Leon L. Gershbein

Hydrocarbons of dogfish and cod livers and herring oil.

Dogfish and cod liver oils and the oil from the whole herring were saponified and the hydrocarbons concentrated by chromatography of the unsaponifiable portion over alumina followed by silica gel treatment of the resulting fractions. Temperature programmed gas chromatography employing a 3% SE-30 packing was applied to the analysis of hydrocarbons of C14 to C32.5. The paraffins comprised two or more groups. Dogfish liver oil gave rise to 7.62% unsaponifiables and pristane, other saturated types, squalene and an additional group, high in unsaturated components, were 193, 325, 308 and 200 mg% in this portion or 15.7, 24.8, 23.5 and 15.3 mg%, respectively, in the oil. Cod liver oil yielded 1.0% unsaponifiables of which the above hydrocarbons in the order stated amounted to 0.30%, 1.15%, 3.29% and 2.27% or 3.0, 11.5, 32.9 and 22.7 mg% in the liver oil. The unsaponifiable material of herring oil (1.35%) was prominent in paraffinic hydrocarbons, the levels of the above specified components being 16.34%, 3.51%, 0.99% and 1.41% as stated or 221, 47.4, 13.4 and 19.1 mg% in the oil. The sterol and alcoholic contents were ascertained for the three marine oils and the glyceryl ether levels found to be highest for dogfish liver oil.

Phospholipids of human hair lipids

Abstract Thin-layer chromatography has been applied to the separation of a mixture of phospholipids from human hair lipids and to its resolution into 8 components. The latter consisted of glycerophosphorylcholine, phosphatidic acid, lysophosphatidylcholine, phosphatidylinositol, phosphatidylcholine, phosphatidylserine and phosphatidylethanolamine; the last three phosphatides made up 65 % of the mixture. The purified phospholipids were submitted individually to hydrolysis and the respective fatty acid esters analyzed by gas chromatography. Sphingomyelin contained the highest level of saturated acids and the ratio of mono- to polyunsaturated acids was close to 1. Phosphatidylethanolamine acids ranged high in unsaturated members and displayed an average double bond number of 1.3/molecule.

Fractionation of hydrocarbons of human hair lipids by chromatographic and thermal diffusion methods

Abstract Hair lipids from adult white and Negro men were individually pooled, the two batches saponified and the unsaponifiable portions in petroleum ether chromatographed over alumina. The column on elution with petroleum ether as such and containing 5 and 10% chloroform, yielded Fractions I–II, inclusive, on removal of the solvents; 100% chloroform eluted another portions. Fraction IV, which in contrast to the latter three, was essentially free of hydrocarbons. Waxy components of C 20 —C 44 , all normal hydrocarbons, were obtained from Fractions I, II and III by treatment with acetone and ranged highest in Fraction I. The residual fluid (Frac. I-Oil) was chromatographed in petroleum ether solution over silica gel and contained olefinic hydrocarbons in addition to paraffinic and naphthenic components. The corresponding oils from Fractions II and III were high in unsaturated hydrocarbons, notably squalene. Good enrichment of the saturated hydrocarbons was accomplished by use of thermal diffusion. The distribution of carbon (C p and C N ) and the mean naphthenic number per molecule ( R N ) for cuts obtained by this criterion were calculated by the n-d-M method; the highest ( R N was in the range of 5.5. Temperature programmed gas chromatography showed the presence of C 15 —C 34 hydrocarbons in the initial liquid mixture and derived cuts, all of the normal paraffins in this range being represented as summarized in Table V.

Isolation and characterization of glycerides in human hair lipids by thin-layer and gas chromatography

Techniques for the quantitative analysis of hair lipids using thin-layer chromatography (TLC) together with a proximate analysis of components in one sample deduced by these criteria are presented. Mono-, di- and triglycerides were separated by TLC using Silica Gel G as adsorbent. The chromatoplates were developed with 98% acetone+2% petroleum ether. Glycerides moved with the solvent front. The requisite portions were scraped off the plates and extracted with acetone and ether. Further TLC, limiting the migration of triglycerides and diglycerides was afforded by use of 95% ethanol as solvent in one direction while monoglycerides moved with the solvent front. For the separation of monoglycerides, chloroform was used as solvent in a second direction. Reference standards and several mixtures were run simultaneously and the spots identified by charring with concentrated sulfuric acid containing dichromate. Additional checking was effected by IR spectra. For determination of glyceride composition, methyl esters of the component fatty acids were prepared by transesterification and submitted to gas chromatography. Comparison of the levels of each of the constituent fatty acids showed no remarkable differences between the three classes of glycerides in one hair lipid pool. Although certain discrepancies in the amounts of a few fatty acid components might be construed for one pool of lipids from hair of white full-headed men (WF-9A) in contrast to findings with two Negro pools, no unequivocal conclusions can be drawn presently.

Hydrocarbons and alcohols of basking shark and pig liver lipids

Four samples of the unsaponifiables of basking shark liver oil were adsorbed on alumina and eluted to yield Fractions 1–5, inclusive. Analyses by temperature programmed GC and by silica gel chromatography showed hydrocarbons in the first four fractions with squalene increasing to Fraction 3 and the pristane level being highest in Fraction 1. Aside from pristane and squalene, other hydrocarbons occurred at levels of 420–750 mg% in the oils on a weight basis, of which about 60% constituted a series of n-paraffins (relative carbon number range: 15.0–38.0) together with smaller amounts of at least one branched chain saturated group. Unsaturated hydrocarbons eluted mainly after squalene. The oils contained up to 460 mg% sterol and 78–270 mg% alcohols of C10 to C30, the ratio of saturated to unsaturated members being about 1.6. The composition of the unsaponifiable lipids of pig liver was quite different from that of the marine oils. It contained 10.6% sterol in addition to 400 mg% alcohols, the latter consisting of 81.8% saturated components (C12 to C31; ratio of saturated: unsaturated members, 4.4). The hydrocarbons comprised 450–700 mg% of the unsaponifiable mixture and squalene, paraffins and additional unsaturated components occurred at levels of 20.6, 24.4 and 11.9 mg%, respectively. The saturated hydrocarbons were high in normal homologs of relative carbon number range, 15 to 36; pristane could not be detected.

In vivo depression of hepatic succinoxidase by sulfhydryl compounds.

Abstract Thioglycolate injected i.v. in dosages of 50 mg/kg, significantly depressed hepatic succinoxidase in immature rats of either sex and adult males whereas, about a five-fold excess was required to elicit a significant response in adult females. Ovariectomy increased and estradiol pretreatment, decreased the sensitivity to thioglycolate. Of a number of sulfur compounds screened on hepatic succinoxidase in males, only 2-mercaptoethanol shared this action with BAL and thioglycolate. The latter two agents were without effect on cytochrome oxidase and diaphorase but both depressed NADH cytochrome c reductase. It is suggested that the mechanism of succinoxidase inhibition involves a combination with Slaters factor, thereby causing a disruption in the normal flow of electrons.

Sub-banding and fine structure of serum lactate dehydrogenase isoenzymes induced by sulfur compounds

Abstract Human serum was incubated 1:1 by volume with solutions of several sulfur compounds of pH 7.2 for at least 24 hours and the total LDH-P determined and the distribution of LDH isoenzymes and fine structure examined by polyacrylamide disc electrophoresis. As with penicillamine, the disulfide but not N-acetyl-penicillamine, caused removal of LDH-5 without affecting the total unitage. The latter was depressed by thiophenoxyacetate (0.10-0.40 M), undiluted dimethyl sulfoxide and levamisole (0.025 M and higher) but not by the last agent at 0.0005-0.001 M which gave rise to fine structure in the gels. As screened in a number of sera of 160–2300 U/L in total LDH, phthalyltetrathioacetate (0.2-1.0 M) elicited 3 unique sub-bands as pre-LDH-2, pre-LDH-3 and pre-LDH-4 to the exclusion of any effect on the total LDH.

Liver lipids of the polar bear,Thalarctos maritimus

The distribution of total and free fatty acids as well as the acids from the glycerides, sterol esters and phospholipids of polar bear liver lipids was ascertained and found to contain somewhat higher levels of unsaturated components as compared to those of such mammals as the pig. Saponification of the liver lipids yielded the hydrocarbons, alcohols and sterols which were analyzed by GLC. The hydrocarbons occurred at an overall level of 55 mg/kg liver or 57.9 mg/100 g total lipids, of which pristane and other saturated hydrocarbons, mainly normal homologs, comprised 2.6 and 5.3 mg/100 g, respectively; the remainder contained squalene (37.7 mg/100 g) and other unsaturated types (12.3 mg/100 g). As based on the total lipids, the levels of fatty alcohols, sterol and glyceryl ethers amounted to 1.65%, 5.9% and 0.03%, respectively. The fatty alcohols displayed about 31 peaks of C12 to C30, of which the hexadecanol and a branched C20 component were prominent.

Secretin inactivation by blood of various animal species

Abstract 1. 1. Sccretin concentrates as well as the natural porcine peptide were incubated with serum from a number of animal species at 37° C. and the extent of inactiation with time ascertained from the ratio of dosages of saline to test mixtures, each eliciting the same submaximal response from the dog. 2. 2. As tested with mouse serum and several concentrates, liberation of ammo-acids accompanied the destruction of hormonal activity. 3. 3. The protease levels of the sera were of that relative order of magnitude as to produce the rates of inactivation determined for each. The rate ranged highest for mouse serum and lowest for species such as the cow. 4. 4. Rat tissue extracts and Conn fractions derived from human plasma and mouse serum also engendered secretin metabolism.

Disc and cellulose acetate electrophoresis of human placental proteins.

Abstract A protein-rich concentrate, PLSR, has been isolated from acetone-dried human term placenta by steps involving extraction with water, dialysis and treatment of the dialysis residue with 95% ethanol and the composition analyzed by electrophoresis on cellulose acetate and polyacrylamide gel. The latter criterion showed the presence of two pre-albumin bands and which could be further enriched by subjecting PLSR to C ohn fractionation procedures. Pre-albumins occurred in only trace amounts in concentrates isolated from late pregnancy and cord sera. As based on electrophoresis on cellulose acetate, splitting of the γ-globulin was observed with several of the fractions. Of a number of media tested, water was quite effective in the extraction of the acetone-dried powder although PLSR obtained by way of a borate buffer of pH 9.0 contained appreciable pre-albumin. Disc electrophoresis of human chorionic gonadotrophin applied as a solution containing 39,000 IU/ml revealed bands analogous to those of PLSR but in contrast to the latter, the zones were of low intensity.