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PubMed1975

Validation of methods for determination of urinary estriol during pregnancy using mass fragmentography.

A specific and accurate method for determination of urinary estriol during pregnancy is described. After acid hydrolysis of 1 ml urine, a fixed amount of [2,4-2H2] estriol (usually 15 mug) is added and the mixture is extracted with diethyl ether. The extracted estriol is converted into the tri-trimethylsily derivative and the amount of unlabeled estriol is determined from the ratio between the recordings at m/e 504 and m/e 506 obtained after analysis with a gas chromatograph-mass spectrometer equipped with an MID (multiple ion detector). The two ions used correspond to the molecular peak in the mass spectrum of unlabeled and (2,4-2H2)-labeled tri-trimethylsilyl derivative, respectively. The relative standard deviation of the method was about 6%. The method was compared with a photometric method using the Kober reaction and the gas liquid chromatographic method used in routine analysis. There was a good correlation in both cases and the regression coefficient was 1.5 and 1.3, respectively.

Estriol↗

PubMed1979

Diurnal variations of estriol and cortisol in serum and urine in late pregnancy with and without fenoterol-therapy.

The serum concentration of unconjugated estriol, total estriol, cortisol in one hour intervals and the urinary excretion of total estrogens and cortisol in three hour intervals was determined in eleven apparently healthy women between 26 and 40 weeks of gestation. They were divided into two groups, in order to study the influence of Fenoterol treatment on the daily variation of the measured parameters. Six patients were treated with Fenoterol and five patients received no medication. Between the two groups no statistical differences could be detected for the measured parameters. Regular and marked diurnal variations in cortisol have been observed, whereas total and unconjugated estriol showed small and irregular daily changes. These daily variations of "estriol" were of the same magnitude as the two-fold methodical standard deviation. For this reason it is not necessary to standardize the sampling time.

Circadian Rhythm↗

PubMed1983

Liquid chromatography with fluorometric detection of unconjugated estriol in serum of pregnant women.

We describe a fluorometric liquid chromatographic assay for the measurement of unconjugated estriol in the serum of pregnant women. Estriol is extracted into methylene chloride/propanol-2 from serum by use of a Clin-Elut extraction column, the extract evaporated, and the residue redissolved in mobile phase. An aliquot is injected onto the liquid chromatograph and the estriol is separated on a reversed-phase octyl column with a mobile phase consisting of acetonitrile/phosphate buffer (23:77, v/v). The effluent is monitored by fluorescence detection. The proposed method offers good reproducibility (CV less than 7%), sensitivity (less than 0.5 micrograms/l), and accuracy. Of many drugs and steroids tested, only 16,17-epiestriol interferes with the estriol analysis.

Chromatography, Liquid↗

PubMed1983

A comparison of serum free estriol with spontaneous urine total estrogen and estrogen/creatinine ratio.

A mathematical comparison was made between the serum free estriol levels, the urine total estrogen, and the total estrogen/creatinine ratio in 83 pregnant women during the last trimester of pregnancy. The blood serum and spontaneous urine samples were obtained simultaneously, and each woman was sampled once. The serum estriol was measured by radioimmunoassay and the urine estrogen levels measured by the aqueous fluorometric Kober technique. The group averages of the three different measurements paralleled each other throughout the third trimester. Linear regression and correlation studies of the measured levels for each subject by the different methods were statistically significant (P less than or equal to 0.0001). The correlation factor between serum free estriol and spontaneous urine total estrogen was +0.5991, and between serum free estriol and the spontaneous urine total estrogen/creatinine ratio was +0.8086. The very high correlation between the two urine and the serum method suggest that spontaneous urine samples may be used to measure estrogen changes during pregnancy.

Adolescent↗

PubMed1984

Effect of estriol treatment on the menstrual cycle and prolactin secretion.

In order to study the biological effects of estriol in women 20 mg estriol was administered daily to 7 young women. Plasma luteinizing hormone (LH), estradiol (E2), progesterone (Pg) and prolactin (Prl) were measured during a treatment and a control cycle every second or third day. Further 3, 6 or 20 mg estriol was administered in a single dose to 5 women and plasma Prl, unconjugated and conjugated estriol (E3) measured over 24 h at 2-3 intervals. In 2 experiments with 20 mg E3, blood samples were taken more frequently, over 6 h. When 20 mg E2 was administered daily, 2 of the 7 young women had anovulatory cycles. The mean plasma E2 was lower during the follicular and ovulatory phases (P less than 0.025) and mean plasma LH was higher (P less than 0.005) during the luteal phase, when E3 was given. Because of the 2 anovulatory cycles the mean Pg value during the luteal phase was lower (P less than 0.05) during treatment. There was a slight decrease in mean Prl in 5 out of 6 women (P less than 0.0005), but in only 1 woman was this decrease substantial (from a mean value of 27.6-18.9 ng/ml; P less than 0.01). When 6 or 20 mg E3 was administered orally in the morning a significant negative correlation (P less than 0.01) between plasma Prl and unconjugated E3 was found. The correlation coefficient was highest (r = -0.74) with 6 mg E3. When 3 mg was administered no obvious effect on Prl section was seen. However, when results from all experiments with identical time schedules were pooled (two with 3 mg, two with 6 mg and one with 20 mg E3) and the mean values for plasma Prl calculated and compared with the mean values obtained in 7 control experiments, it was found that E3 administration in the morning almost abolishes the Prl rise during the following night. There was a statistically significant (P less than 0.0125) decrease in the difference between the maximum value during the night and the minimum value during the day. The minimum value was significantly higher (P less than 0.01) and the maximum value significantly lower (P less than 0.025) after E3 treatment, compared to the control values. It is concluded that long-term administration of 20 mg E3 usually has only a slight but significant decreasing effect on mean plasma Prl concentration measured in the morning, before the next dose is taken.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗

PubMed1984

Preparation of specific antisera to estrone-3-glucuronide and estriol-3-glucuronide.

The preparation and antigenic properties of estrone-3-glucuronide- and estriol-3-glucuronide-bovine serum albumin conjugates in which the hapten is linked to the carrier protein through an (O-carboxymethyl)oxime bridge at the C-6 position on the steroid nucleus, have been described. Antibodies raised against the two immunogens in the rabbit possessed high specificity to estrone-3-glucuronide and estriol-3-glucuronide, respectively, exhibiting little cross-reactivities with other estrogen conjugates and no cross-reactions with related steroids except for free estrogens, their 3-methyl ethers and 3-sulfates. The cross-reactive antibodies were eliminated by partial immunoadsorption on affinity chromatographic media using the estrone-3-methyl ether 17-(O-carboxymethyl)oxime- and estriol-3-methyl ether 16 (or 17)-hemisuccinate-aminohexyl Sepharose conjugates, respectively. The purified antisera exhibited no cross-reactivities with free estrogens and ring A conjugates of estrone and estriol.

Animals↗

PubMed1984

Is serum free estriol measurement essential in the management of hypertensive disorders during pregnancy?

A prospective study of antepartum fetal evaluation of 306 randomized hypertensive pregnancies was carried out. One hundred and fifty-four patients (group 1) were managed in accordance with a protocol (protocol A) which included non-stress test (NST), oxytocin challenge test (OCT), serum unconjugated (free) estriol measurements (E3), and ultrasound measurement of the fetal head-to-abdomen circumference ratio (H/A c.r.). A second group (group 2), composed of 152 patients, was managed using another protocol (protocol B) which included the NST, OCT and ultrasound measurement of the fetal head-to-abdomen circumference ratio (H/A). We found a good correlation between the serial normal tests and the outcome of pregnancies. We obtained good results in patients with abnormal NST-OCT and meconium-stained amniotic fluid and in patients with repeated abnormal NST-OCT with clear amniotic fluid due to active management by early deliveries ignoring fetal lung maturity. This management increased the rate of cesarean section and prematurity. There was a low correlation between patients with abnormal serum free estriol as the only antepartum pathological test and the appearance of intrapartum fetal distress, low birth weight and perinatal morbidity and mortality. The contribution of serum free estriol (E3) measurements in such patients was only of value in cases of intra-uterine fetal growth retardation, but its prediction rate was less than that of ultrasound measurement of head-to-abdomen circumference ratio. Our results indicate that hypertensive pregnancy management without serum free estriol measurement may be valuable and safe.

Estriol↗

PubMed1979

Urinary excretion of estrone glucosiduronate, 17 beta-estradiol-17-glucosiduronate, and estriol-16 alpha-glucosiduronate. Significance of proportionate differences during the menstrual cycle.

The urinary excretion of estrone glucosiduronate, 17 beta-estradiol-17-glucosiduronate, and estriol-16 alpha-glucosiduronate in men and throughout the menstrual cycle in women was measured by specific radioimmunoassay. In 9 men the mean +/- SE excretion of these conjugates was 15.9 +/- 1.4, 2.7 +/- 0.3, and 3.2 +/- 0.2 microgram/24 h respectively. In 15 women studied in the midfollicular phase (day 8) of the menstrual cycle, the excretion was 19.4 +/- 1.7, 2.9 +/- 0.2, and 5.4 +/- 1.3 micrograms/24 h. Excretion of each conjugate was significantly (P less than 0.01) elevated in the midluteal phase (day 22) to 41.9 +/- 3.9, 6.3 +/- 0.8, and 12.2 +/- 1.5 micrograms/24 h respectively (n = 14). The mean excretion of estriol-16 alpha-glucosiduronate was greater than that of 17 beta-estradiol-17-glucosiduronate in the luteal phase (P less than 0.05) but not in the follicular phase or in men (P greater than 0.05). The excretion of each of these specific conjugates measured throughout the menstrual cycle in 7 women was characterized by a sharp midcycle peak and a lower, broader luteal phase peak. The ratios of estriol-16 alpha-glucosiduronate to 17 beta-estradiol-17-glucosiduronate, estrone glucosiduronate to 17 beta-estradiol-17-glucosiduronate, and estriol-16 alpha-glucosiduronate to estrone glucosiduronate throughout the menstrual cycle were analyzed. When the mean ratio during the follicular phase was set at 1, a significant increase (P less than 0.01) occurred in the mean luteal phase ratio in each case: 1.00 +/- 0.03 to 1.66 +/- 0.09, 1.00 +/- 0.04 to 1.30 +/- 0.04, and 1.00 +/- 0.03 to 1.24 +/- 0.04 (mean +/- SE) respectively. The marked alteration in the proportions of these urinary estrogen conjugates may be due to altered metabolism of 17 beta-estradiol, but it more likely reflects a change in the pattern of estrogen secretion or production between the two phases of the menstrual cycle.

Adult↗

PubMedDate not supplied

Production of monoclonal antibodies to estriol and their application in the development of a sensitive nonisotopic immunoassay.

The development of highly specific monoclonal antibodies to estriol and a nonisotopic immunoassay (EIA) for unconjugated estriol based on the use of these monoclonal antibodies have been described. The monoclonal antibodies show little cross reactivity with other steroids and steroid conjugates and can be used directly in immunoassays without any purification. The EIA described here can be performed in 96-well microtiter plates or polystyrene tubes that have been coated with estriol-bovine serum albumin conjugate. In this assay, estriol in the standard or clinical samples (serum or saliva) competes with the immobilized steroid on the plate or the tube for binding with the antibody. The assay shows good agreement with radioimmunoassay (RIA) and is highly sensitive and reliable. Since no prior processing or extraction of the clinical samples is necessary, the method is potentially applicable for routine use in fetal monitoring as well as in a steroid laboratory.

Antibodies, Monoclonal↗

PubMed1995

Review of the endometrial safety during intravaginal treatment with estriol.

To gain more insight into whether intravaginal treatment of local urogenital complaints with the mild-acting oestrogen estriol is capable of inducing proliferation of the endometrium, the results of the clinical studies that have been published over the years have been pooled. Of a total of 19 studies that initially had been selected, four were excluded from the analysis because no baseline biopsies were available, two because endometriae had been evaluated using methods other than with histology, and one study because a sustained-release preparation was used. Pooling of 12 studies (214 subjects) revealed a reasonable amount of long-term data on intravaginal estriol treatment with 61 evaluable biopsies after 6 months and 58 after 12 months. In addition, 13 biopsies were available after 2 years. It appeared that intravaginal estriol treatment using the recommended dosages did not result in endometrial proliferation. All 337 post-baseline biopsies that have been reported in the literature were classified as atrophic. It can be concluded that single daily treatment with intravaginal estriol in the recommended doses in postmenopausal women is safe and without an increased risk of endometrial proliferation or hyperplasia. Consequently, there is no need to add sequential progestogens with these preparations and no withdrawal bleedings will be induced.

Administration, Intravaginal↗

PubMed1988

Measurement of urinary estriol glucuronides during the menstrual cycle by high-performance liquid chromatography.

A simple high-performance liquid chromatographic (HPLC) method for measuring estriol-3-glucuronide (E3-3-G) and estriol-16-glucuronide (E3-16-G) in the urine of non-pregnant women is described. Estriol conjugates were extracted from 4 ml of urine with a small cartridge of graphitized carbon black (Carbopack B). After washing, E3-3-G and E3-16-G were desorbed separately by a two-step elution system. After solvent removal, the two glucuronides were quantified by isocratic ion-suppression HPLC with fluorimetric detection. The analytical recovery of the two estriol metabolites was about 95%. The detection limit of the method was 0.6 ng/ml for both analytes in urine, which is well below the concentrations of clinical interest, and the method is not susceptible to substantial interferences. Data relative to urinary levels of E3-3-G and E3-16-G measured by this method on a daily basis in early morning samples from nine women during their menstrual cycles were compared with those reported in the literature and obtained by radioimmunoassay techniques. Moreover, the potential use of defined changes in the concentrations of the two conjugates for predicting the fertile period of women was assessed.

Adult↗

PubMed1991

Automated direct high-performance liquid chromatographic assay for estetrol, estriol, cortisone and cortisol in serum and amniotic fluid.

An automated direct assay for the simultaneous determination of unconjugated estetrol, estriol, cortisone and cortisol in serum and amniotic fluid, using high-performance liquid chromatography with electrochemical detection and ultraviolet detection, has been developed. The analysis time is ca. 1 h. This system offers good reproducibility with low coefficients of variation (estetrol, 2.3%; estriol, 2.3%; cortisone, 2.6%; cortisol, 1.9%). Detection limits are low enough for routine determinations (estetrol and estriol, 150 pg; cortisone and cortisol, 5 ng). Comparison of the values measured by the present method and by radioimmunoassay revealed significant correlations for estetrol (r = 0.787, p less than 0.01), estriol (r = 0.957, p less than 0.01), cortisone (r = 0.956, p less than 0.01) and cortisol (r = 0.865, p less than 0.01). This system proved to be valuable in monitoring feto-placental function.

Amniotic Fluid↗

PubMed1997

Undetectable maternal serum unconjugated estriol levels in the second trimester: risk of perinatal complications associated with placental sulfatase deficiency.

OBJECTIVE: Our purpose was to determine the prevalence of undetectably low second-trimester maternal serum unconjugated estriol levels and the association with increased perinatal morbidity or mortality in pregnancies at risk for placental sulfatase deficiency. STUDY DESIGN: Nine centers in New England identified singleton pregnancies with undetectably low unconjugated estriol levels. Each unexplained case was matched with four controls; pregnancy outcome information was sought. RESULTS: Among 130,295 pregnancies surveyed, undetectably low unconjugated estriol levels were identified in 167 (13/10,000). Explanations included fetal death (53), overestimated gestational age (50), nonpregnancy (12), and chromosome abnormalities (5). The 41 unexplained cases were compared with 163 matched controls. Male offspring were more frequent (85%) among cases than among controls (55%). Although rates of perinatal complications were not significantly different, primary cesarean sections occurred about twice as often among cases. No perinatal deaths occurred. CONCLUSIONS: Neither severity of symptoms nor perinatal morbidity or mortality currently warrant routine interpretation of unexplained undetectably low unconjugated estriol levels as a marker for placental sulfatase deficiency.

Case-Control Studies↗

PubMed1999

Second-trimester maternal serum marker screening: maternal serum alpha-fetoprotein, beta-human chorionic gonadotropin, estriol, and their various combinations as predictors of pregnancy outcome.

OBJECTIVE: We evaluated the value of all 3 common biochemical serum markers, maternal serum alpha-fetoprotein, beta-human chorionic gonadotropin, and unconjugated estriol, and combinations thereof as predictors of pregnancy outcome. STUDY DESIGN: A total of 60,040 patients underwent maternal serum screening. All patients had maternal serum alpha-fetoprotein measurements; beta-human chorionic gonadotropin was measured in 45,565 patients, and 24,504 patients had determination of all 3 markers, including unconjugated estriol. The incidences of various pregnancy outcomes were evaluated according to the serum marker levels by using clinically applied cutoff points. RESULTS: In confirmation of previous observations, increased maternal serum alpha-fetoprotein levels (>2.5 multiples of the median) were found to be significantly associated with pregnancy-induced hypertension, miscarriage, preterm delivery, intrauterine growth restriction, intrauterine fetal death, oligohydramnios, and abruptio placentae. Increased beta-human chorionic gonadotropin levels (>2.5 multiples of the median [MoM]) were significantly associated with pregnancy-induced hypertension, miscarriage, preterm delivery, and intrauterine fetal death. Finally, decreased unconjugated estriol levels (<0.5 MoM) were found to be significantly associated with pregnancy-induced hypertension, miscarriage, intrauterine growth restriction, and intrauterine fetal death. As with increased second-trimester maternal serum alpha-fetoprotein levels, increased serum beta-human chorionic gonadotropin and low unconjugated estriol levels are significantly associated with adverse pregnancy outcomes. These are most likely attributed to placental dysfunction. CONCLUSION: Multiple-marker screening can be used not only for the detection of fetal anomalies and aneu-ploidy but also for detection of high-risk pregnancies.

Abruptio Placentae↗

PubMed1999

Accuracy of salivary estriol testing compared to traditional risk factor assessment in predicting preterm birth.

OBJECTIVE: The objective was to compare the predictive accuracy (percentage of correct vs incorrect predictions) of salivary estriol levels (SalEst; Biex, Inc, Dublin, Calif) with that of the modified Creasy score for predicting preterm labor followed by preterm delivery. STUDY DESIGN: A triple-blinded prospective trial was conducted at 8 US centers. RESULTS: Among 601 evaluable patients, serial salivary estriol testing correctly predicted the appropriate outcome 91% of the time and the Creasy scoring method correctly predicted the appropriate outcome 75% of the time (McNemar test P <. 001). Among subjects with Creasy scores >/=10 (high-risk group, n = 152), use of salivary estriol testing correctly predicted the end point 87% of the time, compared with only 7.2% correctly predicted by modified Creasy scoring (McNemar test P <.001). CONCLUSION: Salivary estriol assessment was more accurate in predicting outcome than was modified Creasy scoring.

Estriol↗

PubMed1999

Saliva as a fluid for measurement of estriol levels.

OBJECTIVE: The purpose was to establish saliva as a reliable sample for the determination of estriol levels in pregnant women. STUDY DESIGN: Literature and laboratory analyses were used to develop an assembly system to collect and analyze saliva for estriol level. RESULTS: An enzyme-linked immunoassay for estriol has been developed to use whole saliva samples. Analytic performance characteristics for the assay have been determined to be appropriate for clinical determination of preterm risk assessment. CONCLUSION: Saliva can serve as a reliable sample for estriol determination when coupled with an appropriate assay method.

Enzyme-Linked Immunosorbent Assay↗

PubMed1983

Immunoassay of unconjugated estriol in serum of pregnant women monitored by chemiluminescence.

6-Oxoestriol-6-(O-carboxymethyl)oxime-aminobutylethyl- isoluminol conjugate was synthesized. This luminogenic estriol derivative enabled us to develop a solid phase immunoassay method for the determination of unconjugated estriol in serum of pregnant women by the measurement of the bound estriol-isoluminol conjugate upon oxidation with a hydrogen peroxide/microperoxidase system. The sensitivity of the assay was 700 pmol/l. Results obtained by radioimmunoassay and the described method showed good agreement (r = 0.95). The chemiluminescent method is applicable in the routine measurement of unconjugated estriol.

Estriol↗

PubMed1996

Shared reaction in solid-phase immunoassay for estriol determination.

In the search of factors responsible for the experimental difficulties in developing accurate and sensitive solid-phase immunoassay of steroids, an experimental model has been set up for the study of nonspecific interaction of the steroid analyte with the coating protein. Along with the development of a highly sensitive enzyme-linked, solid-phase immunoassay for estriol measurement, we observed evidence of shared reactions. This property, to our knowledge not previously described for monomeric, low-molecular-weight antigens like estrogens, has been attributed to the presence of bovine serum albumin, which is capable of binding estrogens through hydrophobic interactions. The addition of estriol in solution in large excess did not reach a complete inhibition of the binding, so the possibility was excluded that the antibody simply binds to the adsorbed estrogen. The simplest explanation for the occurrence of the reaction is the hypothesis that a family of antigen determinants arises when the estriol is conjugated to a protein carrier. The corresponding antibodies are revealed only when the estrogen participates to the actual analytical system in the form of a steroid-protein conjugate. In the experiment, the estriol has been recognized as being coupled with one or more amino acid side chains present around its site of covalent linkage to the immunogen protein. The discussed results may be of help in developing a solid-phase immunoassay of small antigens as steroids, but also in applying the hybridoma and phage display technologies, the screening methods of which are based on sensitized solid phases.

Antigen-Antibody Reactions↗