Laboratory Tests for Assessing Metabolic Bone Disease


     A. Calcium is present in serum in three forms.  About 50% is
     bound to proteins, predominantly albumin.  About 10% is
     complexed with anions, such as bicarbonate, phosphate,
     citrate, etc., and about 40% is in the free, ionized form. 
     The free, ionized form is the physiologically important
     fraction.
     
     Total calcium is routinely measured in serum or urine
     specimens by a colorimetric method.  The absorption of a dye
     (most commonly cresolphthalein complexone) intensifies upon
     interacting with calcium from acidified specimens in which
     calcium is liberated from bound forms.
     Determination of free calcium is not always readily
     available, but can be measured by ion specific electrode. 
     Total serum calcium concentration is generally proportional
     to free calcium concentration, so long as albumin
     concentration is normal.  When albumin concentration is
     abnormal, total serum calcium concentration is interpreted
     on the basis of what the total calcium concentration would
     be if albumin concentration were normal by adding 0.8 mg/dl
     to the calcium value for each 1 g/dl that albumin is less
     than 4 g/dl, i.e., 
       corrected total calcium = measured value + 
                                 0.8x(4 g/dl - albumin)mg/dl.

     Return to the main document.
     ____________________________________________________________
     
     B. Inorganic Phosphate is routinely measured in serum or
     urine specimens by a colorimetric method.
     ____________________________________________________________
     
     C. The Alkaline Phosphatase isoenzyme from bone becomes
     elevated as a consequence of osteoblast stimulation.  Total
     serum alkaline phosphatase elevations in metabolic bone
     disease are most often only mild, except in Paget's disease
     of bone in which cases values are several tens to hundreds
     times the upper limit of normal.
     ____________________________________________________________
     
     D. Parathyroid Hormone is measured by radioimmunoassay. 
     Until recently only a few commercial, specialty laboratories
     provided reliable results.  Interpretation of results is
     complicated by the fact that the intact, circulating hormone
     is rapidly cleaved by a proteolytic enzyme into N and C
     terminal fragments.  The circulating half life of the N
     terminal fragment is less than an hour.  The C terminal
     fragment has a longer circulating life time of several
     hours.  Assays are available for the intact hormone, for the
     N terminal fragment and for the C terminal fragment.  Values
     for the intact hormone are difficult to interpret because
     the N and C terminal fragments are also included in the
     measurement but with different cross reactivities and is
     therefor not clinically useful.
     Results from the C terminal assay are useful for deriving
     information about the functional status of the parathyroid
     gland.  PTH results must be interpreted along with serum
     calcium results.  Elevated PTH values are found in both
     primary and secondary hyperparathyroidism.  PTH is
     appropriately elevated in secondary hyperparathyroidism in
     which cases serum calcium values are low or low-normal.  PTH
     is inappropriately elevated in primary hyperparathyroidism
     in which cases serum calcium concentrations are elevated. 
     Depressed PTH results are found in both primary
     hypoparathyroidism and in cases of hypercalcemia from excess
     calcium intake and from osteolytic bone destruction.
     The N terminal assay is used when blood specimens are drawn
     from neck veins draining the glands in order to identify the
     location of an autonomously functioning adenoma.

                                     Return to the main document.  
     ____________________________________________________________
     
     E. Indirect Indications of PTH Activity
     
     1. % tubular reabsorption of phosphate ( % TPR ) is
     decreased as a direct effect of PTH on the kidneys and,
     along with the consequently depressed serum phosphate, is
     one of the characteristic findings in primary (or ectopic)
     hyperparathyroidism.  The value is determined from
     measurements of phoshate and creatinine on a serum and a
     spot urine specimen:
            % TPR = 100x[ 1 - (phos.u/phos.s)/(creat.u/creat.s)]
     
     2. The serum Cl-/phosphate ratio increases to 32 - 80 in
     cases of primary (or ectopic) hyperparathyroidism. 
     Decreased serum phosphate, as a consequence of decreased
     renal phosphate reabsorption, is one of the characteristic
     findings in hyperparathyroidism.  A mild to moderate
     metabolic acidosis is also typically associated with
     hyperparathyroidism.  The anion gap is normal so that serum
     Cl- is elevated by the amount that HCO3 is depressed.  The
     ratio provides a simple means to justify a suspicion of
     hyperparathytroidism and to conduct more definitive testing.
     
     3.  Urine cyclic AMP increases as a consequence of the
     interaction of PTH with renal epithelial cell membrane
     receptors and its intracellular formation as the "second
     messenger".  The interaction of PTH-like protein with the 
     receptors has the same effect.

     Cyclic AMP is measured by radioimmunoassay in
     a spot urine specimen and along with values for serum and
     urine creatinine results are expressed with respect to GFR,
     ie:
     
          cyclic AMP excretion rate = 
                        100x(urine cyclic AMP concentraation)/(Cru/Crs)
     
     The test is most useful when PTH is found to not be elevated,
     but all other test results are consistent with hyperPTH. 
     Elevated cyclic AMP excretion rate and depressed or normal
     serum PTH concentration strongly suggests ectopic
     elaboration of a "PTH-like substance".

Return to the main document.


Reviewed: April 3, 2000