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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 TAN1*,
LEE
YOOK
HENG1,2 & 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 Received: 13 April 2018/Accepted:
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 S2-. 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
S2- ions. Keywords: Acrylic microsphere; biosensor; hemoglobin; nitrite determination REFERENCES
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