Evidence for a hypothalamic alteration of catecholamine metabolism in polycystic ovary syndrome.

The role of brain catecholamine (CA) activity in the neuroendocrine regulation of the GnRH-LH system in polycystic ovary syndrome (PCO) was investigated by high-performance liquid chromatography (HPLC) with electrochemical detector. We measured urinary dopamine (DA), noradrenaline (NA), adrenaline (A), vanillylmandelic acid (VMA), homovanillic acid (HVA), 3,4-dihydroxyphenylacetic acid (DOPAC) and total 3-methoxy-4-hydroxyphenylglycol (MHPG) levels in a group of 12 women with PCO before and during peripheral dopa-decarboxylase blockade, by carbidopa. HVA and DOPAC concentrations were significantly lower (P less than 0.001 and P less than 0.005, respectively) in PCO patients compared with twelve control subjects in early follicular phase, whereas total MHPG concentrations and MHPG/VMA ratio were significantly higher (P less than 0.005) in PCO patients. Moreover, HVA and DOPAC concentrations in PCO patients were similar to those of the control subjects in preovulatory phase, while MHPG concentrations remained higher in PCO patients (P less than 0.01). DA, NA, A and VMA concentrations were similar to those of control subjects in both phases of the cycle. During carbidopa administration the concentrations of all urinary CAs and metabolites were unchanged, except those of DA which dropped markedly (P less than 0.001). These data suggest that (1) an altered central catecholamine metabolism consisting of DA deficiency and NA excess is present in PCO, and (2) the site of DA deficiency may be located in the hypothalamus.

Rapid procedure for chromatographic isolation of DOPA, DOPAC, epinephrine, norepinephrine and dopamine from a single urinary sample at endogenous levels.

A three-step procedure has been investigated to extract 3,4-dihydroxyphenylalanine (DOPA), 3,4-dihydroxyphenylacetic acid (DOPAC), epinephrine (E), norepinephrine (NE) and dopamine (DA) from a single urinary sample with the object of obtaining extracts pure enough for specific fluorimetric assay. The procedure described in this paper results from the combination of urine purification on an aluminum oxide column, separation by ion-exchange chromatography of the DOPA-DOPAC fraction from catecholamines, and ether isolation of DOPAC from DOPA. The whole procedure is rapid and easily performed in one work day. Extraction recoveries were 72.4 +- 3.5%, 76 +- 2%, 85.7 +- 3.3%, 85.6 +- 1.4% and 92.4 +- 5.5% for DOPA, DOPAC, E, NE and DA respectively (n=6). The lowest amounts of the five catechols that could be detected in urinary samples by a combination of this extraction procedure and the methods of assay used in our laboratory were 15 ng for DOPA, 40 ng for NE, 20 ng for E, 152 ng for DA and 2.95 micrograms for DOPAC. Urinary volumes convenient for accurate estimation of each compound were 20 ml for healthy human subjects. For pathological or pharmacological purposes, 5 ml of human urine were sufficient. The daily excretion of DOPA, DOPAC, E, NE and DA found by this procedure agrees with data obtained by other authors in healthy subjects. In pathological samples, our three-step procedure led to lower amounts than methods using alumina purification only. The discrepancies between the two methods are discussed in terms of development of internal standards, relative specificity of fluorimetric assays, values of blank eluates, and the possibility of interference from unknown abnormal body metabolites or pharmacological drugs not eliminated by a single-step alumina purification.