Talanta 84 (2011) 169–174 Contents lists available at ScienceDirect Talanta journal homepage: www.elsevier.com/locate/talanta Industrial application of green chromatography—I. Separation and analysis of niacinamide in skincare creams using pure water as the mobile phase Yu Yang a,∗ , Zackary Strickland a , Brahmam Kapalavavi a , Ronita Marple b , Chris Gamsky b a Department of Chemistry, East Carolina University, Greenville, NC 27858, United States b Global Analytical Capability Organization, The Procter & Gamble Company, Cincinnati, OH 45241, United States article info Article history: Received 17 November 2010 Received in revised form 15 December 2010 Accepted 22 December 2010 Available online 8 January 2011 Keywords: Pure water chromatography High-temperature water chromatography Subcritical water chromatography HPLC Niacin Niacinamide 4-Acetamidophenol Skincare cream Industrial application Procter & Gamble abstract In this work, chromatographic separation of niacin and niacinamide using pure water as the sole com- ponent in the mobile phase has been investigated. The separation and analysis of niacinamide have been optimized using three columns at different temperatures and various flow rates. Our results clearly demonstrate that separation and analysis of niacinamide from skincare products can be achieved using pure water as the eluent at 60 ◦ C on a Waters XTerra MS C18 column, a Waters XBridge C18 column, or at 80 ◦ C on a Hamilton PRP-1 column. The separation efficiency, quantification quality, and analysis time of this new method are at least comparable with those of the traditional HPLC methods. Compared with traditional HPLC, the major advantage of this newly developed green chromatography technique is the elimination of organic solvents required in the HPLC mobile phase. In addition, the pure water chromatography separations described in this work can be directly applied in industrial plant settings without further modification of the existing HPLC equipment. © 2011 Elsevier B.V. All rights reserved. 1. Introduction Niacinamide is a chemical compound belonging to the vitamin B group that is commonly found in foods and used in cosmetic skin- care products. Niacinamide is an amide derivative of niacin and is found in bound forms in nicotinamide adenine dinucleotide (NAD), its phosphorylated derivative NAD(P), and their reduced forms NAD(H) and NAD(PH), which are coenzymes important for cellular redox reactions [1,2]. These cofactors have many antioxidant prop- erties and are involved in many enzymatic reactions in the skin [2]. Some of the cosmetic effects of niacinamide include improved skin barrier, reduced skin pore size, facial blotchiness, hyperpigmenta- tion, and skin yellowing, antiwrinkle, and antiacne properties [2]. Niacinamide is also used as a bleaching agent in bleaching cosmet- ics and can control the transfer of melanin from melanocytes [3]. In addition, niacinamide is widely used as a color fixative in meats to maintain fresh color [4,5]. Its antioxidant properties help to reduce the speed of myoglobine oxidation and prolong the red fresh color [5]. ∗ Corresponding author. E-mail address: yangy@ecu.edu (Y. Yang). It is important to be able to accurately analyze niacinamide levels in various products. Quantitative analysis of niacinamide in cosmetics is required for quality control, product release, and reg- ulatory purposes. HPLC with UV or fluorometric detection has been widely used for the determination of niacinamide in pharmaceuti- cals, biological fluids, food, and cosmetics [1,3–6]. However, these traditional HPLC methods require organic solvents in the mobile phase that are hazardous and expensive. With growing awareness about the environment and the increased initiative for the development of “green” technolo- gies throughout the world, reversed-phase liquid chromatography using water as the sole component in the eluent has received increased attention. The ability to eliminate the enormous amounts of hazardous organic solvents that are consumed everyday world- wide and replace them with an efficient separation method that utilizes environmentally benign water offers many benefits both environmentally and economically. Water is too polar to serve as a reversed-phase eluent at ambient temperature using normal RPLC stationary phases such as silica- based C18 and polymer PRP-1 columns. Therefore, there are two ways to achieve LC separation of organics using pure water as the sole mobile phase component. One way is to modify the stationary phase and the other is to heat the water mobile phase. Synovec 0039-9140/$ – see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.talanta.2010.12.044