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TCa 2.2 - 2.6 ACa ?? mmol/L81PRP0PPgPPIPPRPPAP       bW ]l Adjusted Ca?  f[TCa] depends on protein concentration i.e Alb but Ca2+ ions normal: 1g Alb = 0.04 mmol Ca ACa is a correction of mCa to a  normal Alb using appropriate equation Various formulae for adjustment: Marshall: ACa (mmol/L)= [Ca] + (0.02 x [40-Alb]) Tietz: ACa (mg/dL) = [Ca]mg/dL/(0.6 + ) Derived equations: based on the calculation of Barth (Barth et al., Ann Clin Biochem 1996; 33: 55-58)X/P!P/PPIPPPUPW &fl ff  1fH1#M\'How to calculate an adjustment equation((   'How to calculate an adjustment equation((   'How to calculate an adjustment equation((   n5Different Formulae Slope and Intercept for adjustment66 f  mClin Biochem & Met Med, RLBUH f$FFollowing the Barth adjustment Where is this formula coming from? Analysis of 100 samples with normal biochemistry A plot of measured Ca vs Alb concentration A regression line (y = mx + c) A slope = m = 0.0103 RR= 2.20 - 2.60 Re-evaluated in May 2003 on 716 subjects with no sig change (p = 0.602)2BPPHPG>0ioAims fEvaluate formulae for Ca adjustment to be valid at different Alb values particularly at low values < 25g/L Validate against free (ionised) Ca measurement Can we predict free Ca by using a correct adjustment &ZZIpSamples f50 fresh samples from patients with various levels of Alb analysed for TCa (CPC) & Alb (BCG) on a Roche Mod). The samples were analysed within 1 hr with a Bayer C0855 for Ca2+ and pH. values for free Ca2+ were adjusted to pH 7.4 Samples were kept frozen until analysis with FAAS.voJb  N>:7q  VFAAS to confirm the accuracy of the Roche results against a reference method 2OZZZWvx.Is there agreement between Roche Ca and FAAS?// f  y9Is Ca adjustment valid at different Alb concentrations? :: fLNon-linearity of the relationship between serum total Ca and Alb concentration has been reported by many studies e.g Ashby (Ashby et al., Ann Clin Biochem 1986; 23: 533-537) confirmed again in the study of Barth and many other studies (Barth et al., Ann Clin Biochem 1996; 33: 55-58) Is there an optimal formula for adjustment? zZZZ{6=0-Pg@zAdjustment with RLUH equation   "Adjustment with Marshall equation 0#"    Let s compare RLUH adjustment regression line with Marshall equation for the same set of resultscc   |(RLUH ACa vs Leeds and Marshall equations)) f,     So, RLUH equation, Marshall or an alternative equation which give a better adjustment? Make your own lab decision! \/   ,-$Adjustment with a modified equation 0%$      Yy = mx + c iCa/ACa = -0.00004 . [Alb] + 0.4235 iCa = (-0.00004 . [Alb] + 0.4235) / ACa ZZb      !   Summary 2Adjustment of Ca over range of Alb with different equations resulted in significant difference (p < 0.0001). RLUH adjustment equation appears to underestimate the ACa compared with Leeds and Marshall particularly at low alb < 20g/L. To establish consensus on ACa values derivation of local correction formula is recommended Should be part of routine quality control assessment NEQAS Appropriateness of any formula must relate to free Ca (Ca2+) Thus - the ratio of iCa/ACa should be independent of albumin over a wide range of concentrations (10-50+) LHPPP p>_Q Further work  This study is only a pilot study on few samples. For this important issue, a larger sample size is necessary and is currently being investigated. 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'>3___PPT10i.C<+D=' = @B +y  `0(  x  c $8 @   x  c $D9 zq  H  0޽h ? ̙33y___PPT10Y+D=' = @B +]  |p (  x  c $d:   F   0:z `[Alb] [TCa] [Alb] [TCa] , 2EP`6B ~  s *$; , <~     0;  $ Total prot (mg/dL) 18.5% 2%,XB  0D  g XB  0Dfo/XB  0DoXB  0Dfo/XB   0Dfo?XB   0DoXB   0Dfo?H  0޽h ? ffD3fy___PPT10Y+D=' = @B +<  SK =(  x  c $d=    RB  s *Doz RB  s *Do  RB  s *D3o6    0$> )  ; x = [Alb] 2    0>jGxd :y = [Ca] 2   0D? Wg  30 2  0@/ ^ y = mx + c 2 C"? ' 0d@@  Y.* 2ff   ( 0@` P  Y.* 2ff   ) 0d݈  Y.* 2ff   * 0݈@0 W  Y.* 2ff   + 0$ވ '  Y.* 2ff   , 0ވ  7 Y.* 2ff   - 0ވ@ Y.* 2ff   . 0D߈@ 0 7 Y.* 2ff   / 0߈P  Y.* 2ff   0 0@ 0  Y.* 2ff   1 0d0 7 Y.* 2ff   2 0p`G Y.* 2ff   3 0$p` Y.* 2ff   4 00 w Y.* 2ff   5 00 w Y.* 2ff   6 0D  Y.* 2ff   7 0`0  Y.* 2ff   8 0P p  Y.* 2ff   9 0d`  Y.* 2ff   : 0 7 Y.* 2ff   ; 0$p Y.* 2ff   < 00 w Y.* 2ff   = 0  Y.* 2ff  H  0޽h ? ffD3f___PPT10i.C*b+D=' = @B +      (  x  c $d    RB  s *Doz RB  s *Do  RB  s *D3o1 LB  c $Df LB  c $Df LB  c $DLB   c $D    0` Z |y1=TCa8 2fff   0$P J |y2=ACa8 2   0 )  ; x = [Alb] 2    0jGxd :y = [Ca] 2   0$8 `  ^ x 1= [Alb]8 2fff  0r F  ^ x 2= [Alb]8 2  0s n  9a 2f  0ds Wg  30 2  0$t/ b y = mx + c 2 3C"?  0t Sb 2f H  0޽h ? ffD3f___PPT10i.C*b+D=' = @B +    0(  x  c $$'    RB  s *Doz RB  s *Do  RB  s *D3o6 LB  c $Df LB  c $Df LB  c $DLB   c $D    0'`94 |y1=TCa8 2fff   04P9$ |y2=ACa8 2   0 )  ; x = [Alb] 2    0jGxd :y = [Ca] 2   0t `  ^ x 1= [Alb]8 2fff  04 F  ^ x 2= [Alb]8 2RB  s *D X2  0~   0 #  a(x1,y1)` 2fffff  0T Wg  30 2  03/ b y = mx + c 2 3C"?  0Աd b(x2,y2)` 2fffff   04   xy = mx +c (y2 - y1) = m . (x2  x1) + c (ACa  TCa) = m . DAlb + c ACa = TCa + m . D Alb +cc 2            0`~   ttga = m0 2     0TF #   t"Alb=x2-x1D 2  X2  0m X2   0H  0޽h ? ffD3f___PPT10i.C*b+D=' = @B +w  .(  x  c $v ,      dv0e0eA 'U     0v p  `Which equation is a more reliable for adjusting the measured Ca ? Should be locally derived !!$a 2Df  0$w)  U1.940 2   0wC  Y 2.20-2.60 2     0Dx~ L1.94 2  P  0x  lConsidering various forms of calculation can we expect to standardise our reference ACa range in the region?4m 2:1ff,: H  0޽h ? ffD3fy___PPT10Y+D=' = @B +  ](     0$zf     "6 ?x  c $z     0D{f {% ACa = Total Ca + 0.0103 (40.4 - Alb)&& "H  0޽h ? ffD3fy___PPT10Y+D=' = @B +y  0(  x  c $d|    x  c $| "   H  0޽h ? ̙33y___PPT10Y+D=' = @B +E  ld(  ~  s *y j   ~  s *} ?    0 # l!,A l? ? P p& l H  0޽h ? ̙33y___PPT10Y+D=' = @B +  [(    c $䇋 "  "p`Pp  0D }  -Statistical analysis: Data were analysed for normal distribution and then compared with paired t-test. A p value less than 5% (p <0.05) considered significant Lff"zH  0޽h ? ̙33y___PPT10Y+D=' = @B +  G?(    0`  ?Results<fH  0޽h ? ̙33y___PPT10Y+D=' = @B +  E=$(  $x $ c $$     $0 <A m? /T  m x $ <A n??4 ,  nV $ 0 5  Roche Ca FAAS Ca Mean 2.053 2.034 SD 0.18 0.20 SEM 0.029 0.033Z! 2`0<2 ` $ 03I Op =0.28, 233H $ 0޽h ? ffD3fy___PPT10Y+D=' = @B +  ((  (x ( c $䍋 0   ( S ~D0e0e     H ( 0޽h ? ffD3fy___PPT10Y+D=' = @B +$ KC ,(  , , 0$z n2Q. Which equation for adjustment is more reliable? 3(23f , 0ď T  m=Adjustment performed for all samples with different equations> 2>H , 0޽h ? ffD3fy___PPT10Y+D=' = @B +  KC0L(  Lx L c $D @     L 0p xRLUH adjustment of measured Ca (green line) suggests underestimation ?? at low Alb. Slope ideally should approach zero.y 2y L s *du }' ACa = Total [Ca] + 0.0103 (40.4 - Alb)(($08 0  L0  L <A r??0  r` L 0]H L 0޽h ? ffD3fL  @P(  Px P c $Ē R    P s *$z*Jz \Marshall: ACa = Total [Ca] (0.02 . [40-Alb])*/ 2 "d P 0p* Marshall ACa produced a nearly straight regression line (green) which is ideal, however it might overestimate Ca at low Alb leading to false hypercalcaemia  2,  P0 T{ {  A s? V  s  P s *u$w @  H P 0޽h ? ffD3fm PD$(  D D 0䐋  PWhich equation is a better indicator of the adjustment? In relation to free Ca2+4Q(2N$$ Q  D 0NV v  r0To answer this question we modified the equation1 21 1 H D 0޽h ? ffD3fy___PPT10Y+D=' = @B +p   `H(  Hx H c $4P p    H H 0޽h ? ffD3f_  ~p4(  4x 4 c $P P0    40 <A p? :  p E 4 0P  Does RLUH adjustment is underestimating or do the others overestimate!!. Does it matter? If Yes, are they statistically different from each other?& 2TP 4 0Q3& @ p < 0.0001 2 3 4 0R&: y RLBUH LEEDS Marshal Mean 2.309 2.522 2.450 SD 0.18 0.13 0.13 SEM 0.026 0.020 0.020B 2   /  H 4 0޽h ? ffD3fy___PPT10Y+D=' = @B +p   X(  Xx X c $S p=  H X 0޽h ? ffD3f  T(  T T0 <A t? /4 t  T S ~S0e0e  R    T c $TT < 2   T s *T ~( ACa = Total [Ca] + (0.0150 (40 - [Alb])))% T 0Uv rIf the equation corrected by factor of 0.0150 ??? no more overestimation Green line slope approaches zeros 2sH T 0޽h ? ffD3fd \d(  \, \ <U p 8Can we predict Ca2+ based on a correct adjustment ???^9     "`H \ 0޽h ? '>380___PPT10.C@R  `N(  `x ` c $tU   x ` <A u??G7  u ` 0V d  RLUH Marshall Modified m -0.0009 0.0008 -0.00004 r 0.35 0.34 0.017 2+ff ?f D  H ` 0޽h ? ffD3f  h6(  h~ h s *W ,   x h c $X ]  H h 0޽h ? ffD3f  l0(  lx l c $TW    x l c $X `   H l 0޽h ? ffD3f  p0(  px p c $4Y j@    x p c $Y @q `  H p 0޽h ? ffD3f t6(  tx t c $ a  ~ t s * 2   H t 0޽h ? ̙33y___PPT10Y+D=' = @B +] dx\ilT>-f<306aq`l^%vK%"R` Fj~PZEB)6UihH!R&U#Ra"uIHPDVazλzg@Yr?{.w7+.=!kAk)x\nLA9&RtN)QY%ڐhBz>d / ,E.J WhABރEދE("됋!C¿R2r dW>:  l8TQ Ę'}i9Фk7z:3=cAt4.O᪄\a,u3O: C:[1iA4pMGO`ռǰ%g}{Xw]؞iٰd^.+k~q(\/uvfw!z٪1~D4Rӹ . 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