Analisis Kuantitatif Berbasis HPLC untuk Senyawa Antioksidan BHA dan TBHQ pada Margarin

Abstract

Margarin merupakan produk pangan berbasis lemak yang rentan terhadap oksidasi lipid akibat kandungan asam lemak tak jenuh yang tinggi. Antioksidan sintetis seperti butylated hydroxyanisole (BHA) dan tert-butylhydroquinone (TBHQ) umumnya ditambahkan untuk mencegah oksidasi, namun kadarnya perlu dipantau sesuai regulasi keamanan pangan. Penelitian ini bertujuan untuk memvalidasi metode HPLC-UV dalam penentuan kadar BHA dan TBHQ pada margarin berdasarkan AOAC Official Methods of Analysis (2016). Preparasi sampel dilakukan melalui ekstraksi metanol:asetonitril (1:1), sonikasi, sentrifugasi, dan filtrasi. Validasi meliputi uji kesesuaian sistem, linearitas, akurasi, presisi, LOD, LOQ, dan robustness. Nilai %RSD kesesuaian sistem sebesar 0,72% (BHA) dan 1,07% (TBHQ) memenuhi kriteria =2%, dengan linearitas R² = 0,9999 pada rentang 1–50 mg/L. Perolehan kembali rata-rata sebesar 101,64% dan 102,24% memenuhi kriteria AOAC. Kadar BHA dan TBHQ dalam sampel margarin masing masing sebesar 89,04 mg/kg dan 1,26 mg/kg, di bawah batas maksimum 200 mg/kg. Metode ini dinyatakan valid dan layak digunakan untuk analisis rutin.

Margarine is a fat-based food product susceptible to lipid oxidation due to its high unsaturated fatty acid content. Synthetic antioxidants such as butylated hydroxyanisole (BHA) and tert-butylhydroquinone (TBHQ) are commonly added to prevent oxidation, but their concentrations must be monitored in accordance with food safety regulations. This study aimed to validate an HPLC-UV method for the determination of BHA and TBHQ in margarine based on AOAC Official Methods of Analysis (2016). Sample preparation was performed by extraction using methanol–acetonitrile (1:1), followed by sonication, centrifugation, and filtration. Validation parameters included system suitability, linearity, accuracy, precision, LOD, LOQ, and robustness. System suitability %RSD values of 0.72% (BHA) and 1.07% (TBHQ) met the =2% criterion, with linearity R² = 0.9999 over 1–50 mg/L. Mean recoveries of 101.64% and 102.24% met AOAC criteria. BHA and TBHQ concentrations in margarine were 89.04 mg/kg and 1.26 mg/kg, both below the 200 mg/kg maximum limit. The method is valid and suitable for routine analysis.