Method for determining content of conjugated estrogens in pregnant mare urine

Abstract

The invention relates to the technical field of chemical detection, and provides a method for detecting the content of conjugated estrogens in pregnant mare urine. Carbonate in the pregnant mare urinecan form neutral salt, water and carbon dioxide gas when encountering inorganic acid; the column efficiency is improved, so that the problems that 30 samples cannot be measured each time, the columnpressure is increased, the column efficiency is reduced and the like when the content of conjugated estrogens in pregnant mare urine is measured by HPLC can be solved, the measurement accuracy of thesample content is ensured, and reliable data is provided for production. The pH value is adjusted to 7.0-8.0 by using an alkaline substance because: a, the existence state of bound estrogen is relatively stable under an alkaline condition; b, the pH value of the pregnant mare urine is about 7.5-8.5, and the pH value of the pregnant mare urine is adjusted to be slightly alkaline (7.0-8.0), so thatthe pregnant mare urine is closer to the original conditions of the pregnant mare urine; and c, the authenticity and the reproducibility of a detection result have advantages when the sample is slightly alkaline.

Description

technical field [0001] The invention relates to the technical field of chemical detection, in particular to a method for determining the content of conjugated estrogen in urine of pregnant horses. Background technique [0002] Conjugated estrogens are a natural blend of estrogens extracted from the urine of pregnant horses. Clinically, it is mainly used for hormone replacement therapy to relieve clinical symptoms caused by estrogen deficiency. The average content of estrogen rich in urine per liter of pregnant horse is 70-130mg, and each pregnant horse can form 5-100g of estrogen during pregnancy. [0003] However, pregnant horse urine contains a large number of soluble impurities, such as inorganic salts, urea, sugars, flavonoids and isoflavones, phenols and polyphenols, etc. These components are metabolites of pregnant horses, of which phenols and polyphenols Classes have an impact on the separation degree of HPLC detection of combined estrogen components, and also conta...

AI Technical Summary

Problems solved by technology

[0003] However, pregnant horse urine contains a large number of soluble impurities, such as inorganic salts, urea, sugars, flavonoids and isoflavones, phenols and polyphenols, etc. These components are metabolites of pregnant horses, of which phenols and polyphenols Classes have an impact on the separation degree of HPLC detection of combined estrogen components, and also contain insoluble impurities, such as insoluble salts in urine, specifically such as the precipitates that have occurred and the precipitates produced during storage: calcium carbonate, bicarbonate Calcium, calcium phosphate, calcium hydrogen phosphate and other calcium salts such as carbonate, oxalate, etc., the current treatment methods include the following: 1 centrifugation, after centrifugation, dilution and sample injection, this method can only be used for mechanical Impurities can remove most of the insoluble impurities; 2 Natural sedimentation method, take the sedimented clarified pregnant horse urine supernatant to filter, dilute, and inject samples, this method can only remove large mechanical impurities and some other insoluble impurities; 3 Organic dissolution Sedimentation method, use a certain multiple of methanol, ethanol, and acetone to settle impurities in pregnant horse urine, dilute, and inject samples. This method can remove most insoluble impurities and some salt impurities in urine; 4. Filtration method, direct filtration of pregnant horse urine , diluted injection, this method can only remove most of the insoluble impurities

The test product obtained by the above method is used for HPLC detection, and there are problems of poor resolution and low accuracy

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