J. Sep. Sci. 2014, 00, 1–8 1 Lei Zhou 1 Wen Qi 1 Cong Xu 1 Toshiaki Makino 2 Dan Yuan 1 1 Department of Traditional Chinese Medicine, Shenyang Pharmaceutical University, Shenyang, China 2 Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan Received June 18, 2014 Revised August 17, 2014 Accepted August 20, 2014 Research Article A rapid method for simultaneous determination of 52 marker compounds in Xiao-Qing-Long-Tang by ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry † Xiao-Qing-Long-Tang (XQLT) is a classical Chinese medicine formula. It is generally used for the treatment of common cold, bronchial asthma, and allergic rhinitis in Asia. In this study, a multicomponent quantification fingerprinting approach based on ultra high perfor- mance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry has been developed for the analysis of compounds in XQLT in 14.5 min. A total of 52 com- pounds were identified by co-chromatography of sample extract with authentic standards and comparing the retention time, UV spectra, molecular ions and characteristic fragment ions with those of authentic standards, or tentatively identified by MS E determination along with Mass Fragment software. Moreover, the method was validated for the simultaneous quantification of 16 components in XQLT commercial products. The method is practical for comprehensive standardization of XQLT and holistic comparison of its commercial products from different manufacturers. Keywords: Mass spectrometry / Quantification / Traditional Chinese medicine / Ultra high performance liquid chromatography / Xiao-Qing-Long-Tang DOI 10.1002/jssc.201400669  Additional supporting information may be found in the online version of this article at the publisher’s web-site 1 Introduction The formulated traditional Chinese medicines (TCMs) con- tain hundreds or thousands of characteristic phytochemicals, and their efficacy is closely related with formulated herbs and their constituents. Usually, QC utilizes absolute quantitation of a few specific marker compounds derived from major for- mulated herbs [1,2]. Nowadays, there is an increasing interest in using comprehensive fingerprint patterns to characterize more components [3]. Xiao-Qing-Long-Tang (XQLT) is a classical Chinese medicine formula for the treatment of common cold, bronchial asthma, and allergic rhinitis in Asia [4–6], mentioned in Chinese Pharmacopoeia (CP) and Japanese Correspondence: Professor Dan Yuan, Department of Tradi- tional Chinese Medicine, Shenyang Pharmaceutical University, Shenyang 110016, China E-mail: yuandan_kampo@163.com Fax: +86 024 23986502 Abbreviations: CP, Chinese Pharmacopoeia; JP, Japanese Pharmacopoeia; TCM, traditional Chinese medicine; XIC, ex- tracted ion chromatography; XQLT, Xiao-Qing-Long-Tang Pharmacopoeia (JP) [7, 8]. The formula of XQLT consists of eight herbs, namely, Ephedrae Herba, Cinnamomi Ramulus, Asari Radix et Rhizoma, Zingiberis Rhizoma, Paeoniae Radix Alba, Schisandrae Chinensis Fructus, Clycyrrhyzae Radix et Rhizoma Praeparata Cum Melle and Pinelliae Rhizoma Praeparatum. Many pharmacological studies reported its anti-inflammatory, antiviral, and antiallergy activities [5, 6, 9]. Ephedrae Herba and Cinnamomi Ramulus are the key for- mulated herbs of XQLT according to theory of TCM [10], and ephedrine, pseudoephedrine, and glycyrrhizin should be responsible for these aforementioned activities. According to the Chinese and Japanese Pharmacopoeias, ephedrine, pseudoephedrine, paeoniflorin, and glycyrrhizin are used as marker compounds for the QC of XQLT [7, 8]. During the past few years, holistic chemical profiling meth- ods of the formulated TCMs have been developed by using many modern hyphenated techniques, such as UHPLC–ESI- Q-TOF-MS profiles of 47 compounds in methanol extract of Danggui San [11]. Cheng and co-workers [12] reported † This paper is included in the virtual special issue on traditional chinese medicine available at the Journal of Separation Science web- site. C  2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.jss-journal.com