Calibration method of reference extracts and application of calibration method

Abstract

The invention relates to a calibration method of reference extracts. Each of n batches of reference extracts comprises m components. The calibration method comprises the following steps: preparing the extracts into solutions, and separating and detecting the solutions by chromatography; injecting samples of different volumes and performing linear regression on each component to obtain a regression equation; computing a signal mass ratio of the same component in the reference extracts of different batches according to the equation to obtain a m-ary linear equation set of n lines, wherein the equation set have Cn<m> roots; and counting the roots, wherein when relative standard deviation is lower than an acceptable discrimination error R, the calibration is successful. The calibration method can be used for successfully solving the problem that a reference substance is not easily available, provides a scientific and reasonable standard setting thought for traditional Chinese medicine and natural medicine, also provides a mixed reference substance for accurately quantifying related substances in chemical medicine, and further remedies the defect of operation of defining the purity of reference substances by a normalization method.

Description

technical field

[0001] The invention relates to the field of analysis, in particular to a calibration method and application of a control extract. Background technique

[0002] In the process of drug development and testing, reference substances play a very important role, but the separation and purification of reference substances is a tedious and difficult task. The technical requirements for reference substances for content determination in the current new drug registration regulations are: the purity requirement for general chemical drug reference substances is 99.5%, the purity requirement for related substance reference substances is 95%, and the purity requirement for traditional Chinese medicine reference substances is 98%. Most of the purity detection methods in the world use the normalization method. In order to achieve this purity, researchers have exhausted various separation methods such as extraction, column chromatography, high-speed countercurrent chromatogra...

Examples

Embodiment 1

[0191] In the present embodiment, n=4, m=2, four batches of control extracts all contain A 1 、A 2 Two components, the detection method is HPLC, the detection wavelength is 275nm, and the mobile phase is methanol: water = 40:60.

[0192] When the current HPLC-UV measures the substance content, within a certain range, it is basically a linear relationship. In this example, the signal value S 1 , S 2 is A 1 、A 2 The peak area of ​​the component in the chromatogram, the specific linear range [D 11 ,D 12 ] and [D 21 ,D 22 ] and the one-to-one correspondence function relation are determined again in step two, and the one-to-one correspondence function of the signal value and the signal quality of each component in the control extract can be initially drafted as follows:

[0193] S 1 =f 1 (N 1 ) = a 1 *N 1 +b 1

[0194] S 2 =f 2 (N 2 ) = a 2 *N 2 +b 2

[0195] Step 1. Relational expression of each component in each batch of control extracts

[0196] Four batche...

technical field

[0001] The invention relates to the field of analysis, in particular to a calibration method and application of a control extract. Background technique

[0002] In the process of drug development and testing, reference substances play a very important role, but the separation and purification of reference substances is a tedious and difficult task. The technical requirements for reference substances for content determination in the current new drug registration regulations are: the purity requirement for general chemical drug reference substances is 99.5%, the purity requirement for related substance reference substances is 95%, and the purity requirement for traditional Chinese medicine reference substances is 98%. Most of the purity detection methods in the world use the normalization method. In order to achieve this purity, researchers have exhausted various separation methods such as extraction, column chromatography, high-speed countercurrent chromatogra...

Examples

Embodiment 1

[0191] In the present embodiment, n=4, m=2, four batches of control extracts all contain A 1 、A 2 Two components, the detection method is HPLC, the detection wavelength is 275nm, and the mobile phase is methanol: water = 40:60.

[0192] When the current HPLC-UV measures the substance content, within a certain range, it is basically a linear relationship. In this example, the signal value S 1 , S 2 is A 1 、A 2 The peak area of ​​the component in the chromatogram, the specific linear range [D 11 ,D 12 ] and [D 21 ,D 22 ] and the one-to-one correspondence function relation are determined again in step two, and the one-to-one correspondence function of the signal value and the signal quality of each component in the control extract can be initially drafted as follows:

[0193] S 1 =f 1 (N 1 ) = a 1 *N 1 +b 1

[0194] S 2 =f 2 (N 2 ) = a 2 *N 2 +b 2

[0195] Step 1. Relational expression of each component in each batch of control extracts

[0196] Four batche...