Distinct Differences in Lipoprotein Particle Number Evaluation between GP-HPLC and NMR: Analysis in Dyslipidemic Patients Administered a Selective PPARα Modulator, Pemafibrate
Open accessAbstract
AIM: We established a method to evaluate the lipid concentrations, size and particle numbers (PNs) of lipoprotein subclasses by gel permeation chromatography (GP-HPLC). Nuclear magnetic resonance (NMR) is widely used to analyze these parameters of lipoprotein subclasses, but differences of the two methods are unknown. Current study compared the PNs of each lipoprotein subclass measured by GP-HPLC and NMR, and assessed the effect of a selective PPARα modulator, pemafibrate. METHODS: Lipoprotein profiles of 212 patients with dyslipidemia who participated in the phase 2 clinical trial of a selective PPARα modulator, pemafibrate, were analyzed by two methods, GP-HPLC and NMR, which were performed with LipoSEARCH (Skylight Biotech) and LipoProfile 3 (LabCorp), respectively. GP-HPLC evaluated the PNs of 18 subclasses, consisting of CM, VLDL1-5, LDL1-6, and HDL1-6. NMR evaluated the PNs of 9 subclasses, consisting of large VLDL & CM, medium VLDL, small VLDL, IDL, large LDL, small LDL, large HDL, medium HDL and small HDL. RESULTS: Three major classes, total CM&VLDL, total LDL and total HDL were obtained by grouping of corresponding subclasses in both methods and PNs of these classes analyzed by GP-HPLC were correlated positively with those by NMR. The correlation coefficients in total CM&VLDL, total LDL and total HDL between GP-HPLC and NMR was 0.658, 0.863 and 0.798 (all p<0.0001), respectively. The PNs of total CM&VLDL, total LDL and total HDL analyzed by GP-HPLC was 249.5±51.7nM, 1,679±359 nM and 13,273±1,564 nM, respectively, while those by NMR was 124.6±41.8 nM, 1,514±386 nM and 31,161±4,839 nM, respectively. A marked difference in the PNs between the two methods was demonstrated especially in total HDL. The number of apolipoprotein (Apo) B molecule per one ApoB-containing lipoprotein particle, total CM&VLDL plus total LDL, was 1.10±0.05 by GP-HPLC, while 1.32±0.18 by NMR. The number of ApoA-I per one HDL particle was 3.40±0.17 by GP-HPLC, but only 1.46±0.15 by NMR, much less than reported previously.From the phase 2 clinical trial, randomizing 212 patients to pemafibrate 0.025-0.2 mg BID, fenofibrate 100 mg QD, or placebo groups, pemafibrate reduced the PNs of CM, large VLDL1-VLDL3 and medium VLDL4, but not small VLDL5 by GP-HPLC. It significantly decreased the PNs of smaller LDL and larger HDL particles, but increased those of larger LDL and smaller HDL particles. In contrast, NMR showed marked variations in the effect of pemafibrate on lipoprotein PNs, and no significant size-dependent changes. CONCLUSIONS: GP-HPLC evaluates the lipoprotein PNs more accurately than NMR and can be used for assessing the effects of lipid-lowering drugs on lipoprotein subclasses.
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Self-citationYokoyama, S., Remaley, A. T., Sampson, M. L., Ai, M., & Okazaki, M. (2021) · Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids · doi:10.1016/j.bbalip.2021.158986 · Cited by 4 · see citing works
Self-citationOgino, M., Kameda, T., Mutsuda, Y., Tanaka, H., Takahashi, J., Okazaki, M., Ai, M., & Ohkawa, R. (2022) · Bioscience Reports · doi:10.1042/bsr20220291 · Cited by 3 · see citing works
Self-citationTanigawa, R., Nagai, S., Takahashi, H., Suganami, H., & Yamashita, S. (2025) · Journal of clinical lipidology · doi:10.1016/j.jacl.2025.07.006 · Cited by 2 · see citing works
Self-citationIkezaki, H., Furusyo, N., Ai, M., Okazaki, M., Kohzuma, T., Hayashi, J., Shimono, N., & Schaefer, E. J. (2023) · Clinica Chimica Acta · doi:10.1016/j.cca.2023.117521 · Cited by 2 · see citing works
Self-citationTakenaka, Y., Inoue, I., Ikeda, M., & Kakinuma, Y. (2025) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2025.04.26.650752
Takenaka, Y., Inoue, I., Ikeda, M., & Kakinuma, Y. (2025) · Biomedicines · doi:10.3390/biomedicines13102560
- Authors
- Yamashita, S., Okazaki, M., Okada, T., Masuda, D., Yokote, K., Arai, H., Araki, E., & Ishibashi, S.
- Affiliations
- Rinku General Medical Center (JP)
- Tokyo Medical and Dental University (JP)
- The University of Osaka (JP)
- Chiba University (JP)
- National Center for Geriatrics and Gerontology (JP)
- Kumamoto University (JP)
- Jichi Medical University (JP)
- ORCID
- Venue
- Journal of Atherosclerosis and Thrombosis
- Publisher
- Japan Atherosclerosis Society
- Published
- 2021-09-01
- Volume
- 28
- Issue
- 9
- Pages
- 974-996
- ISSN
- 1340-3478 (ISSN-L)
- 1880-3873
- PMC
- PMC8532064
- OpenAlex
- W3126552794
- Cited by
- 14
- References
- 78
- Citations by year
- 2025: 3 · 2024: 2 · 2023: 2 · 2022: 4 · 2021: 3
- FWCI
- 2.58
- Type
- journal article
- Language
- en
- Open access
- diamond
- doi.org
- Licence
- cc-by-nc-sa
- Locations
- Journal of Atherosclerosis and Thrombosis (publishedVersion, cc-by-nc-sa)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
- PubMed Central (publishedVersion, cc-by-nc-sa)
- PubMed (publishedVersion)
- Funders
- Amgen
- Sanofi
- Abbott Japan
- Kowa Company
- Astellas Pharma
- Amgen Astellas Biopharma
- Japan Society for the Promotion of Science (20K17149, 18K08482, 21K08533)
- Daiichi Sankyo Europe
- Topics
- Health Sciences › Medicine › Surgery › Lipoproteins and Cardiovascular Health
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Lipid metabolism and disorders
- Health Sciences › Medicine › Surgery › Cholesterol and Lipid Metabolism
- Keywords
- lipoprotein particle number, size exclusion chromatography, NMR lipoprotein profiling, method comparison, dyslipidemia, pemafibrate, PPARα modulator, lipoprotein subclasses, chylomicrons, VLDL, low-density lipoprotein, HDL cholesterol, lipoprotein particle size, lipid concentration, lipid-lowering therapy
- MeSH terms
- Adult, Aged, Benzoxazoles, Butyrates, Chromatography, Gel, Chromatography, High Pressure Liquid, Female, Humans, Lipoproteins, Male, Middle Aged, Magnetic Resonance Spectroscopy, Placebo Effect, PPAR alpha, Dyslipidemias, Young Adult
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