Pengembangan Metode Penetapan Kadar Campuran Pemanis, Pengawet dan Pewarna Secara Simultan dalam Sirup Esens dengan Menggunakan Kromatografi Cair Kinerja Tinggi

Abstract

Syrup is concentrated sugar solution with or without the addition of food additives. Sweeteners, preservatives and dyes can be hazardous to health if they are over consumed. The purpose of this research was to develop a method for the simultaneous determination of sodium saccharin, sodium cyclamate, sodium benzoate, potassium sorbate, tartrazine and sunset yellow in essence syrup. This research used high performance liquid chromatography reverse phase, three wavelengths with UFLC 1290 DAD (Agilent), column C18 100 mm x 4.6 mm x 3.5 μm (Agilent), spectrophotometer UV Probe 1800 (Shimadzu), standard material sodium saccharin, sodium cyclamate, sodium benzoate, potassium sorbate, tartrazine and sunset yellow. Optimization parameters were include void volume, wavelengths, pH of mobile phase, composition of mobile phase, flow rate and column temperature. Validation parameters were include linearity, limit of detection, limit of quantitation, accuracy, precision and selectivity. The method obtained was used for the simultaneous determination of sodium saccharin, sodium cyclamate, sodium benzoate, potassium sorbate, tartrazine and sunset yellow in essence syrup samples. A total of six essence syrup samples were obtained from wholesale and supermarket in Medan City and given code H, I, J, K, L and M. The results of reseach showed that the optimum conditions were void volume 30% with three-wavelength analysis, i.e: 200 nm for sodium cyclamate; 220 nm for sodium saccharin, sodium benzoate and potassium sorbate; 450 nm for tartrazine and sunset yellow. Mobile phase was phosphate buffer pH 4.5 and methanol 75: 25 (v/v), flow rate was 1.0 ml/min, column temperature was 30oC. The range of retention time was 0.941 minute - 8.583 minute. The results of validation method showed that the ranges of linearity, limit of detection, limit of quantitation, accuracy, percent RSD of the repeatability and reproducibility methods were 0.99945 - 0.99999; 0.03634 ppm - 2.66306 ppm; 0.12113 ppm - 8.87687 ppm; 92.92% - 105.72%; 0.11% - 1.53% and 0.04% - 1.94%, respectively. The selectivity test of methods showed good results. The result showed that potassium sorbate is not contained in essence syrup samples. The ranges of levels of sodium saccharin, sodium cyclamate, sodium benzoate, tartrazine and sunset yellow in the samples of essences syrup were 37.952 mg/kg - 533.990 mg/kg; 2753.140 mg/kg - 5329.890 mg/kg; 464.456 mg/kg - 1615.360 mg/kg; 41.957 mg/kg - 108.048 mg/kg and 31.084 mg/kg - 145.399 mg/kg, respectively. Samples of essences syrup contain of sodium saccharin for the I code and sodium benzoate for the H, J and K codes exceed the maximum usage limit. All samples syrup contain of sodium cyclamate essences exceed the maximum usage limit.

Sirup merupakan larutan gula pekat dengan atau tanpa penambahan bahan tambahan pangan. Bahan pemanis, pengawet dan pewarna dapat mengganggu kesehatan bila dikonsumsi berlebihan. Tujuan penelitian adalah untuk mengembangkan metode penetapan kadar campuran natrium sakarin, natrium siklamat, natrium benzoat, kalium sorbat, tartrazin dan sunset yellow dalam sirup esens. Penelitian menggunakan kromatografi cair kinerja tinggi fase terbalik, tiga panjang gelombang dengan instrumen UFLC 1290 DAD (Agilent), kolom C18 100 mm x 4,6 mm x 3,5 μm (Agilent), spektrofotometer UV Probe 1800 (Shimadzu), bahan baku natrium sakarin, natrium siklamat, natrium benzoat, kalium sorbat, tartrazin dan sunset yellow. Parameter optimasi meliputi volume void, panjang gelombang, pH fase gerak, komposisi fase gerak, laju alir dan suhu kolom. Parameter validasi meliputi linearitas, batas deteksi, batas kuantitasi, akurasi, presisi dan selektivitas. Metode yang diperoleh digunakan untuk penetapan kadar campuran natrium sakarin, natrium siklamat, natrium benzoat, kalium sorbat, tartrazin dan sunset yellow secara simultan dalam sampel sirup esens. Sebanyak enam sampel sirup esens diperoleh dari grosir dan supermarket di Kota Medan dan diberi kode H, I, J, K, L dan M. Hasil penelitian menunjukkan bahwa kondisi optimum pengujian adalah volume void 30% dengan tiga panjang gelombang analisis, yaitu: 200 nm untuk natrium siklamat; 220 nm untuk natrium sakarin, natrium benzoat dan kalium sorbat; 450 nm untuk tartrazin dan sunset yellow. Fase gerak buffer fosfat pH 4,5 dan metanol 75 : 25 (v/v), laju alir 1,0 ml/menit, suhu kolom 30oC. Rentang waktu retensi adalah 0,941 menit - 8,583 menit. Hasil validasi metode menunjukkan bahwa rentang linearitas, batas deteksi, batas kuantitasi, akurasi, persen RSD keterulangan dan ketertiruan metode masing-masing 0,99945 – 0,99999; 0,03634 ppm - 2,66306 ppm; 0,12113 ppm - 8,87687 ppm; 92,92% - 105,72%; 0,11% - 1,53% dan 0,04% – 1,94%. Uji selektivitas metode menunjukkan hasil yang baik. Hasil penelitian menunjukkan bahwa kalium sorbat tidak terkandung dalam sampel sirup esens. Rentang kadar natrium sakarin, natrium siklamat, natrium benzoat, tartrazin dan sunset yellow dalam sampel sirup esens masing-masing 37,952 mg/kg – 533,990 mg/kg; 2753,140 mg/kg – 5329,890 mg/kg; 464,456 mg/kg – 1615,360 mg/kg; 41,957 mg/kg - 108,048 mg/kg dan 31,084 mg/kg – 145,399 mg/kg. Sampel sirup esens mengandung kadar natrium sakarin untuk kode I dan natrium benzoat untuk kode H, J dan K melebihi batas penggunaan maksimum. Semua sampel sirup esens mengandung natrium siklamat melebihi batas penggunaan maksimum.