Simultaneous Determination of Cysteamine and Cystamine in Cosmetics by Ion-Pairing Reversed-Phase High-Performance Liquid Chromatography

Abstract

Cysteamine has been used in cosmetics as an antioxidant, a hair straightening agent, and a hair waving agent. However, recent studies indicate that cysteamine can act as an allergen to hairdressers. The objective of this study was to develop and validate a simple and effective reversed-phase high-performance liquid chromatography (RP-HPLC) method for the measurement of cysteamine and its dimer, cystamine. Sodium 1-heptanesulfonate (NaH-pSO) was used as an ion-pairing agent to improve chromatographic performance. Separation was performed on a Gemini C18 column (250 mm × 4.6 mm, 5 µm particle size) using a mobile phase composed of 85:15 (v/v) 4 mM NaHpSO in 0.1% phosphoric acid:acetonitrile. UV absorbance was monitored at 215 nm. The RP-HPLC method developed in this study was validated for specificity, linearity, limit of detection, limit of quantitation, precision, accuracy, and recovery. Cysteamine and cystamine were chromatographically resolved from other reducing agents such as thioglycolic acid and cysteine. Extraction using water and chloroform resulted in the recovery for cysteamine and cystamine ranging from 100.2-102.7% and 90.6-98.7%, respectively. This validated RP-HPLC method would be useful for quality control and monitoring of cysteamine and cystamine in cosmetics.

Challenges for cysteamine stabilization, quantification, and biological effects improvement

“…The quantification of cys was also assessed with the addition of a surfactant into the mobile phase which interacts with cys, affecting its retention through a reversed-phase column. The addition of sodium dodecyl phosphate (40 mM) using micellar chromatography and sodium 1-heptanesulfonate (4 mM) using the ion pair chromatography to the mobile phase were able to separate and quantify cys and cystamine ( Table 6 [ 16 , 64 ]).…”

“… Method Surfactant Mobile phase Stationary phase Flow rate (mL/min) Temperature (℃) LOD (μM) Refs. Micellar chromatography Sodium dodecyl phosphate (40 mM) Water: acetonitrile: methanol (38:30:32) with phosphoric acid, and sodium dodecyl sulfate C 18 (5 μm; 4.6 mm × 250 mm) 1.6 50 4.15 [ 16 ] Ion pair chromatography Sodium 1-heptanesulfonate (4 mM) Acetonitrile: water (0.1% phosphoric acid + Sodium 1-heptanesulfonate) (85:15) C 18 (5 μm; 4.6 mm × 250 mm) 1 25 12.9 [ 64 ] …”

Cysteamine Eye Drops in Hyaluronic Acid Packaged in Innovative Single-Dose Systems: Stability and Ocular Biopermanence

“…The quantification of cysteamine and cystamine in the compounded formulations was analyzed by ultra-high-performance liquid chromatography (UHPLC) using a modified version of the method proposed by Kim et al [ 38 ]. The analysis was performed with a UHPLC coupled with photodiode array detection (PDA), using an ACQUITY UPLC H-Class System (Waters ® ) with ACQUITY PDA detector (Waters ® ).…”

Recent Research in Ocular Cystinosis: Drug Delivery Systems, Cysteamine Detection Methods and Future Perspectives

“…In this sense, another important barrier for the development of cysteamine topical formulations is the fact that its physicochemical properties combined with its low molecular weight (77.15 g/mol) make its quantification a difficult issue, which is still trying to be solved nowadays [64].…”

“…Derivatization agents, conditions and elution times are summarized in Table 3. Among methods depicted in this table, it is worth highlighting the one recently published by Kim et al It is a simple and effective analytical method for the quantitation of cysteamine using reversed-phase HPLC for simultaneous determination of cysteamine and cystamine, in which the previous derivatization process is not necessary [64].…”