Hassan, G E

Kabiel, A M

Youssef, Y S

IACS Institutional Repository

Spectrographid estimation o f Fe, Mg, Ca, Zn, Sb and Pbin blood serumA. M. K a b i e l , G. E. H a s s a n *, Y. S. Y o u sse e  a n d  F. I. N ashBiophysics Department, Medical Research Institute,Alexandria University, Alexandria, (E. A. R.)[Received 26 March 1974, revised 13 March 1975)For the dotermination of Ca, Mg, Zn, Fo, Sb and Pb in blood serum simultanoously, a method of diiect spootroohemica] analysis of sennu using a synchronized high voltage copper spaik technique is des­cribed, which gave sufficient sensitivity for most of the olomonts.Tho method was calibrated by synthetic standoi’d samples avoiding errors by imitating the organic contents of the serum with a condensa­tion product of glucose uioa and glycine. The anajysis was carried out on human serum for the determination of the elements exot^pt Pb and Sb which were beyond our detection limit. These two elements were determined in two kinds of experimental animals which were previously injected by salts of the elements.1. I k teo d u o t io nTl\o particular advantages of the analytical spoctrographic methods, in general, are speed, remarkable sinsitivity, precision, unequivocal identification. An important field, where samples of limited size occur, is the field of biological losearch. Organs, tissues and bod.y fluids are analyaod for i^aiions metals either to establish limits of normal distribution to ascertain departuies from them under pathologic conditions, or to obtain information of diagnostic value. Many of the mineral elements found in blood ai*o essential to health, and the loiowlodgo of their concentration is very important (Comar Bronner 1968). The chief role of calcium is the blood clotting, essential for contractility of involuntary muscles, acting antagonistically to sodium and potassium, formation of bones and teeth, influence on excitability of nerve fibres and centrers. Under normal conditions the magnesium intake is always adequate, so that effects comparable to calcium deficiency are not seen. The effects of magnesium deficiency are severe if the calcium intake is increased. Other observations also suggest an interrelationship between calcium and magnesium. Difficiency of iron which is a paxt of the haemoglobin molecule, results in anaemia. Also, iron is an impor­tant intercellular catalyst of oxidation-reduction i.e., in oxidases.* Department of Physics, University of Tanta, E.A.E.P resen t A d d re ss  : Department of Physios, Faculty of Engineering, University of Riyadh, Kingdom of (Saudi Arabia,Indian J .  Fhy»- « ,  418-422 (1976)413416 A. M. Kabiel, G. E. Hassan et alwere oai*ofnlly selected free from interferences and coincidence. The lines were densitomotered without background correction. The transmittanoes obtained were converted to relative intensities tising the emulsion calibration curve of the photographic plate. Intensity ratios were calculated for each pair of lines.C. Selection of the Electrode Diameter : (Kabiel et al 1970)The variation of the absolutes intensity of the analysis lines of the analysed elements was studied using the different copper electrodes of diameters varying from 2 to 7 mms. For every diameter, sixtfien replicates—^using the same stan- dai'd solution—wore prepared. Results obtained for the intensity (I) of the lines together with the corresponding coofOicient of variance {C.V.) arc given in table 2 and illustrated in figure 1 . From the table, it is remarkable that the electrode diameter plays an important role with respect to absolute intensity and the standard moan deviation in percent. Best results were obtained usint the most wide electrode as this latter will cause less mechanical loss of thi> somion duo to sputtering when loading tho electrodes. tFig. 1, Variation of tho intensity with tho electrode diametor.D. Sparking ConditionsExcitation in the spark is largely dependent on the spark power. If no losses occur, tho power in KVA is computed from the equation (Nachtriob);P - O V /whore C is the capacitance in Farads, Fq is the initial potentials in volte and /  is the frequency in o/s.Various sparking conditions of excitation concerning the initial sparking voltage, capacitance and inductance were tried to obtain a higli reproducibility and reasonable accuracy of the aoialysis. Invesitigations showed that the inten­sity for all lines was greatly enhanced and became suitable to bo measured using the following conditions :Initial sparking voltage 17 KCapacitance 6 nfInductance 0.3 mhwith such a somewhat high inductance, the spectrum was similar to an arc one which was preferred to brighten tho spectrum background and obtain bettor emiasion linos of the sougl^t elements.E. Volatilisation StudiesExtensive moving-plate studios with one of the standards wore tried. I t  was found, as it appears from figure 2, that each element has a characteristic manner of volatilisation with a peak omis&ion different from the others, andEstimation of Fe, Mg, Ga, 8b and Pb in blood serum 417Fig. 2. Intonsity-timo curves for Mg, Ca, F o f  Z n , Al, Ni, Sb and Pb.that all the elements are volatilised at the first 100 secs, of exposure with remark­able sharp decrease in their corresponding line intensity for more than 120 secs, accompanied by an increase in the darkness of the background. Accordingly, 100 secs were selected to be the suitable time of excitation.418 A- M. K abiel, G. E- H assan et alDaSc8os,CQo015301si$oI  ■CO onI I■§^  o9 ®Ul Oc3g °iis134^C<lH1—i p H p H 0 5 CM p Ht>o CD CO pHC3u I—* I—•N ICM CM O OO oo 0 5 t - M5 CM oMr - l > □ 0 QO 0 0 0 00 5 CM p H CO CM CMt > CO 0 5 Mb p H p Hb O dI-Hla I-H O O Or—1 o o o CO oHH0 0 I > t p l > OOCO CO GO CO Mb p H0 5 QO o o 'd ' CMp Ob0 2»-< CM p H p H OOCO CO O o oMM5 Mb * d><M CM PH PH CM COo 6 kO CO o p HbCID o o o o» o CM CM o i p H1— (o o 0 5 0 5 p H o oPH p H p HCO p H CM CO p H COCO CO p H CO Mb CMp£J d 1—1 p HPM05 CM CO r pi-H CM M5 CO OO CMHHCO CO CO CO cd05 p H CO 05 PHh > 0 0 l O CM p H p H• P*« bCO o Mb CM p Hia> CO 05 o CM COHHp—t p H PH CM CM CMo CM p H p H COCM CM o o cd p HrS b»—1MO CM Mb Mb Mb oCM 0 0 CM CO CM CMMT (i M5 cd t > 00*f—I p H pH p H p H p HKwp H QD CO CMO O l > cd cd p HcB b p HO e<> O o Mb »o CD CM p-HMCD l> 05 C5 05 CDpS^ s CM CO Mb CO r-o Ho 3i) Human SerumThe blood was drawn from the intercubital foaaa vein, in the arm, using a very clean and dry 20 mm hypodermic syringe fitted with special Pt-Ru alloy needle. The blood was then transferred immediately to a very clean contiifuge polyethylene bottle, noting that the blood must run on the wall of the tube to avoid any sort of haemolysis. The blood was centrifuged at 3000 xG  for 15 minutes. The serum fraction was withdrawn with a similar hypodermic syringe. Every one ml of serum was mixed with one ml of solution containing Ni and A1 as internal standards, and the resulting solution was shaked by hand for complete mixing. Then 0.2 ml of this solution was trajisferred on the clean electrode top, treated as the standards and tested spootiographically. Using the constructed working curves, shown in figure 3, the concentrations of the sought elements vero determined. Results obtained are given in table 3.Estimation of Fe, Mg, Ca, 8b and Pb in blood serum 4193. ApplicationMICROGRAMS*pb wij/ai kmiFig. 3. Analytical copper spark working curves for Mg, Ca, Fe, Zn, Pb and Sb.Table 3, Concentration range and average values obtained for normal human serum, cairied on a group of 29 volunteers.Element Concentration range in y/mlAverage concentration in y/mlFe 0 .9 — 1.3 1.15Mg 17 — 36 24Ca 90 — 110 106Zn 0 .6 — 2.1 1 .3Sb N.D. N.D.Pb N.D. N.D.420 A. M. Kabiel, G. E. Hassan et alii) Animal SerumThe determination of the elements sought in the serum of two experimental animals, was performed. As Pb and Sb are not normally present in normal blood, we performed some injection treatments with solutions oontaining Pb and Sb.a) Estimation of Pb : A group of six rabbits was selected, and 5 mis of blood from each was withdi^awn. The serum was obtained as described before, treated in the same way and tested spectrographieally. Results obtained are given in table 4. Five mis. of lead sub-acetate wore then injected in the marginal vein of each rabbit. After 24 hours, again 5 mis of blood wore withdrawn, serum obtained, treated as previously t^xplainod and tested spootrographioally. Results obtained are given in table 4.Table 4. Averages conccjntration of the elements sought belore and after injec tion with load subacetate—in group of 0 rabbits. \KlomoiitAverage concentration before injection in y/mlAverage conceni^ration after injection in y/mlFo 1.41 0.99Mg 22.40 19.00Ca 136.60 210.00Zn 1.69 1.11Sb N.D. N.D.Pb N.D. 1.5b) Estimation of Sb : Two mis of blood, withdrawn from the tail of a group of six rats wore troattxl as dcsc.’'ibed before and the serum was tested spectro- graphically. Results obtained are given in table 5. Two mis of Tartaaemetic wore injected through the largo vein in the tail of each rat. After 24 hours 2 mis of blood wore withdrawn, serum ol)tained and tested spectrogiaphically. Results are given in tabic 5.Table 5. Average concentration of the sougb.t elements, before and after injection with Tartaremetic—in a group of 6 rats.ElementAverage concentration before injection in y/mlAverage concentration after injection in y/rnlFe 1.10 1.6Mg 15.10 13.2Ca 119.20 125.2Zn 0.70 N D .Sb N.D. 3 .6Pb N.D. N.D.Estimation of Fe  ^ Mg  ^ Ca, Sb and Pb in blood serum 4214. D iscussionAoouraoy is one of the broad objects of spectroolu^mical analysis wheir improvement by any order of magnitude would result in vast new fields of applica­tion. In biology and medicine bettor accuracy, could result in a now diagnostic tool. Accuracy in quaaititative spoctrographic woik depends on several factors such as the source of excitation, sort and shape of electrodes and the photometric calculations.To attain best accuracy, the spark excitation source was used which had proved, in routine quantitative work, to bo most suitable compared with the d.c. arc. Generally, spark has roltatively poor concentrational sensitivity, piimorily because of relatively high background radiation. The use of high voltage and capacitance, in order to reach maximum sonsitiWty, had provided us with a spectra which wore aic-like. On the otherhand, the use of a somewhat relatively high inductance, was purposely chosen in order to brighten the arc-like spectra background,A course approaching high accuracy and in certain cases surpassing the sensitivity of the d.c. arc, is the copper spark olootrodo technique. A comparison between excitation of both carbon and cojDpor electrodes was done. Tt was found that, under strictly selected excitation conditions, both electrodes yielded high sensitivity and relatively good precision, but that of copper w^ as more higli in sensitivity. I t  was ajso found that the diameter of the electrodes played an important role with sensitivity. I t  was observed that the sensitivity of the determinations increasfxi as the electrode diameter increased, owing to the fact that the sample loss from the electrode due to the sputtering was leas.The measuromont of the intensity of the spootial lines using the microphoto- meter, was accurately done. The shouldei, of the working curves, at low trans­mittance values which resulted partially from the scattered light that entered the mioiophotometer slit from portions of the spectrograms around the lino itself, had been somewhat corrected by measuring 2/3 of the magnified line.For internal standardisation, different elements were selected such as Al, Ni, and Mn. Unfortunately, it was found that the quantity of Mn added as internal standards was somewhat small with respect to the amount already present in blood which might introduce a source of error during the application of the method. However, Al and Ni wore tested, and the standard mean devia­tion for each sought element with respect to both Al and Ni with and without B.G. correction was calculated. Results are givexr in table 6.Finally, we have to state clearly that wo had not strictly used the control lines (Rais Ultimas) of the analysed elements. I t  was found by Naohtrios and also was observed by us (Kabiel et al 1970) that other lines than Rais Ultimas 2422 A. M. Kahiel, G. E. HaBsan et aJbecame more enhAQoed using the ooppez' spark teohnique and was found to besatisfactory to mako use of especially at low cojioentratiojis.Table 6. The ntandard maaai duration in percent (c.r.) calculated for each aixalysod element with respect to AI and Ni. Results are mean of 8 determinations.AnalysedelementN1 NiWithout B.Q. correctionWith B .a  correctionWithout B.G. correctionWith B.G. correctionFo 4 .4 6 .6 6 .8 9 .9Ca 6 .3 8.4 9 .6 10.3Mg 6.4 8.1 6 .8 7.7  ,iZn 9.4 10.1 6 ,318.6,\.Pb 8.7 10.1 9.1 9 .9  ;vSb 8 .6 11.9 9.3 11.2___________ J-__5. Co n c l u sio nIti is soon from the. previous studies that there is an improvement in the V apidity of the determination and also much accuracy and sensitivity. Also, the validity and superiority of the proposed method for the estimation of micro- grams of Zn, Fe, Ca, Mg, Pb and Sb in biological fluids can be easily  ^ascertained.The results obtained are extraordinary sensitive and exactly similar to that in literature.R e f e r e n c e sComer C. L. & Bronner F. 1968 Mineral Metabolism, Vol. II, Part B, Academic Press Inc.,N e w  Y o rk .Kabiel A. M. ct a l 1970 J. o f A p p lie d  S p e d .,  22, No. 3.Nachirieb N. H. P rin c ip les  a n d  Prewtice o f  Spectroch/emical A n a ly s is ,  M eO ta w -U iU  B ookC om panyi In c .,  N e w  Y o rk .Pfoilaticker K. 1970 Spectroch im  A c ta . 4 , 100.Seidel N« A. Tables q f S p e c tru m  L in e s , V  E  B , Verlag T e c h n ik , Berlin.

Spectrographic Estimation of Fe Mg  Ca, Zn, Sb and Pb  in Blood Serum

http://arxiv.iacs.res.in:8080/jspui/bitstream/10821/6189/1/Spectrographic%20estimation%20of%20Fe%20Mg_A%20M%20Kabiel.pdf

Spectrographic Estimation of Fe Mg Ca, Zn, Sb and Pb in Blood Serum

Abstract

Similar works

Full text

Available Versions

IACS Institutional Repository