Saran

BAB V PENUTUP

5.2 Saran

Perlunya penambahan variasi waktu inkubasi diatas 5 jam untuk mendapatkan hasil penandaan yang lebih optimal.

42 DAFTAR PUSTAKA

[FDA] Food and Drug Association. 2018. Gadolinium Based - Contrast Agent. [diunduh pada 20 Feb 2019]. Tersedia pada: http:www.fda.gov/Drugs/

DrugSafety/PostmarketDrugSafetyInformationforPatientandProviders/ucm1 42882.htm.

[IAEA] International Atomic Energi Agency. 2003. Manual Reactor Produced Radioisotop (IAEA TECDOC 1340). Austria: Vienna.

[IAEA] International Atomic Energi Agency. 2018. Quality Control in the Production of Radipharmaceuticals. (IAEA-TECDOC-1856). Austria: Vienna.

Abbasi E, Milani M, Aval SF, Kouhi M, Akbarzadeh A, Nasrabadi HT, Samiei M. 2016. Silver Nanoparticles: Synthesis Methods, Bioapplications and Properties. Critical Reviews in Microbiology. 42:173–180.

Ahmed H. 2004. Principles and reactions of protein extraction, purification, and characterization. CRC press.

Aime S, Caravan P. 2009. Biodistribution on Gadolinium-Based Contrast Agents, Including Gadolinium Deposition. NIH Public Access. 30(6):1259–1267. Alirezapour B, Jalilian AR, Bolourinovin F, Moradkhani S. 2013. Production and

Quality Control of [⁶⁷Ga]-DOTA-Trastuzumab for Radioimmunoscintigraphy. Iranian Journal of Pharmaceutical Research. 12(2):355–366.

Bharali DJ, Kalil M, Gurbuz M, Simone, TM, Mousa SA. 2009. Nanoparticles and Cancer Therapy : A Concise Review with Emphasis on Dendrimers. International Journal of Nanomedicine. 4:1–8.

Brechbiel, MW, Star RA, Kobayashi H. 2005. Methods for Functional Kidney Imaging Using Small Dendrimer Contrast Agents. United States of America: US Departement of Health and Human Services.

Briley-sæbø K, Hustvedt S, Haldorsen A, Bjørnerud A. 2004. Long-term Imaging Effects in Rat Liver After a Single Injection of an Iron Oxide Nanoparticle Based MR Contrast Agent. Journal of Magnetic Resonance Imaging. 20:622– 631.

Burshberg J. 2002. The Essential Physics of Medical Imaging. California: Lippincot Williams & Wilkins.

Bushong SC & Clarke G. 2013. Magnetic Resonance Imaging-E-Book: Physical and Biological Principles. Elsevier Health Sciences.

43 Caminade Anne-Marie, Turrin C, Laurent R, Ouali A, Delavaux-nicot B. 2011. Dendrimers: Towards Catalyctic, Material, and Biomedical Uses. United Kingdom: John Wiley & Sons, Ltd.

Chakeres DW, Schmalbrock P. 1992. Fundamentals of Magnetic Resonance Imaging. New York: Raven Press, Ltd.

Chappell LL, Milenic DE, Garmestani K, Venditto V, Beitzel MP, Brechbiel MW. 2003. Synthesis and Evaluation of Novel Bifunctional Chelating Agents Based on 1,4,7,10-Tetraazacyclododecane-N,N'',N"",N''"-Tetraacetic Acid for Radiolabeling Proteins. 30:581–595.

Charles S, Vasanthan N, Kwon D, Sekosan G, Ghosh S. 2012. Surface Modification Of Poly(amidoamine)(PAMAM) Dendrimer as Antimicrobial Agents. Tetrahedron letters, 53(49):6670-6675.

Cotton F, Murillo C, Bochmann M. 2007. Advanced Inorganic Chemistry. Advanced Inorganic Chemistry. New Delhi: Wiley India.

Crooks RM, Zhao M, Sun LI, Chechik V, Yeung LK. 2001. Dendrimer-Encapsulated Metal Nanoparticles: Synthesis, Characterization, and Applications to Catalysis. Accounts of Chemical Research. 34(3):181–190. D’Souza MS, Sarkar AB. 2018. Radiological Contrast Agents and

Radiopharmaceuticals. In Side Effects of Drugs Annual. 40:579-594. Elsevier.

Decristoforo C, Zolle I, Rakias F, Imre J, Janoki G, Hesslewood S. 2007. Quality Control Methods of 99mTc Pharmaceuticals. Springer, Berlin, Heidelberg. Depkes RI. 2000. Parameter Standar Umum Ekstrak Tumbuhan Obat. Jakarta:

Departemen Kesehatan Republik Indonesia. 3-30.

Enger SA, Fisher DR, Flynn RT. 2013. Gadolinium-153 as a Brachytherapy Isotope. Physics in Medicine & Biology. 58(4):957.

Fatimah. 2012. Pembuatan dan Uji Sitotoksisitas Nanopartikel Emas-Dendrimer Poliamidoamin (PAMAM) Generasi 4 Terhadap Sel Kanker Payudara MCF-7 [skripsi]. Depok: Universitas Indonesia.

Gandjar IG, Rohman A. 2007. Kimia Farmasi Analisis. Yogyakarta: Pustaka Pelajar. 224, 228.

Garg T, Singh O, Arora S, Murthy Rsr. 2011. Dendrimer - A Novel Scaffold for Drug Delivery. Internasional Journal of Pharmaceutical Sciences Review and Research. 7:211-220.

44 Ghahramani MR, Garibov AA & Agayev TN. 2015. Determination of Radiochemical Purity of Radioactive Microspheres by Paper Chromatography. Journal of Chromatography & Separation Techniques. 6(1):1.

Giblin MF, Sieckman GL, Shelton TD, Hoffman TJ, Forte LR, Volkert WA. 2006. In Vitro and In Vivo Evaluation of 177Lu-and 90Y-Labeled E. Coli Heat-Stable Enterotoxin for Specific Targeting Of Uroguanylin Receptors On Human Colon Cancers. Nuclear medicine and biology. 33(4): 481-488. Ghai A, Singh B, Hazari PP, Schultz MK, Parmar A, Kumar P & Mishra AK. 2015.

Radiolabeling Optimization and Characterization of 68Ga Labeled DOTA– Polyamido-amine Dendrimer Conjugate–Animal Biodistribution and PET Imaging Results. Applied Radiation and Isotopes. 105:40-46.

Greenwood NN & Earnshaw A. 2012. Chemistry of the Elements. Elsevier.

Guerbet LLC. 2013. FDA Advisory Committee Briefing Document Medical Imaging Drugs Advisory Committee Meeting New Drug Application Gadoterate meglumine Injection (Dotarem^®). 204-781

Gunawan AH, Mutalib A, Aguswarini S & Lubis H. 2006. Akumulasi dan clearance dari contrast agents MRI Gd-DTPA yang disimulasikan dengan 153Gd-DTPA dalam hewan mencit. Jurnal Kimia Indonesia, 1(2), 78-81.

Hagel L. 1998. Desalting and buffer exchange with Sephadex^® G-25. Sweden: Amersham Biosciences.

Haryuni RD, Ramli M, Triningsih, Sutari. 2017. Radiolabeled Antibody Fragment for Preparation of (¹⁷⁷Lu-DOTA)m-PAMAM G3-F(ab’)2 Trastuzumab as a Radiopharmaceutical for Cancer Therapy. Atom Indonesia. 43(1):19–25. Healthcare Bio-Sciences. 2007. Instructions 52-1308-00 BB - PD-10 Desalting

Columns. AB Björkgatan 30 751 84 Uppsala Sweden.

Healthcare Bio-Sciences. 2009. Data file 28-9267-48 AB - PD-10 Desalting Columns, PD MidiTrap™ 25, PD MiniTrap™ 25, PD SpinTrap™ G-25, and PD MultiTrap™ G-25. AB Björkgatan 30 751 84 Uppsala Sweden. Hermanson GT. 2008. The Chemistry of Reactive Groups. In Bioconjugate

Techniques. 169–212. London: Elsevier.

Hermanson GT. 2013. Isotopic Labeling Techniques 1. In Bioconjugate Techniques. 507–534. London: Elsevier.

Humani TS, Sutari, Ramli M, Ritawidya R, Haryuni RD. 2017. Preparation of (¹⁷⁷Lu-DOTA)n-PAMAM-[Nimotuzumab-F(ab’)2] as a Therapeutic

Radio-45 immunoconjugate for EGFR Overexpressed Cancer Treatment. Journal of Mathematical and Fundamental Sciences. 49(3):258–268.

Inside Radiology. 2020. Gadolinium Contrast Medium (MRI Contrast Agents). [diunduh pada 3 April 2020]. Tersedia pada: https://www.insideradiology.com.au/gadolinium-contrast-medium/.

Jain K, Kesharwani P, Gupta U, Jain NK. 2010. Dendrimer toxicity : Lets Meet The Challenge. International Journal of Pharmaceutics. 394(1–2):122–142. Kobayashi H, Sato N, Saga T, Nakamoto Y, Ishimori T, Toyama S, Brechbiel MW.

2000. Monoclonal Antibody-Dendrimer Conjugates Enable Radiolabeling Of Antibody With Markedly High Specific Activity With Minimal Loss Of Immunoreactivity. European journal of nuclear medicine, 27(9):1334-1339. Kumar S, Jyotirmayee K., Sarangi M. 2013. Thin Layer Chromatography: A Tool of Biotechnology for Isolation of Bioactive Compounds From Medicinal Plants. International Journal of Pharmaceutical Sciences Review and Research. 18(1):126-132.

Laurent S, Henoumont C, Stanicki D, Boutry S, Lipani E, Belaid S & Vander Elst L. 2017. MRI contrast agents: From molecules to particles (Vol. 5). Springer Singapore.

Lide DR. 2004. Handbook of Chemistry and Physics: A Ready-Reference Book of Chemical and Physical Data, 86th ed. CRC Press, New York. 14-17.

Lehenberger S, Barkhausen C, Cohrs S, Fischer E, Grünberg J, Hohn A, Zhernosekov K. 2011. The low-energy β− and Electron Emitter 161Tb as an Alternative to 177Lu for Targeted Radionuclide Therapy. Nuclear medicine and biology. 38(6): 917-924.

Loreti V, Bettmer J. 2004. Determination of the MRI Contrast Agent Gd-DTPA by SEC–ICP–MS. Anal Bioanal Chem. 1050–1054.

Lumiprobe. 2019. NHS Esters in Reactive Groups [diunduh pada 20 Feb 2019]. Terdapat pada: https:/www.lumiprobe.com/t/reactive-groups/nhs-ester. Magerstadt M, Gansow OA, Brechbiel MW, Colcher D, Baltzer L, Knop RH,

Naegele M. 1986. Gd(DOTA):A n Alternative to Gd(DTPA) as a T1,2 Relaxation Agent for NMR Imaging or Spectroscopy. Magnetic Resonance In Medicine. 808–812.

Markatou, E. 2007. Molecular Interactions Between Dimethoxycurcumin and PAMAM Dendrimer Carriers. Int. J. Pharm. 339: 231-236.

46 Menjoge AR, Kannan RM, Tomalia DA. 2010. Dendrimer-Based Drug and Imaging Conjugates: Design Considerations for Nanomedical Applications. Drug Discovery Today. 15(5–6):171–185.

Millar AM, Beattie LA, Craig F, O'Broen LM. 2009. An Evaluation of GMCP-SA as a Replacement for ITLC-SG When Measuring The Levels of Radiochemical Impurities in ^99mTc Radiopharmaceuticals by Thin-Layer Chromatography. Journal of Labelled Compounds and Radiopharmaceuticals. 52: 538-542.

Mitsumori LM, Bhargava P, Essig M, Haki JH. 2014. Magnetic Resonance Imaging Using Gadolinium-Based Contrast Agents. Topics in Magnetic Resonance Imaging. 23(1):51–69.

Mohseni M, Shojaei S, Mehravi B & Mohammadi E. 2020. Natural Polymeric Nanoparticles as a Non-Invasive Probe for Mesenchymal Stem Cell Labelling. Artificial Cells, Nanomedicine, and Biotechnology. 48(1):770-776.

Nanjwade BK, Bechra HM, Derkar GK, Manvi FV, Nanjwade VK. 2009. Dendrimers: Emerging Polymers for Drug-Delivery Systems. European Journal of Pharmaceutical Sciences. 38:185–196.

Nwe K, Bernardo M, Regino CA, Williams M, Brechble MW. 2011. Comparison of MRI Properties Between Derivatized DTPA and DOTA Gadolinium-Dendrimer Conjugates. Bioorganic Med Chem. 18(16):5925–5931.

Oliveira IMD, Martins PDA, Silva J, Ramos MP, Lima JA, Pujatti PB, Matsuda M. 2014. Alternative Methods For Radiochemical Purity Testing In Radiopharmaceutical.

Olson B, Markwell J. 2007. Assays for Determination of Protein Concentration. Curr Protoc Protein Sci. Chapter 3:Unit 3.4

Pubchem. 2020. Gd-DOTA [diunduh 17 Mei 2020]. Terdapat pada: https://pubchem.ncbi.nlm.nih.gov/compound/Gd-DOTA

Radiation Dosimetry. 2019. What is Count per Second-CPS-Definition [diunduh 18 Juli 2020]. Terdapat pada https://www.radiation-dosimetry.org/what-is-count-per-second-cps-definition/.

Radiologyinfo.org. 2020. Magnetic Resonance Imaging (MRI) [diunduh 8 Mei 2020]. Terdapat pada: https://www.radiologyinfo.org/en/submenu .cfm?pg=mri

47 Rahmania H, Mutalib A, Ramli M, Levita J. 2014. Synthesis and Stability Test of Radiogadolinium(III)-DOTA-PAMAM G3-Trastuzumab as SPECT-MRI Molecular Imaging Agent for Diagnosis of HER-2 Positive Breast Cancer. Journal of Radiation Research and Applied Sciences. 3:1–9.

Ramalho J, Ramalho M, Jay M, Burke LM, Semelka RC. 2016. Gadolinium toxicity and treatment. Magnetic resonance imaging. 34(10): 1394-1398.

Reimer P, Parizel PM, Meaney JF & Stichnoth FA. 2010. Clinical MR imaging. Springer-Verlag Berlin Heidelberg.

Ritawidya R, Setyowati SRI, Rustendi CT, Ramli M, Hidayat B, Subur M, Mashyur J. 2014. Metoda Penyiapan Konjugat (DOTA)n-[Dendrimer PAMAM]-(Trastuzumab)m sebagai Bahan Radiofarmaka Radioimunoterapi (177 Lu-DOTA)n-[Dendrimer PAMAM] - (Trastuzumab)m. Jurnal Ilmu Kefarmasian Indonesia. 12(2):222–231.

Ryzhikov VD, Grinev,BV, Pirogov EN, Onyshchenko GM, Ivanov AI, Bondar VG, Kostyukevych SA. 2005. Use of Gadolinium Oxyorthosilicate Scintillators In X-Ray Radiometers. Optical Engineering, 44(1): 016403.

Saha BG. 2004. Fundamentals of Nuclear Pharmacy. Cleveland, USA: Springer. Sastrohamidjojo H. 1991. Kromatografi, Edisi II. 26-36. Yogyakarta: Liberty. Sastrohamidjojo H. 1992. Spektroskopi Inframerah. Penerbit Liberty, Yogyakarta. Seiler H, Sigel A & Sigel H. 1994. Handbook on metals in clinical and analytical

chemistry. CRC Press.

Skoog DA, West DM, Holler FJ & Crouch SR. 2013. Fundamentals of analytical chemistry. Nelson Education.

Stahl E. 1985. Analisis Obat Secara Kromatografi dan Mikroskopi. Penerjemah: Padmawinata, K. dan I. Sudiro. Penerbit ITB, Bandung.

Svenson S, Tomalia DA. 2005. Dendrimers in Biomedical Applications— Reflections on The Field. Advanced Drug Delivery. 57:2106–2129.

Synchem. 2017. DOTA-NHS-Ester [diunduh 10 Feb 2019]. Terdapat pada: https://www.synchem.de/product/dota-nhs-ester/.

Tafsir Web. 2020. Quran Surat Al-Anbiya Ayat 83. [diunduh 29 Februari 2020]. Terdapat pada: https://tafsirweb.com/5595-quran-surat-al-anbiya-ayat-83.html.

Tekade R, Kumar P, Jain NK. 2009. Dendrimers in Oncology : An Expanding Horizon. Chem Rev. 110:49–87.

48 Thermofisher Scientific. 2020. Dye-Based Protein Detection [diunduh pada 11 Mei 2020]. Tersedia pada: https://www.thermofisher.com/id/en/home/life- science/protein-biology/protein-assays-analysis/protein-assays/bradford-assays.html.

Thermofisher Scientific. 2020. Fourier Transform Infrared (FTIR) Spectroscopy. [diunduh pada 2 Juni 2020]. Tersedia pada: https://www.thermofisher.com/id /en/home/industrial/spectroscopy-elemental-isotope-analysis/molecular-spectroscopy/fourier-transform-infrared-ftir-spectroscopy.html

Thomsen, HS. 2006. Contrast media. Springer-Verlag Berlin Heidelberg.

Tomalia DA, Christensen JB & Boas U. 2012. Dendrimers, dendrons, and dendritic polymers: discovery, applications, and the future. Cambridge University Press.

Weast RC. 1987. Handbook of Chemistry and Physics. RC Weast.

Yenni K, Wira N, Mutalib A, Anggraeni A, Ramli M, Fauzia RP, Bahti H H. 2018. Analysis and Characterization of Complex Compound Gadolinium-(1, 4, 7, 10-Tetraazacyclododecane, 1-4-7-10-Tetraacetic Acid) n-Poliamidoamine Generation 3-Trastuzumab as a Novel Contrast Agent for Magnetic Resonance Imaging. Research Journal of Chemistry and Environment. 22(2): 249-254.

Yurt A, Kazanci N. 2008. Investigation of magnetic properties of various complexes prepared as contrast agents for MRI. Molecular Structure. 892:392–397.

49 LAMPIRAN

Lampiran 1. Perhitungan Bufer Fosfat 0,1 M pH 7,2 sebanyak 100 mL

Mr NaH2PO4 : 120 g/mol Mr NaH2PO4.H2O : 138 g/mol Mr Na2HPO4 : 142 g/mol Mr Na2HPO4.2H2O : 178 g/mol

C asam : NaH2PO4^- C garam : Na2HPO4

pH = pKa – log ^{C asam} C garam 7,2 = 7,2 – log ^Ca Cg Log^Ca Cg= 0 ^Ca Cg= 100 Ca = Cg NaH2PO4 0,05 mol 0,05 mol =^Massa_Mr 0,05 mol = ^g 120 g/mol Massa = 6 g Mr NaH2PO4.H2O

Mr NaH2PO4 x gram= ^{138 g/mol}

120 g/mol x 6 gram=6,9 gram

Na2HPO4 0,05 mol 0,05 mol =^Massa Mr 0,05 mol = ^g 142 g/mol Massa = 7,1 g Mr Na2HPO4.2H2O

Mr Na2HPO4 x gram= ^{178 g/mol}

142 g/mol x 7,1 gram=8,9 gram

1 L 100 mL

NaH2PO4.H2O 6,9 gram 0,69 gram

50 Lampiran 2. Perhitungan Bufer Fosfat 0,05 M pH 7,2 yang mengandung EDTA 5 mM sebanyak 1 L.

Mr NaH2PO4 : 120 g/mol Mr NaH2PO4.H2O : 138 g/mol Mr Na2HPO4 : 142 g/mol Mr Na2HPO4.2H2O : 178 g/mol Mr EDTA : 372,74 g/mol

pH = pKa – log ^{C asam} C garam 7,2 = 7,2 – log ^Ca Cg Log^Ca Cg= 0 ^Ca Cg= 100 Ca = Cg NaH2PO4 0,025 mol 0,025 mol =^Massa_Mr 0,025 mol = ^gram 120 g/mol Massa = 3 gram Mr NaH2PO4.H2O

Mr NaH2PO4 x gram= ^{138 g/mol}

120 g/mol x 3 gram = 3,45 gram

Na2HPO4 0,025 mol 0,025 mol =^Massa Mr 0,025 mol = ^g 142 g/mol Massa = 3,55 g Mr Na2HPO4.2H2O

Mr Na2HPO4 x gram= ^{178 g/mol}

142 g/mol x 3,55 gram=4,45 gram

EDTA 5 mM

51 Lampiran 3. Perhitungan Larutan Bufer Fosfat 0,01 M pH 7,4 sebanyak 1 L pH = pKa – log ^{C asam}

C garam 7,4 = 7,2 – log ^Ca_Cg Log^Ca Cg= 7,2 − 7,4 ^Ca Cg= −0,2 Ca Cg= Log − 0,2 Ca Cg= 0,63 Ca = 0,63 na Cg = 0,63 ng

Konsentrasi 0,01 M ~ dalam 1 L terdapat 0,01 mol na + ng = 0,01 0,63 ng + ng = 0,01 1,63 ng = 0,01 ng = 6,13 x 10-3 mol = 6,13 mmol na = 0,63 ng = 0,63 x 6,13 = 3,86 mmol NaH2PO4.H2O (na = 3,86 mmol)

mmol = ^mg Mr 3,86 mmol = ^mg

120 g/mol mg = 463,2 mg

Massa NaH2PO4.H2O = ^138g/mol

120 g/molx 463,2 mg = 532,68 mg = 0,53268 gram Na2HPO4.2H2O (ng = 6,13 mmol) mmol = ^mg Mr 6,13 mmol = ^mg 142 g/mol mg = 870,46 mg Massa Na2HPO4.2H2O =^{178 𝑔/𝑚𝑜𝑙} 142 𝑔/𝑚𝑜𝑙𝑥 870,46 𝑚𝑔 = 1091,14 𝑚𝑔 = 1,09114 𝑔𝑟𝑎𝑚

52 Lampiran 4. Perhitungan Larutan BSA 5% (^w/v) sebanyak 20 mL

BSA 5% = ^{5 𝑔𝑟𝑎𝑚}

100 𝑚𝐿 𝑥 20 𝑚𝐿 = 1 𝑔𝑟𝑎𝑚

Lampiran 5. Perhitungan larutan buffer amonium astetat pH 5,5 1M sebanyak 500 mL Mr Am.As = 77,08 g/mol M = ^{𝑔𝑟𝑎𝑚} 𝑀𝑟 𝑥 ¹⁰⁰⁰ 𝑉 1 = ^{𝑔𝑟𝑎𝑚} 77,08 𝑔/𝑚𝑜𝑙 𝑥 ¹⁰⁰⁰ 500

Gram = 38,54 gram. Disesuaikan sampai pH 5,5.

Lampiran 6. Pembuatan EDTA 0,05M sebanyak 10 mL Mr EDTA = 372,24 g/mol M = ^{𝑔𝑟𝑎𝑚} 𝑀𝑟 𝑥 ¹⁰⁰⁰ 𝑉 0,05 = ^{𝑔𝑟𝑎𝑚} 372,24 𝑔/𝑚𝑜𝑙 𝑥 ¹⁰⁰⁰ 0,01 Gram = 0,18612 gram.

Lampiran 7. Perbandingan mol DOTA:PAMAM 96:1

NHS DOTA PAMAM

Berat molekul (gram/mol) 761.5 6909

µmol 96 1

Massa (µg) 73104 6909

Konsentrasi (µg/µl) 200

Ditimbang (mg) 73.104

Dipipet (µl) 34.545

Lampiran 8. Penandaan konjugat DOTA-PAMAM dengan GdCl3 Perbandingan mol Mol (µmol) BM (g/mol) massa (µg) ^Konsentrasi Dipipet (µL) DOTA-PAMAM 1 0,16 GdCl3 2 0,33 263,61 87,87 0,1 (µg/µl) 878,7 EDTA 0,05M 2 0,33 292,24 97,41 0,05 (µmol/µl) 6,67

53 Lampiran 9. Radiokromatogram 1 Jam

54 Lampiran 10. Radiokromatogram 2 Jam

55 Lampiran 11 . Radiokromatogram 3 Jam

56 Lampiran 12 . Radiokromatogram 4 Jam