| Abstract: It has proved that the total naphthoquinones in Arnebia euchroma (Royle) Johnst are antineoplastic components by both in vivo and in vitro experiment of modern medicine. But there have been no reported publicly to shikonin emulsion. This research studies the preparation of shikonin emulsion and antineoplastic assay in vitro.The paper includes five major contents as follows:1. Extraction, purification, identification and content determination of the total naphthoquinones in Arnebia euchroma (Royle) Johnst. We can acquire the total naphthoquinones using petroleum ether as a solvent with Soxhlet's extraction and purification with deposition of acid and alkali; Qualitative identification by TLC, there is a same spot between laevogyrate shikonin reference substance and shikonin extraction, Rf is 0.47. The determination of total naphthoquinones in extraction is measured to 83.20% by UV. The determination of shikonin in extraction is measured to 56.25% by HPLC.2. The preparation technology of shikonin lipid emulsion. Using F-68 and emulsificating temperature and speed of dispersing emulsificating machine as changing factors in orthogonal experimental design, adopting the stability of calefaction and centrifugation and particle size as the coindicator, we can analyze stepwise. The optimal prescription are as follows: oil of injection is percent 15, F-68 is percent 2.0, soybean lecithoid is percent 1.2, emulsificating temperature is 70 centigrade, speed of dispersing emulsificating machine is 22000rpm, assistant surfactant is appropriate. The content of total naphthoquinones in emulsion are 0.69mg/mL.3. Checking physical stability of shikinon lipid emulsion. The emulsion has no obvious changes after15 minutes centrifugation with speed of 4000rpm. The average particle size of shikonin emulsion is 2.8 μ m, it accords with demand of injection The viscosities of double distilled water, soybean oil and shikonin emulsion are 3.0 mPa s , 83.2 mPa s and 7.5 mPa s respectively, the results show that the emulsion accords with the demand of injection. There is no obvious difference between emulsion and Intralipid, and three batches samples' zeta potential is 49,48 and 46mV, Intralipid is 42 mV. It proves that the emulsion is steady.4. Control of shikinon emulsion in quality. Shikinon emulsion is fuchsia and O/W emulsion, its pH is 6.5.To identify qualitatively shikonin emulsion by TLC. There is a same spot between laevogyrate shikonin reference substance and shikonin emulsion, and Rf is 0.47. The content of total naphthoquinones in shikonin emulsion can be determined by UV. The results show that the labeled amounts of shikonin emulsion is 97.81 % , RSD 为 0.70%, and it accords demand of preparation.5. Antineoplastic experiment of shikonin lipid emulsion in vitro. The results show that 3.5×10-1 mg/mL shikonin injection have significant antineoplastic effect on EJ, Hela, Bcap and H08910 cells in vitro, and the antineoplastic rate are 73.3%, 61.2%, 81.1% and 63.5% respectively; 3.5×10-2 mg/ml shikonin emulsion have faintish antineoplastic effect on EJ, Hela, Bcap and H08910 cells in vitro, and the antineoplastic rate are 53.6%, 22.0%, 57.8% and 3.0% respectively. Other dose groups have all no antineoplastic effect on EJ, Hela, Bcap and H08910 cells in vitro.
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