Estimation of ZYKR1 in human urine and plasma utilizing LC-MS/MS positive electrospray ionization; a kappa opioid receptor (KOR) agonist

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 1185; p. 122982
• Main Authors: Ghoghari, Ashok, Bhatt, Chandrakant, Patel, Kuldip, Jha, Anil, Patel, Harilal, Jain, Mukul, Momin, Taufik, Parmar, Deven, Patel, Pankaj
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.11.2021
• Subjects: agonists; analgesics; Analgesics - blood; Analgesics - urine; Chromatography, High Pressure Liquid - methods; electrospray ionization mass spectrometry; formic acid; Humans; kappa opioid receptor (KOR); LC-MS/MS; Limit of Detection; methanol; pain; peptides; Peptides - blood; Peptides - urine; pharmacokinetics; Plasma - chemistry; Receptors, Opioid, kappa - agonists; solid phase extraction; Tandem Mass Spectrometry - methods; urine; Urine - chemistry; Validation; ZY17258; ZYKR1; Validation; CC; IS; ZY17258; kappa opioid receptor (KOR); SRM; K2EDTA; LLQ; mcg; LC-MS/MS; QC; ZYKR1; DQC; MQC-H; HQC; MQC-L; LQC
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2021.122982

•ZYKR1 is a short-chain peptide based, selective and peripherally restricted KOR agonist.•The chromatographic separation was performed using gradient elution and column Kinetex C8, 100 A°, 5 µ, 100 mm × 4.6 mm.•ZYKR1 and IS were extracted from human urine and plasma using solid phase extraction.•The linearity range was 0.300–300 ng/ml and 0.500–500 ng/mL for human urine and plasma respectively.•The developed method was used for estimation of ZYKR1 for clinical samples of first in human study. ZYKR1, a short chain novel peptide with selective kappa opioid receptor agonist activity used as analgesics for the treatment of pain management. A sensitive and selective LC-MS/MS assay was developed and validated for estimation of ZYKR1 in human urine and plasma. ZY17258, an analogue compound was used as an internal standard. ZYKR1 was quantified using a selective reaction monitoring in electrospray ionization positive mode. The chromatographic separation was performed using mobile phase consisted of 0.05% v/v formic acid in water and methanol in gradient elution by analytical column Kinetex C8, 100 A°, 5 µm, 100 × 4.6 mm with 8.0 min analytical run time. Solid Phase extraction technique was used for purification of ZYKR1 and IS from human urine and plasma. The calibration curves were linear over range of 0.300 ng/mL to 300 ng/mL and 0.500 ng/mL to 500 ng/mL for human urine and plasma, respectively. No matrix effect and no significant carryover were observed. The extraction recovery was consistent and ranged from about 85% to 93% in human urine and in plasma respectively. Inter-day and intra-day accuracy (bias, %) and precision (CV, %) was −11.11 to 5.91 % and −2.25 to 6.65 % in human urine and −2.74 to 7.17 % and 2.24 to 15.18 % in plasma respectively were well within the acceptance criteria. Both the assays were devoid of endogenous matrix interference and commonly used concomitant drug interference. The validated assays were used for estimation of ZYKR1 from clinical pharmacokinetic study sample bioanalysis in healthy human subjects.