Development of a validated UPLC method for simultaneous estimation of both free and entrapped (in solid lipid nanoparticles) all-trans retinoic acid and cholecalciferol (vitamin D3) and its pharmacokinetic applicability in rats.
A sensitive ultra-performance liquid chromatography (UPLC) method was developed for simultaneous estimation of all-trans retinoic acid (ATRA) and cholecalciferol (vitamin D3) in rat plasma. The method was validated over the linear range of 1.0-5000ng/ml (r(2)=0.999) for both vitamins with a limit of detection of 0.5ng/ml. Chromatographic separation was achieved using liquid-liquid extraction (LLE) on an Acquity BEH RP 18 column (2.1mm×50mm, I.D. 1.7μm), with mobile phase comprising of acetonitrile:methanol:water (90:8:2, v/v/v), at a flow rate of 0.20ml/min and a total run time of 5min. Intra and inter-day variability (RSD) was ≤3.1%, and the accuracy varied between 95.4-99.9% and 95.3-101.1% respectively, for ATRA and 98.5-100.8% and 99.3-101.7%, respectively for vitamin D3. High recovery of ≥96.0% for ATRA and ≥87.80% for vitamin D3 was achieved. ATRA and vitamin D3 were stable in plasma under different storage and processing conditions. The method was applied to estimate the total drug content and entrapment efficiency of ATRA and vitamin D3 loaded solid lipid nanoparticles (SLNs). Concentration of these two agents was determined in rat plasma after simultaneous subcutaneous administration in free form or when loaded into SLNs thus establishing pharmacokinetic application of the developed procedure. Results indicated an improvement in AUC0-∞ by 5.4 times and 29.4 times for ATRA and vitamin D3, respectively, upon their incorporation into SLNs. Simultaneous administration of these two vitamins and their improved and prolonged bioavailability has scope for their use in treatment and control of tuberculosis.
Department of Pharmaceutics, University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh 160014, India.
Manoj
M
Kumar
Department of Pharmaceutics, University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh 160014, India.
Gaurav
G
Sharma
Panacea Biotec Ltd, Lalru, Punjab 140501, India.
Dinesh
D
Singla
Panacea Biotec Ltd, Lalru, Punjab 140501, India.
Sukhjeet
S
Singh
Panacea Biotec Ltd, Lalru, Punjab 140501, India.
Sudhir
S
Sahwney
Panacea Biotec Ltd, Lalru, Punjab 140501, India.
Anurag S
AS
Chauhan
Panacea Biotec Ltd, Lalru, Punjab 140501, India.
Gagandeep
G
Singh
Department of Pharmaceutics, University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh 160014, India. Electronic address: indupalkaur@yahoo.com.
Indu Pal
IP
Kaur
D000818:Animals; D001682:Biological Availability; D002762:Cholecalciferol / Q000737:chemistry* / Q000493:pharmacokinetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057230:Limit of Detection; D008055:Lipids / Q000737:chemistry*; D059625:Liquid-Liquid Extraction / Q000379:methods; D008297:Male; D053758:Nanoparticles / Q000737:chemistry*; D010949:Plasma / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D014212:Tretinoin / Q000737:chemistry*
D008055:Lipids; D002762:Cholecalciferol; D014212:Tretinoin
ATRA; Cholecalciferol; Plasma; Solid lipid nanoparticles; UPLC