Methods for monitoring changes in the core of lipoprotein particles in metabolism and disease

A method for measuring the dynamical, mobility and/or fluidity properties of protein and lipoprotein elements in a sample, including but not limited to lipoprotein particle core mobility and relative core composition, the method comprising:

• placing between about 0.2 and 0.6 mL of a sample into a NMR instrument tuned to measure a nucleus selected from the group consisting of hydrogen-1, carbon-13, nitrogen-15, fluorine-19, and phosphorous-31;
• applying a series of radiofrequency pulses with intermittent delays in order to measure spin-spin and/or spin-lattice relaxation time constants from the time-domain decay of the signal, without the use of chemical shifts and without converting data into the frequency domain by Fourier transform or other means.
• partially suppressing the water signal by including a 180-degree inversion pulse followed by a delay to partially relax the water signal prior to the beginning of a sequence used to record relaxation time constants in the time domain;
• utilizing a relaxation contrast agents to perturb the solvent water or other elements of the sample;
• analyzing the exponentially decaying NMR signal in the time domain using multi-exponential analysis, including an inverse Laplacian transform;
• and
• comparing differences in the relaxation time constants for lipoprotein- or protein-specific elements within a single human subject, or between subjects, to assess normal and abnormal metabolism reflective of increased disease risk or active disease.

A method of identifying the cardiovascular risk factors in patients comprising:

• measuring the pulsed time domain NMR spin-spin relaxation time for a plurality of LDL samples selected from the group consisting of LDL and other individually fractionated lipoprotein or lipoprotein remnant samples;
• normalizing the viscosity of the same samples;
• plotting the product of spin-spin relaxation time and viscosity for the samples against an axis defined by spin-spin relaxation time to thereby develop a database of T2V statistics for the original samples;
• measuring the classic lipoprotein profile for the same samples;
• and
• correlating known risks of cardiovascular disease based upon the classic lipoprotein testing with the results as determined by the T2V statistics to thereby correlate the T2V statistics with the known risks of cardiovascular disease in a statistically acceptable manner.

A diagnostic kit comprising:

• a pulse time domain NMR instrument;
• a sample selected from the group consisting of serum and plasma;
• and
• a database of T2V data that correlates with known cardiovascular risk statistics.

A method of determining cardiac risk factors based upon blood samples, the method comprising:

• measuring the pulse time domain NMR spin-spin relaxation time of a sample selected from the group consisting of serum and plasma;
• measuring the lipid profile of the same sample;
• combining the spin-spin relaxation time and viscosity of the sample to produce a T2V value for the blood sample;
• comparing the T2V value to the lipid profile of the sample to identify the cardiac risk measured by the T2V value based upon the cardiac risk measured by the lipid profile.