CN103558325A - Method for directly determining enantiomer excess of chiral compound in mixed sample - Google Patents
Method for directly determining enantiomer excess of chiral compound in mixed sample Download PDFInfo
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Abstract
本发明公开了一种用于混合样品中手性化合物对映体过量值直接测定的方法，首先利用第一维色谱单元将待测对映体过量值的手性化合物与其他杂质分离并收集至定量环中，再通过两个十通阀的切换，将收集在定量环中的手性对映体转移至配备有手性色谱柱的第二维色谱单元中，利用第二维色谱单元测定得到定量环中的手性化合物的对映体过量值。本发明能够实现混合样品中手性化合物对映体过量值的直接测定，而不需要经过前期复杂的样品纯化过程，从而大大降低了生产成本，缩短了分析时间，同时提高了色谱系统分离能力，应用范围广，尤其适合于不对称合成粗产物中手性药物对映体过量值的测定。
The invention discloses a method for directly measuring the enantiomeric excess value of a chiral compound in a mixed sample. Firstly, the chiral compound with the enantiomeric excess value to be measured is separated from other impurities by using a first-dimensional chromatographic unit and collected in the In the quantitative loop, through the switching of two ten-way valves, the chiral enantiomer collected in the quantitative loop is transferred to the second-dimensional chromatographic unit equipped with a chiral chromatographic column, and is determined by the second-dimensional chromatographic unit. The enantiomeric excess value of the chiral compound in the quantitative loop. The invention can realize the direct determination of the enantiomeric excess value of the chiral compound in the mixed sample without going through the complex sample purification process in the early stage, thereby greatly reducing the production cost, shortening the analysis time, and improving the separation ability of the chromatographic system at the same time. It has a wide range of applications, and is especially suitable for the determination of the enantiomeric excess value of chiral drugs in crude products of asymmetric synthesis.
Description
Technical field
The present invention relates to the analysis of high performance liquid chromatography and chipal compounds, specifically a kind of two-dimensional highly effective liquid phase chromatographic method that realizes the direct mensuration of enantiomers of chiral drugs excessive value in biased sample.
Background technology
In human body, as the biomacromolecule of vital movement important foundation, as protein, polysaccharide, nucleic acid and enzyme etc., be almost chirality entirely, and chiral drug to be brought into play drug effect in human body, just must with biomacromolecule effect.And containing the enantiomorph of the chemicals of chirality factor there is significant difference in pharmacologically active, metabolic process and the toxicity in human body: thus often mapping physical efficiency well with the pharmacological action of biochirality macromolecule interaction performance expection, another enantiomorph often can not interact well, not only itself does not have drug effect, also can partial offset drug effect, even also can produce poisonous metabolic product or cause serious spinoff.Drug evaluation and the research and development centre (CDER) of U.S. food in 1992 and medicine Supervision Bureau (FDA) have promulgated chiral drug governing principle, require all racemic new drugs in U.S.'s listing, its producer all needs to illustrate enantiomorph contained in medicine pharmacological action, toxicity and clinical effectiveness separately, and when two kinds of isomeride have obvious drug effect and toxicological effect difference, must be with the medicine form listing of individual isomer.The states such as EU member country and Japan, Canada have also worked out similar rules subsequently.
Therefore, the medicine of single configuration is very important for medicine research and development, thereby has greatly promoted the development of asymmetric syntheses.And in asymmetric syntheses, of paramount importance is exactly the mensuration of the enantiomeric excess value of target chiral drug in asymmetric syntheses product, therefore in the urgent need to finding the method for high flux Fast Measurement enantiomeric excess value, to shorten the screening time of asymmetric catalyst and the R&D cycle of asymmetric medicine in asymmetric syntheses, promote the development of asymmetric syntheses.Existing assay method is by after the pre-service of COMPLEX MIXED sample solution, carry out again one dimension efficient liquid phase chromatographic analysis, step generally comprises: first by traditional column chromatography chromatogram, carry out separation and purification, remove the impurity such as reactant, accessory substance, intermediate product, solvent, catalyzer, then the testing sample after purifying is carried out to the mensuration of the conventional enantiomeric excess values such as high performance liquid chromatography, gas chromatography.This classic method complex steps of measuring enantiomeric excess value is consuming time, and the quantity of solvent expending large, have greater environmental impacts.
The one dimension high performance liquid chromatography that contrast conventional determining adopts, online two-dimentional chromatographic technique has the advantages such as efficient, accurate, quick, at the aspects such as analyzing and testing of medicinal herb components analysis, complex sample, is widely used.Due to the mensuration that can realize from mixed solution to stratographic analysis single step enantiomeric excess value, online two-dimentional method for chromatographic determination has higher accuracy simultaneously.Therefore utilize online two-dimentional chromatographic technique can greatly improve the efficiency of asymmetric syntheses for the mensuration of biased sample enantiomers of chiral drugs excessive value, shorten the R&D cycle of chirality new drug.
Summary of the invention
Technical matters to be solved by this invention be to provide a kind of simple to operate, high flux Fast Measurement for the direct method for measuring of biased sample chiral compound enantiomer excessive value.
In order to solve the problems of the technologies described above, provided by the invention for the direct method for measuring of biased sample chiral compound enantiomer excessive value, comprise the first dimension chromatogram unit, the second dimension chromatogram unit and two ten-way valves, the first described dimension chromatogram unit comprises the first dimension infusion pump, the first dimension chromatographic column, automatic sampler and DAD detecting device; The second described dimension chromatogram unit comprises the second dimension infusion pump, the second dimension chromatographic column; Quantitatively ring and two ten-way valves are tieed up the translation interface of liquid phase as the first dimension liquid phase and second, wherein: entrance point and the endpiece of the first dimension chromatographic column are connected with No. 1 interface of automatic sampler and the first ten-way valve respectively, automatic sampler is connected with the first dimension infusion pump; Entrance point and the endpiece of the second dimension chromatographic column are connected with No. 5 interfaces and No. 9 interfaces of the first ten-way valve respectively; Quantitatively two end interfaces of ring respectively with being connected with No. 6 interfaces for No. 9 of the second ten-way valve; DAD detecting device is connected with No. 7 interfaces of the second ten-way valve; The second dimension infusion pump is connected with No. 4 interfaces of the first ten-way valve; No. 2 interfaces of the first ten-way valve are connected with waste liquid cylinder; No. 10 interfaces of the first ten-way valve are connected with No. 8 interfaces of the second ten-way valve; No. 3 interfaces of the first ten-way valve are connected with No. 10 interfaces of the second ten-way valve; No. 6 interfaces of the first ten-way valve are connected with No. 5 interfaces of the second ten-way valve; The first described ten-way valve and other interfaces of the second ten-way valve are without connection; On-line determination step is as follows:
(1) the first dimension chromatographic column and the second dimension chromatographic column are installed on high performance liquid chromatograph;
(2) mixed solution of need being measured to enantiomeric excess value is added in chromatogram sample injection bottle;
(3) by automatic sampler sample introduction, now the first ten-way valve position is that 10-1 is that No. 10 interfaces are communicated with No. 1 interface, and the second ten-way valve position is that 1-2 is that No. 1 interface is communicated with No. 2 interfaces;
(4) treat the impurity process DAD detecting device that polarity is less, and chirality enantiomorph to be detected is about to when the first dimension chromatographic column elutes, the first ten-way valve position remains 10-1, and the second ten-way valve position switches to 10-1, and now chirality enantiomorph to be detected is collected in quantitative ring;
(5) when chirality enantiomorph to be detected completely elutes and enters after quantitative ring from the first dimension chromatographic column, the first ten-way valve position remains 10-1, the second ten-way valve position switches to 1-2, continues the first dimension separated, and the impurity that polarity is larger elutes from the first dimension chromatographic column;
(6) after the larger impurity of the first dimension chromatographic column Semi-polarity is complete by wash-out, the first ten-way valve position switches to 1-2, and the second ten-way valve position remains 1-2, to carry out the separation of the second dimension chromatogram; In the separation of the second dimension chromatogram, (R) of chipal compounds to be detected, (S) configuration is separated in the second dimension chromatographic column, and successively through DAD detecting device, acquisition chromatogram, by (R) in computer chromatography figure, (S) peak area of configuration obtains enantiomeric excess value.
The first described dimension chromatographic column is positive nh 2 column.
The second described dimension chromatographic column is positive chiral column.
High performance liquid chromatograph used is Dionex Ultimate3000 system.
In above-mentioned steps (2), need are measured to the mixed solution of enantiomeric excess value and crossed after film is processed and be directly added in chromatogram sample injection bottle through liquid phase, the filter opening of film is 0.45 μ m.
Adopt technique scheme for the direct method for measuring of biased sample chiral compound enantiomer excessive value, the online two-dimensional highly effective liquid phase chromatographic of choice for use replaces traditional one dimension high performance liquid chromatography, and this online two-dimensional highly effective liquid phase chromatographic system has comprised the first dimension chromatogram unit, the second dimension chromatogram unit, quantitatively encircled and two ten-way valves.This first dimension high performance liquid chromatography is equivalent to the chromatographic column separation and purification process of chiral drug in traditional enantiomeric excess values determination method, utilize without the nh 2 column of Chiral Separation Ability the impurity such as reactant, accessory substance, catalyzer, solvent is separated with target chiral drug, and the collection (being collected in quantitative ring) of the switching realize target chiral drug by two ten-way valves.This two-dimensional highly effective liquid phase chromatographic is equivalent in traditional assay method and utilizes one dimension chiral chromatogram to measure the step of enantiomeric excess value, adopt suitable chiral column to carry out chiral separation analysis to target chiral drug, by (R) on chromatogram, the chromatographic peak that (S) configuration is corresponding calculates enantiomeric excess value.
The present invention can directly carry out efficient liquid phase chromatographic analysis to including the mixed solution of enantiomers of chiral drugs, without measuring necessary sample pre-separation in enantiomers of chiral drugs excessive value conventional method, purification step, can directly obtain the enantiomeric excess value of chiral drug in mixed solution.Contrast conventional determining method, the present invention can save solvent, without the purifies and separates before sample feeding, greatly saves analysis time.Meanwhile, the present invention is simple to operate, easily is automated, and in sample introduction process, ten-way valve all can automatically switch, and setting after condition can be in enormous quantities, carry out continuously the mensuration of chiral compound enantiomer excessive value in mixed solution.
In sum, the present invention be a kind of simple to operate, high flux Fast Measurement for the direct method for measuring of biased sample chiral compound enantiomer excessive value.
Accompanying drawing explanation
Fig. 1 is the present invention first dimension chromatogram unit, the second dimension chromatogram unit, the quantitative connection diagram of ring and two ten-way valves.
Fig. 2 is the general analysis step of the present invention's online two-dimentional chromatographic process of measuring chiral compound enantiomer excessive value in biased sample, and the switching of valve and being connected of pipeline in different analytical procedure process.
Fig. 3 is asymmetric catalysis synthesis equation in embodiment.
Fig. 4 is general chromatogram and the three dimensional chromatogram of online two-dimentional stratographic analysis in embodiment.
Embodiment
Below in conjunction with embodiment, the invention will be further described, thereby so that advantages and features of the invention can be easier to be it will be appreciated by those skilled in the art that, protection scope of the present invention is made to more explicit defining.
Embodiment 1:
The parts connect Please of liquid phase is with reference to Fig. 1, for the online two-dimentional chromatographic system of measuring, comprise the first dimension chromatogram unit, the second dimension chromatogram unit and two ten-way valves, its connection is as follows: the first dimension chromatogram unit comprises the first dimension infusion pump 11, the first dimension chromatographic column 13, automatic sampler 12 and DAD detecting device 20; The second dimension chromatogram unit comprises the second dimension infusion pump 16, the second dimension chromatographic column 14; Quantitatively 18 and two ten-way valves of ring are as the translation interface of the first dimension liquid phase and the second dimension liquid phase, wherein: entrance point and the endpiece of the first dimension chromatographic column 13 are connected with No. 1 interface of automatic sampler 12 and the first ten-way valve 15 respectively, automatic sampler 12 is connected with the first dimension infusion pump 11; Entrance point and the endpiece of the second dimension chromatographic column 14 are connected with No. 5 interfaces and No. 9 interfaces of the first ten-way valve 15 respectively; Quantitatively two end interfaces of ring 18 respectively with being connected with No. 6 interfaces for No. 9 of the second ten-way valve 19; DAD detecting device 20 is connected with No. 7 interfaces of the second ten-way valve 19; The second dimension infusion pump 16 is connected with No. 4 interfaces of the first ten-way valve 15; No. 2 interfaces of the first ten-way valve 15 are connected with waste liquid cylinder 17; No. 10 interfaces of the first ten-way valve 15 are connected with No. 8 interfaces of the second ten-way valve 19; No. 3 interfaces of the first ten-way valve 15 are connected with No. 10 interfaces of the second ten-way valve 19; No. 6 interfaces of the first ten-way valve 15 are connected with No. 5 interfaces of the second ten-way valve 19; The first described ten-way valve 15 and other interfaces of the second ten-way valve 19 are without connection;
In the present embodiment, the crude product that comes from asymmetric syntheses for the biased sample of testing, its synthetic reaction step is as follows: 1.0mmol2-aminoacetophenone, 1.5mmol benzaldehyde and 0.1mmol chiral catalyst Hua Cat are joined in 1.0mL methyl alcohol, normal temperature lower magnetic force stirs 48h, reaction generates 2-phenyl-2 with asymmetric carbon atom, 3-dihydro-quinolone (reaction equation as shown in Figure 3), reaction solution is required mensuration (R in the present embodiment, S)-2-phenyl-2, the mixed solution of 3-dihydro-quinolone enantiomeric excess value.
In the present embodiment, the first dimension chromatographic column 13 is positive nh 2 column (150mm * 4.6mm, 5 μ m), and the second dimension chromatographic column 14 is Daicel positive chirality Chiralpak OD-H post (250mm * 4.6mm, 5 μ m).Quantitatively the volume of ring is 5.0mL.The first dimension chromatogram and the second dimension chromatogram are all used chromatographically pure normal hexane and chromatographically pure isopropyl alcohol as mobile phase, and the first dimension liquid phase is used gradient elution (0-7min: the volume ratio of normal hexane and isopropyl alcohol is 98:2; 7-7.9min: the volume ratio of normal hexane and isopropyl alcohol is from 98:2 to 70:30; 7.9-12min: the volume ratio of normal hexane and isopropyl alcohol is 70:30), the second dimension liquid phase is used isocratic elution (volume ratio of normal hexane and isopropyl alcohol is 70:30), and flow velocity is 1.0mL/min, and detection wavelength is 365nm, and UV scanning scope is 200-400nm.
In the present embodiment, (R, S)-2-phenyl-2 in mixed solution, the operation steps of the online two-dimentional chromatographic determination of enantiomeric excess value of 3-dihydro-quinolone is as follows:
(1) nh 2 column as the first dimension chromatographic column 13 and the second dimension chromatogram chirality Chiralpak OD-H post are installed on highly effective liquid phase chromatographic system respectively.Directly from asymmetric catalysis synthesis solution, sample, will be containing (R, S)-2-phenyl-2, the mixed solution of 3-dihydro-quinolone is added in chromatogram sample injection bottle after film (filter opening is the film of 0.45 μ m) is processed.
(2) by the first ten-way valve 15 set positions, be 10-1, the second ten-way valve 19 positions are 1-2, and open the first dimension infusion pump 11 (1.0mL/min), DAD detecting device 20(365nm), column oven (25 ℃ of constant temperature).The baseline of device to be detected steadily after, by ready sample by automatic sampler sample introduction.
(3) because the first dimension chromatographic column 13 is positive nh 2 column, therefore the material reserve capability that polarity is little is poor, first be eluted, treat the impurity process DAD detecting device 20 that polarity is less, and (R to be detected, S)-2-phenyl-2,3-dihydro-quinolone is about to when the first dimension positive nh 2 column elutes (when retention time is 7.9min), the first ten-way valve 15 positions remain 10-1, the second ten-way valve 19 positions switch to 10-1, now, and (R, S)-2-phenyl-2,3-dihydro-quinolone is collected in the quantitative ring 18 of 5.0mL.
(4) treat (R, S)-2-phenyl-2,3-dihydro-quinolone is completely after the first dimension positive nh 2 column elutes and enter quantitative ring 18 (when retention time is 10.0min), the first ten-way valve 15 positions remain 10-1, the second ten-way valve 19 positions switch to 1-2, continue the first dimension separated, now the larger impurity of polarity is eluted from the first dimension chromatographic column 13.
(5) after the larger impurity of the first dimension chromatographic column 13 Semi-polarities is complete by wash-out (when retention time is 12.0min), the first ten-way valve 15 positions switch to 1-2, the second ten-way valve 19 positions remain 1-2, and open the second dimension infusion pump 16(1.0mL/min), close the first dimension infusion pump 11, to carry out the separation of the second dimension chromatogram.In the separation of the second dimension chromatogram, (R)-2-phenyl-2,3-dihydro-quinolone and (S)-2-phenyl-2,3-dihydro-quinolone is separated on ChiralpakOD-H post, and pass through successively DAD detecting device 20, by (R)-2-phenyl-2 in computer chromatography figure, 3-dihydro-quinolone and (S)-2-phenyl-2, it is 35.83% that the chromatographic peak area of 3-dihydro-quinolone obtains enantiomeric excess value.
Chromatogram under its 365nm detection wavelength and the three dimensional chromatogram of 200-400nm scanning are as shown in Figure 3, as can be seen from the figure eluent during 7.9-10.0min is collected in quantitative ring, therefore the response of detecting device is 0, and when carrying out two-dimension analysis (retention time is 12.0-35.0min), be collected in (R)-2-phenyl-2 in quantitative ring before, 3-dihydro-quinolone and (S)-2-phenyl-2, 3-dihydro-quinolone is by the separated detecting device that also passes through successively of Chiralpak OD-H post, and present good degree of separation, the present embodiment has illustrated the feasibility of this assay method well.
Measure the method for enantiomeric excess value in asymmetric syntheses crude product compares with tradition, the present invention does not need complicated time-consuming column chromatography purification step before sample introduction, can realize the direct mensuration that includes liquid chiral compound enantiomer excessive value in interior mixed solution at the bottom of reactant.Simultaneously owing to can directly sample and measure, so can realize the Real-Time Monitoring of enantiomeric excess value and productive rate in asymmetric catalysis synthesis from reaction solution.
The foregoing is only embodiments of the invention; not thereby limit the scope of the claims of the present invention; every equivalent structure or conversion of equivalent flow process that utilizes instructions of the present invention and accompanying drawing content to do; or be directly or indirectly used in other relevant technical fields, be all in like manner included in scope of patent protection of the present invention.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105181910A (en) * | 2015-10-21 | 2015-12-23 | 武汉理工大学 | Method for measuring enantiomer excess value of monosaccharide |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001242181A (en) * | 2000-02-29 | 2001-09-07 | Shimadzu Corp | Auto injector |
| CN1785476A (en) * | 2004-12-10 | 2006-06-14 | 清华大学 | Full bidimensional capillary liquid chromatography separation system |
| WO2006127717A1 (en) * | 2005-05-24 | 2006-11-30 | Chevron Phillips Chemical Co Lp | An integrated on-line two-dimensional method and device for synchronized analytical temperature rising elution fractionation and gel permeation chromatography |
| CN101169391A (en) * | 2006-10-25 | 2008-04-30 | 中国科学院大连化学物理研究所 | A two-dimensional high performance liquid chromatography system and its application |
-
2013
- 2013-10-31 CN CN201310532913.3A patent/CN103558325A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001242181A (en) * | 2000-02-29 | 2001-09-07 | Shimadzu Corp | Auto injector |
| CN1785476A (en) * | 2004-12-10 | 2006-06-14 | 清华大学 | Full bidimensional capillary liquid chromatography separation system |
| WO2006127717A1 (en) * | 2005-05-24 | 2006-11-30 | Chevron Phillips Chemical Co Lp | An integrated on-line two-dimensional method and device for synchronized analytical temperature rising elution fractionation and gel permeation chromatography |
| CN101169391A (en) * | 2006-10-25 | 2008-04-30 | 中国科学院大连化学物理研究所 | A two-dimensional high performance liquid chromatography system and its application |
Non-Patent Citations (3)
| Title |
|---|
| QI LIU 等: "On-line two-dimensional LC: A rapid and efficient method for the determination of enantiomeric excess in reaction mixtures", 《JOURNAL OF SEPARATION SCIENCE》, vol. 36, no. 19, 21 August 2013 (2013-08-21), pages 3158 - 3164 * |
| T. WELSCH 等: "A comprehensive chemoselective and enantioselective 2D-HPLC set-up for fast enantiomer analysis of a multicomponent mixture of derivatized amino acids", 《ANAL BIOANAL CHEM》, vol. 388, 10 July 2007 (2007-07-10), pages 1717 - 1724, XP019537576, DOI: doi:10.1007/s00216-007-1399-4 * |
| 田宏哲 等: "全二维液相色谱（NPLC×RPLC）接口及其应用", 《高等学校化学学报》, vol. 28, no. 4, 30 April 2007 (2007-04-30), pages 630 - 634 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105181910A (en) * | 2015-10-21 | 2015-12-23 | 武汉理工大学 | Method for measuring enantiomer excess value of monosaccharide |
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