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New finding of nalbuphine metabolites in men: NMR spectroscopy and UPLC–MS/MS spectrometry assays in a pilot human study

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Abstract

A safe and efficient semi-synthetic narcotic nalbuphine (NAL) which was broadly applied in analgesic therapy has long been considered to eliminate from human body via phase II conjugation. However, up to the present, neither the complete metabolic pathways nor the identified metabolites of NAL have been clarified in documented reports. In this study, four novel metabolites were discovered by incubating NAL with human liver microsomes. These metabolites were later quantified in blood samples from human volunteers treated with NAL. An accurate and precise new method for simultaneously determining NAL and its metabolites was also established. Their chemical structures were elucidated on the basis of one- and two-dimensional NMR spectroscopic analyses including 1H–1H correlation spectroscopy, nuclear overhauser enhancement spectroscopy, heteronuclear single-quantum correlation, and heteronuclear multiple bond correlation, and further confirmed by mass spectrometry. The analytical method was validated and applied successfully to a pilot human study with ultra-high performance liquid chromatography–tandem mass spectrometry employed with positive ion electrospray ionization via multiple reaction monitoring mode. This is the first report on the qualitative and quantitative analysis of NAL coupled with its two hydroxylated (3′-hydroxynalbuphine and 4′-hydroxynalbuphine) and two conjugated metabolites (nalbuphine-3-β-d-glucuronide and nalbuphine-6-β-d-glucuronide). The present method offers a rapid and simple way of performing pharmacokinetic studies of NAL, and assists in elucidating its metabolic pathway in humans.

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Abbreviations

UGTs:

Uridinyl diphosphate glucuronosyltransferases

P450s:

Cytochrome P450s

UPLC–MS/MS:

Ultra-high performance liquid chromatography-tandem mass spectrometry

1H NMR:

Proton nuclear magnetic resonance

NAL:

Nalbuphine

NAX:

Naloxone

N3G:

Nalbuphine-3-β-d-glucuronide

N6G:

Nalbuphine-6-β-d-glucuronide

3′-OH NAL:

3′-Hydroxynalbuphine

4′-OH NAL:

4′-Hydroxynalbuphine

COSY:

Correlation spectroscopy

NOESY:

Nuclear Overhauser enhancement spectroscopy

HSQC:

Heteronuclear single-quantum correlation

HMBC:

Heteronuclear multiple bond correlation

QC:

Quality control

LLOQ:

Low limit of quantification

HLMs:

Human liver microsomes

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Acknowledgments

This study was funded by Department of Health, Executive Yuan of Taiwan (No. DOH97-TD-I-111-DD002).

Conflict of interest

No conflicts of interest are declared by all of the authors.

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Authors and Affiliations

  1. Graduate Institute of Life Sciences, National Defense Medical Center, No. 161, Sec. 6, Minquan E. Rd., Neihu Dist., Taipei City, 114, Taiwan

    Hong-Jaan Wang & Oliver Yoa-Pu Hu

  2. School of Pharmacy, National Defense Medical Center, No. 161, Sec. 6, Minquan E. Rd., Neihu Dist., Taipei City, 114, Taiwan

    Cheng-Huei Hsiong, Li-Heng Pao, Wen-Liang Chang, Min-Jen Lin, Pei-Wei Huang & Oliver Yoa-Pu Hu

  3. Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, No. 261, Wen-hwa 1st Rd., Kwei-shan, 333, Taoyuan, Taiwan

    Li-Heng Pao

  4. National Research Institute of Chinese Medicine, Ministry of Health and Welfare, No. 155-1, Sec 2, Li Nung St., Beitou Dist., Taipei City, 112, Taiwan

    Li-Jie Zhang

  5. Department of Anesthesiology, Taipei Veterans General Hospital, No. 201, Sec. 2, Shipai Rd., Beitou Dist., Taipei City, 112, Taiwan

    Shung-Tai Ho

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  1. Hong-Jaan Wang
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Correspondence to Oliver Yoa-Pu Hu.

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Wang, HJ., Hsiong, CH., Pao, LH. et al. New finding of nalbuphine metabolites in men: NMR spectroscopy and UPLC–MS/MS spectrometry assays in a pilot human study. Metabolomics 10, 709–718 (2014). https://doi.org/10.1007/s11306-013-0605-y

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  • DOI: https://doi.org/10.1007/s11306-013-0605-y

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