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"Glukosamin HCl merupakan obat yang digunakan dalam pengobatan osteoarthritis. Saat ini semakin banyak penggunaan obat Glukosamin HCl secara transdermal, dimana obat ini akan melewati sirkulasi sistemik, sehingga kadarnya di dalam darah perlu dipantau. Metode analisis menggunakan Kromatografi Cair Kinerja Tinggi (KCKT) dengan detektor fluorosensi telah dikembangkan dan dioptimasi untuk analisis glukosamin HCl dalam plasma manusia in vitro. Glukosamin HCl harus diderivatisasi terlebih dahulu dengan orto-ftalaldehida/ 2- merkaptoetanol untuk mendapatkan gugus kromofor sehingga dapat terdeteksi pada detektor fluorosensi. Glukosamin HCl diekstraksi dari plasma dengan menggunakan asetonitril. Kromatografi dilaksanakan menggunakan kolom faseterbalik Lichrospher® 100 RP-18 (5μm, Merck), fase gerak asetonitril-air yang mengandung 0,25% tetrahidrofuran (11:89) pada kecepatan alir 1,0 mL/menit dan dideteksi pada panjang gelombang eksitasi 335 nm dan emisi 445 nm. Kondisi optimum ini membutuhkan waktu analisis 19.948 menit. Pada rentang konsentrasi 0,0502-10,0400 μg/ml dihasilkan kurva kalibrasi yang linier dengan koefisien korelasi (r) 0,9997. Akurasi (% diff) dari metode ini antara -3,72 hingga 4,39 % dengan presisi (KV) antara 2,23 hingga 3,09%, dan uji perolehan kembali relatif antara 96,28% sampai 104,39%.

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Glucosamine HCl is a drug which is used in osteoarthritis treatment. Recently the use of transdermal?s Glucosamine HCl is increasing, wether glucosamine HCl will pass the systemic circulation, thereby monitoring the blood drug level is necessary. A method using high-performance liquid chromatography (HPLC) with fluorosence detector has been developed for analysis of glucosamine HCl in human plasma in vitro. Before Glucosamine HCl can be detected by fluorosence detector, it must be derivatived with orthophtalaldehyde/2-mercaptoethanol, in order to get chromophore group. Glucosamine HCl was extracted from plasma using acetonitrile. The chromatography was carried out by a reversed-phase Lichrospher® 100 RP-18 (5μm, Merck) with mobile phase consisted of acetonitrile-water containing tetrahydrofuran 0,25% (11:89) at flow rate 1,0 mL/minute and detection was performed at excitation wavelength of 335 nm and emission wavelength of 445nm. This optimum condition was take 19.948 minutes for analysis. Linearity was established for range concentration of 0,0502-10 μg/ml with coefficient correlation (r) was 0,9997. Accuracy (% diff) ranged from -3,72 to 4,39 % , precision (CV) ranged."

"Glukosamin adalah suatu zat yang dapat disintesis di dalam tubuh yang berguna untuk mempertahankan dan memulihkan kinerja sendi. Seiring bertambahnya usia, kemampuan tubuh untuk mensintesis glukosamin menurun sehingga menyebabkan penyakit osteoartritis. Oleh karena itu, telah berkembang suplemen makanan yang mengandung glukosamin yang telah diakui oleh Food and Drug Administration (FDA) untuk pengobatan osteoartritis. Analisis glukosamin HCl dilakukan untuk memperoleh volume, temperatur, waktu, dan waktu kestabilan reaksi yang optimum pada derivatisasi glukosamin HCl dengan FMOC-Cl menggunakan detektor fluoresensi. Larutan standar glukosamin HCl 1 μg/ml ditambah 50,0 μL 0,2 M dapar dinatrium tetraborat dekahidrat pH 8, kemudian divorteks selama 10 detik, ditambah 360,0 μL pereaksi FMOC-Cl 1 mg/ml, campuran divorteks selama 10 detik, diinkubasi menggunakan termomixer pada 1400 rpm dan temperatur 25°C selama 15 menit, selanjutnya disuntikkan sebanyak 20,0 μL ke alat KCKT. Pemisahan dengan KCKT menggunakan kolom Kromasil® C18 (5 μm; 250 x 4,6 mm) dengan komposisi fase gerak air-asetonitril (40:60) dengan kecepatan alir 1,0 ml/menit. Linieritas pada konsentrasi 100-1000 ng/ml dengan koefisien korelasi (r) 0,9995. Nilai batas deteksi (LOD) sebesar 21,98 ng/ml dan batas kuantitasi (LOQ) sebesar 73,26 ng/ml.

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Glucosamine is a synthesized substance in the human body useful for maintaining and restoring the joint's function. Body's capacity to synthesize glucosamine declines with age thus can cause osteoarthritis. There was development of dietary supplement that contains glucosamine which has been approved by the Food and Drug Administration (FDA) for treatment of osteoarthritis. Glucosamine HCl analysis was performed in order to get optimal volume, temperature, time, and reaction stability time in glucosamine HCl derivation with FMOC-Cl using fluorescence detector. Standard solution of Glucosamine HCl added by 50.0 μl 0.2 M disodium tetraborate decahydrate buffer with pH 8 were homogenized for 10 seconds, then the mixed solution was added by 360.0 μl of 1 mg/ml FMOC-Cl reagent and homogenized for 10 seconds. It was then incubated using termomixer at 1400 rpm and a temperature of 25°C for 15 minutes, then as many as 20.0 μl injected into the High Performance Liquid Chromatography (HPLC) instrument. Separation by HPLC using one column of Kromasil® C18 (5 μm; 250 x 4.6 mm) with mobile phase composition of water-acetonitrile (40:60) and flow rate 1.0 ml/minute. Linearity at concentrations of 100-1000 ng/ml with a correlation coefficient (r) 0.9995. The limit of detection (LOD) value was 21.98 ng/ml and the limit of quantitation (LOQ) was 73.26 ng/ml."

"Glukosamin memilki efek terapi farmakologis alternatif untuk pengobatan osteoarthritis. Pemakaian glukosamin secara transdermal merupakan salah satu upaya untuk mengatasi bioavaibilitas yang baik. Bentuk sediaan nanoemulsi dapat meningkatkan kelarutan obat, stabil secara termodinamika dan memiliki penampilan yang transparan dengan ukuran partikel kurang dari 100 nm. Senyawa peningkat penetrasi perkutan diantaranya etanol dan asam oleat ditambahkan ke dalam masing-masing formula nanoemulsi glukosamin, Uji daya penetrasi secara in vitro dengan sel Difusi Franz menggunakan membran abdomen tikus Rattus norvegicus menghasilkan jumlah kumulatif glukosamin yang terpenetrasi setelah 8 jam dari sediaan formula A (etanol 6%) dan formulasi B (asam oleat 6%) secara berturut turut sebanyak 1147,30± 27,45 μg/cm2; 708,72 ± 10,35 μg/cm2. Laju penetrasi atau fluks dari sediaan formula A dan formula B berturut - turut sebesar 218,41 ± 2,68 μg/cm2.jam; 88,59 ± 2,23 μg/cm2.jam. Uji kestabilan fisik dilakukan melalui pengamatan seperti organoleptis, homogenitas, pH, Viskositas, sentrifugasi dan ukuran partikel. Ukuran rata ? rata partikel yang diperoleh pada formulasi A 4,5 nm dan formulasi B 12,3.

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Glucosamine have an alternative pharmacologic theraphy for osteoarthritis. Oral glucosamine does not show any good bioavaibility, for that reason transdermal route is one wich was developed to overcome the problem. Nanoemulsi dosage form can improve drug solubility, thermodynamically stable and has a transparant appearance with particle size less than 100 nm. Percoutaneous penetration ethanol and oleic acid were added to each nanoemulsion formula glucosamine. Penetration test in vitro with Franz diffusion cell using Rattus norvegicus rat abdomen skin as membrane diffusion. Cumulative amount of glucosamine penetrated after 8 hours from formula A (ethanol 6%) and formula B (6%) were 1147,30±27,45 μg/cm2; 708,72±10,35 μg/cm2 respectively. Penetration rate or flux of glucosamine from formula A and formula B, 218,41 ± 2,68 μg/cm2.hour; 88,59 ± 2,23 μg/cm2.hour. Physical stability test is done through organoleptic, homogenaity, pH, viscosity, centrifugation and particle size. The average particle size obtained from formula A and formula B were 4,5 nm and 12,3 nm respectively."

"Pendahuluan. Glukosamin, Kondroitin sulfat dan Methylsufonylmethane (MSM) merupakan suplemen yang sering diberikan pada pasien osteoarhritis (OA) derajat I dan II. Organisasi kedokteran seperti AAOS, OARSI, EULAR memberikan rekomendasi yang berbeda tentang penggunaannya. Studi ini bertujuan untuk menilai efektivitas Glukosamin-Kondroitin sulfat (GK) dan GKM terhadap perbaikan klinis pasien OA sendi lutut.Bahan dan Cara Kerja. Studi ini merupakan uji klinis acak tersamar ganda pada 147 pasien dengan OA lutut derajat Kellgren-Lawrence I atau II. Subjek dipilih dengan metode randomisasi dengan blok permutasi ke tiga kelompok yaitu GK (n=49), GKM (n=50) dan plasebo (n=48). Obat-obatan ini diberikan sekali sehari selama 3 bulan berturut-turut. Skor VAS dan WOMAC dinilai sebelum pemberian suplemen, kemudian minggu ke 4, 8, dan 12.Hasil. Rata-rata usia pasien adalah 61 tahun, dimana 67,3% pasien adalah perempuan. Bila dibandingkan terhadap Skor WOMAC pada kelompok Plasebo, kelompok GKM menurunkan skor WOMAC secara signifikan (perbedaan rerata 7.15, IK 12.06-2.23, p=0.005). Kelompok GK menurunkan Skor WOMAC secara signifikan (perbedaan rerata 8.17, IK 13.49-2.84, p=0.003). Sementara itu pada penilaian Skor VAS, kelompok GKM menurunkan skor secara signifikan terhadap dua kelompok lain, yaitu terhadap kelompok GK secara signifikan (perbedaan rerata 0.68, IK 1.18-0.19, p=0.007), dan Plasebo (perbedaan rerata 0.86, IK 1.37-0.35, p=0.001).Simpulan. Kombinasi suplemen GKM lebih efektif dalam menurunkan nyeri dan meningkatkan fungsi pada pasien OA sendi lutut derajat I dan II dibandingkan dengan GK dan plasebo. Sedangkan suplemen GK secara keseluruhan tidak lebih baik dibandingkan dengan plasebo dalam memberikan perbaikan klinis pada pasien OA sendi lutut derajat Kellgren Lawrence I-II.

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Introduction. Glucosamine-Chondroitin sulfate-Methylsufonylmethane (MSM) as combination are the most popular supplements for patients with knee osteoarthritis (OA). There are emerging controversies and also different recomendation from many medical associations such as AAOS, OARSI, and EULAR regarding the effectiveness of these supplements. This current study evaluated the efficacy of Glucosamine-Chondroitin sulfate (GC), Glucosamine-Chondroitin-MSM (GCM), and placebo on clinical improvement of patients with knee OA Kellgren Lawrence gr I-II.

Material and Methods. This study was a double blind, randomized controlled clinical trial on 147 patients with knee OA Kellgren-Lawrence grade I-II. Subjects were allocated by permuted block randomization to three groups, either GC (n=49), or GCM (n=50), or placebo (n=48). VAS and WOMAC score were measured before treatment, then at 4 th, 8 th and 12th week after treatment.

Result. Mean age of patients was 61 years, and 67,3% were woman. As compared with Placebo group, WOMAC score in GCM group was significantly lower (MD 7.15, CI 12.06-2.23, p=0.005), and in GC group WOMAC Score was also lower (mean difference 8.17, CI 13.49-2.84, p=0.003). Whereas VAS score in GCM group was significantly lower compared to that in GC group (MD 0.18, CI 1.18-0.19, p=0.007) and also compared with Placebo group (MD 0.86, CI 1.37-0.35, p=0.001).

Conclusions. Combinations of GCM made a clinical improvement in patients with knee OA Kellgren Lawrence gr I-II compared with GC and Placebo. GC did not make clinical improvement in overall groups of patients with knee OA Kellgren Lawrence gr I-II."

"ABSTRAKGlukosamin hidroklorida dan kondroitin sulfat merupakan senyawa glikosaminoglikan (GAGs) yang merupakan komponen struktural utama dari tulang yang akan membentuk proteoglikan. Kedua senyawa ini dapat merawat kesehatan tulang dengan menstimulasi sintesis cairan sinovial dan menghambat degradasi kartilage persendian, sehingga dapat digunakan untuk terapi osteoartritis. Penelitian ini bertujuan untuk memperoleh metode analisis yang selektif untuk penetapan kadar glukosamin hidroklorida dan kondroitin sulfat dalam sediaan tablet dan krim. Setelah diderivatisasi menggunakan pereaksi ortoftalaldehida dan 2-merkaptoetanol (OPA/2-ME), sampel dianalisis menggunakan kromatografi cair kinerja tinggi (KCKT) dengan detektor fluoresensi pada panjang gelombang eksitasi 335 nm dan panjang gelombang emisi 445 nm. Glukosamin mempunyai gugus amin primer sehingga dapat diderivatisasi dengan OPA/2-ME, sedangkan kondroitin mempunyai gugus asetil pada gugus amin, sehingga perlu dilakukan deasetilasi menggunakan natrium hidroksida untuk memutus gugus asetil. Fase gerak yang digunakan tetrahidrofuran 0,25% dalam air-asetonitril (87:13) dengan laju alir 1,5 mL/menit. Kondisi analisis yang telah dioptimasi kemudian divalidasi mencakup akurasi, presisi, linieritas, selektivitas, batas deteksi, dan batas kuantitasi. Hasil menunjukkan kadar rata-rata glukosamin hidroklorida dan kondroitin sulfat pada sediaan tablet dan krim adalah 92,76%; 96,11% dan 101,15%; 100,33% memenuhi syarat keberterimaan.

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ABSTRAKGlucosamine hydrochloride and chondroitin sulphate are glycosaminoglycans (GAGs) compound which is a major structural component of bone that form proteoglycans. Both of these compounds can take care of bone health by stimulating the synthesis of synovial fluid and inhibit the degradation of joint cartilage, so it can be used for the treatment of osteoarthritis. The aimed of this study were obtain selective analytical method for the determination of glucosamine hydrochloride and chondroitin sulphate levels in tablet and cream dosage forms. After derivatization using orthophtalaldehyde and 2-mercaptoethanol (OPA/2-ME), samples were analyzed using high performance liquid chromatography (HPLC) with fluorescence detector at excitation wavelength of 335 nm and emission wavelength of 445 nm.. Glucosamine has a primary amine group that can be derivatized with OPA/2-ME, while chondroitin having an acetyl group at the amine group, so we needed deacetylation using natrium hydroxide to break the acetyl group. The mobile phase used tetrahydrofuran 0.25% in water-acetonitrile (87:13) with a flow rate 1.5 mL/min. Analysis conditions have been optimized, validated in terms of accuracy, precision, linearity, selectivity, limit of detection, and limit of quantitation. The results showed average levels of glucosamine hydrochloride and chondroitin sulphate in tablet and cream dosage forms were 92.76%; 96.11% and 101.15%; 100.33% and fulfilled the acceptance criteria."

"Background: Glucosamine, chondroitinsulfate are frequently used to prevent further joint degeneration in osteoarthritis (OA). Methylsulfonylmethane (MSM) is a supplement containing organic sulphur and also reported to slow anatomical joint progressivity in the knee OA. The MSM is often combined with glucosamine and chondroitin sulfate. However, there are controversies whether glucosamine chondroitin sulfate or their combination with methylsulfonylmethane could effectively reduce pain in OA. This study is aimed to compare clinical outcome of glucosamine chondroitin sulfate (GC), glucosamine chondroitin sulfate methylsulfonylmethane (GCM), and placeboin patients with knee osteoarthritis (OA) Kellgren Lawrence grade I II. Methods: a double blind, randomized controlled clinical trial was conducted on 147 patients with knee OA Kellgren Lawrence grade I II. Patients were allocated by permuted block randomization into three groups: GC (n=49), GCM (n=50), or placebo (n=48) groups. GC group received 1500 mg of glucosamine + 1200 mg of chondroitin sulfate + 500 mg of saccharumlactis; GCM group received 1500 mg of glucosamine + 1200 mg of chondroitin sulfate + 500 mg of MSM; while placebo group received three matching capsules of saccharumlactis. The drugs were administered once daily for 3 consecutive months VAS and WOMAC scores were measured before treatment, then at 4th, 8th and 12th week after treatment. Results: on statistical analysis it was found that at the 12th week, there are significant difference between three treatment groups on the WOMAC score (p=0.03) and on the VAS score (p=0.004). When analyzed between weeks, GCM treatment group was found statistically significant on WOMAC score (p=0.01) and VAS score (p<0.001). Comparing the score difference between weeks, WOMAC score analysis showed significant difference between GC, GCM, and placebo in week 4 (p=0.049) and week 12 (p=0.01). In addition, VAS score also showed significant difference between groups in week 8 (p=0.006) and week 12 (p<0.001). Conclusion: combination of glucosamine chondroitinsulfate methylsulfonylmethane showed clinical benefit for patients with knee OAK ellgren Lawrence grade I II compared with GC and placebo. GC did not make clinical improvement in overall groups of patients with knee OA Kellgren Lawrence grade I II."

"ABSTRAKLatar belakang: Cancer stem cells CSC merupakan sel kanker yang memiliki karakteristik sel punca. Kepuncaan CSC berperan dalam menjaga kestabilan jaringan tumor, sifat resisten terhadap terapi dan memicu kejadian relaps. Kepuncaan CSC diduga dapat ditarget secara non-protein specific dengan mengganggu berbagai jalur pensinyalan yang berperan dalam mempertahankan kepuncaan sekaligus menghambat microenvironment. Proses glikosilasi protein berperan dalam kestabilan, transportasi, dan maturasi protein. Glukosamin diduga dapat berpengaruh terhadap interaksi CSC dengan Cancer-associated fibroblast CAF melalui penghambatan glikosilasi. CAF merupakan sel fibroblast di microenvironment yang direkrut oleh sel kanker ke jaringan tumor. Penelitian ini bertujuan untuk menganalisis efek glukosamin terhadap penurunan sifat kepuncaan sel punca kanker payudara ALDH dan hubungannya dengan penanda CAF pada stroma.Metode: Sel punca kanker payudara ALDH ditumbuhkan dalam medium yang mengandung glukosamin selama 24 jam. Conditioned medium yang diperoleh dari sel ALDH CSC-CM atau sel ALDH yang diberi perlakuan glukosamin CSC-CM G digunakan untuk menumbuhkan sel stroma payudara selama 48 jam. Nilai ekspresi gen relatif ALDH1, Oct-4, dan IGF-1 pada CSC, dan gen penanda CAF, ?-SMA dan FAP pada sel stroma diperiksa menggunakan Quantitative Real-Time Polymerase Chain Reaction qPCR .Hasil: Perlakuan glukosamin konsentrasi 4 mM selama 24 jam menyebabkan penurunan ekspresi ALDH1, gen marker CSC pada sel ALDH dan ekspresi Oct-4, gen karakteristik kepuncaan. Ekspresi gen Oct-4 tetap menurun walaupun glukosamin telah dikeluarkan dari medium kultur. Conditioned-medium CM yang diperoleh dari sel punca kanker payudara ALDH dapat memicu peningkatan ekspresi ?-SMA pada sel stroma. Peningkatan ekspresi gen ?-SMA dan FAP pada sel stroma yang diinduksi oleh CSC-CM dapat ditekan oleh CSC-CM G.Kesimpulan: Penghambatan N-glikosilasi oleh glukosamin menyebabkan penurunan ekspresi gen penanda CSC dan gen karakteristik kepuncaan pada sel punca kanker payudara ALDH Perlakuan glukosamin dapat mempengaruhi pensinyalan parakrin CSC-CAF melalui ekspresi gen penanda CAF.

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ABSTRACTBackground Cancer stem cells CSC is known as a subpopulation of cancer cells with stem cell like characteristics. CSC stemness is responsible for tumor maintenance and relapse. A therapeutic agent that is non protein specific yet intensely uptaken by CSC can be a good approach to disrupt the coordinated network in stemness maintenance and simultaneously affect corrupt stromal cells in tumor microenvironment. Glucosamine inhibits post translational glycosylation, essential for sustaining protein stability, trafficking, and maturation. Cancer associated fibroblast CAF differentiation and recruitment to tumor microenvironment is induced by CSC derived growth factors. This study investigated the effect of glucosamine on stemness in ALDH breast cancer stem cell and its ability to interact with stromal cells.Methods ALDH breast cancer stem cell were cultured in medium containing glucosamine for 24 h. Breast stromal cells were culture in conditioned medium obtained from ALDH cells CSC CM or glucosamine treated ALDH cells CSC CM G . Relative expression of ALDH1, Oct 4, and IGF 1 gene in CSC, and SMA and FAP gene in stromal cells were analyzed using Quantitative Real Time Polymerase Chain Reaction qPCR .Results Upon treatment with 4 mM glucosamine for 24 h, ALDH breast cancer stem cell showed significant decrease in expression of ALDH1, a marker of breast cancer stem cell. Under similar condition, Oct 4 stemness gene was also found to be downregulated. Downregulation of Oct 4 expression was maintained after removal of glucosamine. Stromal cells showed increased expression of SMA myofibroblast marker upon cultured in CSC CM. This upregulation was cancelled in CSC CM G exposed stromal cells.Conclusion N linked glycosylation inhibition by glucosamine results in downregulation of stem cell marker and stem cell gene expression in ALDH breast cancer stem cell. CSC rsquo s stemness influences paracrine interaction between CSC dan CAF via expression of CAF marker in stromal cells "