Method for chromatographic finger printing and standardization of single medicines and formulations

• US 7,662,638 B2
• Filed: 11/17/2008
• Issued: 02/16/2010
• Est. Priority Date: 02/08/2001
• Status: Expired due to Fees

• Current Assignee: Council of Scientific & Industrial Research
• Original Assignee: Council of Scientific & Industrial Research
• Inventors: Dadala, Vijaya Kumar, Raghavan, Kondapuram Vijaya
• Primary Examiner(s): Gakh; Yelena G
• Application Number:
  US12/313,068
• Publication Number:
  US 20090232706A1
• Time in Patent Office: 456 Days
• Field of Search: None
• US Class Current: 436/161
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 30/8675 Evaluation, i.e. decoding of the signal into analytical information for analysis of specific compounds see also G01N30/88 and subgroups of G01N33/00; chemical libraries per se C40B
• CPC Class Codes: G01N 30/8686 Fingerprinting, e.g. without prior knowledge of the sample components

Abstract

The present invention provides a method for the chromatographic fingerprinting, chemical and therapeutic standardization, bar-coding of the fingerprints and preparation of a data base for Enterprise Resource Planning (ERP) and Customer Relationship Management (CRM) machines and applications of medicines in general and traditional medicines in particular; this invention includes a software based instrumental method and a novel method of fingerprinting and standardization is proposed for the above purpose and the said method for the chromatographic finger printing which facilitates to correlate the traditional therapeutic standardization methods with the chemical properties of the medicines and humors and provides a rational basis to understand the methods used for the said purpose.

Claims

1. A computer-readable medium storing a computer program for interpreting and manipulating 2-D contour and 3 D chromatograms of an ingredient, said computer program operative to:

i) analyze the colored contour image based on the selection of various colors denoting the concentrations of the various constituents eluted with time, and polarity based on retention time;

ii) analyze the 2-D and 3-D chromatograms of the medicine using all its 3 dimensional properties of the image;

iii) generate a chromatogram having peaks at various retention times along with conjugative properties of the molecules eluted with time in a specified order of polarity;

iv) identify the compounds in the said molecules by the UV-Vis absorptive properties of the various constituents in the image;

v) correlate the reported biological, therapeutic activity of the of various constituents present in the medicines understudy based on the polarity and the conjugative properties of the molecules by dividing the fingerprint into therapeutic zones on X and Y axis;

vi) generate a barcode for a selected peak(s) using the image coordinates viz., X for retention time, Y for wavelength, R for number of red pixels, G for number of green pixels and B for number of blue pixels, provided by the proposed software;

vii) generate a database of fingerprints and barcodes for the samples, facilitating all kinds of database utilities like Enterprise Resource Planning (ERP) and Customer Resource Management (CRM) applications; and

viii) generate a database of the ‘

display windows’

for all the samples to be used by the ENTERPRISE RESOURCE PLANNING (ERP) and CUSTOMER RELATIONSHIP MANAGEMENT (CRM) type of business applications.

2. A computer-readable medium storing a computer program as claimed in claim 1, wherein the solvents used for extraction is selected based on the polarity, hydrophilic and hydrophobic nature of the constituents, sample and its constituents under study.

3. A computer-readable medium storing a computer program as claimed in claim 1, further including an HPLC and wherein the HPLC apparatus used is selected from any commercially available HPLC apparatus with the Photo Diode Array detector, preferably with a gradient or ternary system of pumps.

4. A computer-readable medium storing a computer program as claimed in claim 1, wherein the polarity of the mobile phase of a non-aqueous and an aqueous solvent of a specific pH is controlled by varying the ratio of the mobile phase from 0% to 100% of an aqueous solvents like water or a buffer of a known pH, along with a non-aqueous solvent or vice-versa.

5. A computer-readable medium storing a computer program as claimed in claim 1, wherein on analysis of 3-D and contour chromatograms the computer software is operative to generate a chromatogram with retention time and wavelength on its X and Y-axis.

6. A computer-readable medium storing a computer program as claimed in claim 1, wherein, on analysis of 3-D and contour chromatograms the computer software is operative to generate data having indicated the balancing of the physico properties and the analyte sample qualitatively and quantitatively in a percentage ratio.

7. A computer-readable medium storing a computer program as claimed in claim 1, wherein a single solvent ethanol is used for extraction of the constituents and wherein the same analytical conditions and instrumental parameters are used for all samples to bring the therapeutic generalizations thus achieving therapeutic standardization.

8. A computer-readable medium storing a computer program as claimed in claim 1, wherein the software is operative to:

(a) open chromatographic fingerprint images in different Formats (extensions) like .BMP, JPEG, TIE, GIF from the file folders and analyze it for different colors present in the image with single pixel sensitivity;

(b) displaying the pixel information in the form of a graph having a scale of X (0—

(min. time scale) and Y (200-800 nm) coordinates and a Pie diagram with individual values of each peak (Automatic and Manual) in two separate columns beside the graph;

(c) print all the data generated after analysis using PRINT Icon;

(d) open the page setup for printing using PAGE SETUP Icon;

(e) select a part of the image and analyze using RESIZE Icon;

(f) any number of image analysis windows for different images, and display of status in WINDOW icon;

(g) divide the image in to three Zones at 20 mm interval, using ZONE icon;

(h) invert the selected image using INVERT icon;

(i) switch over to Notepad, Word pad and MS Word, using EDITOR icon;

(j) provide operational information about various features of the Software using, the HELP icon; and

(k) save the data generated using SAVE AS icon as JPEG file format.

9. A computer-readable medium storing a computer program for interpreting and manipulating chromatographic separation data comprising one or more digital color image files of contour chromatograms of the constituent ingredients of an extracted organic or organo-metallic compound separated on the basis of pH and in a specified order of polarity using High Pressure Liquid Chromatography (HPLC), ultraviolet-visible, 3-D chromatography, the computer program operative to analyze a predefined set of colors for each of the one or more digital color image files with single pixel sensitivity in order to determine for the extracted compound a “

fingerprint”

including at least (a) the proportional concentrations of the constituent ingredients and (b) their polarity on a predetermined scale, based upon both the number of pixels and the colors of the constituent ingredients present at various retention times represented in the chromatographic separation data.

10. The computer-readable medium storing a computer program of claim 9, wherein the computer program is further operative to generate a numerical code for any position within any peak in any of the one or more digital color image files, the numerical code comprising Cartesian coordinates in which X represents retention time and Y represents wavelength-absorbance, and further comprising the number of color pixels at given X-Y coordinates for at least one of the colors in the predefined set of colors.

11. The computer-readable medium storing a computer program of claim 10, wherein the numerical code comprises Cartesian coordinates in which X represents retention time and Y represents wavelength-absorbance, and further comprising the number of color pixels at given X-Y coordinates for at least each of the colors red, green and blue.

12. The computer-readable medium storing a computer program of claim 10, wherein the computer program is further operative to generate a database of numerical codes for a plurality of extracted compounds.

13. The computer-readable medium storing a computer program of claim 9, wherein the chromatographic separation data further comprise one or more digital color image files of 3-D chromatograms and the computer program is further operative to generate a numerical code for any position within any peak in any of the one or more digital color image files, the numerical code comprising Cartesian coordinates in which X represents retention time and Y represents wavelength-absorbance, and further comprising the number of color pixels at given X-Y-Z coordinates for at least one of the colors in the predefined set of colors.

14. The computer-readable medium storing a computer program of claim 13, wherein the numerical code comprises Cartesian coordinates in which X represents retention time and Y represents wavelength-absorbance, and further comprising the number of color pixels at given X-Y-Z coordinates for at least each of the colors red, green and blue.

15. The computer-readable medium storing a computer program of claim 13, wherein the computer program is further operative to generate a database of numerical codes for a plurality of extracted compounds.

16. The computer-readable medium storing a computer program of claim 9, wherein the computer program is further operative to generate a database of fingerprints for a plurality of extracted compounds.

17. The computer-readable medium storing a computer program of claim 9, wherein the computer program is further operable to generate a graphical representation of an extracted compound'"'"'s “

fingerprint”

in the form of a Cartesian graph in which the X-axis represents retention time and the Y-axis represents wavelength-absorbance.

18. The computer-readable medium storing a computer program of claim 17, wherein the computer program is further operable to divide the X axis into a plurality of zones corresponding to a predetermined polarity scale.

19. The computer-readable medium storing a computer program of claim 18, wherein said plurality of zones include each of a high-polar zone, a medium polar zone, and a low/non-polar zone.

20. The computer-readable medium storing a computer program of claim 19, wherein said high-polar zone, medium polar zone, and low/non-polar zone are defined at 20 minute intervals along the X-axis.

21. The computer-readable medium storing a computer program of claim 9, wherein the computer program is further operable to generate for the extracted compound a graphical depiction of the relative number of pixels for each of the proportional concentrations of the constituent ingredients.

22. The computer-readable medium storing a computer program of claim 21, wherein the graphical depiction is in the form of a pie chart.

23. The computer-readable medium storing a computer program of claim 16, wherein the computer program is further operative to correlate extracted organic or organo-metallic compounds on the basis of one or more commonalties between their respective fingerprints in the database.