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Sampel HIDROLISIS PATI HIDROLISIS PROTEIN pH 6.0, Termamyl, 95-100 oC, 30 menit pH 7.5, protease, 60 oC, 30 menit HIDROLISIS PATI pH 4.0-4.6, amyloglukosidase, 60 oC, 30 menit PRESIPITASI DF ETOH 95%, Suhu kamar, 60 menit FILTRASI Porositas 2, celite 0.5 g Residu PENCUCIAN RESIDU PENGERINGAN BERAT KONSTAN Total DF 3 x 20 ml ETOH 78% 2 x 10 ml ETOH 95% 1 x 10 ml Acetone 70 oC vacum, semalam atau 105 oC , 5 jam. Koreksi kadar protein & abu DIAGRAM ALIR PENENTUAN TOTAL DF (AOAC, 1995) Sampel + guargum HIDROLISIS PROTEIN pepsin HCl, 37 oC, 30 menit HIDROLISIS PATI buffer asetat, 37 oC Invertase, amylase, Amyloglukosidase INKUBASI 20 MENIT PEMISAHAN GLUKOSA ETOH 66% Centrifugasi PENENTUAN GLUKOSA INKUBASI 120 MENIT PEMISAHAN GLUKOSA ETOH 66% Centrifugasi PENENTUAN GLUKOSA HIDROLISIS PATI GELATINISASI RS HIDROLISIS RS INAKTIVASI ENSIM PEMISAHAN GLUKOSA PENENTUAN GLUKOSA RAG G120 100 oC , 30 menit 2M KOH, 0 oC, 30 menit as. asetat, amyloglukosidase 70 oC, 30 menit 100 oC , 10 menit dingin, pengenceran sentrifugasi Total Glukosa (TG) RS = 0.9 (TG-G120) DIAGRAM ALIR ANALISIS RS (Englyst et. al., 1992) Sampel EKSTRAKSI GULA ETOH 80% EKSTRAKSI LEMAK Petroleom ether HIDROLISIS PATI (I) 95-100 oC , 30 menit Amylase HIDROLISIS PATI (II) INAKTIVASI ENSIM 37-40 oC , semalam Amyloglukosidase, pullulanase mendidih, 15 menit PRESIPITASI TDF & RS sentrifugasi dingin Endapan (TDF & RS) Supernatan (glukosa/pati) HIDROLISIS RS (I) 2 M KOH, 30 menit Amylase HIDROLISIS RS (II) INAKTIVASI ENSIM 37-40 oC , semalam Amyloglukosidase, pullulanase mendidih, 15 menit PRESIPITASI TDF sentrifugasi dingin Supernatan (glukosa) Endapan (TDF) PENENTUAN GLUKOSA Spektrofotometri Glukosa (RS) DIAGRAM ALIR PENENTUAN RS (Englyst & Cummings, 1987) Sampel basis: 1 g total pati PENGUNYAHAN 15 kali, 15 detik INKUBASI I pepsin, pH 1.5 (+HCl) 37 OC, 30 menit INKUBASI II pankreatin, amyloglukosidase pH 5.0; 40 OC , 16 jam PRESIPITASI ETOH 95%, 4 x volume 60 menit FILTRASI porositas 2(40-90 um) 0.5 g celite Filtrat PENENTUAN GLUKOSA Endapan ANALISIS N & abu Pati % Protein & abu GELATINISASI RS 2M KOH, 30 menit HIDROLISIS RS Amyloglukosidase PENENTUAN GLUKOSA RS DIAGRAM ALIR ANALISIS RS (Akerberg et al., 1998) Sampel HIDROLISIS PATI HIDROLISIS PROTEIN pH 6.0, Termamyl, 95-100 oC, 15 (30) menit pH 7.5, protease, 60 oC, 30 menit HIDROLISIS PATI pH 4.0-4.6, amyloglukosidase, 60 oC, 30 menit PENCUCIAN SDF 2 x 10 ml air 2 x 10 ml ETOH 95% 1 x 10 ml Acetone Residu (IDF) PENGERINGAN BERAT KONSTAN IDF 70 oC vacum, semalam atau 105 oC , 5 jam. Koreksi kadar protein & abu DIAGRAM ALIR PENENTUAN INSOLUBLE DIETARY FIBRE (IDF) , AOAC, 1995 Sampel HIDROLISIS PATI MES-TRIS Buffer (pH 8.2, α- amilase, 95-100 oC, 15 (35) menit HIDROLISIS PROTEIN HIDROLISIS PATI FILTRASI pH 7.5, protease, 60 oC, 30 menit pH 4.0-4.7, amyloglukosidase, 60 oC, 30 menit Filtrat Residu (IDF) SDF Air cucian PENCUCIAN I 2 x 10 ml air 70 oC PENCUCIAN II 2 x 15 ml ETOH 78% 2 x 15 ml ETOH 95% 2 x 15 ml aseton 70 oC vacum, semalam atau 105 oC , 5 jam. PENGERINGAN BERAT KONSTAN IDF Koreksi kadar protein & abu DIAGRAM ALIR PENENTUAN SDF DAN IDF, AOAC, 1995