Development of a new HPLC method for in vitro and in vivo studies of haloperidol in solid lipid nanoparticles

Abstract

A simple and sensitive HPLC method was developed and validated for the quantification of haloperidol in solid lipid nanoparticles (SLNs). The developed method was used for detection of shelf life of haloperidol in SLNs. Calibration curve of haloperidol was also constructed in rat plasma using loratidine as internal standard. In vivo studies were performed on rats and concentration of haloperidol in brain and blood was measured for the determination of various pharmacokinetic and hence brain targeting parameters. Chromatogram separation was achieved using C18 column as stationary phase. The mobile phase consisted of 100 mM/L potassium dihydrogen phosphate-acetonitrile-TEA (10:90:0.1, v/v/v) and the pH was adjusted with o-phosphoric acid to 3.5. Flow rate of mobile phase was 2 mL/minute and eluents were monitored at 230 nm using UV/VIS detector. The method was validated for linearity, precision, accuracy, reproducibility, limit of detection (LOD) and limit of quantification (LOQ). Linearity for haloperidol was in the range of 1-16 µg/mL. The value of LOD and LOQ was found to be 0.045 and 0.135 μg/mL respectively. The shelf life of SLNs formulation was found to be 2.31 years at 4 o C. Various parameters like drug targeting index (DTI), drug targeting efficiency (DTE) and nose-to-brain direct transport (DTP) were determined for HP-SLNs & HP-Sol administered intranasally to evaluate the extent of nose-to-brain delivery. The value of DTI, DTE and DTP for HP-SLNs was found to be 23.62, 2362.43 % and 95.77% while for HP-Sol, values were 11.28, 1128.61 % and 91.14 % respectively.Uniterms: Haloperidol/quantification. Haloperidol/pharmacokinetic. Haloperidol/in vitro study. Haloperidol/in vivo study. High performance liquid chromatography/solid lipid nanoparticles. Plasma. Validation. Brain targeting.

Assessment of brain‐to‐blood drug distribution using liquid chromatography

“…Suitable extraction solvents, such as ethyl acetate (Baldessarini et al, 1993;Castel-Branco et al, 2003;Greig et al, 1988;Md et al, 2014;Milane et al, 2009;Minocha et al, 2012;Oberoi et al, 2013;Perin et al, 2017;Wang et al, 2011;Xia et al, 2004), ethers like diisopropyl ether (Yang et al, 2015;Zhang et al, 2007bZhang et al, , 2007c, diethyl ether (Jiang et al, 2008;Shimokawa et al, 2005) and methyl tert-butyl ether (Li et al, 2013;Turnpenny & Fraier, 2009), chlorinated solvents like chloroform (Nakashima et al, 2002;Nakashima et al, 2003;Somani & Khalique, 1985) and methylene dichloride (Aravagiri et al, 1998;Aravagiri et al, 1999), hexane (Tsuneizumi et al, 1992) and isopropyl alcohol (Yasir et al, 2017) and solvent combinations such as diisopropyl ether-methyl chloride (Zhang et al, 2008), ethyl acetate-diisopropyl ether (El Zahar et al, 2018, heptane-isoamyl alcohol (Shibanoki et al, 1984) and benzene-isoamyl alcohol (Lister et al, 1983), have all been successfully applied to various analytes. The method often involves a pH adjustment to improve transfer of analytes into the organic phase.…”

“…The most common methods utilize C 18 reversed phase (RP) columns with varying ratios of mobile phases, as shown in Table 1. The columns used are: Waters Atlantis (El Yang et al, 2015;Zhang et al, 2007bZhang et al, , 2007c, Ultrasphere (Shibanoki et al, 1984), Agilent Eclipse Plus (Liang et al, 2012), Hypersil (Couet et al, 1990;Jiang et al, 2008;Nicot et al, 1986;Weigmann et al, 1999), μBondpack (Colovic et al, 2004;Lister et al, 1983;Somani & Khalique, 1985), Phenomenex Kinetex (Dalvi et al, 2018), Develosil (Nakashima et al, 2002;Nakashima et al, 2003), Xbridge (Nirogi et al, 2013), XTerra ® MS (Ligneau et al, 2007;Minocha et al, 2012), Cosomil (Yasir et al, 2017), LiChrospher (Castel-Branco et al, 2003Respaud et al, 2012), Intersil ODS-2 (Ohmori et al, 2000), Wakopak Handy ODS (Tomita et al, 2007), LiChroCART (Fortuna et al, 2013;Haque et al, 2012;Singh et al, 2011), Zorbax Eclipse (Oberoi et al, 2013;Roma nski et al, 2014;Wang et al, 2011), Gemini (Perin et al, 2017;Zheng et al, 2015), Radial-PAK (Workman et al, 1984), Partosil 10 (Greig et al, 1988Sweeney et al, 1985), Luna (Guo et al, 2007;Saigusa et al, 2011), Agilent SB …”

“…Moreover, several detectors have been used for the assessment of brain-to-blood drug level distribution, as shown in Table 1. UV detection is commonly used where compounds are detected at a wavelength where they strongly absorb (Baldessarini et al, 1993;Castel-Branco et al, 2003;Chang et al, 2005;Colovic et al, 2004;Colovic & Caccia, 2003;Couet et al, 1990;Dalvi et al, 2018;Fortuna et al, 2011;Fortuna et al, 2012;Fortuna et al, 2013;Gaudreault et al, 1996;Greig et al, 1988;Haque et al, 2012;Jiang et al, 2008;Labuzek et al, 2010;Liang et al, 1999;Lister et al, 1983;Milane et al, 2007;Milane et al, 2009;Nakashima et al, 2002;Nakashima et al, 2003;Nicot et al, 1986;Perin et al, 2017;Saigusa et al, 2011;Shimokawa et al, 2005;Singh et al, 2011;Somani & Khalique, 1985;Teoh et al, 2010;Wang et al, 2008;Weigmann et al, 1999;Workman et al, 1984;Yasir et al, 2017). Coupling HPLC…”

“…Hence they could be stored for a prolonged period of time before final analysis (Ayoub et al, 2018;El Zahar et al, 2018;Li et al, 2012;Li et al, 2013;Md et al, 2014;Minocha et al, 2012;Nirogi et al, 2013;Ohmori et al, 2000;Perin et al, 2017;Roma nski et al, 2014;Singh et al, 2011;Turnpenny & Fraier, 2009;Wang et al, 2008;Wang et al, 2011;Yang et al, 2015;Zheng et al, 2015). Some drugs such as ziprasidone, chlorpromazine and haloperidol were stable after three freeze-thaw cycles and in the autosampler for at least 24 h, but in the case of bench-top stability, spiked samples were stable for only 2 h (Yasir et al, 2017;Zhang et al, 2007bZhang et al, , 2008. Therefore, fresh brain homogenate should be analyzed within 2 h or frozen immediately as the storage of brain homogenate at room temperature over 8 h resulted in an increase in the viscosity of brain homogenate samples, which reduced the recovery, as observed for ziprasidone.…”

“…and through i.v. administration as controls (Yasir et al, 2017). Male albino Wistar rats were dosed for 0.5 h and the brain-to-plasma drug concentrations ratio was reported using HPLC-UV.…”