In the present communication results obtained by extending the technique of Jeans et al. to the resolution of a mixture

containing five sngars (lactose, maltose, dextrose, galactose and arabinose each sugar at 1 per cent level) are give

Pillai, V K

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LETTER S TO TIfE EDITOR Frc, . 3 - - S UPER~ ."' TA "'T .-\FTER TRI' •. \.T\I£).:T OF THE RED C ELL :'JE)!HRAXE WI TH P,'.T1E:-<"T'S S£Rt:'M · .'\~"n CC;"'Tl{IF\)'r.i);"G X 11,700 (SIZr. 1.300-3,300 .\.f'.) The photographs show that. there arc particles which sef'm t o be atin.ch~d to senull-rcactcu. cf' ll membrane surface and also those which ar(' free in the supernatant fl uid. On the other hanel, no sHch part icles arc observed in the supernatant or on the cell surfaces reacted \vith normal serum. Further work is being carried out to establish the relationship h('t ween these particles and haemaggllltina ting agent of the disease. Qur th anks are cl1lP to the Director, ::\!ahonal Physical Laboratory of India, for permission to USC' the electron microscope and to .Mr. T\.ullw:.n Bahadur, Junior Scienti-fic Assistant , for technical help ill electron microscopy. B. }l. GnH S. S. MI S 1L\ 5.\:\11 H.-Dun A. PX~JnE Celltral Drug Research IJistitut e] Luck II uw K illg George's Nledical College Luchl/ow I Ileiian CUlwcil of Jf edical Research Neu' Delhi and Nt/Ilol/al Ph\'siwl l. n/Jorafo1y N f W Dillin: "' 24 Jail. 1953 1. CUPT.\, 11 . ]\f., l'j'ISR,\, S. S. & 1 LnuD, S., Clirr. Sci .. 21 ( 1952 ), 78. 2. DOUXCE, A. L. & LA~, T . H ., SCiCHCf, 97 ( 19+3). 584. 3. DOW50~, I. \L & ELFORD, \V. J. , J. gr.l/. ;1,[i(;l'O-binl. 3 (2) ( 1949).298. ASCENDING (PAPER) C HROMATOGRAPHY OF SUGARS RY THE I\UILTlPU; DEVELOPMt<:NT T ECIINIQUt<: JEANES ef (1/.1 H.-WE l)ESCHlHED .\ Ml:LTI PLE clc\'elopment technique for the ascfnding ( paper ) chronwtography of sugars. They llsed two-componcnt sugar mixtures and wen.' able to dr-rive equations on a theoretical bas is from whicli the numher of ru ns neces-sa ry for the max imum distance of separation of two sug:n spots could be calcula ted. I n the present comm nnicJtio!l, results obt a ined by extending the technique of Jc;} nes I!t at. to t he resolution of a mixture containing five sngars (lactose, maltose, dextrose, g::dactosc and arabinose; each sugar at 1 per cent levcl) are given. T he procedure adopted was essen t ially the samf', but rect angula r str ips of filter paper (38 X 4 em" \Yhatman \'0. 1 ) \vere used instead of cyhn drical sheets; and the apparatu:;2 was considerably simpler, being cas it~ ' assembled from a specimen jar, a heaker and a few b its of glass tubing, plasticine and wire cl ip. Certain d ifficulties inherent in the handling of large wpt ftlte r papcr chromatograms could thus be {\\'oided and the solvent loss was also m inimiz('(1. Thc soh'cnt syst.em lIsed was a mixture of n-bllranol, ethanol A P IG. 1 - PROCHE SS OF SC: P :\ RATl O:-': FRO:>,I .\ :>,IIXTt;RE OF FIVE Sl!GARS FOR n {}'!' ERE;":T R U :-: S 0 1' T H E 5()LVE~T [ A, 2 rlln~; R , 3 nlns; C, 4 rllns ; and D,5rllns] 181 J. SCI. INDUSTR. RES., VOL. 12B, 1953 TABLE t - DISTANCE OF SEPARATION BETWEEN SUGAR SPOTS (eM.) SUGAR MIXTl:RE No. 0, 10;:'15" 1 2 3 4 5 6 7 Lactose-JIlalto~e 0-62 Q-SO 1'10 l' 20 1 ·40 I-fiO I-50 Lactose-dextrose 1'74 3·30 '1-40 5 -10 5 '40 0-00 5-30 Laclose-galactose ) -{lO 4·00 f,- 20 ti · ~O f, - ;;0 (;·70 6-40 Lactose-arabirrose 3· 40 r,- 30 ; · \0 7-S0 1'-]0 8-00 7-00 Maltose-dextrose l' l~ 2· ;lfl ;j·;JU 3· 9() ·j'OO ·1- 00 3 ' 80 Maltose-galactose ] -28 3 -20 4-]0 f,· 00 :,"10 :;-20 4-£10 )[aitos.c-arabinose 2· 78 4-,,0 fHlO u·{)(l (; -70 (\.;,0 (j · 20 Dextrose -galactose (I-HI 0'70 O'SO HlO 1· 10 1'20 1'10 Dextrose-arabinose l-tiO 2-0{) 2· 70 2 · 7f) 2· 70 2· 50 2· 40 Galac[ose-ara binose 1-60 1 · 30 1-90 1· (jO 1· 30 1· 30 1·30 " Total disbncc travelled hy the sohTnt ~ n X 26 em ., where n is the number of ru ng. TABLE 2 - NUMBER OF RUNS REQUIRED FOR MAXI:\1UM DISTANCE OF SEPARATION 'OF SUGAR SPOTS C.\LCl;LATED YUt·f.S L '1 0 G Lact{)se (L) 7 0 6 !I[altose (.\I) , 6 Dextrose (0) 6 6 4 Galactose (G) " " " Arahinosc (A) 5 3 3 !>O 100 150 200 TOTAL DISTANCE TRAVEllED 8Y SOLVENT, em. FIG. 2 - DISTANCE T RAVE'LLED BY INDIVIDUAL SUGAR SPOTS ~'S. TOTAL DISTAKCE TRAVE LLED BY SOLVENT [1, arabinose; 2, galactose ; 3, dextrose; 4, maltose ; 5, lactose] and \vater3 , and the spray reagent was made up of benzidine and trichloracetic acid as recomm ended bv Bacon and Erlelmann4• The progress of separation of the sugars from the mixture is indicated in Fig. 1. In Fig. 2 the distances travelled by the individual sugar spots have been plotted agains t the distances travelled byl the solvent. For determining the number of runs (nmax .) in which the distance between the centres of any two sugar spots is a maximum, the respective distances were 182 OIlSER\'ED VAI.UES ~ A L M D G A 5 6 6 6 5 f) 6 5 ti 5 , 6 5 6 " " 6 6 6 3 5 , 4 3 read off from Fig. 2 for a constant. distance (26 em.) travelled by the solvent per nm (TABLE 1) . These observed values of 11ma:-:. have been given in Table 2 together with the theoretical values calculated from the formula log Rf,- log Rf_, _ _ _ log ( l-Rd-log (l-Rf,) given by Jeanes et al. It is seen from Table 2 that there is good agreement between the observed and cal-culated values. This agreement is of added interest since hitherto values of nrnax. do not appear to have been determined for mixtures containing as many as five sugars. The enhanced utility of the Illultiple develop-ment technique, especially in quantitative analysis, is also evident. I am grateful to Dr. N. K . Panikkar, Chief Research OJ-licer, Central :J'larinc Fisher-ies Research Station, for the interest that he has taken in the work. l\fv thanks are also due to ~Ir. R . Viswanatlu.{n for helpful suggestions. V. KRISHNA PILLA! Ccnlral .tfari'll c F isheries R esearch Station ]\1 andapam 3 Oct. 1952 1. JEANES, A., WISE, C. S. & DnILER , R. T., Analyt. Chem., 23 ( 1951),415. . 2. VrswANATHAN , R., Proc. 40th Indian Sci. Congo 1155 .,1953, Pt. III, 281. • 3. BRYSON. J. L. & lIIrTCHELL, T. J., Nature, 167 ( 1950). 864. 4. BACO:-l', J. S. D. & EDEDIA~N, J., Biochem. J., 48 (1951). 115. 

Ascending (paper) chromatography of sugars by the multiple development technique

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Ascending (paper) chromatography of sugars by the multiple development technique

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