Evaluation of Extraction Buffers for Protein Identification in Fish Paste with Chicken Blood Plasma (CBP) Spikes: A Preliminary Mass Spectrometry Study

Nurhazirah Azmi; Siti Aimi Sarah Zainal Abidin; Nurkhurul Ain Zakaria; Mohd Syarafuddin Abdul Shukor; Saiful Anuar Karsani; Low Kim Fatt; Mohd Hafis Yuswan

doi:10.22146/ijc.100755

Evaluation of Extraction Buffers for Protein Identification in Fish Paste with Chicken Blood Plasma (CBP) Spikes: A Preliminary Mass Spectrometry Study

https://doi.org/10.22146/ijc.100755

Nurhazirah Azmi⁽¹⁾, Siti Aimi Sarah Zainal Abidin^(2*), Nurkhurul Ain Zakaria⁽³⁾, Mohd Syarafuddin Abdul Shukor⁽⁴⁾, Saiful Anuar Karsani⁽⁵⁾, Low Kim Fatt⁽⁶⁾, Mohd Hafis Yuswan⁽⁷⁾

(1) Faculty of Applied Sciences, Universiti Teknologi MARA, Perak Branch, Tapah Campus, Tapah Road, Perak 35400, Malaysia; Faculty of Applied Sciences, Universiti Teknologi MARA, Shah Alam, Selangor 40450, Malaysia
(2) Faculty of Applied Sciences, Universiti Teknologi MARA, Shah Alam, Selangor 40450, Malaysia; Malaysia Institute of Transport (MITRANS), Universiti Teknologi MARA, Shah Alam, Selangor 40450, Malaysia
(3) Faculty of Applied Sciences, Universiti Teknologi MARA, Shah Alam, Selangor 40450, Malaysia
(4) Faculty of Applied Sciences, Universiti Teknologi MARA, Shah Alam, Selangor 40450, Malaysia
(5) Institute of Biological Sciences, Faculty of Science, Universiti Malaya, Kuala Lumpur 50603, Malaysia; Universiti Malaya Center for Proteomics Research (UMCPR), Universiti Malaya, Kuala Lumpur 50603, Malaysia
(6) Faculty of Applied Sciences, Universiti Teknologi MARA, Perak Branch, Tapah Campus, Tapah Road, Perak 35400, Malaysia
(7) Halal Products Research Institute, Universiti Putra Malaysia, Serdang, Selangor 43400, Malaysia
(*) Corresponding Author

Abstract

Blood plasma is commonly used as a protease inhibitor in surimi production to improve product quality and stability during processing. However, its use in food is prohibited in Islam and classified as najs al-mutawasitah, necessitating the detection of blood plasma adulteration to uphold halal standards and food safety. This study compared the effectiveness of different extraction buffers—ultrapure water, 0.05 M Tris-HCl, 0.05 M Tris-Urea, and ultrapure water followed by acetone precipitation—on protein and peptide yield from chicken blood plasma (CBP) using liquid chromatography–quadrupole linear ion trap mass spectrometry (LC-QTRAP-MS). Ultrapure water and acetone precipitation yielded the highest protein content, prompting further proteome profiling of CBP, fish paste, and surimi spiked with CBP (0.5, 1, and 1.5%) via liquid chromatography–quadrupole time-of-flight mass spectrometry (LC–QTOF-MS). Apolipoprotein AI (Apo AI) and fibrinogens emerged as key proteins in CBP. Apo AI was detected in all spiked surimi samples, demonstrating its potential as a marker for blood plasma contamination. The proposed method enhances extraction and detection protocols, using mass spectrometry to provide a reliable tool for addressing halal compliance and mitigating food safety risks associated with blood-derived adulterants in surimi products.

Keywords

mass spectrometry; chicken blood plasma; spiked sample; Apolipoprotein AI; fibrinogen

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DOI: https://doi.org/10.22146/ijc.100755