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  3. Preconcentration of raloxifene drug by liquid phase microextraction method using hollow fiber and drug measurement by HPLC method in trace amounts

Preconcentration of raloxifene drug by liquid phase microextraction method using hollow fiber and drug measurement by HPLC method in trace amounts

Number of pages: 102 File Format: word File Code: 32103
Year: 2013 University Degree: Master's degree Category: Medical Sciences
Tags/Keywords: antagonistic - breast cancer - Cancer treatment - Halofiber - Hollow fiber - HPLC - Liquid phase microextraction - Raloxifene - Raloxifene drug
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  • Summary of Preconcentration of raloxifene drug by liquid phase microextraction method using hollow fiber and drug measurement by HPLC method in trace amounts

    Dissertation for M.Sc

    Treatment: Applied Chemistry

    Persian Summary

    Raloxifene is used to treat breast cancer and infertility related to oligomenorrhea or secondary amenorrhea. Raloxifene is a new treatment that has recently become available for the prevention of osteoporosis of the spine. This medicine is taken as one pill a day. In some ways, this drug acts like estrogen, but unlike it, it does not cause vaginal bleeding or increase the risk of breast cancer.

    In fact, there is evidence that this drug protects women from breast cancer in at least the first three years of starting treatment. Raloxifene does not help to solve women's menopausal problems such as hot flushes, hot flashes, and night sweats.

    Effect mechanism: Raloxifene belongs to the group of SERM drugs (selective estrogen receptor modulators). These drugs have effects similar to estrogen (agonist) in some tissues, and have relative similar effects or prevent the effect of estrogen (antagonistic) in other tissues. Raloxifene is prescribed in the treatment of hormone-responsive cancers and in this tissue as an antagonist prevents the activation of the receptor by endogenous estrogens.

    Side effects: hot flushes, vaginal bleeding, cessation of menstruation, vulva itching, digestive disorders, tumor inflammation, decrease in the number of platelets, fluid retention, baldness, uterine fibroids, vision disorders (corneal changes, cataracts and Retinopathy) reduction of platelets or white blood cells, rarely reduction of neutrophils and changes in liver enzymes are side effects of the drug.

         In this study, a liquid phase microextraction method using hollow fiber along with high performance liquid chromatography (HPLC) and UV detector was used to preconcentrate and identify raloxifene in plasma. Raloxifene was extracted from 15 ml alkaline sample solution with pH = 11 into an organic solvent (octanol) which was located in the pores of the fiber wall. After that, this drug was introduced from the organic solvent into the water receiving phase with an acidic nature with pH = 2.5, which was inside the fiber. In the following, the effective factors in microextraction, including the pH of the donor phase and the receptor phase, the type of organic solvent, the ionic strength of the donor phase, the extraction time, the stirring speed, were investigated and optimized. After extracting the drug with optimal conditions, the pre-concentration factor was 108%, the recovery was 81%, the detection limit was 0.3 ng/ml, the linear limit was 1-100 ng/mL with R2=0.99 and RSD = 3.35%. Keywords: microextraction, halofiber, preconcentration, hollow fiber, raloxifene.

     

    Introduction

    Analytic chemistry offers a variety of methods for quantitative and qualitative analysis of substances. Nowadays, the separation methods have made it possible to separate species in complex tissues with a very low detection limit (femtogram). In addition to the separation methods, the sample preparation stage is also one of the most important stages in the analysis process. This step involves converting the texture of a real sample into a state suitable for analysis by a separation technique or other methods. It can be said that the sample preparation stage is designed for the following purposes:

    Removing disturbances from the sample in order to increase the selectivity of the method

    Pre-concentrating the desired analyte and increasing its concentration in such a way that it can be measured with analytical devices.

    Converting the analytes into a form that is suitable for identification be suitable with the analyzer.

    The most basic method of sample preparation is the extraction method. The efforts of analytical chemists to invent and develop measurement methods with high precision and accuracy, as well as to eliminate manual steps that cause low reproducibility in analytical methods, have led to the invention of new extraction methods. Figure (1-1) categorizes the different methods of extraction and microextraction, about which detailed explanations are given in the references.

    In general, liquid-liquid microextraction with drops of extraction methods based on the use of porous hollow fibers will be briefly discussed

    . Statement of the problem

    In order to measure the trace amounts of raloxifene in body fluids, a method should be used that helps the medical team adjust the dose of the drug without the need for blood sampling and through a non-invasive method, and has the power to identify and separate this drug. Using the drug pre-concentration method with liquid phase microextraction with the help of halofiber, very small amounts of this drug can be concentrated and extracted in plasma, then measured by HPLC. Since the excretion of this drug is hepatic and only 6% is excreted through urine, in people with liver failure, the drug dose is 2.5 times, in people with kidney failure, the clearance increases by 15%, and considering its half-life, the plasma sample of people can be used for analysis. This method is very new and has not been used to preconcentrate and measure this drug.

    1-2. Objectives

    Inventing a new and non-invasive method to determine the amount of this drug by optimizing the effective parameters in pre-concentration and measurement of this drug and finally achieving the appropriate prescription dose in those who receive the drug.

     

     

     

     

     

     

     

     

    Chapter Two

    Examining the texts and studies of others in this field

     

     

    2-1. An overview of liquid-liquid extraction and liquid-liquid microextraction methods[1]

    Liquid-liquid extraction is based on the distribution of species between two immiscible phases, usually one of which is water and the other is an organic solvent. In this method, the basis of separation is distribution [12].

    The effective factors in liquid-liquid extraction are:

    The type of extracting solvent, volume of organic solvent for extraction, pH, extraction agent, masking agent and ionic strength of the environment.

    Liquid-liquid extraction is one of the oldest separation methods, simplicity and practicality in scale Analytical and industrial has been one of the survival factors of this method until today. One of the most important disadvantages of this method is the high consumption of organic solvent and environmental pollution, the cost of organic solvent and the creation of toxicity. In this regard, with the progress made in this method and by reducing the amount of organic solvent extracted, micro-extraction methods with solvent were used, and in these methods, the amount of organic solvent used is significantly reduced [29].

  • Contents & References of Preconcentration of raloxifene drug by liquid phase microextraction method using hollow fiber and drug measurement by HPLC method in trace amounts

    List:

    Persian Summary .. 1

    Introduction .. 3

    Chapter One: Generalities

    1-1. statement of the problem 6

    1-2. Goals. 6

    Chapter Two: Examining the texts and studies of others in this field

    2-1. An overview of liquid-liquid extraction and liquid-liquid microextraction methods. 8

    2-1-1. Liquid phase microextraction. 9

    2-1-1-1. Single-drop liquid phase microextraction 9

    2-1-1-2. Hollow fiber liquid phase microextraction (HF-LPME) 11

    2-1-1-2-1. Extraction principles and different systems using HF-LPME. 12

    2-1-1-2-2. Practical aspects and different configurations of HF-LPME. 15

    2-1-1-2-3. Hollow fiber headspace liquid phase microextraction. 18

    2-1-1-3. Liquid phase microextraction using the freezing of extractant solvent 21

    2-1-1-4. Dispersive liquid-liquid phase microextraction (DLPME) 21

    2-1-2. Solid phase extraction. 22

    2-2. Chromatography. 22

    2-2-1. Classification of chromatography methods. 23

    2-2-1-1. High Performance Liquid Chromatography (HPLC) 23

    2-2-1-2. Liquid chromatography devices. 25

    2-2-1-2-1. Mobile phase tank. 26

    2-2-1-2-2. Pumping systems 26

    2-2-1-2-3. Sample injection systems. 27

    2-2-1-2-4. Liquid chromatography columns. 28

    2-2-1-2-5. Piston temperature. 29

    2-2-1-2-6. Detectors 29

    2-3. Review of HF-LPME studies. 33

    2-4.  Review of the study drug. 35

    2-4-1. Pharmacokinetics of the drug 35

    2-4-2. Mechanism of drug action 35

    2-4-3. Uses of medicine 36

    2-4-4. Contraindications and precautions. 36

    2-4-5. side effects 36

    2-4-6. Interferences. 36

    2-4-7- Dosage. 36

    2-5. The importance of measuring raloxifene. 37

    Chapter Three: Materials and Methods

    3-1. Chemicals and equipment. 39

    3-1-1. Chemicals, standards and real samples. 39

    3-1-2. Device equipment. 39

    3-2. Extraction method. 40

    3-2-1. Briefly, the extraction was done in the following steps: 41

    3-2-2. Optimization steps. 42

    3-2-2-1. Optimization of isolation conditions. 42

    3-2-2-2. Optimizing extraction conditions. 42

    3-2-2-2-1. Type of organic solvent. 43

    3-2-2-2-2. The pH effect of the donor phase 43

    3-2-2-2-3. The pH effect of the receptor phase 43

    3-2-2-2-4. The effect of ionic strength of phase donor 43

    3-2-2-2-5. The effect of stirring the analyte solution. 43

    3-2-2-2-6. Effect of extraction time. 43

    3-2-2-2-7. Effect of temperature on extraction. 43

    3-2-3. Evaluating the efficiency of the extraction method. 44

    3-2-3-1. Grading curve. 44

    3-2-3-2. Determination of preconcentration factor (PF) 44

    3-2-3-3. Determination of repeatability (RSD) 45

    3-2-4. Real sample analysis. 45

    Chapter Four: Results

    4-1. Three-phase microextraction based on the use of porous hollow fiber. 47

    4-1-1. Principles of theory. 47

    4-2. Optimization steps. 50

    4-2-1. Optimization of isolation conditions. 50

    4-2-2. Optimizing extraction conditions. 51

    4-2-2-1. Type of organic solvent. 51

    4-2-2-2. The pH effect of the receiver phase and the donor phase 53

    4-2-2-3. The effect of stirring speed of the analyte solution. 55

    4-2-2-4. The effect of ionic strength of phase giver 57

    4-2-2-5. Effect of extraction time. 58

    4-2-2-6. Effect of extraction temperature. 59

    4-3. Determining the analytical parameters of the extraction method. 60

    4-3-1. Preparation of grading curve. 61

    4-3-2. Pre-concentration factor (PF) and recovery percentage (R%) 61

    4-3-3. Limit of detection (LOD) 63

    4-3-4. Reproducibility of method (RSD) 63

    4-4- Real sample analysis. 64

    Chapter Five: Discussion and Conclusion

    5-1. Comparison of the extraction method with the reported methods of other sources. 66

    5-2. conclusion 69

     

    Resources.. 70    

    English summary.. 83

    Appendices.

    Source:

     

    Anthony, J., Bertram, J., Katzung, S. B., Arjamand, M., Amin Asanafi, G., 2017, Pharmacology of Katzong and Trevor, 8th edition, Arjamand, p. 135-140.

    Skoog, D. A., Haller, F. J., Nieman, T. E., Selajgeh, A. R., 1383, Principles of Systematic Analysis, Academic Publishing Center, Volume II, p. 538 to

     

    Abulhassani, J., Manzoori, J. L., Amjadi, M, 2010, Hollow Fiber Based-Liquid Phase Micro Extraction Using Ionic Liquid Solvent for Pre-Concentration of Lead and Nickel From Environmental and Biological Samples Prior to Determination by ElectroThermal Atomic Absorption Spectrometry, J. Hazardous Materials, vol. 176, P. 481-486.

    Andersen, S., Halyorsen, T. G., Pedersen-Bjergaard, S., Rasmussen, K. E., Tanum, L., Refsum, H., 2003, Stereospecific Determination of Citalopram and Desmethylcitalopram by Capillary Electrophoresis and Liquid Phase Micro Extraction, J. pharm. Biomed., No. 33, P. 263.

    Barahona, F., Gjelstad, A., Pedersen-Bjergaard, S., Rasmussen, K. E., 2010, Hollow Fiber Liquid Phase Micro Extraction of Fungicide from Orange Juices, J. Chromatogr. A., vol. 1217, P. 4781.

    Borman, SA, 1983, Diode Array Detection in HPLC, Chem., vol. 55, p. 836.

    Brown, PR., 1973, High pressure Liquid Chromatography: Biochemical and Biomedical Applications, New York: Academic press; p. 91-112.

    Brown, PR., Hartwick RA., 1988, High Performance Liquid Chromatography, New York: Wiley, Chapter 6, p. 300-309.

    Chang, C. H., Tsen, C. W., Hsien, D. W., 2009, Determination of Perchlorate in River by Ion Pair Hollow Fiber liquid phase Microextraction Coupled with Electrospray Ionization Tandem Mass Spectrometry, Talanta, vol. 79, p. 442.

    Chia, K. J., Huang, S. D., 2006, Simultaneous Derivatization and Extraction of Primary Amines in River Water with Dynamic Hollow Fiber LPME Followed by GC/ME, J. Chromatogr A., ??vol. 1103, p. 158-161.

    Cummings, J., Lefevre, G., Small, G., Appel-Dingemanse, S., 2007, Pharmacokinetic rationale for the raloxifene, 69(4 Suppl 1), p. 15–23.

    Dean, J. R., 2010, Extraction Techniques in Analytical Sciences, John Wiley and Sons, p. 201-236.

    Dorsey, J. G., Cooper, W. T., 1994, Retention mechanism of bonded-phase liquid chromatography, J. Anal. Chem., vol. 66, p. 857.

    Ebrahimzadeh, H., Yamini, Y., Firozjaei, H. A., Kamarei, F., Tavassoli, N., Rouini, M.R., 2010, Hollow Fiber-Based Liquid Phase Micro Extraction Combined with High Performance Liquid Chromatography for the analysis of gabapentin in Biological Samples, J. Anal. Chim. Acta, vol. 665, p. 221.

    Ebranimzade, H., Asgharinezhad, A. A., Abedi, H., Kamarei, F., 2011, Optimization of Carrier Mediated Three Phase Hollow Fiber Micro Extraction Combined with HPLC-UV for Determination of Propylthiouracil in Biological Samples, Talanta, vol. 85, p. 1043.

    Emidio, E.S., Prata, V. D., de Santana F. J., Dorea, H. S., 2010, Hallow fiber-based liquid phase micro extraction with factorial design optimization and gas chromatography-tandem mass spectrometry for determination of cannabinoid in human hair, J. Chromatogr. B, p. 2175.

    Engelhardt, H., 1986, Practice of High Performance Liquid Chromatography: Applications, Equipment and Quantitative Analysis, New York: Springer verlag; Chapter 1. p. 195-206.

    Frankfort, S.V., M. Ouwehand, M.J. van Maanen, H., Rosing, C.R. Tullner, J.H. Beijnen, 2006, Rapid Communications in Mass Spectrometry, vol. 20, p. 3330-3336

    Freitas, D. F., Porto, C. E. D., Viera, E. P., Siquera, M. E. P. B., 2010, Three Phase, Liquid Phase Micro Extraction Combined with HPLC-Fluorescence Detection for the Simultaneous Determination of Fluoxetine and NorFluoxetine in Human Plasma, J. Pharma and Biomed Analysis, vol. 51, p. 170.

    Ghambarian., M., Yamini, Y., Esrafili, A., 2011, Three Phase Hollow Fiber Liquid Phase Micro Extraction Based on Two Immiscible Organic Solvents for Determination of Tramadol in Urine and Plasma Samples, J. Pharmaceutical and Biomedical Analysis, vol. 56, p.1041.

    Ghasemi, E., Najsfi, N. M., Raofie, F., Ghassempour, A.

Preconcentration of raloxifene drug by liquid phase microextraction method using hollow fiber and drug measurement by HPLC method in trace amounts
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