Sains Malaysiana 47(9)(2018): 2027–2033

http://dx.doi.org/10.17576/jsm-2018-4709-10

Biosensor Nitrit Optik Berasaskan Hemoglobin Terpegun pada Mikrosfera Poliakrilat

(Optical Nitrite Biosensor based on Hemoglobin-Immobilized Polyacrylate Microspheres)

SYAMIMI HASLAN¹, LING LING TAN^1*, LEE YOOK HENG^1,2 & GOH CHOO TA¹

¹Pusat Kajian Bencana Asia Tenggara (SEADPRI-UKM), Institut Alam Sekitar dan Pembangunan (LESTARI), Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan,Malaysia

²Pusat Pengajian Sains Kimia dan Teknologi Makanan, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

Diserahkan: 13 April 2018/Diterima: 18 Mei 2018

ABSTRAK

Biosensor optik berasaskan hemoglobin (Hb) terpegun pada mikrosfera poli(n-butil akrilat-co-N-akriloksisuksinimida) [poli(nBA-NAS)] telah dibangunkan bagi mengesan kepekatan ion nitrit (NO₂-). Kompleks HEM ferum dalam Hb memangkinkan tindak balas penurunan ion NO₂- kepada nitrik oksida (NO) lalu bergabung dengan deoksihemoglobin (HbFe²⁺) membentuk kompleks ferum-nitrosil-hemoglobin (HbFe²⁺-NO) yang berwarna hijau kekuningan. Spektrofotometer pantulan gentian optik digunakan untuk memantau kepekatan ion NO₂- secara kuantitatif berdasarkan perubahan warna Hb terpegun pada mikrosfera poliakrilat daripada perang kemerahan ke hijau kekuningan pada panjang gelombang pantulan maksimum 668 nm. Pencirian terhadap biosensor nitrit reflektometrik melibatkan ujian kesan pH, kesan kepekatan Hb, julat rangsangan linear, kebolehasilan, jangka hayat dan kesan gangguan ion telah dijalankan. Biosensor ion NO₂- optik terbangun memaparkan julat linear dinamik daripada 5 hingga 50 mg mL^-1 (R²=0.9894) pada pH7.0 dengan had pengesanan (LOD) sebanyak 3.3 mg mL^-1 dan nilai sisihan piawai relatif (RSD) kebolehasilan biosensor sebanyak 5.8%. Jangka hayat biosensor optik nitrit tersebut adalah selama 36 hari dan majoriti ion asing yang sering wujud bersama ion NO₂- dalam sampel air semula jadi tidak menunjukkan kesan gangguan yang bererti terhadap penentuan ion NO₂- menggunakan biosensor optik terbangun kecuali ion Hg²⁺, Ag⁺, Br^- dan S^2-.

Kata kunci: Biosensor; hemoglobin; mikrosfera akrilik; penentuan nitrit

ABSTRACT

Optical biosensor based on hemoglobin (Hb) immobilized on the poly(n-butyl acrylate-co-N-acryloxysuccinimide) microspheres has been developed for the detection of nitrite (NO₂-) ion concentration. The iron HEM complex in the Hb catalyzed the reduction of NO₂- ion to nitric oxide (NO) followed by binding with deoxyhemoglobin (HbFe²⁺) to form the yellowish green iron-nitrosyl-hemoglobin (HbFe²⁺-NO) complex. Fiber optic reflectance spectrophotometer was used to monitor the NO₂- ion concentration quantitatively based on colour change of the immobilized Hb on the polyacrylate microspheres from reddish brown to yellowish green at the maximum reflectance wavelength of 668 nm. Characterization of the reflectometric nitrite biosensor encompassed pH effect, Hb concentration effect, linear response range, reproducibility, shelf life and ion interference effect studies had been conducted. The developed optical NO₂- ion biosensor exhibited dynamic linear range from 5 to 50 mg mL^-1 (R²=0.9894) at pH7.0 with a limit of detection (LOD) of 3.3 mg mL^-1 and reproducibility relative standard deviation (RSD) value of the biosensor at 5.8%. The shelf life of the optical nitrite biosensor is about 36 day and majority foreign ions that are often co-exist with NO₂- ion in the natural water sample did not show significant interference effect towards the determination of NO2- ion using the developed optical biosensor except Hg²⁺, Ag⁺, Br^- and S^2- ions.

Keywords: Acrylic microsphere; biosensor; hemoglobin; nitrite determination

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 *Pengarang untuk surat-menyurat; email: lingling@ukm.edu.my