| ObjectiveJiao-Tai-Wan (JTW) was a famous prescription which has been used for centuries to treat insomnia. Protoberberine-type alkaloids were the main active ingredients in JTW. The purpose of our study was to explore pharmacokinetics and PK-PD combination modeling of berberine, palmatine, coptisine, epiberberine and jatrorrhizine between normal and insomnic animals by brain microdialysis.Methods1. Simultaneous quantification of berberine, palmatine, coptisine, epiberberine and jatrorrhizine in rat plasma by UPLC-MS/MS.The separation of the five compounds was carried out on a C18 column using a mobile phase consisting of acetonitrile and water (containing 5 mmol/L ammonium acetate adjusted to pH 5.0 with formic acid). The detection was performed by multiple reaction monitoring mode via electrospray ionization source operating in the positive ionization mode.2. Pharmacokinetics study of five protoberberine-type alkaloids in normal and insomnic rats after single and multiple oral administration of Jiao-Tai-WanThe insomnic rats were intraperitoneal injection of one-dose para-chlorophenylalanine acid (PCPA) to induce insomnic model. Quantifucation of five protoberberine-type alkaloids in rat plasma was achieved by using a rapid LC-MS/MS method. Plasma samples were constructed pharmacokinetic profiles using plotting drug concentration versus time and estimate pharmacokinetic parameters. The SPSS 17.0 was used for comparisons by an unpaired student’s t test.3. Simultaneous determination of berberine, coptisine, epiberberine, palmatine and jatrorrhizine in rat microdialysate by HPLC-MS/MS.The five protoberberine-type alkaloids were separated on a BDS Hypersil C18 column (2.1×50 mm i.d.,2.4 μm, Thermo Scientific) with a Phenomenex C18 guard column (3×4 mm i.d.).The elution gradient for LC-MS/MS analysis consisted of two solvent compositions:5 mmol ammonium acetate adjusted to pH 5.0 (A) and acetonitrile (B). Gradient elution was carried out according to the following program:0-3.7 min, 75%→25% mobile phase A; 3.7-4.0 min,25% mobile phase A. Accuracy, precision, carry-over, cross talk, matrix effects and recovery of five analytes were all satisfactory.4. PK-PD combination modeling of five protoberberine-type alkaloids in normal and insomnic rats after administration of Jiao-Tai-WanHeads of the rats were shaved before placing them in a stereotaxic apparatus. An microdialysis guide cannula was stereotaxically inserted in a cranial burr hole made by a skull drill using the following coordinates, in relation to the bregma (hippocampus:AP-5.2 mm, ML+4.5 mm, DV-3.8 mm). The microdialysis guide cannula was implanted to the rats, the denture base polymers type II was poured on the cathead and rivets. The rats in insomnic and normal groups were given intraperitoneal injection of one-dose PCPA (300 mg/kg) and the same volume of physiological saline, respectively. JTW extracts were administered orally after a 2 h equailibration time. The microdialysis samples were collected every 30 min for 10 h. The collected samples were kept at-20℃ and analyzed by LC-MS/MS.Results1. Simultaneous quantification of berberine, palmatine, coptisine, epiberberine and jatrorrhizine in rat plasma by HPLC-MS/MS.It is successfully established simultaneous determination of berberine, palmatine, coptisine, epiberberine and jatrorrhizine in rat plasma by HPLC-MS/MS method.The method is sensitive and high throughput. Accuracy, precision, stability and extraction recovery of five protoberberine-type alkaloids were all satisfactory.2. Pharmacokinetics study of five protoberberine-type alkaloids in normal and insomnic rats after single and multiple oral administration of Jiao-Tai-WanThe five protoberberine-type alkaloids of single-dose normal groups had low bioavailability and delay of peak time, as well as slow absorption. Either normal groups or model groups, the pharmacokinetic behavior of multiple-dose had signicant differences comparing with the single-dose; Both single-dose and multiple-dose, the pharmacokinetic behavior of insomnic rats had significant differences comparing the normal rats. Multiple dosing may increase the bioavailability, which will improve the absorption of JTW in insomnic rats and bring into active role in therapeutical effect.3. Simultaneous determination of berberine, palmatine, coptisine, epiberberine and jatrorrhizine in rat microdialysate by HPLC-MS/MS.It is successfully established simultaneous determination of berberine, palmatine, coptisine, epiberberine and jatrorrhizine in rat microdialysate by UPLC-MS/MS method. The developed method was convenient, sensitive and specific, which needed no complicated sample processing and small amount of dialysis samples. Linearity, accuracy, precision, stability, cross talk, carry-over and matrix effect of five analytes were all satisfactory.4. PK-PD combination modeling of five protoberberine-type alkaloids in insomnic and normal rats after administration of Jiao-Tai-WanInsomnic rats showed the better absorption and bioavailability than normal rats. In addition, the plasma pharmacokinetics was not similar to brain pharmacokinetics. The Tmax of normal rat was shorter than those in plasma pharmacokinetics. Protoberberine-type alkaloids could have a direct action on neuron in the hippocampus. The Emax of DA of five protoberberine-type alkaloids in insomnic rats was higher than that in normal rats,and the ECe50 of five protoberberine-type alkaloids in insomnic rats was lower than that in normal rats,which suggested that the DA efficacy of five protoberberine-type alkaloids in insomnic rats was higher than that in normal rats.ConclusionBetween normal and insomnic rats, there were significant differences in plasma and brain pharmacokinetics. Either single or multiple dosing, pharmacokinetic behaviors of model rats were different compared with the normal group, whether normal groups or model groups, the pharmacokinetic behavior of multiple dosing had significant differences comparing with the single dosing. The plasma pharmacokinetics was not similar to brain pharmacokinetics, and protoberberine-type alkaloids could have a direct action on neuron and accumulate in the hippocampus. Protoberberine-type alkaloids could readily penetrate the blood-brain-barrier, and help improve the insomnia state. The DA efficacy of five protoberberine-type alkaloids in insomnic rats was higher than that in normal rats. Jiao-Tai-Wan produced the sedative hypnotic effect to treat insomnia, which implements its mechanism by adjusting the concentration of DA. For the six other neurotransmitters influence is relatively complex, which may be its complex pharmacokinetic behavior. |
PDF Full Text Request