Research Article, Issue 1 Analytical Methods in Environmental Chemistry Journal Journal home page: www.amecj.com/ir AMECJ ------------------------ 1. Introduction Fe 2+ into Fe 3+ which was bonded to transferrin [2]. Copper enter to biochemical reactions and human physiology. In biological matrix such as plasma, copper complex to proteinsas caeruloplasmin and albumin or amino acids [3]. The defect of central nervous system caused to multiple sclerosis (MS) disease in human due to demyelination of nerve fibers of the brain and spinal cord. Based on previous studies, the genetic, immunological and environment factors have main source for creating MS in humans with disability. But, the environment factors such as heavy metals such as copper, zinc, cadmium and lead has significant role in MS [5, 6]. The deadline of time exposure of heavy metals in Jamileh Esmaeili a,b , Samira Shirooei c and Azam Bakhtiarian d,* a Ph.D Student in Department of Biology, School of Basic Science, Science and Research Branch, Islamic Azad University, Tehran , Iran b Department of Pharmacology, Tehran University of Medical Sciences C Sciences Research center, Health Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran *,d Department of Pharmacology Tehran University of medical Sciences In-vitro extraction and separation of copper ions from human blood samples based on amoxicillin/clavulanic acid by ultrasound assisted- dispersive centrifuge liquid-liquid micro extraction Copper (Cu) acts as a main co-factor in humans and enter into cuproenzymes that catalyze electron transfer reactions (ETR) required for cellular respiration, iron oxidation, neurotransmitter biosynthesis, antioxidant, peptide amylation. The Cu intake has been associated with toxic effects in humans include influenza-like syndrome , hemolysis and kidney failure due to Cu sulfate intake [1]. Cu exists in blood plasma in Cu bound to caeruloplasmin (65–71%) and convert the iron *Corresponding Author: Azam Bakhtiarian Email: bakhtiar12@yahoo.com https://doi.org/10.24200/amecj.v3.i01.94 A R T I C L E I N F O: Received 18 Nov 2019 Revised form 9 Jan 2020 Accepted 12 Feb 2020 Available online 30 Mar 2020 Keywords: Copper, Separation, Human blood, Amoxicillin/clavulanic acid, Ultrasound assisted-dispersive centrifuge liquid-liquid micro extraction A B S T R A C T The low concentration of copper (Cu 2+ ) can be effected on the central nervous system (CNS) and caused to multiple sclerosis (MS). Although many antibiotics can treat the bacterial infections but some of antibiotics decrease essential metal concentrations in human body and must be controlled by determining. In this study, in-vitro extraction of copper (Cu 2+ ) with amoxicillin/clavulanic acid (AMOXC) has been studied due to interacting with metals. By procedure, Cu 2+ ions were separated from blood samples by ultrasound assisted-dispersive centrifuge liquid-liquid micro extraction (USA-DC-LLME). The mixture of AMOXC (0.01 g), ionic liquid ([BMIM][PF6]) and acetone injected to 10 ml of serum blood sample at human pH=7.2. After extraction, the concentration of Cu 2+ ions was determined by flame atomic absorption spectrometry (F-AAS). The LOD, enrichment factor (EF), linear range (LR) and working range (WR) were obtained 6 μg L -1 , 9.92, 0.018-0.5 mg L -1 and 0.02-2.58 mg L -1 , respectively (RSD<1.1%). The validation of technology was confirmed by ICP and spiking samples. Anal. Method Environ. Chem. J. 3 (1) (2020) 55-62