Simultaneous determination of niacin and its metabolites—nicotinamide, nicotinuric acid and N‐methyl‐2‐pyridone‐5‐carboxamide—in human plasma by LC‐MS/MS and its application to a human pharmacokinetic study

Abstract

An LC-MS/MS method for the simultaneous quantitation of niacin (NA) and its metabolites, i.e. nicotinamide (NAM), nicotinuric acid (NUA) and N-methyl-2-pyridone-5-carboxamide (2-Pyr), in human plasma (1 mL) was developed and validated using nevirapine as an internal standard (IS). Extraction of the NA and its metabolites along with the IS from human plasma was accomplished using a simple liquid-liquid extraction. The chromatographic separation of NA, NAM, NUA, 2-Pyr and IS was achieved on a Hypersil-BDS column (150 x 4.6 mm, 5 microm) column using a mobile phase consisting of 0.1% formic acid : acetonitrile (20:80 v/v) at a flow rate of 1 mL/min. The total run time of analysis was 2 min and elution of NA, NAM, NUA, 2-Pyr and IS occurred at 1.37, 1.46, 1.40, 1.06 and 1.27 min, respectively. A detailed validation of the method was performed as per the FDA guidelines and the standard curves were found to be linear in the range of 100-20000 ng/mL for NA; 10-1600 ng/mL for NUA and NAM and 50-5000 ng/mL for 2-Pyr with mean correlation coefficient of ≥ 0.99 for each analyte. The method was sensitive, specific, precise, accurate and suitable for bioequivalence and pharmacokinetic studies. The developed assay method was successfully applied to a pharmacokinetic study in humans.

Nicotinuric Acid

“…Nicotinuric acid is the major catabolic product of nicotinic acid and regarded as a good index for assessing nicotinic acid biotransformation in the liver (2). The structure of nicotinuric acid is a type of acylglycine, which participates in various coenzyme tasks, including glycolysis, gluconeogenesis, the citric acid cycle, and oxidation, for all tissues as well as in forming long-chain fatty acids (3).…”

N-methyl-2-pyridone-5-carboxamide (2PY)—Major Metabolite of Nicotinamide: An Update on an Old Uremic Toxin

“…A number of analytical methods (such as high-performance liquid chromatography with ultraviolet or fluorescence detection, and liquid chromatography with tandem mass spectrometry (LC-MS/MS)) can be used to determine the concentration of 2PY [10,11,12,13,14]. Mullangi et al have discussed the drawbacks and the advantages of the various methods reported [15].…”

“…The advantages of the present method include a short run time, a simple sample preparation procedure (blood sample was deprotonated by acetonitrile), and a lower limit of quantification (50 ng/mL, equivalent to 0.32 µmol/L) compared to other techniques [10,11]. Our measurements confirmed the progressive rise in 2PY concentrations with successive CKD stages; the highest concentration was observed in hemodialysis patients (3100 ± 1870 ng/mL, equivalent to 20.37 ± 12.29 µmol/L) and greatly exceeded the value determined for healthy subjects (140 ± 20 ng/mL, equivalent to 0.92 ± 0.13 µmol/L) (Figure 2).…”

Simultaneous determination of atorvastatin and niacin in human plasma by LC‐MS/MS and its application to a human pharmacokinetic study

“…Quadrupoles Q1 and Q3 were set on unit resolution. As earlier publications have discussed the details of fragmentation patterns of AT (Nirogi et al ., ), NA (Inamadugu et al ., ) and IS (Chi et al ., ), we are not presenting the data pertaining to this. The analysis data obtained were processed using Analyst software™ (version 1.4.2).…”

“…As per the literature, several LC‐MS/MS methods have been reported for the determination of AT (Bullen et al ., ; Borek‐Dohalsky et al ., ; Ghosh et al ., ; Jani et al ., ; Liu et al ., ; (Ma et al ., ; Macwan et al ., ; Nirogi et al ., ; Novakova et al ., ) and NA (Inamadugu et al ., ; Peoples et al ., ; Pfuhl et al ., ), individually or with other drugs (Pilli et al ., ; Wang et al ., ) in biological samples. The major disadvantages of the all these methods include complicated and expensive extraction procedures and long chromatographic run time.…”