Abstract
Objective: A simple, Rapid, and sensitive HPLC method utilizing UV detection was developed and validated for the simultaneous estimation of Fluticasone propionate (FP) and Salmeterol xinafoate (SX) in solutions and in vitro human plasma.
 Methods: Chromatographic analysis was done on SUPELCO® RP-C18 column (150 x 4.6 mm, 5 μm particle size) with an isocratic mobile phase composed of methanol, acetonitrile, and water (50:20:30, v/v) mixture while flow rate was set to 1 ml/min. Detection with UV at maximum absorbance wavelength (ʎmax) values of 236 and 252 for FP and SX, respectively. Spiked plasma samples were liquid-liquid extracted by diethyl ether and reconstituted using methanol.
 Results: Method was accurate and precise over a linear (R2>0.995) range of (0.067-100 µg/ml) and (0.0333-50 µg/ml) for FP and SX, respectively. LOD/lOQ values were 0.13/0.6 and 0.06/0.3 µg/ml for FP and SX, respectively.
 The developed method was successfully applied for the analysis of FP and SX in spiked human plasma samples. The method is considered to be accurate and precise over a linear (R2>0.9969) range of (6.67-66.67 µg/ml) and (3.33-33.3 µg/ml) for FP and SX, respectively. Extraction efficiency was approved by recovery values of (94.98–102.46 %) and (96.54–102.62 %) for FP and SX, respectively.
 Conclusion: This validated method revealed simple and cheap extraction procedures and detectors, non-buffered mobile phase, and short retention times with excellent resolution.
Highlights
Glucocorticosteroids and corticosteroids are classes of steroid hormones naturally synthesized in the adrenal cortex from cholesterol; regulate many aspects of metabolism and immune functions [1]
Method optimization λ-max values were determined to be (236, 252, and 202 nm) for Fluticasone propionate (FP), Salmeterol Xinafoate (SX), and βE, respectively. λmax values are very similar to those found in the literature for FP and SX [26, 29, 32]
When (FP and SX) were combined with βE as IS, the detection wavelength was programmed according to the timeprofile shown in
Summary
Glucocorticosteroids and corticosteroids are classes of steroid hormones naturally synthesized in the adrenal cortex from cholesterol; regulate many aspects of metabolism and immune functions [1]. 1a), a new generation of glucocorticosteroids, is considered as one of the important medications for asthma diseases. It has a strong therapeutic effect against bronchi inflammation. 1b) is a new long-acting β2-agonist used in the treatment of nocturnal airway obstruction and has proved to be highly effective in this aspect as well [2, 3]. The two drugs are formulated as dry powder inhalers or pressurized metered-dose inhalers individually or in the combined formulation. It is not yet known whether using Salmeterol xinafoate (SX) alone or in combination with Fluticasone propionate (FP) constitutes the best treatment [4, 5] Salmeterol Xinafoate (SX) (fig. 1b) is a new long-acting β2-agonist used in the treatment of nocturnal airway obstruction and has proved to be highly effective in this aspect as well [2, 3].
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