The method for determination of both free and esterified cholesterol, depending upon the precipitation of free cholesterol as the digitonide, has been most widely accepted and used, although it was time-consuming and technically difficult procedure. Recently the chromatographic procedure using silica gel column has become available for the same purpose, but the methods reported previously require relatively large amount of sample. This paper describes a micromethod using silica gel column for separation of free cholesterol from its esters.
The procedure is as follows: -To approximately 8ml of ethanol-ether (3:1) in a 10ml volumetric flask is added 0.5ml serum. The suspension is brcught to boiling, colled, and made to 10ml with the same solvent, A 5ml aliquot of the filtrate is evaporated to dryness and red is solved in 5ml of hexane-ether (99:1).
The solution is placed upon the chromatographic column (100mm by 5mm), containing 1mg of activated sillica gel. The ester fraction is completely eluted with 20ml cf hexaneether (50:50). The ester fraction is saponified by Abell-Pearson's procedure.
The contents of cholesterol in both fractions are determined with Liebermann-Burchard reaction.
The method was found to be rapid, simple and quantitative.

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The quantitative Estimation of urinary 17-ketosteroids (KS) is generally based on the Zimmermann's reaction in the routine laboratory and still presents a number of problems. Among the rest, we must be careful about the nonketonic chromogens in the final urinary extract, In order to remove these nonketonic chromogens, Talbot et al, have conducted the application of Girard's reagent T, but this method is not feasible for the routine clinical laboratory because of the complexity of its operation, Therefore, empirical correction equations were studied for deducting the interfering color absorption, but they were found unreliable by various authors. On the other hand, various solvents were used to isolate the urinary 17-KS but they were not so selective, However, Cahen and Salter's Method with chloroform is relatively selective to isolate the urinary 17-KS, so we examined the method of Cahen and Salter and improved it as follows
1) We have pointed out the nonketonic chromogens still included in the chloroform extract by Cahen and Salter's method and elevated the accuracy of the value of quantitative estimation of urinary 17-KS, deducting the urinary blank value from the value by Cahen and Salter's method, The value of urinary blank indicates that of the color absorption of chloroform extract of the urinary dried ether extract added only with alcohol and KOH.
2) The methods deducting the interfering color by empirical correction equations were studied and found unreliable on account of giving plus error in case of the specimens whose blank held purple color in the ether extract.
3) In the procedure of laboratory operation, it was devised to carry out all processes i. e. hydrolysis and extraction using a small volume of urine, washing, color development and chloroform extraction, with only one centrifuge tube with glass stopper. Therefore, the hydrolysis by the customary method employing a reflux condenser and the complicated manipulation using a separatory funnel were all saved. Consequently this simplified method enabled to spare time for the operation and moreover ceased bringing about the error that results from the increase of density of 17-KS due to a loss in quantity of ether during the process of extraction and washing.
4) Turbidity due to emulsion particles in the chloroform layer, giving consequently high absorption values, becomes perfectly clear by the addition of small volume of absolute ethanol.
5) The values of urinary 17-KS obtained by our method tally well with those obtained by the Girard's reagent T, and recovery of dehydroisoandrosterone added to urine is satisfactory averaging 95 percent.

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Authors recognized that a complex acid mucopolysaccharide was histochemically found in the walls of capillaries and arterioles within some leprous skin lesions.
This substance is to be recognized as mucoitin sulfuric acid from the staining attitude with Hempelmann stain.
From a viewpoint of the classification of leprosy, such substance appears to be found in lepromatous type, especially at the stage of acute ENL reaction, but not in tuberculoid type of leprosy.

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