Trad. Med. J., May - August 2017 Vol. 22(2), p 124-130 ISSN-p : 1410-5918 ISSN-e : 2406-9086 124 Traditional Medicine Journal, 22(2), 2017 Submitted : 01-02-2017 Revised : 24-07-2017 Accepted : 16-08-2017 Antituberculosis Activity of Brotowali (Tinospora crispa) Extract and Fractions against Mycobacterium tuberculosis using Microplate Alamar Blue Assay Method Aktivitas Antituberkulosis Ekstrak dan Fraksi Batang Brotowali (Tinospora crispa) terhadap Mycobacterium tuberculosis dengan Metode Microplate Alamar Blue Assay Retno Wahyuningrum 1,2* , Ritmaleni 1 , Tatang Irianti 1 , Subagus Wahyuono 1 , Takushi Kaneko 3 , Titik Nuryastuti 4 1 Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Indonesia 2 Faculty of Pharmacy Universitas Muhammadiyah Purwokerto, Indonesia 3 Tuberculosis Alliance, New York, United State 4 Faculty of Medicine Universitas Gadjah Mada, Yogyakarta, Indonesia ABSTRACT Tuberculosis (TB), in which caused by pathogenic bacteria, Mycobacterium tuberculosis, has become the major causes of death among all of infectious diseases. The increasing incidence of multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) has created a need to discover a new antituberculosis drug candidate. The aim of this study was to screen extract and fractions of Tinospora crispa for activity against Mycobacterium tuberculosis H37Rv using the Microplate Alamar Blue Assay (MABA) method. T. crispa extract was prepared by maceration in ethanol (96%) and antituberculosis activity was carried out using MABA method. The result of this study showed that ethanolic extract of T. crispa exhibit antituberculosis activity with minimum inhibition concentration of 12.5 mg/ml. ABSTRAK Tuberkulosis (TB) adalah penyakit yang disebabkan oleh infeksi Mycobacterium tuberculosis dan menjadi penyakit infeksi yang menyebabkan paling banyak kematian di dunia. Daruratnya penyakit TB dan meningkatnya kasus multidrug-resistant tuberculosis (MDR-TB) dan XDR-TB ini mendorong dilakukannya banyak penelitian dan pengembangan obat antiTB baru. Penelitian ini bertujuan untuk mengetahui aktivitas antituberkulosis batang Brotowali. Batang Brotowali kering diekstraksi dengan metode maserasi menggunakan pelarut etanol 96%. Uji aktivitas antituberkulosis secara in vitro terhadap M. tuberculosis H37Rv dilakukan dengan metode Microplate Alamar Blue Assay (MABA). Hasil penelitian menunjukkan bahwa ekstrak etanol batang Brotowali memiliki aktivitas antituberkulosis terhadap M. tuberculosis H37Rv dengan konsentrasi hambat minimum (KHM) sebesar 12,5 mg/ml. INTRODUCTION Tuberculosis (TB) is an infectious disease and the second leading cause of death from a single infectious disease in the world after human immunodeficiency virus (HIV) (Prabu et al., 2015). In 2014, TB had caused 1.5 million of deaths (World Health Organization, 2015). It remains a major global health problem. This disease is caused by Mycobacterium tuberculosis, an intracelluler obligate and aerobic bacillus that multiplies within macrophage. TB treatment is generally consist of combination of four first line drugs: isoniazid, rifampicin, ethambutol, and pyrazinamide for two Correspondence author: Retno Wahyuningrum Email : retno_aulady@yahoo.com months and followed by four additional months of isoniazid and rifampicin therapy (Nguta et al., 2015). Unfortunately, in the last few decades, multi-drug resistant (MDR-TB) was arised. MDR- TB is caused by strain of M. tuberculosis that are resistant to isoniazid and rifampicin (Keshavjee and Farmer, 2012). In 2014, there were 480.000 MDR-TB cases and estimated 190.000 people died because of this disease (World Health Organization, 2015). Approximately 9.6% of the total MDR-TB cases are classified as extensively drug-resisten tuberculosis (XDR-TB) (Nguta et al., 2015). In this cases, M. tuberculosis strain is not only resistant to isoniazid and rifampicin, but also resistant to any fluoroquinolones and to at least first line injectable antituberculosis drug, i.e amikacin, capreomycin, and kanamycin. The