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Melamine ubedo piny ma nyayu two i cam ma copo bedu i kit cam moko ma rwinyo man ma rwinyo. Lembe ma jutimo i somo eni ubedu mi ketho cing ikum nwangu man kwanu pa melamine i cam pa juma nyathin man cak ma buru. Wel cam 40 ma jucwalo, ikinde cak pa juma nyathin man cak, kud i kabedu matung tung mi Iran jutimo sayusac ikumgi. Kite ma melamine utiye kudu i lanyuth juporo ku tiyo ku high-performance liquid chromatography-ultraviolet (HPLC-UV) system. Calibration curve (R2 = 0.9925) juyike pi nwangu melamine ikind 0.1–1.2 μg mL−1. Kabedo mi pimo man nwangu ubedo 1 μg mL−1 man 3 μg mL−1, kubang wadi. Melamine jupimu i cam pa juma nyathin man cak ma jumaku man adwogi ne unyutho nia melamine i cam pa juma nyathin man cak ma jumaku ubedo 0.001–0.095 mg kg−1 man 0.001–0.004 mg kg−1, kubang wadi. Lembe maeni ucungo iwi cik pa EU man Codex Alimentarius. Ber ni ngeyo ni camu cak maeni ma melamine migi nok nyayu ngo peko madit ikum yot kum pa ju ma camu. Eni de jukonyo ku adwogi pa pimo peko.
Melamine ubedo piny ma kwo ma nyinge C3H6N6, ma wok kud i cyanamide. Etiye ku kero manok mi nyai i pii man etiye ku 66% pa nitrogen. Melamine ubedo piny ma jutiyo kudu lee i tic mi yiku piny ma jutiyo kudu pi yubu plastik, yath, man jam tic mi yubu cam (ma uketho i iye jam tedo cam man jam tedo)1,2. Melamine de jutiyu kudu calu yath pi remo two. Nitrogen madwong in melamine copo telo wi tic marac ku jamb tedu man miyo kit pa molecules pa protein i cam3,4. Pieno, medu melamine i cam, ikinde cak, medu nitrogen. Pieno, jumoko giparu niya protein ma nwang i cak ubedo madwong uloyu ma ubedu kudu.
Pi gram acel acel pa melamine ma ju medo, wel pa protein ma i cam bi medere ku 0.4%. Ento, melamine utiye piny ma nyai i pii man ecopu nyayu peko madit. Medu 1.3 grams pa melamine i piny ma pii calu cak copo medu rwom pa cak ku 30%5,6. Kadi bende ju medo melamine i cam pa lei man pa dhanu pi medu rwom pa protein7, Codex Alimentarius Commission (CAC) man judongo pa ngom gi yiyu ngo melamine ni piny ma ju medu i cam man gi ketho ni piny ma rac ka ju mwonyo, ju yuyu, nyo ju mako i adila. I 2012, dilo mi yot kum mi ng'om zoo (WHO) mi ng'om zoo pi sayusac iwi two kanca uketho melamine ni piny ma nyayu two kanca i rwom mir 2B pi en ecopu bedu rac ni yot kum pa dhanu8. Bedo ku melamine pi kare malac copo nyayu two kanca nyo ret pa adhi2. Melamine i cam copo rwate ku cyanuric acid mi yubu yellow crystals ma copo nyai ungo i pii ma copo nyayu peko i kum adhi man ic, man bende two kanca mi ic man dwoko pek pa kum piny9,10. Ecopu nyayu tho pa cam man, ka etiye madwong, tho, asaga ikum juma nyathin man awiya ma fodi gitie aradu.11 Kanisa mi yot kum mi ng'om zoo (WHO) de uketho wel pa melamine ma dhanu camu kubang ceng (TDI) ni 0.2 mg/kg pa pek pa kum dhanu kubang ceng nimakere ku cik pa CAC.12 The US Food and Drug Administration (US) uketho wel ma malu pa melamine pi dhanu nimakere ku cik pa CAC. 1 mg/kg i cak pa juma nyathin man 2.5 mg/kg i cam mange.2,7 I Dwi mir abongwen 2008, jukoro nia ju mapol ma yiku cak pa juma nyathin gi medo melamine i cak mi cak pi medo rwom pa cak migi, ma ukelo tho pa cak man ukelo tho pa melamine i ngom zoo ma unyai awiya ma kadhu 2004 man 2004 uketho dhanu ma kadhu 50,000 i ot yat. 13
Caku cak epe kare ku kare pi the lembe matung tung calu peko mi kwo mi adhura, two pa min ot nyo nyathin, ma ucwalu tic ku cak cak pi miyo cak. Macalu adwogi, juyike factory mi yubu cak pa juma nyathin ma ceng ku cak pa min ic14. Cak pa juma nyathin ma jucwalu i soko pol kare juyike ku cak pa dhyeng man pol kare juyike ku cak moko ma juyiko ku moo, proteins, carbohydrates, vitamins, minerals man piny mukende mapol. Pi bedu ceng ku cak min ot, protein man moo ma nwang i formula ne lokore, man nimakere ku kit cak, ju medo ku piny ma calu vitamins man minerals calu iron15. Calu awiya ma fodi gitie aradu gitiye dul ma rwinyo man gitiye ku lworo mi nwangu two, bedo ber pa madhu cak ma buru ubedo piny ma pire tek pi yot kum. I ngei lembe mi melamine poisoning ikind awiya mi China, ngom mapol i ng'om zoo gi ketho wigi ikum lembe eni, man rwom pa lembe eni de umedire. Pieno, piretek ni medo kero pa gwoko cam pa juma nyathin pi gwoko yot kum pa juma nyathin. Tye yore matung tung mi nwangu melamine i cam, ikinde utiye high-performance liquid chromatography (HPLC), electrophoresis, sensory method, spectrophotometry man antigen-antibody enzyme-linked immunosorbent assay16. I 2007, US Food and Drug Administration (FDA) uketho man ugoro yore mi HPLC pi nyang ikum melamine man cyanuric acid i cam, ma en e yore maber loyo mi nyang ikum kadhiri pa melamine17.
Kit pa melamine i cak pa juma nyathin ma jupimu ku yore manyen mi infrared spectroscopy ubedo 0.33 ku 0.96 milligrams pi kilogram acel (mg kg-1). 18 Somo moko i Sri Lanka unwangu niya wel pa melamine in cak ma nyai ubedo ikind 0.39 ku 0.84 mg kg-1. Mi medo, cam pa juma nyathin ma jucwalu kudi ngom mawoko ubedo ku rwom ma malu pa melamine, i 0.96 man 0.94 mg/kg, kubang wadi. Kite eni utiye piny pa rwom ma juketho (1 mg/kg), ento yub mi lubo tok jeni mitere pi bedo ber pa juruwil. 19
Somo madwong udaru ngiyo kadhiri pa melamine i cam pa awiya mi Iran. Ikind 65% pa lanyuth ubedo ku melamine, ku rwom pa 0.73 mg/kg man rwom ma malu pa 3.63 mg/kg. Somo mange onyuthu nia kadhiri pa melamine i cam pa juma nyathin ubedo 0.35 ku 3.40 μg/kg, ku rwom pa 1.38 μg/kg. Ikum zoo, bedo man kadhiri pa melamine i cam pa awiya mi Iran jutimo i lembe matung tung, ku lanyuth moko ma melamine ukadhu wel ma judongo pa gavumente uketho (2.5 mg/kg/camu).
Ka waparu pi tic ma lee pa cak ma jumaku i tic mi cam man beru pa cak ma jumaku i cam pa juma nyathin, somo eni umitu ni ketho yore mi nwangu melamine i cak ma jumaku man cam ma jumaku. I andha, lembakeca mir acel mi somo eni ubedu mi yubu yore ma nyap, mayot man ma atira mi nwangu melamine i cam pa juma nyathin man cak ma juyubu ku tiyo ku high performance liquid chromatography (HPLC) man ultraviolet (UV) detection; Mir aryo, lembakeca mi somo eni ubedu mi nyang ikum melamine ma nwangre i cam pa juma nyathin man cak ma jucwalu i soko mi Iran.
Jam tic ma jutiyo kudu pi ngiyo melamine lokore ni makere ku kama juyubu iye cam. Yore mi ngiyo HPLC-UV ma rwinyo man ma gen jutiyo kudu pi pimo melamine residues i cak man cak pa juma nyathin. Piny ma wok kud i cak utiye ku proteins man moo matung tung ma copo nyayu peko i pimo melamine. Pieno, calu ma Sun et al. 22, yore maber man maber mi yweyu piny mitire mapodi jutimo sayusac ikum jam tic. I lembe maeni, watiyo ku filter mi syringe ma jubolo wang acel acel. I lembe maeni, watiyo ku C18 column mi poko melamine i cam pa juma nyathin man cak ma buru. Cal mir acel nyutho chromatogram pi nwangu melamine. Mi medo, dwoko pa lanyuth ma tye ku 0.1-1.2 mg/kg melamine ucaku 95% ku 109%, equation mi dwoko piny ubedo y = 1.2487x − 0.005 (r = 0.9925), man wel mi apoka poka (RSD) ucaku �0 ku 8%. Lembe ma nwang nyutho nia yore ne utiye maber in rwom ma juponjo (Table 1). Kabedo mi nwangu (LOD) man rwom mi kwanu (LOQ) pa melamine ubedo 1 μg mL−1 man 3 μg mL−1, kubang wadi. Mi medo, UV spectrum pa melamine unyutho absorption band i 242 nm. Yore mi nwangu lembe utiye ma rwinyo, ma gen man ma atira. Yore maeni jucopo tiyo kudu pi nyang ikum kadhiri pa melamine.
Adwogi marom rom juwodho kud i bang jugor mapol. Yore mi tiyo maber pa liquid chromatography-photodiode array (HPLC) juyike pi nyang ikum melamine i piny ma juyubu ku cak. Kabedo ma piny pa wel ubedo 340 μg kg−1 pi cak ma buru man 280 μg kg−1 pi cak ma jumaku i 240 nm. Filazzi ku juwadi. (2012) onyuthu nia melamine unwangre ngo i cam pa juma nyathin ku HPLC. Ento, 8% pa lanyuth pa cak ma jumaku ubedo ku melamine i kadhiri pa 0.505-0.86 mg/kg. Tittlemiet et al.23 gitimo somo marom man ginwangu nia melamine ma nwang i cam pa juma nyathin (namba pa lanyuth: 72) ku high-performance liquid chromatography-mass spectrometry/MS (HPLC-MS/MS) ubedo 0.0431–0.346 mg kg−1. I lembe ma juponjo pa Venkatasamy et al. (2010), yore mi green chemistry (ma epe ku acetonitrile) man reversed-phase high-performance liquid chromatography (RP-HPLC) jutiyo kudu pi pimo melamine i cak man cak. Kit ma piny ma jucaku kudu ubedo 1.0 ku 80 g/mL man adwogi ne ubedo linear (r > 0.999). Yore ne unyutho dwoko pa 97.2-101.2 iwi rwom pa 5-40 g/mL man dwoko piny ubedo piny pa 1.0% pa apoka poka. Bende, LOD man LOQ ma juneno ubedo 0.1 g mL−1 man 0.2 g mL−124, kubang wadi. Lutter et al. (2011) unwango nya melamine utiye i cak pa dhyeng man cak pa juma nyathin ku tic ku HPLC-UV. Kit pa melamine ubedo ikind < 0.2 ku 2.52 mg kg-1. Kite ma HPLC-UV ubedo kudo ubedo 0.05 ku 2.5 mg kg-1 pi cak pa dhyeng, 0.13 ku 6.25 mg kg-1 pi cak pa juma nyathin ku dul pa protein pa <15%, man 0.25 ku 12.5 mg kg-1 pi cak pa juma nyathin ku dul pa protein pa 15%. Adwogi pa LOD (man LOQ) ubedo 0.03 mg kg−1 (0.09 mg kg−1) pi cak pa dhyeng, 0.06 mg kg−1 (0.18 mg kg−1) pi cak pa nyathin <15% pa protein, man 0.12 mg kg−1 (0.36 mg kg11) pi cak pa nyathin <15% protein, pi nyutho nya ratio pa 3 ku 1025 pi LOD ku LOQ, kubang wadi. Diebes et al. (2012) utimo sayusac ikum kadhiri pa melamine i cam pa juma nyathin man cak ma jumaku ku HPLC/DMD. I cam pa juma nyathin, kadhiri ma piny man ma malu ubedo 9.49 mg kg−1 man 258 mg kg−1, kubang wadi. Kabedo mi nwangu (LOD) ubedo 0.05 mg kg−1.
Javaid et al. onyuthu nia melamine residues in cam pa awiya ubedo ikind 0.002-2 mg kg-1 ku Fourier transform infrared spectroscopy (FT-MIR) (LOD = 1 mg kg-1; LOQ = 3.5 mg kg-1). Rezai et al.27 uketho HPLC-DDA (λ = 220 nm) yore mi pimo melamine man ginwangu LOQ pa 0.08 μg mL−1 pi cak, ma ubedu piny uloyu ma junwango i somo eni. Sun et al. uketho RP-HPLC-DAD pi nwangu melamine i cak ma pii ku tic mi kwanyo piny ma tek (SPE). Ginwangu LOD ku LOQ pa 18 ku 60 μg kg−128, kubang wadi, ma etiye lembe ma pire tek uloyu ma sagu somo ma kawoni. Montesano et al. unyutho tic pa yore mi HPLC-DMD pi pimo kadhiri pa melamine i cam ma ju medo ku rwom pa 0.05-3 mg/kg, ma ubedu tek ungo ni yore ma jutiyo kudu i somo eni29.
Labongo jiji, kabedo mi sayu lembe timo tic ma pire tek i gwoko kwo pa piny ni kadhu kud i lubo tok piny ma rac i sampol matung tung. Ento, tic ku wel madwong pa reagents man solvents i saa mi ngiyo copo kelo yik pa residues ma rac. Pieno, green analytical chemistry (GAC) juyike i 2000 mi jwigo nyo daru adwogi marac pa yore mi ngiyo ikum jutic man kabedu26. Yore macon mi nwangu melamine macalo chromatography, electrophoresis, capillary electrophoresis, man enzyme-linked immunosorbent assay (ELISA) jutiyu kudu pi nwangu melamine. Ento, ikind yore madwong mi nwangu, electrochemical sensors ukelo nen pa dhanu madwong pi bedo migi maber, yero, sawa mi ngiyo piny pio pio, man bedo yot ni jutic30,31. Green nanotechnology tiyo ku yore mi kwo pi yubu nanomaterials, ma copo jwigo piny ma rac man tic ku kero, ma copo ketho tic ku yore maber. Nanocomposites, pi lapor, ma juyike ku piny ma nyayu peko i kwo pa dhanu, jucopo tiyo kudu i biosensors pi nwangu piny ma calu melamine32,33,34.
Somu eni nyutho nia solid-phase microextraction (SPME) jutiyu kudu maber pi tic maber pa kero man bedo maber ka juporo ku yore macon mi kwanyo piny. Bedo ber pa kabedu man tic maber pa SPME miyo ebedo yore maber pa yore macon mi kwanyo piny i analytical chemistry man emiyo yore maber man maber mi yiko lanyuth35.
I 2013, Wu et al. uketho piny ma tiyo ku rwom ma malu pa plasmon resonance (mini-SPR) ma tiyo ku dul pa melamine ku anti-melamine antibodies pi nwangu melamine pio pio i cam pa juma nyathin ku kony pa immunoassay. SPR biosensor ma juporo ku immunoassay (ma jutiyo kudu ku melamine-conjugated bovine serum albumin) utiye piny ma yot ni tiyo kudu man ma wel pare lapiny ku rwom pa 0.02 μg mL-136 kende.
Nasiri ku Abbasian gitiyo ku jam tic ma tek ma jutingo ku graphene oxide-chitosan composites (GOCS) pi nwangu melamine i lanyuth ma jucwalo37. Yore eni unyutho bedo maber mandha, bedo atira, man dwokowang lembe. Jam tic mi GOCS unyutho rwom ma malu (239.1 μM−1), rwom ma lageng pa 0.01 ku 200 μM, rwom ma rwate pa 1.73 × 104, man LOD ma romo 10 nM. Bende, somo moko ma Chandrasekhar et al. i 2024 utingo yore ma ber man ma ber pa piny. Gitiyo ku papaya peel extract calu piny ma jwigo piny pi yubu zinc oxide nanoparticles (ZnO-NPs) i yore ma ber pi kwo pa lei. I ngeye, yore moko ma pire tek mi micro-Raman spectroscopy juyubu pi nyang ikum melamine i cam pa juma nyathin. ZnO-NPs ma wok kud i lum mi poto onyuthu kero migi calu jam tic ma pire tek mi ngiyo two man diro ma gen, ma wel pare lapiny pi lubo man nwangu melamine38.
Alizadeh et al. (2024) otiyo ku highly sensitive metal-organic framework (MOF) fluorescence platform pi nyang ikum melamine in milk powder. Kabedo ma rwom pare piny man rwom ma piny pa sensor, ma juporo ku 3σ/S, ubedo 40 ku 396.45 nM (ma rom ku 25 μg kg−1 ku 0.25 mg kg−1) man 40 nM (ma rom ku 25 μg kg−1), kubang wadi. Kabedo eni utiye piny pa rwom ma malu (MRLs) ma juketho pi ngeyo melamine i cam pa juma nyathin (1 mg kg−1) man cam/camu mukende (2.5 mg kg−1). Jam tic ma ler (terbium (Tb)@NH2-MIL-253(Al)MOF) unyutho bedo maber man kero mi pimo maber loyo HPLC39 i nwangu melamine i cak. Biosensors man nanocomposites in green chemistry gi medo kero mi nwangu piny kende ngo ento gi dwoko piny peko mi kabedu ma lubu cik mi dongo ngom.
Cik mi chemistry macol jutiyo kudu i yore matung tung mi ngiyo melamine. Yore acel utiye mi yubu yore mi kwanyo piny ma nyai ma nyai ma tiyo ku polar polymer β-cyclodextrin ma juporo ku citric acid pi kwanyo melamine 40 kud i lanyuth macalo cak pa juma nyathin man pii ma lyet. Yo mange tiyo ku Mannich reaction pi nyang ikum melamine in cak. Yore eni yot ungo, eber ku kabedu, man eber mandha ku rwom ma lapiny pa 0.1-2.5 ppm man rwom ma lapiny mi nwangu 41. Bende, yore maber man ber pi kabedu pi nwangu wel pa melamine i cak ma pii man cak pa juma nyathin juyubu ku tic ku Fourier transform infraspect transmission ku rwom ma malu man rwom ma malu 1 ppm man 3.5 ppm, kubang acel acel 42. Yore eni nyutho tic pa cik mi green chemistry pi dongo yore maber man ma rii pi nyang ikum melamine.
Somo madwong uketho yore manyen mi nwangu melamine, calu tic ku solid-phase extraction man high-performance liquid chromatography (HPLC)43, man fast high-performance liquid chromatography (HPLC), ma mito ngo pre-treatment ma tek nyo ion-pair reagents, ma dwoko piny wel pa kemikol ma rac. Yore eni kende ngo ma mio adwogi maber pi nyang ikum melamine in piny ma juyubu ku cak, ento de lubu cik mi green chemistry, ma jwigo tic ku kemikol ma rac man jwigo adwogi marac ikum kwo pa dhanu.
Jam piny pyer angwen ma tung tung ju temo wang adek, man adwogi ne nen i Table 2. Kite ma melamine ubedo kudu i cak pa juma nyathin man cak ma jumaku ubedo 0.001 ku 0.004 mg/kg man 0.001 ku 0.095 mg/kg, kubang wadi. Aloka loka madit moko ope ma juneno ikind ju adek ma oro migi ubedu ka camu cak pa juma nyathin. Mi medo, melamine junwange i 80% pa cak ma buru, ento 65% pa cak pa juma nyathin ubedu ku melamine.
Melamine ma nwangre i cak ma juyubu i tic ubedo malo uloyu cak pa juma nyathin, man tung tung ne ubedo tek (p<0.05) (lanyut 2).
Adwogi ma junwango ubedo piny pa rwom ma FDA uketho (piny pa 1 ku 2.5 mg/kg). Mi medo, adwogi ne urombo ku cik ma juketho i CAC (2010) man EU45,46, niwacu rwom ma malu ma juyio utiye 1 mg kg-1 pi cam pa juma nyathin man 2.5 mg kg-1 pi cak.
Ni lubre ku lembe ma juponjo i 2023 pa Ghanati et al.47, wel melamine ma nwangre i kit cak matung tung i Iran ubedo 50.7 ku 790 μg kg−1. Adwogi migi ubedu piny pa rwom ma FDA yiyu. Adwogi mwa utiye piny uloyu pa Shoder et al.48 man Rima et al.49. Shoder et al. (2010) unwango ni kadhiri pa melamine in cak (n=49) ma juporo ku ELISA ubedo 0.5 ku 5.5 mg/kg. Rima ku juwadi. timo sayusac ikum melamine ma udong i cak ku fluorescence spectrophotometry man ginwangu niya melamine ma nwang i cak ubedo 0.72–5.76 mg/kg. Somo moko jutimo i Canada i 2011 pi neno kadhiri pa melamine i cam pa juma nyathin (n=94) ku tic ku liquid chromatography (LC/MS). Kit pa melamine junwango ni piny ma juyio (ma kwong: 0.5 mg kg−1). Ecopo bedu ngo nia wel pa melamine ma junwango ubedo yore moko ma jutiyo kudu pi medo wel pa protein. Ento, ecopu ngo ni gonyo ku tic ku yath, loko kabedo pa piny ma i iye, nyo lembe mange ma rom ku meno. Bende, kama melamine uai i cak ma jucwalo i Canada ungiyu ngo50.
Hassani et al. upimo wel pa melamine i cak ma buru man cak ma pii i soko mi Iran i 2013 man ginwangu adwogi marom. Adwogi ne unyutho nia muweko kit cak acel man cak ma pii, lanyuth mukende zo ubedu ku melamine, ku kadhiri nicaku 1.50 ku 30.32 μg g−1 i cak man 0.11 ku 1.48 μg ml−1 i cak. Ma pire tek, cyanuric acid junwange ngo i lanyuth moku ci, ma jwigo peko pa melamine ni juwil. 51 Somo mukadho upimu rwom pa melamine in chocolate ma juyubu ku cak. Ikind 94% pa jam piny ma jucwalo kudi ngom mawoko man 77% pa piny ma jucwalo kud i Iran ubedu ku melamine. Kite ma melamine utiye kudu i lanyuth ma jucwalo kudi ngom mawoko ucaku 0.032 ku 2.692 mg/kg, man ma nwangire i lanyuth pa Iran ucaku 0.013 ku 2.600 mg/kg. Ikum zoo, melamine junwango i 85% pa lanyuth, ento piny acel kende re ma nwango rwom ma malu pa rwom ma juyiyu.44 Tittlemier et al. uketho rwom pa melamine in cak ma nyai nicaku 0.00528 nicidho 0.0122 mg/kg.
Table 3 nyuthu adwogi mi pimo peko pa dul adek mi oro. Peko ubedo piny pa 1 ikind oro zo. Pieno, epe peko mi yot kum ma nyayu two kud i melamine i cak pa juma nyathin.
Kite ma piny ma rac pa piny ma juyubu ku cak copo bedu ni kum nyayu ma rac ma jutimo ngo i saa mi yiko, ento kadhiri ma malu copo bedu ni kum medo piny ma jutimo ku paru. Bende, peko ma copo wok ikum yot kum pa dhanu niai kud i camu piny ma juyubu ku cak ma melamine nok i iye nen ni nok. Ecopu bedu ni camu piny ma tye ku melamine manok epe ku peko moku ikum yot kum pa ju ma camu52.
Ka waparu pi beru pa gwoko leng pa cam i tic mi cak, asaga ikum gwoko yot kum pa dhanu, piretek ni yubu man ni ketho yore moko mi pimo man poro kadhiri pa melamine man residues i cak man cam pa juma nyathin. Yore mayot man maber pa HPLC-UV spectrophotometric juyike pi nyang ikum melamine i cam pa juma nyathin man cak ma buru. Yore ne juyike pi ni neno nia etiye maber man ma atira. Nwangu man kwan pa yore ne unyutho nia etiye maber akeca pi pimo kadhiri pa melamine i cam pa juma nyathin man cak ma buru. Nimakere ku lembe mu'a, melamine junwango i pol pa lanyuth pa Iran. Kabedo pa melamine ma junwango ubedu piny pa rwom ma CAC ungolo, ma nyutho nia camu piny ma juyubu ku cak eni nyayu ngo peko ikum yot kum pa dhanu.
Jam tic ceke ma jutiyo kudu ubedu mi rwom ma malu: melamine (2,4,6-triamino-1,3,5-triazine) 99% maleng (Sigma-Aldrich, St. Louis, MO); HPLC-grade acetonitrile (Merck, Darmstadt, Germany); pii maleng akeca (Millipore, Morfheim, France). Jam tiyo ku syringe (Chromafil Xtra PVDF-45/25, pore size 0.45 μm, membrane diameter 25 mm) (Macherey-Nagel, Düren, Germany).
Ultrasonic bath (Elma, Germany), centrifuge (Beckman Coulter, Krefeld, Germany) man HPLC (KNAUER, Germany) jutiyo kudu pi yiko lanyuth.
Jam tic ma tiyo maber akeca (KNAUER, Germany) ma juketho i iye UV detector jutiyo kudu. Lembe ma jutimo i HPLC ubedo kudu: UHPLC Ultimate system ma juketho i iye ODS-3 C18 analytical column (4.6 mm × 250 mm, particle size 5 μm) (MZ, Germany) jutiyo kudu. HPLC eluent (mobile phase) ubedo TFA/methanol mixture (450:50 mL) ku flow rate pa 1 mL min-1. Bor pa yamo ma junwango ubedo 242 nm. Dwong pa yath ubedo 100 μL, lyethu pa column ubedo 20 °C. Calu ma kare mi gwoko yath ubedo malac (dakika 15), yath mukende umaku jutim ingey dakika 25. Melamine ungiyure ku poro kare mi gwoko man UV spectrum peak pa melamine.
Jam piny ma julwongo ni melamine (10 μg/mL) juyike ku pii man jugwoko i friji (4 °C) mabor ku lero. Dilute stock solution ku mobile phase man iyub tic maber. Piny acel acel ma juyiko juyike i HPLC wang 7. Equation mi calibration 10 ju kwanu ku regression analysis pa peak area ma jumoko man concentration ma jumoko.
Cak dyang ma jucwalo (20 samples) man samples pa cak dyang matung tung (20 samples) juwilo kud i duka madongo man ot yath i Iran pi miyo cam ni awiya ma oro migi tung tung (dwi 0-6, dwi 6-12, man >dwi 12) man jugwoko i friji (4°C) nitundu jutimo sayusac. I ngeye, 1 ± 0.01 g pa cak ma juyiko maber jupimo man ju rubu ku acetonitrile:pi (50:50, v/v; 5 mL). Jam rubu ne juyike pi dakika acel, man juyike i ultrasonic bath pi dakika 30, man i ngeye juyike pi dakika acel. I ngeye, juyiko jamb piny ne i 9000 × g pi dakika 10 i lyethu mi ot man juyiko piny ma udong i 2 ml autosampler vial ma jutiyo kudu ku 0.45 μm syringe filter. Filtrate (250 μl) ju rubu ku pii (750 μl) man ju ketho i HPLC system10,42.
Mi ketho yore ne utiye andha, wa nwango dwoko piny, bedo atira, rwom mi nwangu (LOD), rwom mi kwanu (LOQ), man rwom maber i thenge maber. LOD juloko ni piny ma juporo ku bor ma malu wang adek pa dwand ma kwong. Icing ling acel, piny ma jukwanu ku bor ma malu wang 10 pa signal-to-noise ratio juloko ni LOQ.
Adwogi pa jam tic ne ungiyu ku tiyo ku calibration curve ma uketho i iye data points abiro. Melamine matung tung jutiyo kudu (0, 0.2, 0.3, 0.5, 0.8, 1 man 1.2). Linearity pa yore mi kwano melamine ungiyure. Mi medo, rwom matung tung pa melamine jumedo i lanyuth ma oling. Calibration curve juyike ku medo 0.1-1.2 μg mL-1 pa melamine solution i cak pa juma nyathin man cak ma juyubu man R2 = 0.9925. Ber pa lembe ne jupimu ku timo man dwoko wang lembe man juyubu ku ketho lanyuth i ceng mir acel man adek ma ulubo (i wang adek). Repeatability pa yore ne jupimu ku kwanu RSD % pi rwom adek matung tung pa melamine maju medo. Somo mi dwoko piny jutimo pi ni nyang kudu lemandha. Kite ma judwoko kudu piny ku yore mi kwanyo piny eca ju kwanu i rwom adek pa melamine (0.1, 1.2, 2) i lanyuth pa cam pa juma nyathin man cak ma muthwo9,11,15.
Piny ma jucamu kubang ceng (EDI) juporo ku formula ma e: EDI = Ci × Cc/BW.
Ka Ci utiye wel pa melamine, Cc utiye cak ma jucamu man BW utiye pek pa awiya.
Kwano lembe jutimo ku SPSS 24. Kite ma jutimo ku Kolmogorov-Smirnov test; lembe zo ubedo nonparametric tests (p = 0). Pieno, Kruskal-Wallis test man Mann-Whitney test jutiyo kudu pi nyang tung tung madit ikind ju.
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