MULTIPLE CHROMATOGRAMS FOR DETECTION OF AMINO ACIDS IN BLOOD SERUM SISTER HELENE VEN HORST and YOLANDA CARSTENS Marycrest College, Davenport, Iowa
A SIMPLE piece of apparatus was constructed whereby 2 to 16 chromatograms could be run simultaneously and under identical cond~tions. A modification of the technique of Gin, et al.' was used in the separation and detection of amino acids in pathological and normal blood sera. A metal circular rim of 50-cm. diameter and 2.5-em. height was used as the support for one to eight lengths of one-centimeter-wide Whatman No. 1 or Xo. 4 filter paper. The 0.5-em. groove a t the outer top of the rim contained a heavy piece of cord underneath which the filter paper strips were fastened. By inserting short pieces of cord a t right angles to the rim, it was easy to pull the cord out of the groove and anchor the strips of filter paper beneath it. The strips of filter paper were punched simultaneously in their centers and fastened radially across the frame so that the holes were mounted exactly above ' GIRI, KRISHNAMURTHY, A Y D VEYKITASUBRAVIZ.IAW Lancet, 263, 56%3
(1952).
one another. A semicircular piece of 9-cm. filter paper was cut a t the straight edge to form a brush-like tip. The paper was then rolled cylindrically into a wick and inserted in the punched hole of the filter paper strips. The brush edge of the wick mas immersed about one centimeter into the phenol carrier solution contained in a small evaporating dish. (See the accompanying illustration.) Samples of pathological or normal sera %.ere applied in 0.5-mm. spots a t a 2.5-cm. radius from the wick. The apparatus allowed the use of two samples on each strip, one on either side of the wick. After the spots had dried, the apparatus was placed horizontally in a 55-cm. square box, approximately 15 em. high, furnished with a removable glass top. For Whatman No. 4 filter paper the time required for the carrier t o travel the 22-cm. length of the strip was approximately six hours. The rim and mounted strips mere removed from the box and dried in a forced-draft hood. Spraying with the developing agent was carried out directly on the mounted strips. In some cases the strips were remowd and plnvcd adjnrent to cnrh other O I I a pnpcr rray hcforc spraying. Tht: strips were d ~ . i in d an own at (iO°C.
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.\d\.antu~rsof this appamtus nrc: [ I ) lsrgc llnmbrrs of snrnplcs of irrn may he chn)mutogruphrd aimnlt n " ~, (2) : idrnti~.alc ~ d i r i o n sof .wxirntina the ~, amino acids.. drvine. and develooine . thk chromatograms are insured; and (3) comparison may be made with controls of pure amino acid samples analyzed u~lileri ~ l ~ n t eolditinl~s. i~d .\ rni~lintnlrrmodt.1 I I thi.; ~ :lpprlnitus \ws n w l in the prrliminnry nm11ysi.i of the srntrn. ;\ wirc frame w i s vonsl r ~ ~ c t v31.d d 1111. tilrw p:tpvr t r i p s wrrr folded o \ w t l ~ rrdp. of t l ~ t swirtr. Tllc wine x i ~ kt c v h l ~ i ~ uwas c wed nod thr rntirr appnr;itus w:is i l ~ x l - r t dinro i n cnipty ~lt.siwator. 17ndr~r thr.;r m~ditinnsn n npprosin~:rtvnn:tlysi~wnl(l hr ni:dc withi11R I I how. "
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