[ABSTRAK
Kurkumin merupakan senyawa polifenol yang umumnya terdapat padarimpang kunyit (Curcuma longa L.). Setelah pemberian peroral, kurkumin dalamtubuh akan segera dimetabolisme melalui proses reduksi maupun konjugasi. Olehkarena itu, kadar kurkumin di dalam darah sangat kecil sehingga diperlukan metodebioanalisis yang selektif dan sensitif. Metode Kromatografi Cair Kinerja Ultra Tinggi? Tandem Spektrometer Massa (KCKUT-SM/SM) yang spesifik dan cepat telahdikembangkan dan divalidasi untuk menetapkan kadar kurkumin dalam plasmamanusia menggunkan diazepam sebagai baku dalam. Pemisahan dilakukanmenggunakan kolom C18 Acquity® Waters, UPLC BEH 1,7 μm, 2,1 x 100 mm, fasegerak asam format 0,15% - Asetonitril (50:50), laju alir 0,5 mL/menit dengan metodepreparasi sampel ekstraksi cair-cair menggunakan campuran larutan etil asetatmetanol(95:5). Mode ionisasi yang digunakan adalah multiple reaction monitoring(MRM) dengan mode Electrospray ionization positif dengan nilai m/z berturut-turut369,05 > 176,95 dan m/z 284,95 > 193 untuk kurkumin dan diazepam. Metodebioanalisis menunjukkan presisi dan akurasi yang baik dengan nilai % KV dan % bias< 15% untuk semua konsentrasi (QCL, QCM dan QCH) dengan nilai kurva kalibrasiyang linear (r = 0,999) pada rentang 1 ? 100 ng/mL dan nilai LLOQ untuk senyawakurkumin sebesar 1,0 ng/mL. Metode ini telah diaplikasikan untuk menentukkankadar kurkumin dalam plasma 1 orang sehat yang telah diberi sediaan kurkumin 1800mg. Dari penelitian diperoleh hasil tidak ditemukannya kurkumin dalam bentukbebas, tetapi bentuk kurkumin terglukuronidasi dan tersulfatasi. Perbandingan antarajumlah terglukuronidasi dan tersulfatasi 4:1. Metode analisis yang diperoleh sudahmemenuhi kriteria validitas menurut Guidance EMEA 2011 dengan sensitivitas yangtinggi sehingga dapat diaplikasikan untuk studi in-vivo

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ABSTRACT
Curcumin is a polyphenol, found in the spice turmeric from the rhizome of the herbCurcuma Longa. After oral administration, Curcumin undergoes rapid metabolism byconjugation and reduction. Curcumin levels are generally low so that the requiredbioanalytical method is selective and sensitive. A simple, specific and rapid UPLCMS/MS method has been developed and validated for the estimation of curcumin inhuman plasma, using diazepam as internal standard (IS). The separation using UPLCBEH C18 column 1.7 μm, 2,1 x 100 mm Acquity® Waters; 0.15% formic acid -acetonitril (50:50, v/v) as mobile phase; flow rate 0.5 mL/min; using liquid-liquidextraction with the mixture of ethyl acetate-methanol (95:5) for the samplepreparation. The ionization mode using electrospray ionization (ESI) detection inmultiple reaction monitoring (MRM) in positive ionization mode. The MS/MS iontransitions monitored were m/z 369.05 >176.95 and 284.95 > 193 for curcumin anddiazepam respectively. The method was proved to be precise and accurate (expressedas coefficient of variation, % CV and differentiation, % diif) was < 15% for allconcentration (QCL, QCM and QCH) with a coefficient correlation ( r = 0.999)and linearity range of 1 ? 100 ng/mL, LLOQ for curcumin was 1 ng/mL. TheMethod was applicated to determine the level of curcumin in healthy subject afteroral administration 1800 mg of curcumin dosage form. No Free curcumin wasdetected in plasma sample, but curcumin glucuronides and sulfates were detected inplasma subject. The ratio of glucuronide to sulfate was 4: 1. The analytical methodfullfilthe criteria of validity by the EMEA Guidance 2011 with high sensitivity and itwould be applicable to in-vivo study., Curcumin is a polyphenol, found in the spice turmeric from the rhizome of the herbCurcuma Longa. After oral administration, Curcumin undergoes rapid metabolism byconjugation and reduction. Curcumin levels are generally low so that the requiredbioanalytical method is selective and sensitive. A simple, specific and rapid UPLCMS/MS method has been developed and validated for the estimation of curcumin inhuman plasma, using diazepam as internal standard (IS). The separation using UPLCBEH C18 column 1.7 μm, 2,1 x 100 mm Acquity® Waters; 0.15% formic acid -acetonitril (50:50, v/v) as mobile phase; flow rate 0.5 mL/min; using liquid-liquidextraction with the mixture of ethyl acetate-methanol (95:5) for the samplepreparation. The ionization mode using electrospray ionization (ESI) detection inmultiple reaction monitoring (MRM) in positive ionization mode. The MS/MS iontransitions monitored were m/z 369.05 >176.95 and 284.95 > 193 for curcumin anddiazepam respectively. The method was proved to be precise and accurate (expressedas coefficient of variation, % CV and differentiation, % diif) was < 15% for allconcentration (QCL, QCM and QCH) with a coefficient correlation ( r = 0.999)and linearity range of 1 – 100 ng/mL, LLOQ for curcumin was 1 ng/mL. TheMethod was applicated to determine the level of curcumin in healthy subject afteroral administration 1800 mg of curcumin dosage form. No Free curcumin wasdetected in plasma sample, but curcumin glucuronides and sulfates were detected inplasma subject. The ratio of glucuronide to sulfate was 4: 1. The analytical methodfullfilthe criteria of validity by the EMEA Guidance 2011 with high sensitivity and itwould be applicable to in-vivo study]