{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T14:52:53Z","timestamp":1761663173955,"version":"build-2065373602"},"reference-count":28,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2013,6,13]],"date-time":"2013-06-13T00:00:00Z","timestamp":1371081600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>A reference extractive, containing multiple active known compounds, has been considered to be an alternative to individual reference standards. However, in the Chinese Pharmacopoeia (ChP) the great majority of reference extractives have been primarily used for qualitative identification by thin-layer chromatography (TLC) and few studies on the applicability of reference extractives for quantitative analysis have been presented. Using Salvia miltiorrhiza Bunge as an example in this paper, we first present a preliminary discussion on the feasibility and applicability of reference extractives for the quantitative analysis of TCMs. The reference extractive of S. miltiorrhiza Bunge, comprised of three pharmacological marker compounds, namely cryptotanshinone, tanshinone I and tanshinone IIA, was prepared from purchased Salvia miltiorrhiza Bunge by extraction with acetone under reflux, followed by silica gel column chromatography with stepwise elution with petroleum ether-ethyl acetate (25:1, v\/v, 4.5 BV) to remove the non-target components and chloroform-methanol (10:1, v\/v; 3 BV) to yield a crude reference extractive solution. After concentration, the solution was further purified by preparative reversed-phase HPLC on a C18 column with isocratic elution with 77% methanol aqueous solution to yield the total reference extractive of S. miltiorrhiza Bunge. Thereafter, the reference extractive was applied to the quality assessment of S. miltiorrhiza Bunge using high-performance liquid chromatography (HPLC) coupled with diode array detection (DAD). The validation of the method, including linearity, sensitivity, repeatability, stability and recovery testing, indicated that this method was valid, reliable and sensitive, with good reproducibility. The developed method was successfully applied to quantify seven batches of samples collected from different regions in China and the results were also similar to those obtained using reference standards, with relative standard deviation (RSD) &lt;3%. Preparation of a reference extractive of S. miltiorrhiza Bunge was significantly less expensive and time consuming than preparation of a corresponding reference standard. Quantitative analysis using a reference extractive was shown to be simple, low-cost, time-saving and practical, with high sensitivity and good stability; and is, therefore, a strong alternative to the use of reference standards.<\/jats:p>","DOI":"10.3390\/molecules18066919","type":"journal-article","created":{"date-parts":[[2013,6,13]],"date-time":"2013-06-13T11:30:15Z","timestamp":1371123015000},"page":"6919-6935","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Application of a Rapid and Efficient Quantitative Analysis Method for Traditional Chinese Medicines: The Case Study of Quality Assessment of Salvia miltiorrhiza Bunge"],"prefix":"10.3390","volume":"18","author":[{"given":"Wen-Guang","family":"Jing","sequence":"first","affiliation":[{"name":"Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences,  Beijing 100700, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Zhang","sequence":"additional","affiliation":[{"name":"Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences,  Beijing 100700, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li-Yan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Medicine College of Guiyang College of Traditional Chinese Medicine,  Guiyang College of Traditional Chinese Medicine, Guiyang 550002, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dong-Zhe","family":"Wang","sequence":"additional","affiliation":[{"name":"Medicine College of Guiyang College of Traditional Chinese Medicine,  Guiyang College of Traditional Chinese Medicine, Guiyang 550002, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yue-Sheng","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences,  Beijing 100700, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"An","family":"Liu","sequence":"additional","affiliation":[{"name":"Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences,  Beijing 100700, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2013,6,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.ctim.2005.06.004","article-title":"Are national quality standards for traditional Chinese herbal medicine sufficient? Current governmental regulations for traditional Chinese herbal medicine in certain Western countries and China as the Eastern origin country","volume":"13","author":"Dobos","year":"2005","journal-title":"Complement. Ther. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.aca.2009.10.024","article-title":"Recent analytical approaches in quality control of traditional Chinese medicines\u2014a review","volume":"657","author":"Jiang","year":"2010","journal-title":"Anal. Chim. Acta"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1016\/S1872-2059(06)60022-9","article-title":"Strategy and chromatographic technology of quality control for traditional chinese medicines","volume":"24","author":"Li","year":"2006","journal-title":"Chin. J. Chromatogr."},{"key":"ref_4","first-page":"2388","article-title":"Reference extractive determination of 5 components content of pueraria labta","volume":"37","author":"Xue","year":"2012","journal-title":"Chin. J. Chin. Mater. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.ijcard.2007.01.004","article-title":"Cardiovascular effects of Danshen","volume":"121","author":"Cheng","year":"2007","journal-title":"Int. J. Cardiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1006\/taap.2002.9367","article-title":"Salvia miltiorrhiza Inhibits Biliary Obstruction-Induced Hepatocyte Apoptosis by Cytoplasmic Sequestration of p53","volume":"182","author":"Oh","year":"2002","journal-title":"Toxicol. Appl. Pharmacol."},{"unstructured":"Guo, J.X. (1992). Research and Clinical Application of the Root of Salvia Miltiorrhiza. Bunge, Chinese Medicine and Technology Press.","key":"ref_7"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1348","DOI":"10.1016\/j.carbpol.2011.09.024","article-title":"Extraction, preliminary structural characterization, and antioxidant activities of polysaccharides from Salvia miltiorrhiza Bung","volume":"87","author":"Wu","year":"2012","journal-title":"Carbohydr. Polym."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1177\/0091270005282630","article-title":"Danshen: An overview of its chemistry, pharmacology, pharmacokinetics, and clinical use","volume":"45","author":"Zhou","year":"2005","journal-title":"J. Clin. Pharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/S0304-3835(00)00391-8","article-title":"Salvia miltiorrhiza inhibits cell growth and induces apoptosis in human hepatoma HepG2 cells","volume":"153","author":"Liu","year":"2000","journal-title":"Cancer Lett."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1080\/10715760000301471","article-title":"Tanshinone II-A inhibits low density lipoprotein oxidation in vitro","volume":"33","author":"Niu","year":"2002","journal-title":"Free Radic. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1503","DOI":"10.1080\/10826070600674968","article-title":"Fingerprinting of Salvia miltiorrhiza Bunge by thin-layer chromatography scan compared with high speed countercurrent chromatography","volume":"29","author":"Gu","year":"2006","journal-title":"J. Liq. Chromatogr. Relat. Technol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1248\/cpb.53.481","article-title":"Quality Assessment of Radix Salviae","volume":"53","author":"Ping","year":"2005","journal-title":"Miltiorrhizae. Chem. Pharm. Bull."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.chroma.2004.08.145","article-title":"Fingerprinting of Salvia miltiorrhiza Bunge by non-aqueous capillary electrophoresis compared with high-speed counter-current chromatography","volume":"1057","author":"Gu","year":"2004","journal-title":"J. Chromatogr A"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1365\/s10337-007-0244-4","article-title":"Simultaneous determination of seven active compounds in radix salviae miltiorrhizae by temperature-controlled ultrasound-assisted extraction and HPLC","volume":"66","author":"Qu","year":"2007","journal-title":"Chromatographia"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"368","DOI":"10.1002\/pca.1063","article-title":"Simultaneous determination of ten active components in traditional Chinese medicinal products containing both gegen (Pueraria iobata) and danshen (Salvia miltiorrhiza) by high-performance liquid chromatography","volume":"19","author":"Chang","year":"2008","journal-title":"Phytochem. Anal."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"931","DOI":"10.1002\/bmc.836","article-title":"Improved quality evaluation of Radix Salvia miltiorrhiza through simultaneous quantification of seven major active components by high-performance liquid chromatography and principal component analysis","volume":"21","author":"Ma","year":"2007","journal-title":"Biomed. Chromatogr."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.jchromb.2006.08.026","article-title":"Development and evaluation of an efficient HPLC\/MS\/MS method for the simultaneous determination of pseudoephedrine and cetirizine in human plasma: Application to Phase-I pharmacokinetic study","volume":"846","author":"Ma","year":"2007","journal-title":"J. Chromatogr. B"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2256","DOI":"10.1002\/jssc.200700149","article-title":"Chromatographic fingerprinting analysis of Danshen root (Salvia miltiorrhiza Radix et Rhizoma.) and its preparations using high performance liquid chromatography with diode array detection and electrospray mass spectrometry (HPLC-DAD-ESI\/MS)","volume":"30","author":"Liu","year":"2007","journal-title":"J. Sep. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1002\/bmc.911","article-title":"Determination of fifteen bioactive components in Radix et Rhizoma. Salviae. Miltiorrhizae. by high-performance liquid chromatography with ultraviolet and mass spectrometric detection","volume":"22","author":"Cao","year":"2008","journal-title":"Biomed. Chromatogr."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.jchromb.2006.08.002","article-title":"Development of the fingerprints for the quality of the roots of Salvia miltiorrhiza and its related preparations by HPLC-DAD and LC\u2013MSn","volume":"846","author":"Liu","year":"2007","journal-title":"J. Chromatogr. B"},{"unstructured":"Pharmacopoeia Commission of People\u2019s Republic of China (2010). Pharmacopoeia of the People\u2019s Republic of China.","key":"ref_22"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1158\/1940-6207.CAPR-10-0020","article-title":"Cryptotanshinone inhibits cancer cell proliferation by suppressing mammalian target of rapamycin-mediated Cyclin D1 expression and Rb phosphorylation","volume":"3","author":"Chen","year":"2010","journal-title":"Cancer Prev. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1885","DOI":"10.1093\/carcin\/bgn151","article-title":"anshinone I suppresses growth and invasion of human breast cancer cells, MDA-MB-231, through regulation of adhesion molecules","volume":"29","author":"Nizamutdinova","year":"2008","journal-title":"Carcinogenesis"},{"key":"ref_25","first-page":"431","article-title":"Study on Technical Processes for Extraction and Purification of Total Tanshinones","volume":"31","author":"Zhang","year":"2008","journal-title":"J. Chin. Mater. Med."},{"key":"ref_26","first-page":"1815","article-title":"Study of comparison on different purify technologies of total tanshinones","volume":"39","author":"Wu","year":"2008","journal-title":"J. Chin. Tradition. Herb Drugs"},{"key":"ref_27","first-page":"198","article-title":"Analysis of Cryptotanshinone by Electron Spray Ionization Mass Spectrometry and Electron Ionization Mass Spectrometry","volume":"27","author":"Dai","year":"2006","journal-title":"J. Chin. Mass Spectrum. Soc."},{"key":"ref_28","first-page":"24","article-title":"Elucidation of tanshinone IIA and tanshione I by electron spray ionization mass spectrometry and electron ionization mass spectrometry","volume":"27","author":"Dai","year":"2008","journal-title":"Chin. J. Anal. Lab."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/18\/6\/6919\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:47:19Z","timestamp":1760219239000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/18\/6\/6919"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,6,13]]},"references-count":28,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2013,6]]}},"alternative-id":["molecules18066919"],"URL":"https:\/\/doi.org\/10.3390\/molecules18066919","relation":{},"ISSN":["1420-3049"],"issn-type":[{"type":"electronic","value":"1420-3049"}],"subject":[],"published":{"date-parts":[[2013,6,13]]}}}