經濟部中央標準局貝工消費合作社印製 A7 B7 五、發明説明（） > 本發明係有關類紫杉固醇Μ離心分配層析法之離心*和 更特別地，有關紫杉結（Taxotere)和10 -去乙醯基漿果赤 徽素I之纯化。 自紫杉針葉萃取之10 -去乙醯基漿果赤徽素11[在依例如 歐洲'專利£卩0,253,738或£卩0,336,841號或國際專利申請 荼第PCT W0 92/09,589號中所述之條件下製備紫杉萜上有 用。 以萃取紫杉針葉所.得之10 -去乙醯基漿果赤徽素jf根據 其所源自之種類而含有不純物，其主要為屬於類紫杉固醇 族之產物，例如2cc-苄醯氧基-4-乙醯氧基-5/3, 20-環氧 -1/3，7yS ,10/3，13α ,19 -五經基-9-氧-11-紫杉稀、2α -苄醯氧基-4-乙醯氧基- 5/3 ,20 -環氧- lyS ,7α，1〇召，13« -四羥基-1卜紫杉烯或毒素。 以部分合成自10 -去乙醯基獎果赤徽素Μ所得之紫杉秘 含有為主要不純物之內容於10 -去乙釀基漿果赤激素f之 不纯物之酯化產物Μ及其他源自例如在支键2’位置上之碳 之表異構作用之不純物。 10 -去乙醢基漿果赤徽素®和紫杉萜本身通常可丨乂液相 層析法於管柱內，和更特別地Μ高效液相層析法（HPLC)|$ 矽石管柱内純化。然而，雖然該等方法特別合適於數十克 之純化*但是工業上之應用有其限制.，例如消耗之溶_ 量 和為毒性殘渣所污染之撐體（矽石）之處理及破壊，其@ 成主要的限制。 在很不安定和常帶毒性之類紫杉固醇之情形下，特別重 ~ ~ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） e —I I I I I I I I 裝 I I I I 訂 __ I I i I I 線 (請先閲讀背面之注意事項再填寫本頁) 經濟部中央標準局員工消費合作社印製 A7 _B7 五、發明説明（） » 要的是能使用具良好生產力，不需要高價購買、使用和破 壞之固體撐體，需要使用少量之溶劑和可容易自動化Μ利 連續生產之純化方法。 頃發現且構成本發明之主題者係類紫杉固醇之純化，和 更特別地紫杉萜和1 0 -去乙醢基漿果赤徽素I之純化可Κ 離心分配曆析法（C P C )或高速逆流層析法達成，其原理例 如由 A. Foucault和 P. Rosset於 Analusis, LL (3 )， 157-167 (1988)中記述。 離心分配層析法Μ建立在理論方式上和自動地產生之連 績平衡而使得有效分配混合物之姐成於兩種不相混之液層 中分離成為可能。該方法提供很有效之分配機構、固定相 之強度滯留和移動相之高速*而導致在數小時內之良好分 離。 離心分配層析法需要使用2或多種溶液，Μ提供2個部 分互混相。雖然有無限量之溶劑展現此特色，但工業上特 別重要的是使用非氛化和非毒性工業溶劑。在可工業使用 且招致較低危險性之溶劑中，如甲醇之酵、如甲基第三丁 基醚之醚、如乙酸乙酯之酯、如甲基異丁基嗣之酮和如庚 烷或異辛烷之脂族烴可被提及。 為了進行有效的純化作用，必須選擇快速且完全分開Μ 產生兩相之溶劑混合液，其分配係數係介於0 . 1和10之間 ，和較佳地介於0 . 5和5之間，和更佳地在.1左右。 分配係數（Κ)例如等於10之產物係指，溶於上層相之量 較之溶於下層相者多10倍·。结果，若上層相在移動時*其 ____ - 4 ~ — 本紙張尺度適用中國國家標準（CNS ) Α4規格（210Χ297公釐） C : ----------1------.訂丨------^ (請先閲讀背面之注意事項再填寫本頁) 經濟部中央橾準局貝工消费合作社印製 A7 B7 五、發明説明（） * 將很快地溶析出，若下層相在移動時，在很長的時間後會 有著很差的分離。相對地，當K是1左右時，二相之溶解度 實際上相當，且兩種形式之溶析可被使用，和適當地合併 時Μ獲得最大的分離。 特就10 -去乙醯基漿果赤徽素15而論，脂族酮和水的混 合物，如甲基異丁基酮/丙酮/水（體積比為2:3:2)混合 液為最特別適合的。 再者，必須使固定相中之滯留，其為在平衡時由固定相 所佔有總設備體積之部分，能儘可能地高且等於至少50% ，而固定相之流速則被設定。 再者*為了改進生產力，能在高流速下進行是特別有利 的，其受限於轉子驅動之最大壓力。 在紫杉萜之特別情形下，有利的是使用脂族烴、酯、醇 和水之混合液，如庚烷/乙酸乙酯/甲酵/水（2:4:3:2 ，體積比）糸統，其具80%之滯留和0.5區域之分配係數 ，因此得到快速溶析之窄峰，因此導致在生產力上顯著之 改進。 離心分配層析法可在任何適合此目的之商業來源之設備 中進行，如該等已由Yoichiro Ito所發展者[CRC Crit. Rev. Anal. Chea·，Π_, 65 (1986)]或巳由京都（日本） 之SANKI工程公司所銷售者。 本發明亦有闞純化的紫杉萜和純化的10 -去乙醯基槳果 赤徽素II ，其係自進行根據本發明之方法所獲得者。 Μ下之實施例顯示本發明可如何地實施。 本紙張尺度適用中國國家楳準（CNS ) Α4規格（210X297公釐） e : I . 11· n 訂 線 (請先閱讀背面之注意事項再填寫本頁) 38975S at ______B7五、發明説明（） 經濟部中央橾準局負工消费合作社印«. 宜Jj例1 -紫杉萜（Taxotere)之纯化 紫杉萜（或4-乙醢氧ί-2α-苄醯氧基-5>8,20-環氧 -Ια ,7点，10/3 -三羥基-9-氧-1卜紫杉烯-13α -基（2R, 3S) -3-第三丁氧基羰基胺基-3-笨基-2-羥基丙酸酯）係由 4-乙醯氧基-2α -苄醯氧基-5/3 ,20-環氧-1/3-羥基- 7/9 ，10雙（2,2,2-三氯乙氧基羰氧基）-9-氧-11-紫杉烯 -13α-基（2R, 3S)-3-第三丁氧基羰基胺基-3-苯基-2-羥 基丙酸酯與‘鋅在醋酸之存在下，在歐洲專利ΕΡ0,336,841 號中所述之條件下處理製得。 10克4-乙醢氧基-2α -苄醯氧基-5/3 ,20-環氧- lyS -羥 基-7/3,10/8-雙（2,2,2-三氯乙氧基羰氧基）-9-氧-11-紫杉烯-13α -基（2R，3S)-3-第三丁氧基羰基胺基-3-笨 基-2-羥基丙酸酯溶於150毫升甲基第三丁基醚中，然後 加入10克样。蒸發至乾涠後，獲得灰色粉末，再置入反應 器中。 攪拌後，在5分鐘內加入6毫升醋酸和50毫升乙腈之混 合液。混合液Μ水浴在45 下外在加熱1小時。在冰浴中 冷卻後* 850奄升甲基第三丁基醚Κ激烈撗拌導入，然後 於4號孔洞之燒结玻璃中濾掉鋅鹽和過量之鋅。 濾液經濃縮至乾涸。因而獲得泡沫（12克），其立即Μ 20毫升乙酸乙酯、15奄升甲醇、10毫升庚烷和10毫升水溶 取。 離心分配層析法K SANKI LLI-07裝置進行。離心機由裝 (請先閲讀背面之注意事項再填寫本頁) -裝. e 訂 線 _____~ 6 * 本紙張尺度適用中國國家標準（CNS ) Α4規格（210Χ297公釐） 經濟部中央標準局貝工消費合作社印簟Printed by A7 B7, Shellfish Consumer Cooperative, Central Bureau of Standards, Ministry of Economic Affairs 5. Description of the invention () > The invention relates to centrifugation of taxoid M centrifugal distribution chromatography * and, more specifically, to taxotere (Taxotere ) And 10-purified erythromycin I. Extracted from Yew needles 10-deacetylated erythromycin 11 [Conditions described in, for example, European 'patents £ 0,253,738 or £ 0,336,841 or International Patent Application No. PCT W0 92 / 09,589 Useful for the preparation of docetaxel. Extracted from Yew needles. The 10-deacetylated berry erythromycin jf contains impurities according to the type from which it is derived, which is mainly a product belonging to the taxoid family, such as 2cc-benzidine Oxy-4-ethenyloxy-5 / 3, 20-epoxy-1 / 3, 7yS, 10/3, 13α, 19-pentaethyl-9-oxy-11-taxane, 2α-benzidine Oxy-4-ethoxyl-5 / 3,20-epoxy-lyS, 7α, 10zol, 13 «-tetrahydroxy-1btaxane or toxin. The content of the yew secret that is partially synthesized from the 10-deacetylammonium erythromycin M is the main impurity, and the esterification products of the impurities of the 10-deethyl berry red hormone f and other sources Impurities from, for example, epimerization of carbon at the 2 'position of a branch bond. 10-Deacetylated erythromycin® and docetaxel itself can usually be chromatographed in a column, and more specifically HPLC | silica column Within purification. However, although these methods are particularly suitable for tens of grams of purification *, industrial applications have their limitations. For example, the amount of solvent consumed and the treatment and breaking of the support (silica) contaminated by toxic residues, its @ Into major restrictions. In the case of very unstable and often toxic taxol such as paclitaxel, this paper is particularly heavy ~ ~ This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) e —IIIIIIII Pack IIII order __ II i II (Please read the notes on the back before filling out this page) Printed by the Consumers Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 _B7 V. Description of the invention () »What is needed is good productivity, no need to buy, use and destroy at high prices The solid support requires a small amount of solvent and a purification method that can be easily automated for continuous production. The purification of paclitaxel, and more specifically the purification of docetaxel and 10-desethyl erythromycin I found and constituting the subject of the present invention, can be performed by centrifugal partition calorimetry (CPC) Or high-speed countercurrent chromatography, the principle of which is described, for example, by A. Foucault and P. Rosset in Analusis, LL (3), 157-167 (1988). Centrifugal partition chromatography, M, is based on a theoretical balance and a series of results that are automatically generated to make it possible for the sister of an effective partition mixture to separate into two immiscible liquid layers. This method provides a very efficient distribution mechanism, strength retention of the stationary phase, and high speed of the mobile phase * leading to good separation within hours. Centrifugal partition chromatography requires 2 or more solutions, and M provides 2 fractions that are intermixed. Although an unlimited number of solvents exhibit this feature, it is particularly important industrially to use non-aqueous and non-toxic industrial solvents. In solvents that can be used industrially and incur lower risks, such as methanolic fermentation, ethers such as methyl tert-butyl ether, esters such as ethyl acetate, ketones such as methyl isobutylfluorene, and heptane or Aliphatic hydrocarbons of isooctane can be mentioned. In order to perform effective purification, it is necessary to select a solvent mixture that quickly and completely separates M to produce two phases, the partition coefficient of which is between 0.1 and 10, and preferably between 0.5 and 5, And better around .1. A product with a partition coefficient (K) of, for example, 10 means that the amount dissolved in the upper phase is 10 times more than that in the lower phase. As a result, if the upper phase is moving * its ____-4 ~ — This paper size applies the Chinese National Standard (CNS) A4 specification (210 × 297 mm) C: ---------- 1 ---- -. Order 丨 ------ ^ (Please read the notes on the back before filling out this page) Printed by A7 B7, Shellfish Consumer Cooperatives, Central Bureau of Standards, Ministry of Economic Affairs V. Description of invention () * will soon Dissolution, if the lower phase is moving, there will be poor separation after a long time. In contrast, when K is about 1, the solubility of the two phases is practically equivalent, and the two forms of leaching can be used, and M can achieve the maximum separation when properly combined. As far as 10-deacetylated erythromycin 15 is concerned, a mixture of aliphatic ketone and water, such as methyl isobutyl ketone / acetone / water (volume ratio 2: 3: 2) is the most special suitable. In addition, the stationary phase must be retained, which is the part of the total equipment volume occupied by the stationary phase at equilibrium, which can be as high as possible and equal to at least 50%, and the flow rate of the stationary phase is set. Furthermore * In order to improve productivity, it is particularly advantageous to be able to perform at high flow rates, which is limited by the maximum pressure of the rotor drive. In the special case of docetaxel, it is advantageous to use a mixture of aliphatic hydrocarbons, esters, alcohols, and water, such as heptane / ethyl acetate / formaldehyde / water (2: 4: 3: 2, volume ratio) 糸System, which has a retention of 80% and a distribution coefficient in the 0.5 region, so a narrow peak for rapid dissolution is obtained, which results in a significant improvement in productivity. Centrifugal partition chromatography can be performed in any commercial source equipment suitable for this purpose, such as those developed by Yoichiro Ito [CRC Crit. Rev. Anal. Chea ·, Π_, 65 (1986)] or by Seller of SANKI Engineering Co., Ltd. in Kyoto (Japan). The present invention also includes taurine-purified docetaxel and purified 10-deacetylamylate erythromycin II obtained from performing the method according to the present invention. The examples under M show how the invention can be implemented. This paper size applies to China National Standards (CNS) Α4 size (210X297 mm) e: I. 11 · n Thread (please read the precautions on the back before filling this page) 38975S at ______B7 V. Invention Description () Economy Printed by the Ministry of Health, the Central Bureau of quasi-industry, Consumer Cooperatives «. JJ Example 1-Purification of Taxane (Taxotere) (or 4-Ethyloxyl-2α-benzyloxy-5-> 8,20- Epoxy-1α, 7-point, 10 / 3-trihydroxy-9-oxo-1 taxane-13α-yl (2R, 3S) -3-third butoxycarbonylamino-3-benzyl-2- Hydroxypropionate) is based on 4-ethoxyl-2α-benzyloxy-5 / 3, 20-epoxy-1 / 3-hydroxy-7 / 9, 10 bis (2,2,2-tri (Chloroethoxycarbonyloxy) -9-oxy-11-taxane-13α-yl (2R, 3S) -3-third butoxycarbonylamino-3-phenyl-2-hydroxypropionate And 'zinc in the presence of acetic acid, prepared under the conditions described in European Patent EP 0,336,841. 10 g of 4-ethoxymethyl-2α-benzyloxy-5 / 3,20-epoxy-lyS-hydroxy-7 / 3,10 / 8-bis (2,2,2-trichloroethoxy (Carbonyloxy) -9-oxy-11-taxane-13α-yl (2R, 3S) -3-third butoxycarbonylamino-3-benzyl-2-hydroxypropionate dissolved in 150 ml Methyl tert-butyl ether was then added to a 10 g sample. After evaporation to dryness, a gray powder was obtained and placed in the reactor. After stirring, a mixture of 6 ml of acetic acid and 50 ml of acetonitrile was added over 5 minutes. The mixed liquid M water bath was externally heated at 45 ° C for 1 hour. After cooling in an ice bath, 850 liters of methyl tert-butyl ether K was vigorously mixed and introduced, and then the zinc salt and excess zinc were filtered out in sintered glass with a No. 4 hole. The filtrate was concentrated to dryness. Thus, a foam (12 g) was obtained, which was immediately dissolved in 20 ml of ethyl acetate, 15 ml of methanol, 10 ml of heptane and 10 ml of water. Centrifugal partition chromatography was performed with K SANKI LLI-07 device. The centrifuge is installed (please read the precautions on the back before filling this page)-installed. E Thread _____ ~ 6 * This paper size is applicable to China National Standard (CNS) Α4 size (210 × 297 mm) Central Bureau of Standards, Ministry of Economic Affairs Shellfish Consumer Cooperative
38975S ____B7_____ 五、發明説明（） t 有揚上閥門且限制壓力至55巴之兩個活塞泵所供給。一個 泵藉選擇閥用來供输兩液層之一或另一；另一涸泵則用Μ 注入。 低速旋轉（ 200 rpn)和高流速（110毫升/分）下以固 定相填充裝置後，注入即立刻進行。 · •兩液相之注入係始於水相，而溶析係K己烷/乙酸乙酯 /甲醇/水（2: 4: 3: 2，體積比）糸统之水相在60毫升 /分之流速下進行。.收集固定相之1.9升，相當於71%之 滯留，和然後收集15毫升之區分。區分72至110經集中和 濃縮至140毫升之體積。形成之沈澱Μ過滹分開。如此可 得80. 5%產量之5.2克紫杉萜三水合物*使用内檷準法分 析為99 . 7%。 實-施例2 -紫杉萜之純化 使用在實施例1中所述之程序，但Μ 82克4-乙醯氧基 -2ot -苄醯氧- 5yS ,20-環氧-羥基-7yS ,10々-雙（ 2,2,2-三氯乙氧基羰氧基）-9-氧-11-紫杉烯-13α-基 (2R, 3S)-3-第三丁氧基羰基胺基-3-苯基-2-羥基丙酸酯 、82克鋅和49.2毫升醋酸和328毫升乙腈之150毫升混合液 開始，可獲得理論上含有49 . 69克紫杉萜或53克紫杉萜Η 水合物之115克粗產物。 袓產物Μ 450毫升甲基第三丁基醚溶取且然後Μ 3次總 量450毫升含15克氯化納之水清洗。合併之水相M2次 200毫升甲基第三丁基醚萃取。三次醚萃取液經合併且濃 ------- - ~ 7 ~ 本紙張尺度適用中國國家標準（CNS ) Α4規格（210Χ297公釐） I I I I __ 訂 I . I 線 (請先閲讀背面之注意Ϋ項再填寫本頁)38975S ____B7_____ V. Description of the invention () t Two piston pumps with raised valves and limited pressure to 55 bar are supplied. One pump uses a selection valve for one or the other of the two layers; the other pump is injected with M. After filling the device with a solid phase at low speed (200 rpn) and high flow rate (110 ml / min), the injection was performed immediately. · • The injection system of two liquid phases starts from the water phase, while the dissolution system is Khexane / ethyl acetate / methanol / water (2: 4: 3: 2, volume ratio). The water phase of the system is 60 ml / min. Performed at a low flow rate. Collect 1.9 liters of stationary phase, which is equivalent to 71% retention, and then collect 15 ml. The divisions 72 to 110 are concentrated and concentrated to a volume of 140 ml. The formed precipitate was separated. In this way, 5.2 grams of docetaxel trihydrate with a yield of 80.5% can be obtained using internal quasi-analysis to 99.7%. Example 2-Purification of docetaxel was carried out using the procedure described in Example 1, but M 82 g of 4-ethoxymethyl-2ot-benzylidene-5yS, 20-epoxy-hydroxy-7yS, 10々-bis (2,2,2-trichloroethoxycarbonyloxy) -9-oxy-11-taxane-13α-yl (2R, 3S) -3-third butoxycarbonylamino Starting from a mixture of 3--3-phenyl-2-hydroxypropionate, 82 g of zinc and 49.2 ml of acetic acid and 328 ml of acetonitrile in 150 ml, theoretically 49.69 g of docetaxel or 53 g of docetaxel can be obtained 115 g of crude product of hydrate. The hydrazone product M was dissolved in 450 ml of methyl tert-butyl ether and then washed 3 times in total with 450 ml of water containing 15 g of sodium chloride. The combined aqueous phases were extracted twice with 200 ml of methyl tert-butyl ether. Three times ether extraction solution is combined and concentrated --------~ 7 ~ This paper size is applicable to Chinese National Standard (CNS) A4 specification (210 × 297 mm) IIII __ Order I. I line (Please read the note on the back first (Please fill in this page for the item)
389^5£ a7 B7 五、發明説明（） ) 縮至乾涸。如此可得72克泡沫，其產物Μ 120毫升甲酵溶 取0 在所得之淡黃色溶液中，依序加入1 6 0毫升乙酸乙酯、 80毫升水和80毫升庚烷。 如'此可得總體積為460奄升之兩清徹的液相，上層僅僅 佔總髏積的1/4而非35%，且不含紫杉萜。 離心分配層析法装置在110奄升/分之流速及200 rpm 轉速下Μ有機相填充。當裝置充滿時，轉速升至800 r pm 。水相經注入且在之後有機相注入後，Μ 200毫升有機相 進行潤洗，且然後Μ水相，用60毫升/分之流速溶析。 收集2.5升過量之固定相，.相當於67. 7%之滯留。 在200毫升水性移動相通過後，收集360個12至13毫升 之區分。區分25至110經濃縮至.200毫升體積，得到 87.7%產量之46.5克紫杉萜三水合物之沈澱，其純度為 99.1%。389 ^ 5 £ a7 B7 V. Description of the invention ()) Shrunk to dryness. In this way, 72 g of a foam was obtained, and the product M of 120 ml of formic acid was dissolved. To the obtained pale yellow solution, 160 ml of ethyl acetate, 80 ml of water, and 80 ml of heptane were sequentially added. For example, two clear liquid phases with a total volume of 460 liters can be obtained, and the upper layer only accounts for 1/4 of the total cross-sectional area instead of 35%, and does not contain docetaxel. The centrifugal distribution chromatography device was filled with M organic phase at a flow rate of 110 L / min and a rotation speed of 200 rpm. When the device is full, the speed rises to 800 r pm. After the water phase was injected and after the organic phase was injected, the M 200 ml organic phase was rinsed, and then the M water phase was eluted with a flow rate of 60 ml / min. Collect 2.5 liters of excess stationary phase, which is equivalent to 67.7% of retention. After 200 ml of the aqueous mobile phase had passed, 360 12 to 13 ml divisions were collected. The 25 to 110 fractions were concentrated to a volume of .200 ml, and a precipitate of 46.5 g of docetaxel trihydrate was obtained in a yield of 87.7%, with a purity of 99.1%.
置JUL1 - 1 〇 -去乙醯基漿果赤黴素I 離心分配層析法於總體積245毫升之SANKI HPCPC序列 1000屬析儀中進行。 經濟部中央橾準局貝工消费合作杜印製 {請先閲讀背面之注意事項再填寫本頁) 轉子在100 rpm轉動時，Κ降下方式填充，同時泵入6 毫升/分流速之有機相和6毫升/分流速之甲基異丁基酮 /丙酮/水（2: 3: 2，體積比）混合液之水相。 接著轉速升至1 200 rpm和然後注入含有83.3%10_去乙 醯基漿果赤徽素JI和3.4% 10-去乙醢基-19-羥基漿果赤 「_ - - - ^ -本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） 38975S a7 ____B7 五、發明説明（）Centrifugal partition chromatography with JUL1-10-deacetylsulfate-baccatin I was performed in a SANKI HPCPC Sequence 1000 analyzer with a total volume of 245 ml. Printed by Shelley Consumer Co-operation, Central Bureau of Standards, Ministry of Economic Affairs {Please read the precautions on the back before filling out this page) When the rotor is rotating at 100 rpm, it will be filled by KK lowering mode, and the organic phase and The aqueous phase of a mixed solution of methyl isobutyl ketone / acetone / water (2: 3: 2 by volume) at a flow rate of 6 ml / min. Then the speed was increased to 1 200 rpm and then injected with 83.3% 10_ deacetylammonium erythromycin JI and 3.4% 10-deacetylamyl-19-hydroxyberry erythroline `` _---^-this paper size applies China National Standard (CNS) A4 specification (210X297 mm) 38975S a7 ____B7 V. Description of the invention ()
I 嫌素II之1.5克沈澱之粗10 -去乙醯基槳果赤徽素皿。 溶析Μ下降之方式M3毫升/分流速之水相進行，每2 分鐘收集6毫升之餾分。 10 -去乙醢基-19 -羥基漿果赤徽素ffi係包含於餾分39至 52 中。 ， •在第61餾分時，逆轉溶析之方向，然後使用有機相為移 動相，K上升之方式，流逮為5奄升/分，且每2分鐘收 集10毫升之餾分。 餾分70至84含有10 -去乙醯基漿果赤徽素u，其在減壓 下濃縮。在15毫升乙腈中再结晶後，可得白色結晶以乙腈 (1莫耳/莫耳）溶劑合之1·25克10-去乙醢基漿果赤徼 素ΒΙ ，其純度由HPLC使用内標準法測定為98%。 修正之產率為91 . 2%。 Β. .1 ----------^1-----1Τ------.^ (請先閱讀背面之注意事項再填寫本頁) 經濟部中央標準扃員工消费合作社印裝 丨9 - . 本紙張尺度適用中國國家揉準（CNS > Α4規格（210Χ297公釐）1.5 1.5 grams of precipitated crude II-deacetylin II. The dissolution step M was performed in a water phase at a flow rate of M3 ml / min, and 6 ml of fractions were collected every 2 minutes. The 10-deethylamidine-19-hydroxyberry erythrobin ffi system is contained in fractions 39 to 52. At the 61st fraction, the direction of dissolution was reversed, and then the organic phase was used as the mobile phase and the K was raised, the flow rate was 5 liters / min, and 10 ml of fractions were collected every 2 minutes. Fractions 70 to 84 contained 10-deacetylsulforubin u, which was concentrated under reduced pressure. After recrystallizing in 15 ml of acetonitrile, white crystals can be obtained with acetonitrile (1 mole / mole) as a solvent and 1.25 g of 10-desacetomethylberry erythrobin Β1, whose purity is determined by HPLC using internal standard It was determined to be 98%. The revised yield was 91.2%. Β. .1 ---------- ^ 1 ----- 1Τ ------. ^ (Please read the precautions on the back before filling out this page) Central Standards of the Ministry of Economy 扃 Employee Consumption Cooperative print 丨 9-. This paper size is applicable to the Chinese national standard (CNS > Α4 (210 × 297 mm)