Automation of multiple sephadex LH-20 column chromatography for the simultaneous separation of plasma corticosteroids

Sippell, Wolfgang G.; Lehmann, Paul F.; Hollmann, G.

doi:10.1016/s0021-9673(00)84673-7

Cited by 43 publications

(7 citation statements)

References 9 publications

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“…The micromethod of multisteroid analysis has been previously described and evaluated (4,5). In brief, it included the addition of tracer amounts of each of the eight steroids to a 250-p1 plasma sample, extraction with methylene chloride, and a mechanized Sephadex LH-20 chromatography for the simultaneous separation of the steroids.…”

Section: Methodsmentioning

confidence: 99%

“…For full-term healthy infants, data on plasma steroid levels in the neonatal period are available (2,3), but studies on adrenal steroids in healthy premature infants are still lacking. Our micromethod of multisteroid analysis (4,5) enabled us to measure simultaneously in a single small plasma sample individual mineralocorticoids, glucocorticoids, and progestins in premature infants at birth and longitudinally during the first hours and days of life.…”

Section: P Progesterone 170hp 17-hydroxyprogesteronementioning

confidence: 99%

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Plasma Mineralocorticoids, Glucocorticoids, and Progestins in Premature Infants: Longitudinal Study during the First Week of Life

et al. 1988

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ABSTRACT. Plasma levels of aldosterone, corticosterone, 11-deoxycorticosterone, progesterone, 17-hydroxyproge.sterone, 11-deoxycortisol, cortisol, and cortisone were measured simultaneously by a micromethod of multisteroid analysis in eight vaginally delivered premature infants (PI) of 33-36 wk gestation with uneventful peri-and postnatal course. Mean concentrations (nglml) in umbilical arterial and in peripheral venous or capillary plasma sampled longitudinally at age 2 h to 7 days were compared with the same kind of data obtained from a group of 12 term infants (TI) who served as controls. Mean aldosterone was two to five times higher in PI than in TI (umbilical artery, 2 h to 7 days; p < 0.05), whereas 11-deoxycorticosterone was lower in PI from 2 h ( p < 0.01) until 7 days (NS).Corticosterone was significantly higher in PI than TI at 6 and 24 h after birth, whereas cortisol was slightly lower (NS) in PI in umbilical artery and 2 h after birth, but higher (p c 0.02) at 6 h, showing less variation in PI than in TI. 17-Hydroxyprogesterone levels in PI were two to three times higher ( p c 0.02) during 6 h until 7 days after birth. The data suggest that PI are able to maintain high aldosterone levels in the early neonatal period. Higher levels of the active glucocorticoids (cortisol and corticosterone) seen after delivery point to a more stressful extrauterine adaptation of PI. Furthermore, the data demonstrate that the adrenal cortex is fully functioning in premature infants (33-36 wk gestation) as well as in term infants. (Pediatr Res 23: 525-529,1988) Abbreviations RDS, respiratory distress syndrome UA, umbilical artery Aldo, aldosterone DOC, 11-deoxycorticosterone E, cortisone S, 11-deoxycortisol B, corticosterone F, cortisol P. progesterone 170HP, 17-hydroxyprogesterone Abrupt cessation of steroid supply from the placenta after severing the umbilical cord, and rapid postnatal involution of the fetal zone of the adrenal cortex dramatically changes the hormonal milieu of the fetus after birth. There is increasing evidence that the adrenal cortex plays an important role in the postnatal adaptation of newborns (1). For full-term healthy infants, data on plasma steroid levels in the neonatal period are available (2, 3), but studies on adrenal steroids in healthy premature infants are still lacking. Our micromethod of multisteroid analysis (4, 5) enabled us to measure simultaneously in a single small plasma sample individual mineralocorticoids, glucocorticoids, and progestins in premature infants at birth and longitudinally during the first hours and days of life.Herein we assess in detail the adrenocortical function of healthy moderately premature infants after birth and provide reference data for the physiologically most important corticosteroids. MATERIALS AND METHODSEight premature appropriate for gestational age infants (33-36 wk gestation) of both sexes (three males, five females) with uneventful peri-and postnatal courses were studied longitudinally from birth to day 7. Their mothers (20-36 yr) were heal...

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Section: Methodsmentioning

confidence: 99%

Section: P Progesterone 170hp 17-hydroxyprogesteronementioning

confidence: 99%

Plasma Mineralocorticoids, Glucocorticoids, and Progestins in Premature Infants: Longitudinal Study during the First Week of Life

et al. 1988

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“…Levels of unconjugated aldosterone, corticosterone, 1 1 -deoxycorticosterone, progesterone, 17-hydroxyprogesterone, I 1-deoxycortisol, cortisol, and cortisone were measured from the same small plasma sample. The micromethod of multisteroid analysis has been previously described and evaluated (9,10). In brief, it included the addition of tracer amounts of each of the eight steroids to a 250-p1 plasma sample, extraction with methylene chloride, and a mechanized Sephadex LH-20 chromatography for the simultaneous separation of the steroids.…”

Section: Doerrmentioning

confidence: 99%

Adrenocortical Steroids in Small-for-Gestational-Age Term Infants during the Early Neonatal Period1,2

et al. 1989

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ABSTRACT. We evaluated adrenocortical steroid concentrations a t birth and during postnatal adaptation (2 h until 7 days) in 10 vaginally delivered term small-for-gestational-age (SGA) infants and 1 2 term appropriate-forgestational age infants. Plasma aldosterone, ll-deoxycorticosterone, corticosterone, progesterone, 17-hydroxyprogesterone, 11-deoxycortisol, cortisol, and cortisone were longitudinally measured by specific RIA after Sephadex LH-20 chromatography. Mean aldosterone was significantly higher in SGA than in appropriate-for-gestationalage infants (2 h to 7 days; p < 0.001). In SGA infants, cortisone and cortisol levels were significantly lower in umbilical artery (p < 0.05), and all glucocorticoid levels were significantly lower 12 h after birth (p < 0.05). Thereafter (24 h to 7 days), only 11-deoxycortisol levels remained significantly lower in SGA; corticosterone and cortisol levels were even higher @ < 0.05) in SGA 24 h after birth. The data suggest that SGA infants maintain high aldosterone levels throughout the 1st wk of life. Low cortisol and cortisone levels in umbilical artery a s well a s low glucocorticoid levels a t 2 h and/or 1 2 h compared to term appropriate-for-gestational-age infants may reflect either a less stressful postnatal adaptation or, more likely, a reduced adrenocortical synthesis in term SGA infants. (Pediatr Res 25:115-118, 1989) Abbreviations SGA, small for gestational age UA, umbilical artery AGA, appropriate for gestational age Perinatal problems in neonates with severe intrauterine growth retardation are frequent. They include asphyxia, a high neonatal mortality, electrolyte imbalance, hypoglycemia, cardiorespiratory problems, and neurologic abnormalities (1). Organ weights are significantly reduced (2); in particular, the adrenal cortex is small with respect to size and cell number (3).

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“…MULTIPLE PLASMA STEROIDS IN AF 817 dried extract was subjected to Sephadex LH-20 column chromatography as previously described (12). Glass columns, 1 cm in diameter, were packed with a mixture of 10 g LH-20 and 2% methanol in dichloromethane.…”

mentioning

confidence: 99%

Concentration of 14 Steroid Hormones in Human Amniotic Fluid of Midpregnancy*

Forest¹,

Peretti²,

Lecoq³

et al. 1980

The Journal of Clinical Endocrinology & Metabolism

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Amniotic fluid (AF) levels of all steroids leading from pregnenolone (delta 5Preg) to androgens and estrogens of both the delta 5 and the delta 4 pathways and those of cortisol and cortisone have been determined in 63 normal pregnancies (12-19 weeks gestation). The 12 unconjugated steroids [delta 5Preg, 17 alpha-hydroxypregnenolone, progesterone, 17 alpha-hydroxyprogesterone, dehydroepiandrosterone (DHA), delta 4-androstenedione, delta 5-androstene-3 beta,17 beta-diol (delta 5Adiol), testosterone (T), estrone, 17 beta-estradiol (E2), cortisol, and cortisone] were measured by specific RIAs after appropriate purification by Celite or LH-20 column chromatography, while the sulfates of DHA and delta 5Preg were assayed directly on diluted samples. There were distinct sex differences; T and delta 4-androstenedione levels were higher (P < 0.001) in males than in females, while AF levels of delta 5Preg, 17 alpha-hydroxypregnenolone, 17-hydroxyprogesterone, DHA, and delta 5Adiol were higher (P < 0.05) in females than in males. AF levels of E2 were significantly higher in females only between 15 and 19 weeks gestation. There was no difference between sexes in AF levels of progesterone, estrone, cortisone, and cortisol. AF levels of T, delta 4-androstenedione, and E2 decreased with age in males, and AF levels of DHA increased in females during the period examined. These observed sex differences suggest that T and delta 4-androstenedione may reflect fetal testicular activity, while E2 and 17-hydroxyprogesterone might reflect fetal ovarian activity. Determination of AF levels of T would appear to be a valuable screening test for antenatal diagnosis of sex (predictive error, less than or equal to 15%), but not in the presence of steroidogenic enzyme defects. Elevated levels of 17-hydroxyprogesterone were found in the AF of two fetuses with either a 17-20 desmolase defect or 21-hydroxylase deficiency; AF levels of androgens were low in the former and high in the latter.

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Automation of multiple sephadex LH-20 column chromatography for the simultaneous separation of plasma corticosteroids

Cited by 43 publications

References 9 publications

Plasma Mineralocorticoids, Glucocorticoids, and Progestins in Premature Infants: Longitudinal Study during the First Week of Life

Plasma Mineralocorticoids, Glucocorticoids, and Progestins in Premature Infants: Longitudinal Study during the First Week of Life

Adrenocortical Steroids in Small-for-Gestational-Age Term Infants during the Early Neonatal Period1,2

Concentration of 14 Steroid Hormones in Human Amniotic Fluid of Midpregnancy*

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