STUDIES OF CRYPTOCOCCUS POLYSACCHARIDES BY INFRARED SPECTROPHOTOMETRY
|
|
- Annabel Burns
- 5 years ago
- Views:
Transcription
1 STUDIES OF CRYPTOCOCCUS POLYSACCHARIDES BY INFRARED SPECTROPHOTOMETRY SEYMOUR LEVINE, E. EDWARD EVANS, AND PAUL W. KABLER From the St. Francis Hospital, Jersey City, New Jersey; the University of Alabama Medical Center, Birmingham, Alabama; and the Robert A. Taft Sanitary Engineering Center, U. S. Public Health Service, Cincinnati, Ohio Infrared spectrums of isolated capsular polysaccharides have been used to segregate types within certain groups of encapsulated bacteria. The results of spectral typing of purified pneumococcal polysaccharides (1) and of crude klebsiella polysaccharides (2) have paralleled the results of serological typing. In a few cases the spectral method was less sensitive, so that certain serological types could not be differentiated. In a few cases the spectral method was more sensitive in differentiating strains than standard capsular reactions and agglutination tests, and in these cases the subtypes established by spectral analysis received serological confirmation by the more refined method of capsular reactions with reciprocally absorbed serums (2). In the present study, the technique of infrared spectrophotometry has been applied to the yeast-like fungus, Cryptococcus neoformans. Strains of C. neoformans have been divided into three types by capsular, agglutination, and precipitin reactions (3, 4). The purpose of this report is to compare the infrared spectral analysis of purified cryptococcal capsular polysaccharides with the serological data. MATERIALS AND METHODS Cultures of C. neoformans were grown in neopeptone dialysate broth with constant agitation (5). In one instance, a type A culture was grown in a casein hydrolysate medium (ph 7) of the fol- Received for publication October 28, Supported in part by a research grant from the National Institutes of Health, Public Health Service. 269 lowing composition: Casein hydrolysate-acid (Nutritional Biochemicals Co.) Glucose Yeast extract (dialysate) 1.0% (vjv) 2.0% 0.1% Cell-free filtrates (5) of cultures were acidified by the addition of glacial acetic acid to give a final concentration of 1% (v/v). Following this, sodium acetate crystals (% w/v) were dissolved by stirring. The ph at this point was 5.3 to 5.4. The capsular polysaccharide (S) was precipitated by the addition of ethanol. Varying quantities were necessary to effect precipitation. Type A polysaccharide (SA) generally required from 1.5 to 2.0 volumes of ethanol. Type B (SB) precipitated with 0.6 to 0.9 volumes. Type C polysaccharide (SC) generally precipitated with 0.5 to 0.6 volumes. The S precipitate was then dissolved in water together with additional acetic acid and sodium acetate and the precipitation with ethanol was repeated as before from two to five times. Certain samples were subjected to other procedures before the final ethanol precipitation. A number of samples (see table 1) were freed from traces of protein by Sevag's method (4, 6). One type B preparation was precipitated initially with 2.5 volumes of ethanol (SB8); since this material was contaminated with fraction R it was further purified by precipitation with lead acetate (5). The S fraction precipitated while R remained in the supernate. This lead-precipitated material was also treated by Sevag's method and is referred to as SB1 in the table. The lead was removed by electromigration (5). Most S preparations have been given a final ethanol precipitation in the presence of sodium acetate and acetic acid, then rinsed with ethanol and ether on hard filter paper and desiccated. This procedure resulted in a high ash value (5) and necessitated a correction for sodium content when weighing. More recently, the final aqueous solution of the polysaccharide has been dialyzed against cold distilled water and lyophilized. The samples were dissolved in distilled water and air dried on the surfaces of silver chloride discs that were slightly tilted so as to produce wedge-shaped films of variable thickness (8). The spectrums were recorded automatically by a Perkin-Elmer model 21 double beam infrared Downloaded from at Penn State University (Paterno Lib) on May, 2016
2 270 SEYMOUR LEVINE, E. EDWARD EVANS, AND PAUL W. KABLER TABLE 1 Sample Strain Methods of purification Absorbance Absorbance Absorbance ratio 5.85 I' 6.25 I' 5.851'/6.25 I' SA2 RE E~1. 75) SA3 RE E 1.75)+S SA4 RE E(1. 75) SA6 RE E~1.75)+S SA5 DU E 1.5 )+S SA7 DU E~1.5 ) SA E 1. 75) SA9 732 E(1.5 ) SB E(2.5 ) SBI 1523 E(2.5 )+L+M+S SB E~0.6 )+S SB E 0.6 ) SB E(0.9 ) SB7 L2 E(0.75) SCI LE E(2.5 ) SC2 LE E(0.5 )+S SC3 LE E(0.8 ) Abbreviations: E =ethanol precipitation; figures in parentheses represent volumes of ethanol used; S = Sevag method of deproteinization; L =Iead precipitation; M =electromigration. spectrophotometer with sodium chloride prism. The variable thickness films were adjusted in the sample beam to give 15% transmission at 9.5 microns (p) wavelength. RESULTS AND DISCUSSION All the spectrums showed absorption maximums at nearly the same wavelengths, but there were differences in the intensity of the bands (figure 1). Occasionally a weak band in one spectrum was represented only by an inflection in another spectrum. The strongest band in all the spectrums was a broad absorption from 8.5 IJ. to.5 IJ.. Its deepest point was usually at 9.5 IJ., but especially in type Band C preparations, the band was resolved into two components with absorption maximums at 9.35 and 9.6 J.I.. The descending limb showed an inflection at 8.9 IJ. (and in type Band C preparations a second inflection at 8.7 IJ.), and the ascending limb showed an inflection at.2 IJ.. Klebsiella and pneumococcal polysaccharides, and in fact most carbohydrates, have their broadest and deepest infrared absorption in this region. Strong absorptions at about 6.2 and 7.1 IJ. were present in all the cryptococcal spectrums. These have been described previously in all klebsiella spectrums and in many of the pneumococcal spectrums, and evidence has been presented to indicate that they were due to the carboxylate ion, probably in the form of uronic acid salts (1). Weak absorptions or inflections were present at about.9, 11.2, and 12.5 IJ.. The long wavelengths have been called the "fingerprint region" because identifications of many organic compounds have been based on specific configurations in this portion of the spectrum; this was equally true in identifying closely related klebsiella and pneumococcal types. However, all the cryptococcal polysaccharides showed the same general configuration in this region, with minor differences in band depth that could not be correlated with serological type. All the cryptococcal polysaccharide spectrums had bands at about 5.8 IJ. and IJ.. In contrast to the above, the depths of these two bands varied greatly in the different spectrums. The 5.8 J.I. band was in the carbonyl region of the spectrum, while the 8.0 to 8.1 IJ. band was in a region where esters absorb strongly; it is likely that both bands were due to the ester linkage, probably acetyl groups bound to the hydroxyls of the constituent sugar chains. Also associated with these two bands was a weak absorption at about 7.25 IJ. which Downloaded from at Penn State University (Paterno Lib) on May, 2016
3 CRYPTOCOCCUS POLYSACCHARIDES 271 IIIftl.MIU IICt $ ,f--r-,rr-1-r,-r-r--r----,--,--,---,---, '0.1 r J L---L_L---L_L---L_L---L_L---L---J.' 0 0 to II IAVELElCnllIIIICI ' Type 6.1 J IIL--"---'---"---'---"---'---"---'---"---'.I' 0...,...-,--r-r---.~-~,----r--r-----r---, 5C-3 Type C FIG. I.-Infrared spectrums of purified cryptococcus polysaccharides of Types A, B, and C. The bands at 5.8 JL and at 8.0 to 8.1 JL are strong in SA6, weak in SC3 and intermediate in SBI. may have been due to the terminal methyl of the acetyl group. These bands have been described in many of the klebsiella and pneumococcal polysaccharides (1, 2). Their correlation II with serological typing of cryptococci is described below. As a group, the cryptococcal spectrums resembled the klebsiella more than the pneumococcal spectrums. All the cryptococcal and all the klebsiella spectrums showed strong carboxylate ion bands (6.2 and 7.1 )1.). A minority of the pneumococcal spectrums showed carboxylate bands, while amide bands were very frequently observed; amide bands were never observed ill either cryptococcal or klebsiella spectrums. Effect of method of preparation The growth medium is an important factor when spectrums of whole cells are recorded but IS much less important when purified polysaccharides are examined. Polysaccharide from a type A culture grown in neopeptone dialysate broth was not spectrally different from polysaccharide of the same strain grown in casein hydrolysate medium. Infrared spectrums are additive, so that impurities influence the spectrum in proportion to their amount. For example, the removal of traces of protein from a number of specimens influenced the spectrums very little or not at all. However, several of the spectrums showed evidence of sodium acetate impurity, in the form of broadening and deepening of the 6.2 and 7.1)1. carboxylate bands; this effect could be induced by deliberately adding sodium acetate to a pure preparation. Dialysis of the contaminated preparations resulted in sharpening and decreased intensity of the carboxylate bands. The presence of fraction R could not be detected in the spectrums. The spectrum of SB8, containing R, was identical with those of SB1 and SB2 whose fraction R had been removed. Similarly, SC1 which contained fraction R was spectrally identical with SC2 and SC3 which lacked it. Through the kindness of Mrs. Julia Downloaded from at Penn State University (Paterno Lib) on May, 2016
4 272 SEYMOUR LEVINE, E. EDWARD EVANS, AND PAUL W. KABLER Einbinder we were able to examine capsular polysaccharides prepared from a pathogenic strain of C. neoformans and from a strain of C. neoformans var. innocuous by a completely different method (extraction with 30% potassium chloride and 1% potassium carbonate) (7). The configurations of the spectrums were similar to those illustrated here except that the 5.8 and 7.25 JL bands were absent and the 8.0 to 8.1 JL band was very weak. In contrast, spectrums of whole untreated cryptococcal cells, of both strains, showed that the three bands were present along with bands attributable to other cell constituents. These results suggested the possibility that the method of extraction had destroyed acetyl groups and thus altered the polysaccharide spectrum. In order to test this possibility, aliquot samples of the dried whole cryptococcal cells were subjected to extraction by boiling water, by water at 0 C, by water at room temperature, by 30% potassium chloride at room temperature, and by 30% potassium chloride plus 1% potassium carbonate at room temperature. The polysaccharides were isolated by alcohol precipitation and examined without further purification. The acetyl bands at 5.8, 7.25, and 8.0 to 8.1 JL were present in the first four preparations and almost completely absent from the last. These results indicated that the potassium carbonate was responsible for the loss of acetyl groups and consequent spectral alterations. Potassium carbonate is an alkaline salt, and ester linkages are readily hydrolyzed at alkaline ph. Potassium chloride is a neutral salt and hence inactive in this respect. This experiment was also carried out on a klebsiella strain with identical results. Correlation of spectral results and serological typing Spectral comparison of all the preparations showed large and consistent differences only in the acetyl bands (5.8 JL carbonyl band and 8.0 to 8.1 JL ester band) (figure 1). All the type C preparations had very weak carbonyl and ester bands. All the type A and B preparation had strong or moderate carbonyl and ester bands; those in type A tended to be stronger than those in type B, but there was sufficient overlap to prevent spectral differentiation. In the table the depth of the 5.8 JL carbonyl band of each preparation has been indicated by its absorbance value. It is possible to compare the values taken from different spectrums because each spectrum was recorded in an area of the polysaccharide film whose thickness was the same (8) (15% transmission at 9.5 JL). Therefore the lower absorbances (higher transmissions) recorded for type C preparations probably mean lower carbonyl (acetyl) content. The figures cannot be interpreted quantitatively because of uncontrollable factors such as nonspecific light-scattering. Another approach is the calculation of the ratio between the absorbance at 5.8 JL and the absorbance of the neighboring carboxylate band at 6.2 JL; these absorbance ratios are listed in the table and they may give a slightly more accurate indication of the relative acetyl content of the polysaccharides. Visual comparison of the depths of the 5.8 and 6.2 JL bands (first two bands in each of the spectrums in figure 1) gives essentially the same indication of relative acetyl content. Neither these ratios nor a number of others permitted the spectral differentiation of types A and B due to the overlap of some of the values.* In contrast, type C preparations were easily identified by their low acetyl content. A parallel may be noted with serological results which have demonstrated * It is possible that this differentiation would be facilitated by a more quantitative technique, such as the incorporation of the polysaccharide in potassium bromide peliets. Downloaded from at Penn State University (Paterno Lib) on May, 2016
5 CRYPTOCOCCUS POLYSACCHARIDES 273 a closer relation between types A and B than between type C and either A or B (3,4). In view of the basic similarity of the cryptococcal spectrums, it seems possible that their differences in serological reactivity may depend in part on the degree of acetylation of the polysaccharide chains. The previous studies of the pneumococcal (1) and klebsiella (2) groups showed that many types were easily identified by numerous type-specific spectral bands and configurations. There were only a few instances in which spectral differentiation was difficult or impossible, or in which differentiation had to be based on differences merely in intensity of certain bands (acetyl bands), and these instances occurred in closely related strains. Therefore the present results with cryptococcal polysaccharides have provided additional evidence that they are, as a group, closely related. SUMMARY Infrared spectrums of purified cryptococcal capsular polysaccharides have been presented. Some of the absorption bands were attributed to carboxylate ions and to acetyl groups in ester linkage; these bands have been described previously in klebsiella and some pneumococcal capsular polysaccharides. Klebsiella and pneumococcal polysaccharides were characterized by many type-specific bands and configurations, but polysaccharides from the three cryptococcal types showed generally similar bands indicating the close relationships in this group. There were, however, large differences in the intensity of certain bands. Type C with very weak acetyl bands was easily differentiated from types A and B with strong and intermediate acetyl bands; different strains of types A and B showed some overlap in values. The spectral results thus paralleled the previous demonstration of a closer relation between types A and B than between type C and either A or B. It is possible that differences in degree of acetylation play an important role in determining the serological specificity of cryptococcal capsular polysaccharides. Impurities of protein or fraction R were not detectable in the spectrums. Contamination with sodium acetate was detected in some preparations and was eliminated by dialysis. Evidence is presented to show that the acetyl groups in ester linkage were destroyed if alkaline salts were employed in the preparation of polysaccharides. REFERENCES 1. Levine, S., Stevenson, H. J. R. and Kabler, P. W. 1953, Arch. Biochem. Biophys. 45: Levine, S., Stevenson, H. J. R, Bordner, R H. and Edwards, P. R. 1955, J. Infect. Dis. 96: Evans, E. E. 1950, J. Immuno!' 64: Evans, E. E. and Kessel, J. F. 1951, J. Immuno!' 67: Evans, E. E. and Theriault, R J. 1953, J. Bact. 65: Sevag, M. G. 1934, Biochem. Ztschr. 273: Einbinder, J. M., Benham, R. S. and Nelson, C. T. 1954, J. Invest. Dermat. 22: Riddle, J. W., Kabler, P. W., Kenner, B. A, Bordner, R H., Rockwood, S. W. and Stevenson, H. J. R. 1956, J. Bact. 72: Downloaded from at Penn State University (Paterno Lib) on May, 2016
Received for publication May 9, Since the absolute transmission values at any. disk, the film thickness, scattering of light and
INFRARED SPECTROPHOTOMETRY OF ENTERIC BACTERIA SEYMOUR LEVINE, HEBER J. R. STEVENSON, LESLIE A. CHAMBERS, AND BERNARD A. KENNER Environmental Health Center, United States Public Health Service, Cincinnati,
More informationLEPTOSPIRES. Washington, D. C. preparations since they contained insoluble materials.
INFRARED SPECTROPHOTOMETRY OF CELLULAR CONSTITUENTS OF LEPTOSPIRES MORRIS D. SCHNEIDER AND JOSEPH McLAUGHLIN, JR. Third Army Area Medical Laboratory, Fort McPherson, Georgia, and the Division of Biochemistry,
More informationBIOCHEMICAL STUDIES ON CARBOHYDRATES. XL. Preparation of Mucoitin* from Umbilical Cords.
The Journal of Biochemistry, Vol. 28, No. 3. BIOCHEMICAL STUDIES ON CARBOHYDRATES. XL. Preparation of Mucoitin* from Umbilical Cords. MASAMI BY SUZUKI. (From the Medico-Chemical Institute, Hokkaido Imperial
More information1 Preparation and Characterization of Lignin-Carbohydrate Complexes
1 Preparation and Characterization of Lignin-Carbohydrate Complexes To explain the difficulty in separating lignin from carbohydrates in wood, Erdman (1866) hypothesized that the two combined chemically
More informationRICINOLEATE UPON BACTERIA
A COMPARATIVE STUDY OF THE ACTION OF SODIUM RICINOLEATE UPON BACTERIA From the Division of Laboratories and Research, New York State Department of Health, Albany Received for publication, May 14, 1928
More informationCHEMO-IMMUNOLOGICAL STUDIES ON CONJUGATED CARBOHYDRATE-PROTEINS IV. Tm~ SX~TI~SlS of Tm~ p-amn~obenzx~ ETm~R OF THE SOLUBLE
Published Online: 1 September, 1931 Supp Info: http://doi.org/10.1084/jem.54.3.431 Downloaded from jem.rupress.org on October 31, 2018 CHEMO-IMMUNOLOGICAL STUDIES ON CONJUGATED CARBOHYDRATE-PROTEINS IV.
More informationExperiment 1. Isolation of Glycogen from rat Liver
Experiment 1 Isolation of Glycogen from rat Liver Figure 35: FIG-2, Liver, PAS, 100x. Note the presence of a few scattered glycogen granules (GG). Objective To illustrate the method for isolating glycogen.
More informationPurity Tests for Modified Starches
Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016 Purity Tests for Modified Starches This monograph was also published in: Compendium
More informationPectins. Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016
Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016 Pectins This monograph was also published in: Compendium of Food Additive Specifications.
More informationby both esterification and acetylation of the liver concentrate inorganic salts and a source of energy such as glycerol or
BETA ALANINE AS A GROWTH ACCESSORY FOR THE DIPHTHERIA BACILLUS J. HOWARD MUELLER AND SIDNEY COHEN Department of Bacteriology and Immunology, Harvard University Medical School, Boston, Massachusetts Received
More informationPURPOSE: To synthesize soap from fat and lye. To observe the physical and chemical properties of soap.
FATS AND SAP: SAPNIFIATIN PURPSE: To synthesize soap from fat and lye. To observe the physical and chemical properties of soap. FATTY AIDS AND FATS: Fats and oils are mixtures of complex esters. Fat esters
More informationAZO-XYLAN (BIRCHWOOD)
ASSAY OF endo-1,4-ß-xylanase using AZO-XYLAN (BIRCHWOOD) S-AXBP S-AXBL 10/07 Megazyme International Ireland 2007 PRINCIPLE: This assay procedure is specific for endo-1,4-ß-d-xylanase activity. On incubation
More informationTHE isolation and availability of crystalline
Unidentified Factors in Poultry Nutrition. PROPERTIES AND PRELIMINARY FRACTIONATION OF A GROWTH FACTOR IN CONDENSED FISH SOLUBLES H. MENGE, C. A. DENTON, J. R. SIZEMORE, R. J. LILLIE AND H. R. BIRD Bureau
More informationStudies on the Urea-Dewaxing of Lubricating Oils*
Studies on the Urea-Dewaxing of Lubricating Oils* Naoki Yata** Summery: A behavior of activators and inhibitors of the urea adduct formation was investigated, and these two factors seemed to be of primary
More informationTHE RELATIONSHIP BETWEEN TWO METHODS FOR EVALUATING FIVE-CARBON SUGARS IN EUCALYPTUS EXTRACTION LIQUOR
THE RELATIONSHIP BETWEEN TWO METHODS FOR EVALUATING FIVE-CARBON SUGARS IN EUCALYPTUS EXTRACTION LIQUOR Congcong Chi, a,b* Zeng Zhang, a Weiwei Ge, a and Hasan Jameel b Alkaline pre-extraction and hydrothermal
More informationph Switchable and Fluorescent Ratiometric Squarylium Indocyanine Dyes as Extremely Alkaline Sensors
ph Switchable and Fluorescent Ratiometric Squarylium Indocyanine Dyes as Extremely Alkaline Sensors Jie Li, Chendong Ji, Wantai Yang, Meizhen Yin* State Key Laboratory of Chemical Resource Engineering,
More informationXXVI. STUDIES ON THE INTERACTION. OF AMINO-COMPOUNDS AND CARBOHYDRATES.
XXVI. STUDIES ON THE INTERACTION. OF AMINO-COMPOUNDS AND CARBOHYDRATES. II. THE PREPARATION OF GLUCOSE UREIDE. BY ALEXANDER HYND. From the Department of Physiology, University of St Andrews. (Received
More information3. PRELIMINARY PHYTOCHEMICAL SCREENING
93 3. PRELIMINARY PHYTOCHEMICAL SCREENING 3.1 INTRODUCTION All the drugs- Ayurvedic, Unani and Herbal extracts were subjected to preliminary phytochemical screening to test the presence of alkaloids, carbohydrates
More informationmelanogenum glucose is necessary for growth and
UTILIZATION OF ETHANOL BY ACETIC ACID BACTERIA M. R. RAGHAVENDRA RAO1 AND J. L. STOKES' Department of Bacteriology, Indiana University, Bloomington, Indiana Received for publication June 5, 1953 It is
More information(From Washington University, School of Medicine, Saint Louis)
Published Online: 1 June, 1964 Supp Info: http://doi.org/10.1084/jem.119.6.997 Downloaded from jem.rupress.org on November 11, 2018 IMMUNOCHEMICAL STUDIES ON THE SPECIFIC CARBO- HYDRATE OF GROUP G STREPTOCOCCI*,
More informationBIOCHEMICAL STUDIES ON PEARL FRACTIONATION AND TERMINAL AMINO ACIDS OF CONCHIOLIN. By SHOZO TANAKA, HIROYUKI HATANO AND GINZABURO SUZUE
The Journal of Biochemistry, Vol. 47, No. 1, 1960 BIOCHEMICAL STUDIES ON PEARL VII. FRACTIONATION AND TERMINAL AMINO ACIDS OF CONCHIOLIN By SHOZO TANAKA, HIROYUKI HATANO AND GINZABURO SUZUE (From the Department
More informationEXPERIMENT 4 DETERMINATION OF REDUCING SUGARS, TOTAL REDUCING SUGARS, SUCROSE AND STARCH
Practical Manual Food Chemistry and Physiology EXPERIMENT 4 DETERMINATION OF REDUCING SUGARS, TOTAL REDUCING SUGARS, SUCROSE AND STARCH Structure 4.1 Introduction Objectives 4.2 Experiment 4a: Reducing
More informationTHE ISOLATION OF A MUCOPOLYSACCHARIDE FROM SYNOVIAL FLUID*
THE ISOLATION OF A MUCOPOLYSACCHARIDE FROM SYNOVIAL FLUID* BY KARL MEYER, ELIZABETH M. SMYTH, AND MARTIN H. DAWSON (From the Department of Ophthalmology, College of Physicians and Surgeons, Columbia University,
More informationBRIEFING Assay + + +
BRIEFING Sodium Starch Glycolate, NF 22 page 2933 and page 3202 of PF 22(6) [Nov. Dec. 1996]. The United States Pharmacopeia is the coordinating pharmacopeia for the international harmonization of the
More informationTECHNICAL METHODS GASTRIC CONTENTS AND URINE THE IDENTIFICATION OF BARBITURATE DRUGS IN. aspirated and examined as well.
J. clin. Path. (1950), 3, 152. TECHNICAL METHODS THE IDENTIFICATION OF BARBITURATE DRUGS IN GASTRIC CONTENTS AND URINE BY From the Pathological Depariment, Salisbury General Infirmary (RECEIVED FOR PUBLICATION
More informationEUDRAGIT L 100 and EUDRAGIT S 100
Technical Information EUDRAGIT L 100 and EUDRAGIT S 100 Specification and Test Methods Ph. Eur. Methacrylic Acid - Methyl Methacrylate Copolymer (1:1) Methacrylic Acid - Methyl Methacrylate Copolymer (1:2)
More informationCHEMICAL STUDIES ON BACTERIAL AGGLUTINATION II. THE IDENTITY OF PRECIPITIN AND AGGLUTININ* BY MICHAEL HEIDELBERGER, PH.D., AND ELVIN A.
CHEMICAL STUDIES ON BACTERIAL AGGLUTINATION II. THE IDENTITY OF PRECIPITIN AND AGGLUTININ* BY MICHAEL HEIDELBERGER, PH.D., AND ELVIN A. KABAT (From the Laboratories of the Departments of Medicine and Biological
More informationLutein Esters from Tagetes Erecta
Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016 Lutein Esters from Tagetes Erecta This monograph was also published in: Compendium
More informationESCHERICHIA COLI-MUTABILE1. antiseptics employed "activated" the lactase which was present, "activate" the lactase.
ON THE "ACTIVATION" OF THE LACTASE OF ESCHERICHIA COLI-MUTABILE1 CHARLES J. DEERE Department of Chemistry, University of Tennessee School of Biological Sciences, Memphis Received for publication August
More informationH O. rapidly reduces. They dissolve. because they can hydrogen bond to the water molecules.
3.9 arboxylic Acids and Derivatives Naming arboxylic acids These have the ending oic acid but no number is necessary for the acid group as it must always be at the end of the chain. The numbering always
More information3016 Oxidation of ricinoleic acid (from castor oil) with KMnO 4 to azelaic acid
6 Oxidation of ricinoleic acid (from castor oil) with KMnO 4 to azelaic acid CH -(CH ) OH (CH ) -COOH KMnO 4 /KOH HOOC-(CH ) -COOH C H 4 O (.) KMnO 4 KOH (.) (6.) C H 6 O 4 (.) Classification Reaction
More informationTHE ACTIVE PRINCIPLES OF CANNABIS
XIV. THE ACTIVE PRINCIPLES OF CANNABIS INDICA RESIN. I BY THOMAS SPENCE WORK, FRANZ BERGEL AND ALEXANDER ROBERTUS TODD From the Biochemical Department, Lister Institute, London (Received 24 November 1938)
More informationStudies on the Determination of Bile Pigments I. Standard of Purity for Bilirubin
Studies on the Determination of Bile Pigments I. Standard of Purity for Bilirubin Richard J. Henry, S. L. Jacobs, and Neil Chiamori HE ACCURACY of the standardization for determination of bilirubin is
More informationL 346/6 Official Journal of the European Union
L 346/6 Official Journal of the European Union 9.12.2006 COMMISSION DIRECTIVE 2006/128/EC of 8 December 2006 amending and correcting Directive 95/31/EC laying down specific criteria of purity concerning
More informationThe Presence of Pyruvate Residues i TitleSimilar Polysaccharide (Commemorati Professor Sango Kunichika On the Oc Author(s) Hirase, Susumu; Watanabe, Kyoko Citation Bulletin of the Institute for Chemi University
More informationEXPERIMENTAL. PREPARATION OF MATERIALS.
F FATTY ACIDS AN By L. A. Bhatt and H. E. Watson (with 2. H. Patel). A knowledge of the solidifying point of binary mixtures of fatty acids or of their esters is of value lor determining the proportion
More informationCarboxylic Acids, Esters and Acyl Chlorides
R hemistry A 432 arboxylic Acids, Esters and Acyl hlorides arboxylic Acids, Esters and Acyl hlorides arboxylic acids contain the functional group, attached to an alkyl stem. They are widely found in nature,
More informationCARBOXYLIC ACIDS AND THEIR DERIVATIVES: NUCLEOPHILIC ADDITION-ELIMINATION AT THE ACYL CARBON
CARBOXYLIC ACIDS AND THEIR DERIVATIVES: NUCLEOPHILIC ADDITION-ELIMINATION AT THE ACYL CARBON RED ANT WAS SOURCE OF FORMIC ACID (RCOOH) Lecture 8 ORGANIC CHEMISTRY 2 Introduction The carboxyl group (-CO
More informationTHE ESTIMATION OF TRYPSIN WITH HEMOGLOBIN
THE ESTIMATION OF TRYPSIN WITH HEMOGLOBIN BY M. L. ANSON Am) A. E. MIRSKY (From the Laboratories of The Rockefeller Institute for Medical Research, Princeton, N. J., and the Hospital of The Rockefeller
More informationPetrolatum. Stage 4, Revision 1. Petrolatum is a purified semi solid mixture of hydrocarbons obtained from petroleum.
1 001-1208PDG.pdf Petrolatum Stage 4, Revision 1 Definition Petrolatum is a purified semi solid mixture of hydrocarbons obtained from petroleum. It may contain a suitable antioxidant. Description and Solubility
More informationChapter 18 Carboxylic Acids and Their Derivatives. Nucleophilic Addition- Elimination at the Acyl Carbon
Chapter 18 Carboxylic Acids and Their Derivatives. Nucleophilic Addition- Elimination at the Acyl Carbon Introduction The carboxyl group (-CO 2 H) is the parent group of a family of compounds called acyl
More information» Croscarmellose Sodium is a cross linked polymer of carboxymethylcellulose sodium.
BRIEFING Croscarmellose Sodium, NF 22 page 2856 and page 702 of PF 30(2) [Mar. Apr. 2004]. A modification is made in the test for Degree of substitution to correct the endpoint color to agree with the
More information2. Which of the following is NOT true about carbohydrates
Chemistry 11 Fall 2011 Examination #5 For the first portion of this exam, select the best answer choice for the questions below and mark the answers on your scantron. Then answer the free response questions
More informationARTESUNATE TABLETS: Final text for revision of The International Pharmacopoeia (December 2009) ARTESUNATI COMPRESSI ARTESUNATE TABLETS
December 2009 ARTESUNATE TABLETS: Final text for revision of The International Pharmacopoeia (December 2009) This monograph was adopted at the Forty-fourth WHO Expert Committee on Specifications for Pharmaceutical
More informationCryptococcal Antigen Latex Agglutination Test
Cryptococcal Antigen Latex Agglutination Test Cat. No.:DLAT1109 Pkg.Size:100T Intended use The Cryptococcal Antigen Latex Agglutination Test is a simple and rapid latex agglutination test for the qualitative
More informationEXPERIMENT 8 (Organic Chemistry II) Carboxylic Acids Reactions and Derivatives
EXPERIMENT 8 (rganic Chemistry II) Carboxylic Acids Reactions and Derivatives Pahlavan/Cherif Materials Medium test tubes (6) Test tube rack Beakers (50, 150, 400 ml) Ice Hot plate Graduated cylinders
More information1/3/2011. Chapter 17 Carboxylic Acids and Their Derivatives. Nucleophilic Addition- Elimination at the Acyl Carbon
Introduction The carboxyl group (-CO 2 H) is the parent group of a family of compounds called acyl compounds or carboxylic acid derivatives Chapter 17 Carboxylic Acids and Their Derivatives. Nucleophilic
More informationCH 3 CH 2 CH 2 CH 2 OH
1 The alcohols form a homologous series. The first member is methanol and the fourth is butanol. 3 O methanol 3 2 2 2 O butanol (a) Give two general characteristics of a homologous series. (ii) alculate
More informationMONOGRAPHS (USP) Saccharin Sodium
Vol. 31(4) [July Aug. 2005] HARMONIZATION 1225 MONOGRAPHS (USP) BRIEFING Saccharin Sodium, USP 28 page 1745 and page 612 of PF 31(2) [Mar. Apr. 2005]. The United States Pharmacopeia is the coordinating
More informationBCH302 [Practical] 1
BCH302 [Practical] 1 Carbohydrates are defined as the polyhydroxy aldehydes or polyhydroxy ketones. Most, but not all carbohydrate have a formula (CH 2 O)n (hence the name hydrate of carbon). Sugars ends
More informationProduction of Low Ester (LM) Pectin 307 In this study, attempts were made to determine the experimental procedures required to prepare low ester apple
Scientia Iranica, Vol. 12, No. 3, pp 306{310 c Sharif University of Technology, July 25 Research Note Production of Low Ester (LM) Pectin by De-esterication of High Ester (HM) Apple Pectin I. Alemzadeh,
More informationHAGEDORN AND JENSEN TO THE DETER- REDUCING SUGARS. MINATION OF LARGER QUANTITIES OF XIV. AN APPLICATION OF THE METHOD OF
XIV. AN APPLICATION OF THE METHOD OF HAGEDORN AND JENSEN TO THE DETER- MINATION OF LARGER QUANTITIES OF REDUCING SUGARS. By CHARLES SAMUEL HANES (Junior Scholar of the Exhibition of 1851). From the Botany
More information22. The Fischer Esterification
22. The Fischer Esterification A. Background Esters are an incredibly important functional group in organic chemistry. Esters are typically very pleasant smelling molecules and are therefore frequently
More informationQUALITATIVE TESTS OF CARBOHYDRATE
QUALITATIVE TESTS OF CARBOHYDRATE MACROMOLECULE CARBOHYDRATES Are the key source of energy used by living things. Also serve as extracellular structural elements as in cell wall of bacteria and plant.
More informationEFFECT OF HIGH SALT CONCENTRATIONS ON COLOR PRODUCTION OF THE BIURET REACTION FOR PROTEIN ANALYSIS
EFFECT OF HIGH SALT CONCENTRATIONS ON COLOR PRODUCTION OF THE BIURET REACTION FOR PROTEIN ANALYSIS HAROLD L. ROSENTHAL, PH.D., AND TOYOKO KAWAKAMI, M.T. (ASC1>) Division of Biochemistry, Department of
More informationCRYSTALLINE PEPSIN BY JOHN H. NORTHROP. (From the Laboratories of The Rockefeller Institute for Medical Research, Princeton, iv. J.
CRYSTALLINE PEPSIN III. PREPARATION OF ACTIVE CRYSTALLINE PEPSIN FROM INACTIVE DENATURED PEPSIN BY JOHN H. NORTHROP (From the Laboratories of The Rockefeller Institute for Medical Research, Princeton,
More informationChapter 18. Carboxylic Acids and Their Derivatives. Nucleophilic Addition-Elimination at the Acyl Carbon
Chapter 18 Carboxylic Acids and Their Derivatives. Nucleophilic Addition-Elimination at the Acyl Carbon Carboxylic Acids Organic compounds characterized by their acidity Contains COOH group (must be at
More informationpossibilities occurs. It has been found that the organism acquires addition of vitamin B1 to cells of P. pentosaceum which had
ADAPTATION OF THE PROPIONIC-ACID BACTERIA TO VITAMIN B1 SYNTHESIS INCLUDING A METHOD OF ASSAY M. SILVERMAN AND C. H. WERKMAN Bacteriology Section, Industrial Science Research Institute, Iowa State College,
More informationTHE COMPOSITION OF FLAXSEED MUCILAGE*
THE COMPOSITION OF FLAXSEED MUCILAGE* BY ERNEST ANDERSON AND HARRY J. LOWE (From the University of Arizona, Tucson) (Received for publication, January 20,1947) Flaxseed mucilage contains d-galacturonic
More informationLignin Isolation from Pulp
Lignin Isolation from Pulp Several different enzymatic, chemical and mechanical methods have been developed for the isolation of lignin from wood and pulp. However, due to the heterogeneous nature of wood
More informationCarboxylic Acid Derivatives Reading Study Problems Key Concepts and Skills Lecture Topics: Structures and reactivity of carboxylic acid derivatives
Carboxylic Acid Derivatives Reading: Wade chapter 21, sections 21-1- 21-16 Study Problems: 21-45, 21-46, 21-48, 21-49, 21-50, 21-53, 21-56, 21-58, 21-63 Key Concepts and Skills: Interpret the spectra of
More informationOrganic Molecule Composition of Milk: Lab Investigation
Name: Organic Molecule Composition of Milk: Lab Investigation Introduction & Background Milk & milk products have been a major food source from earliest recorded history. Milk is a natural, nutritionally
More informationIt is recommended to use titanium coils. ph meter to control ph of the process. It is advised to use an automatic dosage for the maintenance products.
CRODA III Decorative Trivalent Chrome Process CRODA III is a hexavalent free chrome process, allowing to obtain slightly blue and bright deposits, optimal for decorative applications. ADVANTAGES Depuration
More informationTHE SOLUBLE SPECIFIC SUBSTANCE OF A STRAIN OF FRIEDLANDER'S BACILLUS. (From the Hospital of The Rockefeller Institute for Medical Research.
Published Online: 1 November, 1925 Supp Info: http://doi.org/10.1084/jem.42.5.701 Downloaded from jem.rupress.org on November 20, 2018 THE SOLUBLE SPECIFIC SUBSTANCE OF A STRAIN OF FRIEDLANDER'S BACILLUS.
More informationON TEA TANNIN ISOLATED FROM GREEN TEA.
70 [Vol. 6 ON TEA TANNIN ISOLATED FROM GREEN TEA. By MICHIYO TSUJIMIIRA. (Received September 8th., 1930). The author(1) has recently isolated Tea catechin from green tea and pro posed the following formula
More informationFacile Isolation of Carotenoid Antioxidants from Solanum lycopersicum using Flash Chromatography
Facile Isolation of Carotenoid Antioxidants from Solanum lycopersicum using Flash Chromatography Jack E. Silver, jsilver@teledyne.com, Paul Bellinghausen, Nancy Fowler, and Ruth Pipes, Teledyne Isco, Inc.,
More informationTHERMALLY OXIDIZED SOYA BEAN OIL
THERMALLY OXIDIZED SOYA BEAN OIL Prepared at the 39th JECFA (1992), published in FNP 52 Add 1 (1992). Metals and arsenic specifications revised at the 55th JECFA (2000). An ADI of 0-3 mg/kg bw was established
More informationIsolation, Separation, and Characterization of Organic Acids*
In Dashek, William V., ed. Methods in plant biochemistry and molecular biology. Boca Raton, FL: CRC Press: pp. 107-113. Chapter 9.1997. Isolation, Separation, and Characterization of Organic Acids* William
More informationRELATIONSHIP TO RESISTANCE IN KLEBSIELLA PNEUMONIAE
THE SIGNIFICANCE OF LACTOSE FERMENTATION AND ITS RELATIONSHIP TO RESISTANCE IN KLEBSIELLA PNEUMONIAE VICTOR J. CABELLI' AND M. J. PICKETT Department of Bacteriology, University of California, Los Angeles,
More informationbelonging to the pseudoglobulins, forming a heat-stable, dialysable vasoconstrictor (Received 2 April 1942)
284 J. Physiol. (I942) IOI, 284-288 6I2.462.1:6I2.I46 PREPARATION AND SOME PROPERTIES OF HYPERTENSIN (ANGIOTONIN) BY P. EDMAN, U. S. VON EULER, E. JORPES AND 0. T. SJOSTRAND From the Physiology Department
More informationELECTROPHORETIC STUDIES OF SONIC EXTRACTS OF PROTEUS VULGARIS
ELECTROPHORETIC STUDIES OF SONIC EXTRACTS OF PROTEUS VULGARIS I. EFFECT OF GROWTH ENVIRONMENT ON ELECTROPHORETIC PATTERNS' SIDNEY D. RODENBERG Laboratory of Microbiology, Division of Biology, University
More informationGLYCOGEN BEFORE THE LAB YOU HAVE TO READ ABOUT:
GLYCGEN BEFRE THE LAB YU HAVE T READ ABUT:. Glycogen structure. 2. Glycogen synthesis and degradation (reactions with structural formulas and enzymes). 3. The role of glycogen in liver and muscles. INTRDUCTIN
More informationISOLATION AND PROPERTIES OF FURTHER COMPONENTS OF THE ANTIBIOTIC MOENOMYCIN. U. Schacht and G. Huber Farbwerke Hoechst AG., Frankfurt am Main, Germany
VOL. XXII NO. 12 THE JOURNAL OF ANTIBIOTICS 597 MOENOMYCIN. VII» ISOLATION AND PROPERTIES OF FURTHER COMPONENTS OF THE ANTIBIOTIC MOENOMYCIN U. Schacht and G. Huber Farbwerke Hoechst AG., Frankfurt am
More informationCarbonyl Chemistry VI + C O C. 1pm In Geology Room 112. Exam is Monday 11am-1pm. Chemistry /06/02
arbonyl hemistry VI Ō - + hemistry 391 11/06/02 Exam is Monday 11am-1pm 1pm In Geology Room 112 The Dibasic Acids h - My - Such - hemistry 391 11/06/02 Good- Apple- Pie- Fischer Esterification Esters can
More informationLieutenant Commander, Medical Corps, United States Naval Reserve,
Published Online: 1 February, 1946 Supp Info: http://doi.org/10.1084/jem.83.2.97 Downloaded from jem.rupress.org on September 3, 2018 STUDIES ON THE CHEMICAL NATURE OF THE SUBSTANCE INDUCING TRANSFORMATION
More informationRebaudioside a From Multiple Gene Donors Expressed in Yarrowia Lipolytica
Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016 Rebaudioside a From Multiple Gene Donors Expressed in Yarrowia Lipolytica This
More informationMost of the ethanol that is used as a biofuel in this country is produced from corn.
Chem 251 Ethanol from Corn Most of the ethanol that is used as a biofuel in this country is produced from corn. In this experiment you will make ethanol from frozen corn kernels using a process similar
More informationASSAY OF using AZO-FRUCTAN S-AZFR5 11/17
www.megazyme.com ASSAY OF endo-fructanase using AZO-FRUCTAN S-AZFR5 11/17 Megazyme 2017 PRINCIPLE: The substrate is the high molecular weight fraction of chicory fructan (DP ~ 20-60) dyed with an azo-dye
More informationBRIEFING. Nonharmonized attributes: Identification, Heavy metals, Characters, Labeling, Bacterial endotoxins, Sterility, Storage.
BRIEFING Citric Acid, Anhydrous, page 872 of PF 28(3) [May June 2002]. The European Pharmacopoeia is the coordinating pharmacopeia for the international harmonization of the compendial standards for the
More informationusually contain somatic protein as the principal antigen, determination of the Heidelberger and Kendall (1932) developed a method for the quantitative
A MOUSE PROTECTION METHOD FOR THE ESTIMATION OF ANTIGENIC PNEUMOCOCCAL POLYSACCHARIDE IN SOLUTION CURTIS SANDAGEl AND ORTON K. STARK Miami University, Oxford, Ohio Received for publication May 5, 1947
More informationChem 263 Dec 1, 2016
Chem 263 Dec 1, 2016 eactivity of Carboxylic acid Derivatives More eactive S 2 ' ' + a ' 2 - M + Less eactive Example: Acid chloride to anhydride Since an acid chloride is more reactive than an anhydride,
More informationISOLATION AND PURIFICATION OF A SEROLOGICALLY ACTIVE PHOSPHOLIPID FROM BEEF HEART
SOLATON AND PURFCATON OF A SEROLOGCALLY ACTVE PHOSPHOLPD FROM BEEF HEART BY MARY C. PANGBORN (From the Division of Laboratories and Research, New York State Department of Health, Albany) (Received for
More informationPelagia Research Library
Available online at www.pelagiaresearchlibrary.com Der Chemica Sinica, 2013, 4(1):67-74 ISSN: 0976-8505 CODEN (USA) CSHIA5 Analysis of Abakaliki Rice Husks N. B. Ekwe Chemical Engineering Department, University
More informationprotein (Eaton 1936 a, 1937; Pappenheimer 1937). If other
COMPARATIVE STUDIES ON THE PURIFICATION OF TETANUS AND DIPHTHERIA TOXINS MONROE D. EATON AND AXEL GRONAU Department of Bacteriology, Washington University School of Medicine, St. Louis, Missouri Received
More information12BL Experiment 2: Extraction & Saponification of Trimyristin from Nutmeg
12BL Experiment 2: Extraction & Saponification of Trimyristin from Nutmeg Safety: Proper lab goggles/glasses must be worn (even over prescription glasses). Heating of organic solvents releases irritating
More informationChem 263 Nov 21, 2013
hem 263 Nov 21, 2013 arbohydrates- emiacetal Formation You know from previous lectures that carbonyl compounds react with all kinds of nucleophiles. ydration and hemiacetal formation are typical examples.
More informationHydroxypropyl Starch (Tentative)
Residue Monograph prepared by the meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 82 nd meeting 2016 Hydroxypropyl Starch (Tentative) This monograph was also published in: Compendium
More informationCarbohydrates. Objectives. Background. Experiment 6
1 of 6 3/15/2011 7:27 PM Experiment 6 Carbohydrates Objectives During this experiment you will look at some of the physical and chemical properties of carbohydrates. Many of the carbohydrates, especially
More informationThe PGA Screen TM HT-96. The kit contains 96 1ml conditions in a deep-well block.
The PGA Screen TM HT-96 MD1-51 A novel precipitant and a totally new crystallization space to explore. A revolutionary new systematic screen from the York Structural Biology Laboratory (YSBL) based on
More informationColour of starch-iodine complex as index of retrogradability of starch pastes
Vol. 8(5), pp. 89-93 May, 2014 DI:10.5897/AJPAC2014.0571 Article Number: 802AA7844885 ISSN 1996-0840 Copyright 2014 Author(s) retain the copyright of this article http://www.academicjournals.org/ajpac
More informationASSAY OF USING BETA-GLUCAZYME TABLETS
ASSAY OF endo-β-glucanases USING BETA-GLUCAZYME TABLETS T-BGZ 12/12 Note: Changed assay format for malt β-glucanase Megazyme International Ireland 2012 SUBSTRATE: The substrate employed is Azurine-crosslinked
More informationJournal of Chemical and Pharmaceutical Research, 2015, 7(8): Research Article
Available online www.jocpr.com Journal of Chemical and Pharmaceutical Research, 215, 7(8):257-261 Research Article ISSN : 975-7384 CODEN(USA) : JCPRC5 Pulping process for rice straw in basic ionic liquid
More informationPECTIN IDENTIFICATION
www.megazyme.com PECTIN IDENTIFICATION ASSAY PROCEDURE (500 Assays per Kit) K-PECID 08/18 Megazyme 2018 INTRODUCTION: Pectins consist of the partial methyl esters of polygalacturonic acid and their sodium,
More informationSimultaneous determination of triglycerides and cholesterol esters in serum by infrared spectrophotometry
Simultaneous determination of triglycerides and cholesterol esters in serum by infrared spectrophotometry NORMAN K. FREEMAN Donner Laboratory of Medical Physics and Biophysics, Lawrence Radiation Laboratory,
More informationAZO-WHEAT ARABINOXYLAN
www.megazyme.com ASSAY OF endo-1,4-b-xylanase using AZO-WHEAT ARABINOXYLAN S-AWAXP S-AWAXL 05/17 Megazyme 2017 PRINCIPLE: This assay procedure is specific for endo-1,4-β-d-xylanase activity. On incubation
More informationNational Standard of the People s Republic of China. National food safety standard. Determination of pantothenic acid in foods for infants and
National Standard of the People s Republic of China GB 5413.17 2010 National food safety standard Determination of pantothenic acid in foods for infants and young children, milk and milk products Issued
More informationTHE JOURNAL OF ANTIBIOTICS. Polyketomycin, a New Antibiotic from Streptomyces sp. MK277-AF1. II. Structure Determination
THE JOURNAL OF ANTIBIOTICS Polyketomycin, a New Antibiotic from Streptomyces sp. MK277-AF1 II. Structure Determination ISAO MOMOSE, WEI CHEN, HIKARU NAKAMURA, HIROSHI NAGANAWA, HIRONOBU IINUMA and TOMIO
More informationTHERMALLY OXIDIZED SOYA BEAN OIL interacted with MONO- and DIGLYCERIDES of FATTY ACIDS
THERMALLY OXIDIZED SOYA BEAN OIL interacted with MONO- and DIGLYCERIDES of FATTY ACIDS Prepared at the 39th JECFA (1992), published in FNP 52 Add 1 (1992). Metals and arsenic specifications revised at
More informationTRANSAMINASES IN SMOOTH BRUCELLA ABORTUS, STRAIN 19
TRANSAMINASES IN SMOOTH BRUCELLA ABORTUS, STRAIN 19 BY ROBERT A. ALTENBERN AND RILEY D. HOUSEWRIGHT (From the Chemical Corps Biological Laboratories, Camp Detrick, Frederick, Maryland) (Received for publication,
More informationCHAPTER 4: RESULTS AND DISCUSSION. 4.1 Structural and morphological studies
hapter 4: Fourier Transform Infrared Spectroscopy (FTIR) HPTER 4: RESULTS N ISUSSION 4.1 Structural and morphological studies 4.1.1 Fourier Transforms Infrared Spectroscopy (FTIR) The scanning of the samples
More information