Dienestrol Diacetate Research Articles

Simultaneous determination of nine estrogens in bullfrogs using filtered solid phase extraction and ultra-performance liquid chromatography-tandem mass spectrometry

建立了基于通过型固相萃取小柱净化的超高效液相色谱-三重四极杆质谱联用(UPLC-MS/MS)同时快速准确测定牛蛙中9种雌激素(雌三醇(E3)、17β-雌二醇(β-E)、17α-雌二醇(α-E)、17α-炔二雌醇(EE2)、雌酮(EI)、己烯雌酚(DES)、己二烯雌酚(DE)、己烷雌酚(HEX)、醋酸双烯雌酚(DD))残留的检测方法。样品经乙腈提取,经PRiME HLB固相萃取柱净化,Waters Acquity UPLC BEH C18柱(100 mm×2.1 mm, 1.7 μm)分离,以0.5 mmol/L氟化铵水溶液-乙腈体系为流动相梯度洗脱,流速为0.3 mL/min,采用电喷雾正负离子切换模式(ESI+/ESI-)和多反应监测(MRM)扫描方式检测,基质匹配外标法定量分析。该研究优化了液相色谱条件,相比于乙酸铵水溶液-乙腈体系和氨水溶液-乙腈体系,0.5 mmol/L氟化铵水溶液-乙腈体系作为流动相时9种雌激素普遍具有更佳的灵敏度。相比于甲醇和乙酸乙酯,乙腈作为提取溶剂时9种雌激素的提取率提高15%~40%。考察了HLB、C18、Silica、PRiME HLB共4种不同类型的固相萃取小柱的基质净化效应,结果表明,PRiME HLB柱具有更好的基质净化能力。经PRiME HLB净化后,所有化合物的回收率均在70%~125%之间。DD的回收率从47%提高到74%, DES的回收率从180%降低到123%,有效减弱了基质效应。在最佳的实验条件下,E3、β-E、α-E、EI、DE、HEX、DD的线性范围为0.5~100.0 μg/L, EE2和DES的线性范围为1.0~100.0 μg/L, 9种雌激素在各自的线性范围内均有良好的线性关系,相关系数为0.9953~0.9994,方法检出限为0.17~0.33 μg/kg,方法定量限为0.5~1.0 μg/kg,在2.0、10.0、80.0 μg/kg 3个加标水平下,9种雌激素的加标回收率为65.1%~128.2%,相对标准偏差为1.9%~17.6%。该方法操作简便、快速、灵敏,重复性好,可用于大批量样品的同时快速准确检测。

Tryptophan Reduces Hepatic Lipid Accumulation Induced Experimentally by Refeeding after Starvation or Estrogen Administration in Broiler Chicks.

Two experiments were conducted to study the effects of dietary L-tryptophan supplementation on hepatic lipid accumulation in broiler chicks. Male broiler chicks were treated with 3 days refeeding after starvation or administrations of dienestrol diacetate (0.01% in diet) for 12 days. Low tryptophan diet (0.12%) was formulated and supplemented to provide the specified concentrations of tryptophan (0.12-0.46%). Supplementation of L-tryptophan beyond the requirement for growth reduced the liver weight and liver lipid content expressed as per unit feed intake in the starved-refed chicks. Either higher or lower dietary tryptophan concentration compared with the requirement decreased the liver weight and liver lipid content in the estrogen-administered chicks. These results demonstrate tryptophan supplementation beyond the requirement reduces the hepatic lipid accumulation induced experimentally in broiler chicks.

Research Note: Performance of Leghorn Type Hens Fed Two Levels of Energy and a Synthetic Estrogen During the Growing Period

Feeding of an estrogen, dienestrol diacetate, at 352 mg/kg of diet to December-hatched White Leghorn type pullets, from 16 to 20 wk of age, caused onset of production to be delayed approximately 3 wk. The dietary dienestrol diacetate also resulted in increased body weights at 30 and 46 wk in one experiment. Hens receiving the estrogen laid significantly (P<.05) heavier but fewer eggs during most of the production year than did those fed diets without the estrogenic compound. The addition of 2.2% fat to diets of pullets from 0 to 20 wk of age failed to influence their performance in the layer house.

Reduction in hepatic lipid and plasma estradiol in estrogenized chicks injected with ascorbic acid.

Injecting White Leghorn chicks every other day with 20 mg ascorbic acid significantly reduced the increase in liver weight and lipids caused by feeding a diet with 0.1% dienestrol diacetate. In chicks fed two different basal diets containing 0.1% dienestrol diacetate, injecting chicks every other day with 20 mg alpha-tocopherol did not significantly reduce liver weight or lipids while the ascorbic acid injections did. Injecting meat-type chicks implanted with estradiol with 10 mg ascorbic acid daily significantly reduced liver weight, liver lipids, and plasma estradiol, but injecting with 8 mg alpha-tocopherol daily had no significant effect.

The influence of environmental temperature, thyroid status and a synthetic oestrogen on the induction of fatty livers in chicks.

1. Hepatic lipid content, lipogenic enzyme activity and plasma lipid concentration were measured in chicks reared at 21 degrees or 34 degrees C and after thyroxine (T4), thiouracil (TU), propylthiouracil (PTU), dienestrol diacetate (DD) or PTU with DD had been given for 14 d. 2. At 34 degrees C there was a significant increase in the total liver lipid and triglyceride content. 3. Injections of T4 decreased liver lipid content whereas it was increased by feeding PTU or DD. The effects of PTU were more pronounced at 21 degrees C while those of DD were more pronounced at 34 degrees C. 4. There were significant interactions between temperature, thyroid status and synthetic oestrogen treatments on total lipid and triglyceride content of the liver. Fatty liver with marked steatosis could be produced through synergic actions of PTU and DD in chicks maintained at 21 degrees C.

Phase Shifting of the Circadian Clock by Diethylstilbestrol and Related Compounds in Neurospora crassa

Phase shifts of the circadian conidiation rhythm in Neurospora crassa were induced by 3-hour treatments of mycelia in liquid medium with diethylstilbestrol (DES), dienestrol (DIE), hexestrol (HEX), diethylstilbestroldipropionate (DESP), and dienestroldiacetate (DIEA). Over a 24-hour period beginning 24 hours after the transition from light to constant dark, maximum phase shifts occurred about 36 hours. DES was the most effective of the drugs tested, giving 10-hour phase advances at 20 micromolar. DIE and HEX caused similar phase shifts as DES at 40 micromolar. The two derivatives of the last, DESP and DIEA, were much less effective in shifting phase; only a few hours of phase advance result from treatments at 80 micromolar concentrations.The activity of isolated plasma membrane ATPase was inhibited by DES and partially by HEX, but not by DIE, DESP, or DIEA. O(2) consumption of the mycelia was inhibited equally by DES, DIE, and HEX, while DIEA and DESP had little effect. Phase-shifts by DES cannot be interpreted as evidence that plasma membrane ATPase is a component of the circadian clock.

Gas-Liquid Chromatographic Determination of Diethylstilbestrol in Molasses-Based Liquid Feed Supplements

A gas-liquid chromatographic (GLC) method is described for the determination of diethylstilbestrol (DES) in molasses-based liquid feed supplements, using dienestrol diacetate as the internal standard. A sample equivalent to 110 mug DES was diluted, acidified with dilute sulfuric acid, extracted with chloroform, and subjected to basic sodium acetate cleanup. The bis-(trimethylsilyl) acetamide derivative was prepared and determined by GLC. Interfering peaks and/or low recoveries were found to be related to emulsions and the procedure incorporates centrifugation to minimize these.

The Effect of Methionine, Thiouracil, Dienestrol Diacetate and Thyroprotein on the Development and Prevention of Fatty Liver in Pullets

The effect of two levels each of methionine (0.0 and 0.07 percent), thiouracil (0.0 and 0.05 percent), dienestrol diacetate (0.0 and 0.007 percent), and thyroactive casein (0.0 and 0.0125 percent) on the performance, organ changes, and liver composition in 640 pullets of two strains was studied in a 24 factorial arrangement of treatments. Egg production, egg characteristics, feed conversion, organ weights, and liver composition were parameters measured.Supplemental methionine increased the phosphorus content of liver fat in strain A, but other parameters in the two strains were not affected by the increase in dietary methionine.The thiouracil increased weight gains, grams of fat per total liver, percent of liver fat, thyroid weight, and heart weight but decreased the phosphorus content of liver fat. Nine typical cases of fatty liver syndrome with large liver hematomas occurred in the thiouracil treated birds and one case occurred in an untreated pullet.Dienestrol diacetate did not affect egg production, egg characteristics, organ weights, and liver composition in the two strains.Thyroprotein decreased weight gain, abdominal fat, liver weight, liver fat, thyroid weight, and percent red cells, but decreased percent blood spots in eggs and adjusted weights of the kidney and heart in both strains.

Effects of Sex, Strain and Estrogens on Quality of Chicken Roasters

This study was conducted to determine the effect of sex, strain and hormone treatment on cooked yields and meat quality of chicken roasters. The two strains were the Hubbard White Mountain and the Penobscot. The three treatments were the injection of estradiol-17 beta monopalmitate, the feeding of dienestrol diacetate, and the control birds which did not receive any hormones.Cooking yields were significantly higher for male roasters than for female roasters. Taste panel scores for breast meat showed a significant difference in flavor and odor between the meat from the two sexes, that of the male chickens obtained higher scores. Panel scores for the thigh samples were higher for flavor of Penobscot birds than Hubbard White Mountain. Texture of thigh meat was preferred to that of breast meat. No other differences were detected in flavor, odor and texture.Shear values obtained from a Kramer Shear press showed highly significant differences between the two types of meat. The samples from the Musculus iliotibialis were more tough than the samples from the Pectoralis major and the samples from the Pectoralis minor. Compression slope values of the Pectoralis minor were significantly higher for the Penobscot strain as compared to Hubbard White Mountain strain. Sex was the only factor that had any effect on shear values of thigh muscle. Male roasters were more tender than the female roasters. The hormone treatments had no effect on any of the measurements obtained from either the Kramer Shear press or taste panel scores.

Evaluation of Dienestrol Diacetate (Lipamone ) Supplementation of Broiler Finisher Feeds on Pigmentation, Growth Characteristics and Market Quality

SINCE as early as 1939 it has been known that estrogenic compounds can cause increased fat content of muscle tissue. Lorenz (1945) showed that the subcutaneous implantation of diethystilbestrol (DES) in three-week old cockerels resulted in a fattening effect. Similar beneficial effects from the use of DES have been reported by other workers, however; as of December, 1959, this product has not been approved for use in poultry. Dienestrol diacetate, an estrogenic compound available for use in poultry, has been investigated by a number of workers, but with contradictory results. Boone et al. (1961) found that the addition of dienestrol diacetate to the feed the last three weeks of the growing period increased body weights and abdominal fat deposits of nine-week old broilers, but that feed efficiency, fleshing and finish were unaffected. Begin and Grainger (1957) found no differences in weight gains of broilers fed dienestrol diacetate from five to…

Influence of Protamone and Dienestrol Diacetate on Bone Fragility of Caged Layers

THE effect of various thyroproteins on laying hens has been studied fairly extensively since 1923 when Crew and Huxley first attempted to demonstrate that thyroid feeding had an effect on egg production. Turner et al. (1945a, b) found that protamone, an iodinated casein with high thyroidal activity, increased egg production when added to the diet. These workers concluded that an optimum level of protamone in the feed ranged between five and ten grams per hundred pounds of feed. Additional work by Turner et al. (1946) demonstrated that protamone fed continuously at ten grams per 100 pounds of feed prevented a decrease in egg production during the summer months. Work by Singh and Shaffner (1950) indicated that thyroprotein elevated the basal metabolic rate. However, Lillie et al. (1952) questioned whether it improved egg production although the hen’s basal metabolism was increased.The use of estrogens to prevent further degeneration and relieve…

Gas chromatographic separation of isomeric dienestrol diacetates and analyses of industrial products

The three geometrical isomers of dinestrol diacetate may be determined quantitatively and simultaneously by gas-liquid chromatography using 3% SE- 30 on Gas-Chrom Z as stationary phase; they can also be determined in the presence of indenestrol diacetate and pinacolone 4,4′-bis( p-hydroxyphenyl)-3-hexanone diacetate. The gas-liquid chromatographic method may be used in analyses of industrial residues and in purity determination of pharmaceutical grade dienestrol and dienestrol diacetate. An auxiliary thin-layer chromatographic procedure is also described.

Gas Chromatographic Method for Determination of Diethylstilbestrol in Feeds

Abstract A GLC method was developed for the determination of diethylstilbestrol in feeds within the 0.0011–0.0022% range, using dienestrol diacetate as an internal standard. A 10 g sample was extracted with 7% ethanol in chloroform and subjected to a modified alkaline cleanup. The bis-(trimethylsilyl) acetamide derivative was then prepared and determined by GLC. No interference was encountered with commercial feed samples.

Influence of Various Dietary Factors on Bone Fragility of Caged Layers ,

BONE fragility of caged layers often results in bone breakage during processing, causing down-grading of ready-to-cook hens. Many processors with batch-type pickers discriminate against or even refuse to buy caged layers. As the number of cage operations increases, bone fragility will become a more important economic factor to the poultry industry.As the hen lays, she depletes her skeletal system of calcium. Common (1938) showed that up to 24% of the total body calcium of hens laying on a low-calcium diet can be withdrawn from the skeleton for shell formation. Taylor and Moore (1954) reported that there was a reduction in percent shell immediately after pullets were placed on a low-calcium diet; the reduction became progressively greater with each egg laid. They also found that the percent bone ash of pullets dropped when they were placed on a low-calcium diet.Jones et al. (1965) reported that administration of dienestrol diacetate .

Influence of Cholesterol on Estrogen Induced Aortic Ruptures in Turkeys.

SummaryA high incidence of aortic ruptures occurred when turkeys were fed diets containing diethylstilbestrol, dienestrol diace-tate, and dienestrol diacetate-cholesterol. A low incidence of mortality occurred when turkeys were fed a diet fortified with both cholesterol and diethylstilbestrol, and there was a marked lowering of total plasma lipid of such treated poults. No mortality occurred in cholesterol fed and control birds. Coronary atherosclerosis was severe in diethyl-stilbestrol, dienestrol diacetate, and dienestrol diacetate-cholesterol fed turkeys, moderately severe in diethylstilbestrol-cholesterol fed turkeys, and mild in cholesterol fed turkeys.

Determination of Dienestrol Diacetate in Mixed Feeds

Abstract Dienestrol diacetate in broiler feeds is isolated by extraction followed by alkaline hydrolysis to yield the parent phenolic compound. Interferences are removed by partition between NaOH and chloroform, in a manner similar to the AOAC procedure for diethylstilbestrol. The dienestrol is acidified, extracted with chloroform, and measured spectrophotometrically at 610 mμ after reaction with anisaldehyde. Recovery of dienestrol diacetate from 4 feeds known to contain 23–92 ppm ranged from 93 to 107% with coefficients of variation of 7.0–16.4%. Values for blank feeds were low enough to exclude a feed blank from the analytical procedure. Amounts of colorimetric reagents, reaction time, and linearity of response were studied. Of 24 common medicaments and 6 solvents tested, only acetone interferes.

Aortic ruptures in turkeys induced by diethylstilbestrol and dienestrol diacetate.

SummaryHigh levels of dienestrol dia-cetate in the feed of turkeys (512, 526, 1024 mg/lb) caused aortic ruptures and decreased weight gains, while a low level (256 mg/lb of feed) caused no mortality and increased weight gains. Comparable mortality, hyper-lipemia, hypercholesterolemia, and hypotension resulted from the feeding of 16 gm diethylstilbestrol/100 lb and 512 or 1024 mg dienestrol diacetate per pound of feed.

The Effect of a Single Subcutaneous Injection of Estradiol-17-Monopalmitate on Growth in Turkeys

IT HAS been well established that synthetic estrogenic compounds such as diethylstilbestrol and dienestrol diacetate when administered to chickens or turkeys will result in increased weight gains and fat deposition (Lorenz, 1944, 1954; and Moreng et al. 1963). Resulting improvement in fat deposition has been verified by many workers and residue studies extensively investigated; excellent reviews of the residue problem have been presented by Umberger et al. (1959) and Umberger and Goss (1959).The present study was carried out to evaluate various levels of estradiol-17-monopalmitate (EMP) in polyethylene glycolic paste with respect to improving gain and carcass qualities of young torn turkeys. In addition, tissue residues were determined. PROCEDUREA preliminary investigation (Trial 1) was designed to provide a reference point of EMP to the action of diethylstilbestrol (DES) in turkeys. Four treatment groups of a commercial line of Broad White turkeys consisting of approximately 21 males each, were individually …

Effects of Dosage Level and Length of Feeding Dienestrol Diacetate on Potentiation of Oxytetracycline

VARIOUS methods of antibiotic potentiation have been reported including lowering of the dietary calcium level (Price and Zolli, 1959), addition of terephthalic acid (TPA) to the diet (Peterson 1958) or injecting chicks with estradiol (Harms et al., 1963).More recently Harms et al. (1964) reported that feeding 35 mg. per kg. of diet of the estrogenic hormone, dienestrol diacetate (DD), was not effective in improving antibiotic utilization; however, a significant increase was obtained when 356 mg. were fed.The study reported herein was conducted to determine the effect of feeding various levels of DD for different periods of time upon oxytetracycline blood serum levels. EXPERIMENTAL PROCEDURETwo successive experiments were conducted as a 3×6 factorial arrangement of treatments with levels of 0, 35, 70, 140, 280 and 560 mg. of DD per kg. of diet.Each treatment was given for 5, 10 and 20 day periods. The hormone was furnished…

THE ORAL PROGESTATIONAL AND ANTI‐OVULATORY PROPERTIES OF MEGESTROL ACETATE AND ITS THERAPEUTIC USE IN GYNAECOLOGICAL DISORDERS

Megestrol acetate alone (47 cases), combined (115 cases) with mestranol, or sequential with dienestrol diacetate, was prescribed for various gynecologic disorders or for studies of its progestational effects. Doses were usually 1 Planovin tablet (5 mg megestrol acetate and .1 mg mestranol) for 20 day cycles, or from 2.5-15 mg megestrol acetate with or without 2.5-5 mg dienestrol diacetate. Results of treatment were 4 of 10 permanent cures of secondary amenorrhea; 10 permanent cures of dysmenorrhea with temporary improvement in 6; 6 of 7 patients with endometriosis were relieved, although 2 had to maintain cyclic treatment after up to 9 months continuous ingestion; 20 of 21 cases of metrorrhagia were relieved; 6 of 7 women with menopausal symptoms were improved; 18 of 21 women used Planovin successfully for contraception. Studies with 21 women who took megestrol acetate before undergoing planned sterilization showed that neither 5 mg megestrol acetate or .1 mg mestranol alone inhibited ovulation in all women, as observed during laparotomy, but the combined tables was 100% effective.