title: Development and validation of a LC-MS/MS method for homocysteine thiolactone in plasma and evaluation of its stability in plasma samples.
abstract: The present study demonstrates the development and validation of a sensitive method for the quantification of homocysteine thiolactone (HCTL) in human plasma using the technique of LC-MS/MS. The gradient elution of HCTL was achieved within 5min using ZIC HILIC column having acetonitrile with 0.1% formic acid and water with 0.1% formic acid. The method was validated for the linearity, sensitivity, accuracy, precision, recovery, matrix effect and stability. A good linearity was found within a range of 0.5-32.5nmol/ml. Quantification was performed using multiple reaction monitoring (MRM) mode based on the molecular/fragment ion transitions for HCTL (118/56) and homatropine (276.1/142.2) as internal standard. Generally, HCTL levels in plasma were found to be highly unstable. In order to verify the stability of the HCTL levels in plasma for a longer period, the samples were extracted immediately and stored at -86°C. Using the above method it was found to be stable for a period of 1 month. The method was well applied for quantification of HCTL in plasma of healthy human volunteers.
authors.affiliation: High Precision Bio-analytical Facility, Department of Ocular Pharmacology and Pharmacy, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India.
authors.forename: Beauty
authors.initials: B
authors.lastname: Arora
authors.affiliation: Department of Biochemistry and Cell Biology, Sankara Nethralaya, 18, College Road, Nungambakkam, Chennai 600006, TN, India.
authors.forename: Angayarkanni
authors.initials: A
authors.lastname: Narayanasamy
authors.affiliation: High Precision Bio-analytical Facility, Department of Ocular Pharmacology and Pharmacy, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India.
authors.forename: Jayabalan
authors.initials: J
authors.lastname: Nirmal
authors.affiliation: High Precision Bio-analytical Facility, Department of Ocular Pharmacology and Pharmacy, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India.
authors.forename: Nabanita
authors.initials: N
authors.lastname: Halder
authors.affiliation: High Precision Bio-analytical Facility, Department of Ocular Pharmacology and Pharmacy, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India.
authors.forename: Santosh
authors.initials: S
authors.lastname: Patnaik
authors.affiliation: High Precision Bio-analytical Facility, Department of Ocular Pharmacology and Pharmacy, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India.
authors.forename: Alok K
authors.initials: AK
authors.lastname: Ravi
authors.affiliation: High Precision Bio-analytical Facility, Department of Ocular Pharmacology and Pharmacy, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India. Electronic address: tvelpandian@hotmail.com.
authors.forename: Thirumurthy
authors.initials: T
authors.lastname: Velpandian
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D005260:Female; D006710:Homocysteine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry; D006801:Humans; D057230:Limit of Detection; D008297:Male; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: D006710:Homocysteine; C007957:homocysteine thiolactone
keywords: HILIC; Homocysteine thiolactone; LC–MS/MS; Plasma; Stability; Validation