PERBANDINGAN ANALISIS KROMATOGRAFI CAIR KINERJA TINGGI ELUASI GRADIEN DENGAN ISOKRATIK PADA PENENTUAN VITAMIN B1, B2 DAN B6 DALAM SEDIAAN SIRUP MULTIVITAMIN SECARA SIMULTAN

Ni Luh Kasih Ariani; Ni Made Suaniti; James Sibarani

Ni Luh Kasih Ariani Magister Kimia Terapan, Universitas Udayana, Denpasar, Bali-Indonesia
Ni Made Suaniti 1. Magister Kimia Terapan, Fakultas Pascasarjana, Universitas Udayana, Denpasar, Bali-Indonesia 2. Jurusan Kimia, FMIPA, Universitas Udayana, Jimbarani, Badung, Bali-Indonesia
James Sibarani 1. Magister Kimia Terapan, Fakultas Pascasarjana, Universitas Udayana, Denpasar, Bali-Indonesia 2. Jurusan Kimia, FMIPA, Universitas Udayana, Jimbarani, Badung, Bali-Indonesia

Abstract

ABSTRAK: Vitamin B terutama B1 (Tiamin), B2 (Riboflavin), dan B6 (Piridoksin) sering terkandung dalam sirup multivitamin sehingga diperlukan analisis untuk mendeteksi secara simultan dalam campuran. Tujuan penelitian ini adalah membandingkan analisis vitamin B1, B2, dan B6 dalam sirup multivitamin secara simultan dengan kromatografi cair kinerja tinggi eluasi gradien dan isokratik, menggunakan kolom C18, panjang 15 cm, dan dengan pelarut campuran metanol: air : asam asetat glasial (10:90:1). Fase gerak adalah campuran natrium heksan sulfonat 5 mM dalam asam asetat glasial 0,5% dengan metanol yang dicampur secara gradien. Kondisi optimum metode gradien diperoleh pada laju alir 2,0 mL/menit, panjang gelombang 280 nm dengan waktu retensi 3,441 menit untuk piridoksin; 4,985 menit untuk riboflavin dan 7,393 menit untuk tiamin dengan resolusi 2,272 antara riboflavin dan piridoksin. Hasil uji presisi riboflavin dan piridoksin menggunakan metode isokratik (campuran natrium heksan sulfonat 5 mM dengan metanol dengan perbandingan 70 : 30) masing-masing dengan RSD adalah 1,377 dan 1,376 sedangkan metode gradien adalah 0,693 dan 0,825. Uji linearitas ketiga vitamin menggunakan dua metode isokratik dan gradien memenuhi persyaratan dengan R² = 0,999.

Kata kunci : Vitamin B, sirup multivitamin, kromatografi cair kinerja tinggi

ABSTRACT: The aim of this research is to compare the simultaneous analytical results of B1 (Tiamin), B2 (Riboflavin), and B6 (Piridoxin) in multivitamin syrup between gradient and isocratic methods using high performance liquid chromatography (HPLC). The separation was performed on 15 cm length of C18 column and a mixture of methanol, water and glacial acetate acid with ratio of 10:90:1 was used as solvent. The eluent was a mixture of methanol and 5 mM sodium hexane sulfonate in 0.5% glacial acetate acid and gradually mixed using 2 different pumps. The optimal analytical conditions for gradient method were found to be 2,0 mL/min of flow rate and 280 nm of wavelength. On this gradient method, the vitamins were completely separated with retention times of 3.441 minutes, 4.985 minutes, and 7.393 minutes for piridoxin, riboflavin and tiamin respectively. Meanwhile, isocratic method (mixture of 5 mM sodium hexane sulfonate with metanol of 70 : 30) on the same conditions can only separate riboflavin and piridoxin. The RSD values for riboflavin and piridoxin were 1.377% and 1.376% respectively on isocratic methods and 0.693% and 0.825% respectively on gradient method. The linearity of both methods have fulfilled the requirement of R² values that are higher than 0.999 for all vitamins.

Keywords: B Vitamins, Multivitamin Syrup, High Performance Liquid Chromatography

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References

] Evert A., 2013, Medline Plus, Vitamins, University of Washington Medical Center Diabetes Care Center, Washington.
[2] Lachman L., Lieberman H.A., Kanig J.L., 1994, Teori dan Praktek Farmasi Industri, UI Press, 942-943.
[3] Savitri M., 2007, Metode Cepat Penentuan Simultan Kadar Kafein, Vitamin B2 dan B6 dalam Minuman Berenergi dengan Teknik Zero-Crossing, IPB, Bogor.
[4] Yantih N., Widowati D., Wartini, Aryani T., 2011, Validation of HPLC Method for Determination of Thiamine Hydrochloride, Riboflavin, Nicotiamide, and Pyridoxine Hydrochloride in Syrup Preparation, Canadian Journal on Scientific and Industrial Research Vol. 2, No. 7.
[5] Poongothai S., Ilavarasan R., Karrunakaran C.M., 2010, Simultaneous and accurate Determination of Vitamins B1, B6, B12 and Alpha Lipoic Acid in Multivitamin Capsule by Reverse – Phase High Perfomance Liquid Chromatographic Method, International journal of Pharmacy and Pharmaceutical Sciences Vol. 2, Suppl 4.
[6] Scoot, Paymond P.W., 1995, Techniques and Practice of Chromatography, Marcel Dekker, Inc, USA 27-65, 284-287.
[7] Snyder L.R., Kirkland J.J., Dolan J.W., 2010, Introduction to Modern Liquid Chromatography, 24-30.
[8] US Pharmacopeial Covention, 2008, The United States Pharmacopeia, 31st rev., The National Formulary, 31st ed., The United States Pharmacopeial Convention, Inc., Rockville, 1034.
[9] Harmita, 2004, Petunjuk Pelaksanaan Validasi Metode dan Cara Perhitungannya, Majalah Ilmu Kefarmasian, Vol. I No. 3, Departemen Farmasi F-MIPA, UI
[10] Ahuja S., Dong M.W., 2005, Handbook of Pharmaceutical Analysis by HPLC, Elsevier Academic Press, New York, 21-35.