JPH04105041A - Method and device for measurement of mirror image isomer surplus rate - Google Patents

Abstract

PURPOSE:To measure the mirror image isomer surplus rate of different type of optical activating bodies included in a mixture specimen by calculating the primary coupling constant of the optical activation amount and the absorptive spectrum of the mixture specimen while the pure specimen is expressed with the quantities specified in Standards. CONSTITUTION:The absorptive spectrum U(lambda) of a mixture specimen 10 is measured by a spectro-photometer 12, and the CD curve V(lambda) is measured by a circle two-color type measuring device 14 and stored in memories 16a, 16b of a microcomputer 16. For each pure specimen, the CD curve and the absorption spectrum specified in Standards on the concentration basis are accommodated in an external memory device 18, and if the operator enters the name of component which he assumes as contained by the specimen 10, a reference data reading part 16c reads the CD curve g1(lambda) and absorption spectrum f1(lambda) of this component out of the external memory device 18 and stores in memory parts 16d, 16e. At this time, a component concentration calculating part 16f determines the primary coupling constant C1+ in Eq. I, while a component difference concentration calculating part 16g determines the primary coupling constant C1- in Eq. II, and a mirror image isomer surplus rate calculating part 16h can calculate the surplus rate from Eq. III.

Description

[Detailed description of the invention] [Industrial application field]

The present invention relates to a method and apparatus for measuring enantiomeric excess.

[Conventional technology]

Taking pharmaceuticals as an example, the substances that make up pharmaceuticals are often optically active, and the medicinal efficacy and
Because side effects, toxicity, etc. often differ, it is necessary to measure enantiomeric excess in drug development and testing. Enantiomeric excess can be determined with an optical rotation needle in the case of pure samples (single component). Furthermore, when the peaks of the talomadogram of a pure sample are completely separated for the d-form and the β-form, the enantiomeric excess can be determined from the ratio of the difference and sum of the respective peak areas. However, there are very few substances that can be separated in this way, and moreover, for a mixed sample consisting of multiple components, separation between each component and optical separation of that component are required at the same time. It is generally almost impossible to determine enantiomeric excess by chromatography. As described above, the technology for separating optical isomers has not been established, and it has been almost impossible to determine the enantiomeric excess of a sample containing a mixture of a plurality of optically active substances.

[Problem to be solved by the invention]

In view of these problems, an object of the present invention is to provide a method and apparatus for measuring the enantiomeric excess rate, which can measure the enantiomeric excess rate of each of a plurality of optically active substances contained in a mixed sample. It's about doing.

[Means of promise to solve problems]

In order to achieve this objective, in the enantiomeric excess measurement method according to the present invention, the spectrum U(λ) of a mixed sample is measured, and the spectrum U(λ) is converted into a pure sample i (i=
For each of 1 to n), the linear coupling constant C3° of the concentration-normalized spectrum fi (λ), Σ+=1 C1°f, (λ) is calculated, and the linear coupling constant C3° is calculated for the mixed sample. The amount of optical activity V(
λ), and the optical activity V(λ) is calculated as the optical activity g+ with respect to the wavelength normalized by the concentration for each of the pure samples i (i=1 to m, m≦n) of the optically active substance.
Calculate the linear bond constant C1- when expressed as the linear bond ΣCi-g +wl (λ) of (λ), and calculate the enantiomeric excess rate %e of component 1 contained in the mixed sample.
, e, (i>= 100 I C+-l /C++ is calculated. Here, the optical activity amount refers to CD (circular dichroism) and ORD.
(optical rotational dispersion), which in the case of a pure sample, has opposite signs and equal absolute values for the d-form and the β-form. In addition, the amount related to the enantiomeric excess refers to the enantiomeric excess itself or an amount from which the enantiomeric excess can be easily determined, such as the ratio of the d-form and the β-form of the component. shall be. In addition, in the enantiomeric excess rate measuring device according to the present invention, in order to carry out the above method, the spectrum U (λ
), an optical activity measuring device that measures the optical activity amount (λ) with respect to the wavelength λ of the mixed sample, and normalization by concentration for each of the pure samples i (i = l to n). The resulting spectrum fi (λ) is stored, and the pure sample i (i=1~
a reference data storage means storing the optical activity amount gt (λ) with respect to the wavelength normalized by the concentration for each of m, m≦n); and the spectrum U(λ) of the mixed sample;
The linear coupling constant of the normalized spectrum for the pure sample is 01. component concentration calculation means for calculating 1=linear coupling constant 61° when expressed as fi (λ), and normalization of the optical activity amount V (λ) with respect to the wavelength λ of the mixed sample with respect to the pure sample. Linear combination of optical activity ΣC, -g
, (λ), a component difference concentration calculation means for calculating a constant C1-, and a linear combination constant 01°, C1 calculated by the component concentration calculation means and the component difference concentration calculation means. - based on the enantiomeric excess of component 1 contained in the mixed sample %e, e, (i)
= 100 C=l/C, enantiomeric excess calculation means.

[Effect]

According to the present invention, it is possible to measure the enantiomeric excess of each of a plurality of types of optically active substances contained in a mixed sample, which could not be measured conventionally.

【Example】

Hereinafter, an embodiment of the method and apparatus for measuring enantiomeric excess rate according to the present station will be described based on the drawings. FIG. 1 shows the configuration of an enantiomeric excess measuring device. The mixed sample 10 contains n components i (i=l~n), among which m components 1 (11~m, m≦
It is assumed that n) is an optically active substance and the others are optically inactive substances. Taking pharmaceuticals as an example, in drug development and testing, these components contained in the mixed sample 10 are known in advance. For example, the absorption spectrum U in the ultraviolet-visible region of the mixed sample 10
(, l) with the spectrophotometer 12, and the mixed sample 1
The CD curve V(λ) of 0 is measured by the circular dichroism measuring device 14 and stored in the absorption spectrum storage section 16a and the CD curve storage section 16b of the microcomputer 16, respectively. The hardware of the microcomputer 16 has a well-known configuration, and the software configuration is shown in FIG. 1 by functional blocks 16a to 16h. On the other hand, the external storage device 18 stores information about various pure samples.
Absorption spectra and CD curves normalized by concentration are stored. The operator operates the keyboard 22 while checking on the display device 20 to manually input the names of components contained in the mixed sample 10 or that are assumed to be contained in the mixed sample 10. In response, the reference data successive section 16c reads the absorption spectrum fr (λ) (i=1 to n) of the component from the external storage device 8 and stores it in the first reference data recording section 1.
6d, and reads out the CD curve g+ (λ) of the component J (1-1 to m) from the external storage device]8 and stores it in the second reference data storage section 16e. The component concentration calculation section 16f includes an absorption spectrum storage section 16a.
U(λ) stored in the first reference data storage al
Express the linear combination of f and (λ) stored in E16d to find the linear combination constant. That is, U(λ)-ΣC++fi(λ)... (1);
; The linear binding constant C8+ at 1 is calculated as the concentration of component 1. This can be obtained by a known multi-component analysis method or by substituting the wavelength λ, (l-1 to n) of n points into the above equation (1).
It can be obtained by solving the original simultaneous equations. Similarly, the component difference concentration calculation section 16g calculates the CD curve storage section 1.
6b is expressed as a linear combination of g(λ) stored in the second reference data storage section 16e, and its linear combination constant is determined. In other words, ■(λ)-Ct-g+(λ)... (2)+
Let the linear coupling constant C,- at =1 be the differential concentration of component 1,
Obtained using the above method. Here, the optically active substance 1 (i
=1~m), the concentration of the d-isomer is Cid, and the concentration of the L-isomer is C.
++, the following formula holds true. C0゜-c, d+ C++...
(3) C+--C,,,IC+,...
- (4) The enantiomeric excess rate calculation unit 16h calculates the optically active substance l based on C+-, C+-(+-1 to m) calculated by the component concentration calculation unit 16f and the component difference concentration calculation unit 16g. Enantiomeric excess %e, e, (1-100I C+-l/CI+...(
5) is calculated and supplied to the display device 20 for display, as well as to the printer 24 for recording. For the enantiomeric excess measuring device configured as described above, an absorption spectrum as shown in FIG. 2 is supplied to the absorption spectrum storage section 16a, and a CD curve as shown in FIG. 3 is supplied to the CD curve storage section 16b. Table 1 below shows the simulation results when supplying

[Brief explanation of drawings]

1 to 3 relate to an embodiment of the method and apparatus for measuring enantiomeric excess of the present invention, FIG. 1 is a block diagram showing the configuration of the enantiomeric excess measuring apparatus, and FIG. 2 is a simulation Fig. 3 is a CD curve diagram of the mixed sample used in the simulation. In the figure, 10 is a mixed sample 16 is a microcomputer

【Effect of the invention】

As explained above, the enantiomeric excess measuring method and device according to the present invention can measure the enantiomeric excess of each of a plurality of optically active substances contained in a mixed sample, which could not be measured conventionally. It has excellent effects.

Claims (1)

[Claims] 1) Measure the spectrum U(λ) of the mixed sample, and compare the spectrum U(λ) with the pure sample i (i=1 to
n) for each of the concentration-normalized spectra f_
Linear bond of i(λ)▲There are mathematical formulas, chemical formulas, tables, etc.▼
Calculate the linear coupling constant C_i_+ when expressed as (λ), measure the optical activity amount V(λ) with respect to wavelength λ of the mixed sample, and calculate the optical activity amount (λ) from Linear combination of optical activity g_i (λ) with respect to wavelength normalized by concentration for each sample i (i=1~m, m≦n)▲There are mathematical formulas, chemical formulas, tables, etc.▼Represented by (λ) Calculate the linear binding constant C_i_- at the time, and calculate the enantiomeric excess rate %e of component i contained in the mixed sample. e. (i)
=100|C_i_-|/C_i_+ A method for measuring enantiomeric excess rate characterized by calculating an amount related to. 2), a spectrum measurement device that measures the spectrum U (λ) of a mixed sample; an optical activity amount measurement device that measures the optical activity amount V (λ) with respect to the wavelength λ of the mixed sample; and a pure sample i (i=1 -n), and the concentration-normalized spectra f_i(λ) are stored for each of the optically active pure samples i (i=1-m, m≦n). Optical activity amount g_i regarding wavelength
(λ), and a linear combination of the spectrum U(λ) of the mixed sample and the normalized spectrum of the pure sample ▲There are mathematical formulas, chemical formulas, tables, etc. ▼_i_+f_i(λ) When expressed as
A component concentration calculation means for calculating a linear coupling constant C_i_+, and a linear combination of the normalized optical activity of the pure sample to calculate the optical activity V(λ) of the mixed sample with respect to the wavelength λ. There is a component difference concentration calculation means for calculating the linear coupling constant C_i_- when expressed as ▼(λ),
Based on the linear coupling constants C_i_+ and C_i_- calculated by the component concentration calculation means and the component difference concentration calculation means,
% enantiomeric excess of component i contained in the mixed sample
e. e. (i)=100|C_i_-|/C_i_+ An enantiomeric excess ratio calculation means for calculating an amount related to (i)=100|C_i_-|/C_i_+. An enantiomeric excess ratio measuring device characterized by having these.