CN106248816B - The method for splitting equol enantiomter and measuring its content in bean product - Google Patents
The method for splitting equol enantiomter and measuring its content in bean product Download PDFInfo
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- CN106248816B CN106248816B CN201610542735.6A CN201610542735A CN106248816B CN 106248816 B CN106248816 B CN 106248816B CN 201610542735 A CN201610542735 A CN 201610542735A CN 106248816 B CN106248816 B CN 106248816B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
Abstract
The present invention discloses a kind of method for splitting equol enantiomter and measuring its content in bean product, includes the following steps：(1) it uses methanol to extract and be added interior calibration method to pre-process bean product to be measured；(2) the equol enantiomter in pretreated bean product to be measured is detached and is detected using high performance liquid chromatography tandem mass spectrum method, wherein high-efficient liquid phase chromatogram condition is：Chromatographic column uses polysaccharide derivates reverse phase coating-type chiral chromatographic column；For mobile phase by A phases and B phase compositions, A phases are ammonium acetate solution, and B phases are methanol.The present invention develops a kind of method splitting equol enantiomter using high performance liquid chromatography tandem mass spectrum, by optimizing liquid chromatogram parameter, preferable analysis condition is determined, to simplify extraction process, the influence of matrix effect is avoided by target mode in addition；This method can effectively avoid the appearance of false positive results, can realize the quick measurement of S equols in bean product.
Description
Technical field
The invention belongs to biochemical analysis detection fields, and in particular to a kind of fractionation equol enantiomter and measurement
The method of its content in bean product.
Background technology
Equol (Equol), chemical name are 7- hydroxyls -3- (4- hydroxyphenyls)-coumaran, and chemical structural formula is such as
Under, it is final metabolite one of of the isoflavones by being generated under certain bacterial metabolisms in human body intestinal canal.Recent study
It was found that equol is the major embodiment person of isoflavones physiological function, with estrogen adjust relevant disease such as breast cancer,
Prostate cancer, climacteric syndrome and angiocardiopathy, osteoporosis etc. have preventive and therapeutic action.But
It was found that only isoflavones can be metabolized as female horse by the human individual of 30%-50% under the effect of its intestinal microflora
Phenol, therefore, exogenous supplement equol are just particularly important.Equol is chipal compounds, and there are two types of enantiomters, divide
Not Wei R- equols and S-equol, since it is different from the affine degree of estrogen receptor, the two have different biologies
Characteristic is learned, and what body metabolism generated is S-equol.Therefore, the technology of exploitation chiral resolution equol enantiomer and foundation
The content assaying method of S-equol has a very important significance in food.
Equol chemical structural formula
From equol in food is usually metabolized by isoflavones biofermentation, isoflavones is flavone
Close object, be primarily present in soybean and bean product, be currently known may the food containing equol be fermented type bean product, Anagyris foetida
Rotten is common fermented type bean product with fermented bean curd.Have the study found that in 138 portions of Taiwan Taibei city bean curd with odor 91% sample
Product examine has gone out S-equol, but carries out qualitative detection to equol only in accordance with high performance liquid chromatography (HPLC) method when measurement,
Accuracy needs further to be confirmed.The assay method of other equols has：Enzyme linked immunosorbant assay, gas chromatography,
Liquid chromatography and liquid chromatography-mass spectrometry.Wherein enzyme linked immunosorbant assay, gas chromatography can not all detect individually
Enantiomter, and high performance liquid chromatography is more demanding for sample pre-treatments, complex steps, thereby increases and it is possible to there are false positives
Interference.
Invention content
According to deficiency in the prior art, the present invention is developed using high performance liquid chromatography-tandem mass (HPLC-MS/MS)
A kind of technology splitting equol enantiomter, passes through and optimizes liquid chromatogram and mass spectrometry parameters, it is determined that optimized analysis item
Part simplifies extraction process, and the influence of matrix effect is avoided, it can be achieved that fermented bean curd and bean curd with odor etc. by target mode in addition
The foundation of the quick measurement of S-equol in fermented type bean product, this method can be safe meal supplement equol and promotion beans system
The quality of product provides technical support.
The technical solution adopted by the present invention is as follows：
A method of it splitting equol enantiomter and measures its content in bean product, include the following steps：
(1) it uses methanol to extract and be added interior calibration method to pre-process bean product to be measured；
(2) use high performance liquid chromatography-tandem mass method by the equol mapping in pretreated bean product to be measured
Isomers is detached and is detected, wherein high-efficient liquid phase chromatogram condition is：Chromatographic column uses polysaccharide derivates reverse phase coating-type hand
Property chromatographic column, for mobile phase by A phases and B phase compositions, A phases are ammonium acetate solution, and B phases are methanol.
Preferably, in step (1), the bean product to be measured are fermented type bean product, such as fermented bean curd, bean curd with odor etc..
Preferably, in step (1), from separation and detection result for, it is described in be designated as chloramphenicol.
Preferably, in step (1), according to the characteristic of specific sample to be tested, the method for sample pretreatment of the invention is：
Bean product to be measured, chloramphenicol and methanol are subjected to mixing homogeneous, ultrasonic extraction, centrifuging and taking supernatant.
It is further preferred that the adding proportion of the bean product to be measured and methanol is 10g：(10~15) mL；
It is further preferred that a concentration of 0.35~0.45 μ of the chloramphenicol in bean product to be measured and chloramphenicol mixture
G/g, preferably 0.40 μ g/g.
It is further preferred that the homogenizing time is 1~3min, preferably 2min.
It is further preferred that the ultrasonic extraction time is 1~3min, preferably 2min.
It is further preferred that the centrifugal condition is：Rotating speed is 7000~8000rpm, and centrifugation time is 8~12min, compared with
It is good to be：Rotating speed is 8000rpm, centrifugation time 10min.
Still more preferably, the specific method is as follows for the sample pretreatment：Accurately weigh bean product solid content to be measured
Appropriate chloramphenicol-D5 internal standards are added in test tube in 10.0g, make its a concentration of 0.40 μ g/g, after chromatography methanol 10mL is added,
Matter 2min, ultrasonic extraction 2min, then with 8000rpm speed centrifuge 10min, take supernatant 0.5mL with chromatography methanol dilution extremely
1.0mL is packed into sample injection bottle, to be measured.
The bean product to be measured of the present invention are extracted by methanol, and operating procedure is simple, and the time is shorter；Chloramphenicol internal standard is added to keep away
The influence of matrix effect is exempted from so that separation and testing result are accurate, reliable.
Preferably, in step (2), the polysaccharide derivates reverse phase coating-type chiral chromatographic column uses CHIRALCEL OJ-
RH chiral columns；The preferred model of CHIRALCEL OJ-RH chiral columns：Grain size is 5 μm, column internal diameter 4.6mm, and column length is
150mm。
In step (2), higher sensitivity in order to obtain enhances mass spectrographic Ionization Efficiency, the ammonium acetate solution
In ammonium acetate content be 8~12mmol/L, preferably 10mmol/L.
It in step (2), is eluted using isocratic elution mode, A phases：The volume ratio of B phases is 70~80：30~20, it is excellent
Select A：B=80:20；Preferable flow rate is 0.8~1.2mL/min, more preferably 1.0mL/min；It is preferred that column temperature is 20~40 DEG C, more
Preferably 40 DEG C, at a temperature of this, equol separation is good, and retention time shortens, and peak area is larger；It is preferred that sample size is 5 μ L.
In step (2), the MS detection parameters include mainly dry temperature degree, fragmentation voltage and collision energy etc., these ginsengs
Number can make sample parent ion obtain maximum transmitted efficiency and daughter ion higher response intensity in mass spectrum, improves detection side
The sensitivity of method directly affects detection sensitivity and accuracy.Preferably Mass Spectrometry Conditions are：Ion source：Electric spray ion source
(ESI), it is detected using negative ion mode.Dry gas：High-purity N2, dry temperature degree：300~400 DEG C, dry gas stream speed：8~
12L/min, atomizing pressure：25~35psi, capillary voltage：3500~4500V.Using multiple-reaction monitoring (MRM) pattern.
It is further preferred that dry temperature degree：350 DEG C, dry gas stream speed：10L/min, atomizing pressure：30psi, capillary
Tube voltage：4000V.
By the Scanning Detction to equol standard solution, m/z 241 [M-H] is obtained-For parent ion, and to fragmentation voltage
(Fragmentor), collision energy, dry gas temperature etc. are optimized, and broken voltage when chloramphenicol is quantitative is 120V,
Collision energy is 15V, and broken voltage when chloramphenicol is qualitative is 120V, broken when collision energy 35V, R- equol is quantitative
Voltage is 60V, and broken voltage when collision energy 6V, R- equol is quantitative is 60V, collision energy 9V, and S-equol is fixed
Broken voltage when amount is 60V, collision energy 6V, and broken voltage when S-equol is quantitative is 60V, collision energy 9V,
It is final to determine that mass spectrometry parameters are shown in Table 1.
A technical solution in above-mentioned technical proposal has the advantages that：
(1) present invention develops a kind of fractionation equol pair using high performance liquid chromatography-tandem mass (HPLC-MS/MS)
The method for reflecting isomers, by optimizing liquid chromatogram parameter, it is determined that preferable analysis condition, to simplify extraction process,
The influence of matrix effect is avoided by target mode in addition；This method can effectively avoid the appearance of false positive results, can
Realize the quick measurement of S-equol in bean product, the foundation of this method can be safe meal supplement equol and promotion bean product
Quality provide technical support.
(2) present invention studies chromatographic column, and test result finds that CHIRALCEL OJ-RH chiral columns can be effective
Equol isomers is split, good separating effect and has good chromatographic peak profile, compared with other chiral chromatographic columns such as Nucleodex β-PM
It detaches excellent.
(3) present invention studies mobile phase, higher sensitivity in order to obtain, enhances mass spectrographic ionization effect
The A phases of rate, mobile phase of the invention select ammonium acetate solution, B phases to select methanol, and using the mobile phase, chromatographic peak profile is symmetrical,
High sensitivity can effectively split equol to isomers, and retention time is longer, and separating degree is higher, can meet Accurate Determining list
The requirement of a enantiomer.
(4) through the invention fractionation equol enantiomter and the method for measuring its content in bean product, detection
Sensitivity is higher, and detection limit and quantitative limit are relatively low.
(5) through the invention fractionation equol enantiomter and the method for measuring its content in bean product, have
The good rate of recovery and precision, the rate of recovery and the precision of equol enantiomter are satisfied by《Good Laboratory control rule
Model food Physico-chemical tests》The requirement of (GB/T 27404-2008).
Description of the drawings
Fig. 1 is that equol splits enantiomter typical case's chromatogram.
Fig. 2A~Fig. 2 C are to split equol enantiomter mass spectrometry parameters optimization figure, wherein Fig. 2A：Optimization
Fragmentor voltages；Fig. 2 B：Optimize collision energy；Fig. 2 C：Optimizing drying temperature degree.
Fig. 3：S-equol equation of linear regression and related coefficient.
Fig. 4：R- equols equation of linear regression and related coefficient.
Specific implementation mode
Embodiment 1
1 experimental section
1.1 instruments and reagent
The key instrument used in experiment has：(Agilent is public for Agilent-1200 type quick separatings high performance liquid chromatography
Department), 6410 type triplex tandem level four bars mass spectrums (Agilent companies).Methanol (chromatographically pure, Tedia companies), ammonium acetate (analysis
It is pure, Chinese medicines group) it is chromatographically pure.(purity is more than purchased from Sigma companies for equol racemic compound, S-equol
98%).Chloramphenicol-D5 (internal standard IS) is purchased from AccuStandard companies (purity 99%, the U.S.)；CHIRALCEL OJ-RH hands
Property column (Japanese Daicel companies).
1.2HPLC-MS/MS condition
1.2.1HPLC condition
Chromatographic column：CHIRALCEL OJ-RH chiral columns (150mm × 4.6mm, 5 μm, Japanese Daicel companies).Mobile phase：
A phases are ammonium acetate solution containing 10mmol/L, and B phases are methanol.Isocratic elution：A：B=80:20.Flow velocity：1.0mL/min.Column
Temperature：40 DEG C, sample size：5μL.
1.2.2MS/MS condition
Ion source：Electric spray ion source (ESI), is detected using negative ion mode.Dry gas：High-purity N2, dry temperature degree：
350 DEG C, dry gas stream speed：10L/min, atomizing pressure：30psi, capillary voltage：4000V.Using multiple-reaction monitoring (MRM)
Pattern.By the Scanning Detction to equol standard solution, m/z 241 [M-H] is obtained-For parent ion, and to fragmentation voltage
(Fragmentor), collision energy, dry gas temperature etc. are optimized, final to determine that mass spectrometry parameters are shown in Table 1.
1.3 sample treatment
The fermented types such as fermented bean curd, bean curd with odor bean product solid content 10.0g accurately is weighed in 50mL cleaning centrifuge tubes, is added
Appropriate chloramphenicol-D5 internal standards, make its a concentration of 0.40 μ g/g, after chromatography methanol 10mL is added, homogeneous 2min, and ultrasonic extraction
2min, then 10min is centrifuged with 8000rpm speed, supernatant 0.5mL chromatography methanol dilutions to 1.0mL are taken, sample injection bottle is packed into,
It is to be measured.
2 results and discussion
The optimization of 2.1 chromatographic conditions
In order to reach the best chromatography separating effect of equol enantiomter, the present invention to chromatographic column type, temperature and
Flow visualizing is optimized and investigates.According to existing result of study, Chiral mobile phase additives and chiral chromatogram are utilized
Column can efficiently separate equol enantiomter.But result above is not suitable for Liquid Chromatography-Tandem Mass Spectrometry technology.It is flowing
The cyclodextrin boiling point added in phase is higher, is easy to block mass spectrum spray needle；Needed when using chiral chromatographic column using n-hexane,
Isopropanol equal solvent does mobile phase, causes mass ions efficiency very low, sensitivity is poor.
The present invention has carried out selection to chromatographic column first and has investigated, and test result finds that polysaccharide derivates reverse phase coating-type is chiral
Chromatographic column CHIRALCEL OJ-RH chiral columns can effectively split equol isomers, good separating effect and have good chromatography
Peak shape, compared with Nucleodex β-PM etc., other chiral chromatogram post separations are excellent, therefore following tests selection polysaccharide derivates reverse phase coating
Type chiral chromatographic column CHIRALCEL OJ-RH chromatographic columns.
Secondly, chromatogram column temperature is optimized.20-60 DEG C of column temperature (60 DEG C of chromatographic column highest tolerable temperature) is set,
As a result, it has been found that at 20 DEG C, 30 DEG C, 40 DEG C with the raising of temperature, equol separation is good, and retention time shortens, peak area
It gradually increases, peak area no longer changes at 40-60 DEG C, but separating degree is deteriorated, therefore following experiment uses 40 DEG C of column temperature.
Finally, it is investigated with respect to acetonitrile-water system and methanol-water solution using Flow Injection Chemiluminescence Method.As a result, it has been found that in second
Equol retention time only 2.5min can not split enantiomter in nitrile-aqueous systems；It is used as flowing using methanol-water solution
Xiang Shi, chromatographic peak profile is symmetrical, high sensitivity, can effectively split equol to isomers, retention time is divided in 8.6-11min
Reach 2.0 from degree.Higher sensitivity in order to obtain enhances mass spectrographic Ionization Efficiency, 10mmol/L is added into mobile phase
Ammonium acetate, under conditions of identical mobile phase ratio, S-equol retention time 8.43min, R- equol retention time
9.68min, separating degree reach 2.8, can meet the requirement of Accurate Determining single enantiomer.Typical chromatogram is shown in Fig. 1.
The optimization of 2.2 Mass Spectrometer Method conditions
The MS detection parameters include mainly dry temperature degree, fragmentation voltage and collision energy etc., these parameters can make sample
Product parent ion obtains maximum transmitted efficiency and daughter ion higher response intensity in mass spectrum, improves the sensitivity of detection method
Detection sensitivity and accuracy are directly affected, therefore the above parameter is optimized one by one.Experimental method is as follows：First, exist
Under the pattern for selecting ion detection (SIM), input sample mother ion mass-to-charge ratio m/z 241, optimization fragmentation voltage (0V~240V),
Ensure the maximum transmitted efficiency of parent ion, i.e., more to reach collision cell as far as possible, specific experiment result is shown in Fig. 2A；Then, son from
Under sub- scan pattern, input the mass-to-charge ratio and different collision energies (0V~40V) of parent ion, investigate different collision energies to mother from
The influence of son and daughter ion peak intensity selects collision energy when daughter ion maximum intensity, while determining best daughter ion matter lotus
Than concrete outcome is shown in Fig. 2 B；In dry 150 DEG C -350 DEG C of the gas temperature range of setting, when investigating optimum sensitivity under MRM patterns
Drying temperature degree, concrete outcome is shown in Fig. 2 C.By testing above, the parameters such as best fragmentation voltage, collision energy such as table 1 is determined
It is shown.
The parameter that the Mass Spectrometry Conditions of chloramphenicol-D5 (IS) refer in GB/T 20756-2006 is configured.
1. internal standard equol of table detects mass spectrometry parameters
2 linear relationship of embodiment and detection limit
Take appropriate equol racemic compound standard solution that blank fermented bean curd sample is added with chloramphenicol-D5 standard solution
In, a series of sample of different quality concentration of 5.0,20,100,200,500,1000 μ g/kg of accurate formulation presses " embodiment 1
In 1.3 " under prescriptive procedure handled, after optimization under conditions of respectively sample introduction measure, it is different with equol mapping respectively
The internal scalar quantity ion peak areas ratio of quota ion peak area average value of structure body is ordinate (Y), respectively with equol pair
It is that abscissa (X) carries out linear regression (linear graph is shown in Fig. 3-Fig. 4) to reflect isomer concentration with internal standard concentration ratio, obtains recurrence side
Journey, related coefficient and the range of linearity；With the corresponding a concentration of detection limit of signal-to-noise ratio (S/N) >=3, (S/N) >=10 corresponding concentration
As quantitative limit, the detection limit and quantitative limit of two equol enantiomters are obtained, the results are shown in Table 2.
Linear equation, the range of linearity, related coefficient, quantitative limit and the detection limit of 2. equol enantiomter of table
3 recovery of standard addition of embodiment
Equol racemic standard solution is added in blank fermented bean curd sample, it is 200 μ g/ to make blank mark-on sample concentration
Kg, prescriptive procedure is handled under " 1.3 ", and replication 5 times calculates its rate of recovery and precision, the results showed that, equol
The rate of recovery of enantiomter is satisfied by with precision《Good Laboratory controls specification food Physico-chemical tests》(GB/T 27404-
2008) requirement, the results are shown in Table 3.
The rate of recovery and Precision test result (n of equol enantiomter of the addition of table 3. in blank fermented bean curd sample
=5)
The detection of 4 actual sample of embodiment
Using the method after the optimization in embodiment 1 to 7 kinds of bean product (4 kinds of fermented bean curd, 2 kinds of bean curd with odors, a kind of beans in the market
It is rotten) it is determined, wherein detect S-equol in 2 kinds of bean curd with odors, a kind of fermented bean curd, content is between 0.48-55.4 μ g/kg, institute
Have and R- equols are not detected in sample, it is almost the same with existing result of study.
Conclusion：
The present invention develops the technology that high performance liquid chromatography splits equol enantiomter, establishes high performance liquid chromatography
The method of equol in series connection quadrupole rod mass spectrum internal mark method determination bean product.This method is easy to operate, and detection sensitivity is high, and ties
Fruit is reliable, it can be achieved that the complete fractionation of two kinds of enantiomters of equol and quickly being detected, the foundation of this method in bean product
Skill can be provided for the quality of safe meal supplement equol (the especially S-equol with health-care efficacy) and promotion bean product
Art is supported.
Claims (10)
1. a kind of method measuring equol enantiomter content in fermented bean products, characterized in that include the following steps：
(1) use methanol extract and be added in calibration method fermented bean products to be measured are pre-processed, it is described in be designated as chlorine mould
Element；
(2) use high performance liquid chromatography-tandem mass method by the equol mapping in pretreated fermented bean products to be measured
Isomers is detached and is detected, wherein high-efficient liquid phase chromatogram condition is：Chromatographic column uses polysaccharide derivates reverse phase coating-type hand
Property chromatographic column, the polysaccharide derivates reverse phase coating-type chiral chromatographic column use CHIRALCEL OJ-RH chiral columns；Mobile phase by
A phases and B phase compositions, A phases are ammonium acetate solution, and B phases are methanol, A phases：Phase=80 B:20.
2. the method as described in claim 1, characterized in that in step (1), the pretreated method is：By fermentation to be measured
Bean product, internal standard and methanol carry out mixing homogeneous, ultrasonic extraction, centrifuging and taking supernatant.
3. method as claimed in claim 2, it is characterized in that：In step (1), the addition of the fermented bean products to be measured and methanol
Ratio is 10g：(10~15) mL；The homogenizing time is 1~3min；The ultrasonic extraction time is 1~3min；The centrifugation
Condition is：Rotating speed is 7000~8000rpm, and centrifugation time is 8~12min.
4. the method as described in claim 1, it is characterised in that the CHIRALCEL OJ-RH chiral column models：Grain size is 5
μm, column internal diameter 4.6mm, column length 150mm.
5. the method as described in claim 1, characterized in that in step (2), ammonium acetate in the ammonium acetate solution contains
Amount is 8~12mmol/L.
6. the method as described in claim 1, characterized in that in step (2), high-efficient liquid phase chromatogram condition：Flow velocity be 0.8~
1.2mL/min；Column temperature is 20~60 DEG C.
7. method as claimed in claim 6, which is characterized in that the high-efficient liquid phase chromatogram condition：Flow velocity is 1.0mL/min,
Column temperature is 40 DEG C.
8. the method as described in claim 1, characterized in that in step (2), Mass Spectrometry Conditions are：Ion source：Electric spray ion source
ESI is detected using negative ion mode；Dry gas：High-purity N2, dry temperature degree：300~400 DEG C, dry gas stream speed：8~12L/
Min, atomizing pressure：25~35psi；Capillary voltage：3500~4500V；Using multiple-reaction monitoring pattern.
9. method as claimed in claim 8, characterized in that the dry temperature degree：350 DEG C, dry gas stream speed：10L/min,
Atomizing pressure：30psi, capillary voltage：4000V.
10. method as claimed in claim 9, characterized in that with m/z 241 [M-H]-for parent ion, when chloramphenicol is quantitative
Broken voltage is 120V, collision energy 15V, and broken voltage when chloramphenicol is qualitative is 120V, and collision energy 35V, R- is female
Broken voltage when horse phenol is quantitative is 60V, and broken voltage when collision energy 6V, R- equol is qualitative is 60V, collision energy
For 9V, broken voltage when S-equol is quantitative is 60V, collision energy 6V, and broken voltage when S-equol is qualitative is
60V, collision energy 9V.
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Citations (3)
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|---|---|---|---|---|
| CN101338294A (en) * | 2008-08-07 | 2009-01-07 | 河北农业大学 | Acinetobacter spp AUH-JLM455 and process preparing S-equol by conversion thereof |
| EP2526940A2 (en) * | 2002-07-24 | 2012-11-28 | Children's Hospital Medical Center | Compositions containing enantiomeric equol, and methods for their making |
| CN102925378A (en) * | 2012-05-11 | 2013-02-13 | 华侨大学 | Proteus mirabilis strain and method for producing S-equol through daidzein conversion by using the same |
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2016
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2526940A2 (en) * | 2002-07-24 | 2012-11-28 | Children's Hospital Medical Center | Compositions containing enantiomeric equol, and methods for their making |
| CN101338294A (en) * | 2008-08-07 | 2009-01-07 | 河北农业大学 | Acinetobacter spp AUH-JLM455 and process preparing S-equol by conversion thereof |
| CN102925378A (en) * | 2012-05-11 | 2013-02-13 | 华侨大学 | Proteus mirabilis strain and method for producing S-equol through daidzein conversion by using the same |
Non-Patent Citations (3)
| Title |
|---|
| Development of chiral liquid chromatography–tandem mass spectrometry isotope dilution methods for the determination of unconjugated and total S-equol in human plasma and urine;Jeffry B. Plomley等;《Journal of Pharmaceutical and Biomedical Analysis》;20101230;第55卷;第125-134页 * |
| Equol, a natural estrogenic metabolite from soy isoflavones:convenient preparation and resolution of R- and S-equols and their differing binding and biological activity through estrogen receptors alpha and beta;Rajeev S. Muthyala等;《Bioorganic & Medicinal Chemistry》;20041231;第12卷;第1561-1566页第2,5节 * |
| 手性固定相高效液相色谱法拆分雌马酚对映体;黄雅燕等;《药物分析杂志》;20131231;第33卷(第6期);第973-976页 * |
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