% index in water at a rate extension of 0.25 cm/ min. After stretching, the fibers were allowed to relax overnight in water and restretched. Comparison of the stress applied (in kg/mm ) versus strain percent was made before and after treatments with collagen hydrolysate. It was found that when collagen hydrolysate is sorbed to the hair there is an increase in its tensile strength, markedly observable on cold-waved hair. 

Figure 24 Size exclusion chromatography patterns of high-temperature (solid Une) and high-salt (dotted line) fractions desorbed form damaged (a) and virgin hair (b) after application of a collagen hydrolysate. A higher amount of large peptides was adsorbed on bleached/waved hair than on virgin hair. Large peptides were found to be less effectively bound than smaller peptides, as they were selectively removed by hot water (32).

The use of protein hydrolysates and quaternized derivatives in shampoo formulations has been demonstrated to reduce the loss of tensile strength caused by anionic tensides on the hair fibers (124,125). This protective effect is higher for quaternized derivatives than for parent hydrolysates. A wheat protein hydrolysate was found to be more effective than a collagen hydrolysate. Since the two tested products had similar isoelectric point and average molecular size, the difference could be explained by the higher hydrophobicity of the vegetable derivative. 

A specific heavy-metals contamination (chromium) was detected in collagen hydrolysates prepared from trimming/cuttings of chromium-tanned hides up to 100 ppm of chromium was found in samples of powdered collagen hydrolysates on the market at the end of the 1970s (135). The oxidation state of chromium was not determined, but probably it was Cr ", less toxic than Cr, as in the tanning industry only basic salts of Cr " are used. 

Turowski A, Adelmann-GiiU BC. Substantivity to hair and skin of I-labelled collagen hydrolysates under application simulating conditions. Int J Cosmet Sci 1985 7 71-84. 

Mintz GR, Reinhart GM, Lent B. Relationship between collagen hydrolysate molecular weight and peptide substantivity to hair. J Soc Cosmet Chem 1991 42 35-44. 

Work from the Wilson/Inolex companies on collagen hydrolysates (78-80) utilized a staining procedure based on the ninhydrin test for hydroxyproline to demonstrate the adsorption (and penetration) of these materials on keratin substrates. This approach was reported in some detail by Stem and Johnsen (81), who demonstrated the highest substan-tivity for hydrolysates of molecular weight around 1000. 

Maybrook scientists have reported the use of a radiotagging procedure (tritium labeling) to demonstrate the substantivity of collagen hydrolysates to hair (82). 

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