Fingerprint Analysis of Flos Carthami by Pressurized CEC and LC By:

A fingerprint analysis of Flos Carthami was performed using a standardized capillary electrochromatography (CEC) procedure. This procedure was first used to establish the electrochromatographic profile of genuine Flos Carthami from Tacheng, Xinjiang, China. This Flos Carthami fingerprint was then used to identify and assess the consistency of raw herbs from different sources in China. The study of a limited number of samples from ten different sources demonstrated a reasonable consistency among their CEC fingerprints relative to that of the genuine sample. Using this technique, we can readily distinguish the fingerprint of Flos Carthami from that of Stigma Croci, a possible substitute in traditional Chinese medicine, and Flos Hemerocallis, a commercial adulterant. A method based on high performance liquid chromatography (HPLC) is described to establish fingerprints of Flos Carthami simultaneously. The fingerprints obtained with HPLC consist of 21 common peaks within 65 min while 43 common peaks obtained with CEC. CEC showed better performance on fingerprinting of hydroxysaffloryellow A and its neighboring peaks, which contained more chemical information than that of HPLC. It was proven that CEC could be a feasible and effective method for development of fingerprint of TCM based on the comparison with HPLC. Article: INTRODUCTION Fingerprint analysis has been introduced and accepted by the US Food and Drug Administration (FDA) as one of the requirements for botanical preparations [1] and by the European Agency for the Evaluation of Medicinal Products for herbal preparations [2]. Furthermore, fingerprint analysis has been introduced and accepted by the World Health Organism (WHO) as a strategy for assessing herbal medicines [3]. Recently, fingerprinting has also been required by the State Food and Drug Administration Bureau of China for the standardization of injections made from TCMs or their raw materials [4]. For herbs, fingerprinting is primarily used for identification and assessment of the stability of the plants [5]. High-performance liquid chromatography (HPLC) fingerprint analysis has reportedly been used for similar purposes with some botanical medicines and their raw materials [5, 6]. Flos Carthami, the dried flower of Carthamus tinctorius L., has long been used as a food colorant, dye, and flavoring agent [7] in oriental countries. Now, it is being used as an herbal medicine in China for the treatment of uterine congestion, cardiovascular diseases, thrombosis, and high cholesterol [8]. Some of its constituents are known to exhibit pharmacological and biological activities. For example, hydroxysaffloryellow has been shown to restrain the conglomeration of platelet, promote blood circulation, remove blood stasis, anti-oxidation, and promote metabolism [9]. According to the Chinese Pharmacopoeia, TLC is required for the identification of raw herbs and the UV measurement of safflower yellowand red-pigments in Flos Carthami [10]. The quantitation of safflower yellowand red-pigments, adenosine, rutin, and quercetin in Flos Carthami using UV, CE, and HPLC has been reported previously [11–17]. Stigma Croci (also known as Fan Honghua, Saffron) is known to be a possible substitute for Flos Carthami, and Flos Hemerocallis has been found as a common adulterant in the market. The application of capillary electrophoresis (CE) in fingerprint analysis was limited in China; although, the CE fingerprint of ethanol--water extracts of Flos Carthami were reported [18]. There were only 15 peaks observed in the HPLC fingerprint of water extracts from Flos Carthami been previously reported [19]. HPLC has high precision, sensitivity and reproducibility for fingerprinting TCM, but HPLC is not suitable for analysis of some highly viscous samples. While CE has high speed, efficiency, ultra small sample volume, and minimal consumption of solvent, but the reproducibility and the selectivity are not as good as in HPLC. Capillary electrochromatography (CEC) is a hybrid technique that combines the selectivity of HPLC and the separation efficiency of CE. CEC can provide more chemical information of the raw herbs and which will be more powerful in differentiating geographical origins. Now CEC has become an attractive technique for pharmaceutical analysis because it provides highly efficient and rapid separations. So we intend to develop a characteristic fingerprint of Flos Carthami by CEC that can be used to identify and control the quality of the raw herbs. This fingerprint can help to distinguish between Flos Carthami and its substitutes or adulterants and can be used to assess variations in Flos Carthami that are grown in the different areas of China.