Raman spectroscopy for detection of glycated analytes

• US 10,006,922 B2
• Filed: 12/21/2012
• Issued: 06/26/2018
• Est. Priority Date: 12/22/2011
• Status: Active Grant

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Dasari, Ramachandra, Barman, Ishan, Dingari, Narahara Chari, Kang, Jeon Woong
• Primary Examiner(s): Nur, Abdullahi
• Application Number:
  US14/367,633
• Publication Number:
  US 20140349337A1
• Time in Patent Office: 2,013 Days
• Field of Search: 356301, 356 72- 73
• CPC Class Codes: G01N 21/65 Raman scattering
• CPC Class Codes: G01N 2440/38 addition of carbohydrates, e.g. glycosylation, glycation
• CPC Class Codes: G01N 2800/042 Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism
• CPC Class Codes: G01N 33/49 Blood chemical methods for determining blood cell populations G01N33/5094; chemical analysis of blood groups or blood types G01N33/80
• CPC Class Codes: G01N 33/723 Glycosylated haemoglobin
• CPC Class Codes: G01N 33/726 Devices

Abstract

The present invention relates to the optical measurement of blood analytes, such as glycated hemoglobin (HbA1c) and serum albumin as a functional metric of mean blood glucose in the diagnosis of diabetic patients. Non-enhanced Raman spectroscopy is employed as the analytical method for quantitative detection of blood analytes. Using processing techniques, non-enzymatic glycosylation (glycation) of the analytes results in measurable and highly reproducible changes in the acquired spectral data, which enable the accurate measurements and classification of glycated and unglycated analytes.

Claims

1. A method for detecting of one or more analytes in a sample using Raman shifted light, comprising the steps of:

obtaining a solution including the sample from a patient, the sample comprising a glycated analyte;

concentrating the sample from the patient to form a concentrated sample;

illuminating the concentrated sample with light from a light source such that at least a portion of the sample emits Raman shifted light;

detecting the Raman shifted light and generating Raman spectral data for the glycated analyte in the concentrated sample; and

analyzing the Raman spectral data to determine a level of at least the glycated analyte in the sample.

2. The method of claim 1 wherein the one or more analytes are proteins.

3. The method of claim 2 wherein the proteins are selected from the group consisting of glycated hemoglobin and glycated serum albumin.

4. The method of claim 1 wherein the sample is selected from the group consisting of plasma, serum and whole blood.

5. The method of claim 1 further comprising illuminating the sample with infrared light and detecting a Raman shifted signal from the glycated analyte.

6. The method of claim 1 further comprising the steps of:

obtaining a solution of a whole blood sample comprising hemoglobin from the patient;

separating blood components of the whole blood sample into serum and cellular components;

depositing the cellular components onto a surface and drying to provide a dried solution;

acquiring Raman data from the dried solution; and

using principal component analysis to determine a level of glycated hemoglobin.

7. The method of claim 1 further comprising the steps of:

obtaining a serum sample comprising serum albumin;

delivering of the solution onto a surface and drying the solution to obtain a dried solution;

acquiring Raman spectra from the dried solution; and

using principal component analysis to determine levels of glycated serum albumin.

8. The method of claim 1 further comprising detecting an image of the analyte.

9. The method of claim 1 further comprising imaging the sample with an imaging detector.

10. The method of claim 9 further comprising generating a quantitative phase image of the sample.

11. The method of claim 1 further comprising drying a fluid containing the analyte.

12. The method of claim 1 further comprising performing a non-enhanced Raman spectral measurement of the sample.

13. The method of claim 1 further comprising suspending the sample to concentrate the analyte in a sampled region.

14. The method of claim 13 further comprising inverting the sample.

15. The method of claim 1 further comprising delivering light from a laser onto the sample such that a quantitative phase image is obtained.

16. The method of claim 1 further comprising processing the Raman data with a data processor.

17. The method of claim 1 further comprising determining a concentration of the glycated analyte in the sample.

18. The method of claim 1 further comprising displaying spectral data on a display.

19. The method of claim 1 forming a ring shaped sample and scanning light across the sample.

20. A system for detecting a glycated analyte in a sample using Raman shifted light comprising:

a sample processing concentrator device to concentrate a sample from a patient that includes a glycated analyte;

a light delivery system to illuminate the concentrated sample with light;

a detector that detects Raman shifted light from the glycated analyte in the concentrated sample, the detector generating Raman spectral data; and

a data processor that processes the Raman spectral data, the data processor being programmed with instructions to determine a quantitative concentration level of at least the glycated analyte in the sample.

21. The system of claim 20 wherein the one or more analytes are proteins.

22. The system of claim 20 wherein the proteins are selected from the group consisting of glycated hemoglobin and glycated serum albumin.

23. The system of claim 20 wherein the sample is selected from the group consisting of plasma, serum and whole blood.

24. A system of claim 20 wherein the system simultaneously detects glycated hemoglobin and glycated serum albumin in the sample.

25. The system of claim 20 further comprising processing the data using principal component analysis to determine levels of glycated hemoglobin.

26. The system of claim 20 further comprising processing the data using principal component analysis to determine levels of glycated serum albumin.

27. The system of claim 20 further comprising an imaging system to image the sample.

28. The system of claim 20 wherein the system comprises a light source to deliver light onto a blood sample and thereby generate non-enhanced Raman spectra to determine a level of glycation in a diabetic patient.

29. The system of claim 20 wherein the processing device comprises a hydrophobic substrate.

30. A method for simultaneous detection of glycated hemoglobin and glycated serum albumin comprising the steps of:

obtaining a solution including a sample from a patient, the sample comprising hemoglobin and serum albumin;

depositing a portion of the solution onto a surface and drying the solution to form a dried concentrated sample;

acquiring Raman spectral data from the dried sample; and

analyzing the Raman spectral data to determine levels of glycated hemoglobin and serum albumin.

31. The method of claim 30 wherein the depositing step comprises positioning the solution on a hydrophobic surface.

32. The method of claim 30 further comprising inverting the surface to dry the sample.

33. The method of claim 30 further comprising concentrating the sample.

34. The method of claim 30 further comprising forming a ring shaped sample.

35. The method of claim 30 further comprising illuminating the sample with light from a light source.

36. The method of claim 30 wherein the analyzing step further comprises processing the Raman data with a data processor to determine a level of glycation in a diabetic patient.

37. The method of claim 30 further comprising generating a Raman image of at least a portion of the sample.

38. The method of claim 30 further comprising forming at least one of a reflectance image, a confocal image or a quantitative phase image of the sample.

39. The method of claim 30 further comprising acquiring the Raman data by detecting Raman light from the sample with a detector.