42171 8 4 A7 B7 五、發明説明（1 ) 發明領域 本發明係揭示一種分析光學異構物之再循環高效液相層 析裝置與方法，其特徵在於應用自動切換閥控制器，控制 對掌異構物層析管柱和一般層析管柱，於適當的時機，在 高效液相層析裝置中自動切換，以達到再循環分析樣品之 目的。結果顯示，對於無法以對掌異構物層析管柱得到良 好分離之具有光學異構物的樣品，藉本發明之裝置及方法 ，可以得到良好的分離效果。 發明背景 在醫藥品中，有相當多的藥品是以光拳異構物的混合物 型態被使甩，而在該混合物中，通常只有其中的一.個成分 對治療疾病有效，其他成分往往具有負面的影響而造成副 作用，但是，礙於該混合物爲光學異構物，其物理化學性 質十分相似，不易分離，因此，只好以混合物型態被使 用。 , 目前，有關光學異，構物的研.'究，係以f不對稱合成與光學 分割二種方法進行。在光學異構物合成的過程中，一 ^固可 以分離光學異構物的分析方法是很重要的，藉此分析方法 經濟部中央標隼局員工消費合作社印製 (請先閲讀背面之注意事項再填寫本頁) \ ' ，可以追蹤評估反應的結果。 ， 爲了分離光學異構物，有許多高效液相層析管柱的製造 商，紛紛推出各種具有分離光學異構物功能的對掌異構物 層析管柱。對有些光學異構物而言，這些層析管柱確實能 得到良好的分離效果，但是，仍有許多光學異構物，無法 在這些現有的對掌異構物層析管柱中，得到良好的分離效 4 72 78 .UOC ~ 4 - 本紙張讀適用肀國國家標準（CMS ) A4規格（210X297公釐）42171 8 4 A7 B7 V. Description of the invention (1) Field of the invention The present invention discloses a recycling high performance liquid chromatography device and method for analyzing optical isomers, which is characterized by applying an automatic switching valve controller to control the difference The structural chromatography column and the general chromatography column are automatically switched in a high-performance liquid chromatography device at an appropriate timing to achieve the purpose of recycling the analysis sample. The results show that for the samples with optical isomers that cannot be separated well with the palm isomer chromatography column, the device and method of the present invention can obtain a good separation effect. BACKGROUND OF THE INVENTION In pharmaceuticals, there are quite a few medicines that are dumped in the form of a mixture of optical isomers. In this mixture, only one of them is usually effective. The other ingredients are often effective in treating diseases. Negative effects cause side effects, but because the mixture is an optical isomer, its physical and chemical properties are very similar, and it is not easy to separate. Therefore, it has to be used as a mixture. At present, the research on optical heterogeneity and structure. 'Research is carried out by two methods: f asymmetric synthesis and optical segmentation. In the process of optical isomer synthesis, the analytical method that can separate optical isomers is very important. This analysis method is printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the precautions on the back first) Fill out this page again) \ ', you can track the results of the evaluation response. In order to separate optical isomers, there are many manufacturers of high-performance liquid chromatography columns, which have launched various palm isomer chromatography columns with the function of separating optical isomers. For some optical isomers, these chromatographic columns can indeed achieve good separation results. However, there are still many optical isomers that cannot be obtained in these existing palm isomer chromatography columns. Separation efficiency 4 72 78 .UOC ~ 4-This paper reads the national standard (CMS) A4 specification (210X297 mm)
42171 R 經濟部中央樣準局貞工消費合作社印製 五、發明説明（2 果。本發明即是爲解決在一般對掌異構物層析管柱中，無 法得到良好分離效果的樣品而設計。 在鬲效液相層析儀中，應用自動切換閥控制器來切換層析管柱’以達到特別之使用目的，早在1975年即被應用。 在以往的文獻中，有關切換層析管柱在高效液相層析儀上 的應用，主要著重於一般化合物的定量，而且，主要爲利 用不同層析管柱或移動相的性質，以達到分離的效果。從 未有文獻報導應用切換層析管柱在高效液相層析儀上，以 重覆循環分析的原理，達成光學異構物分離之方法。 發明概诚 本發明目的爲提出一種新穎分析光學異構物之再循環高 效液相層析裝置及方法，其可克服無法經一般對掌異構物 層析管柱而得到良好光學異構物分離效果的問題。 本發明王要是由具有時間事件event)輸出功能的液壓 泵、樣mr輸入裝置、六閥式自動切換閥控制器、對掌異構 物層析管柱、一般層析管柱、偵測器裝置及積分儀所组合 而成的自動重覆循環分析之高效液相層析裝置。利用液壓 泵輸出時間事件的訊號，控制六閥式自動切換閥控制器， 在適當的時機作切換的動作，配合對掌異構物層析管柱與 一般層析管柱之間特別的管路連接方式，以達到再循環分 析樣品的效果。 圖式簡單説明 圖1爲再循環高效液相層析裝置之結構圖。 圖2爲液壓泵控制六閥式自動切換閥控制器之訊號時序 本紙張尺度適用中國國家標準（CNS ) A4規格（210X29?公釐） (請先閲讀背面之注意事項再填寫本頁) ο 裝- 訂_ -線 42171 8 A7 B7 經濟部中央標率局貝工消費合作社印製 五、發明説明（3 ) 圖。 圖3爲六閥式自動切換閥控制器背面連接口（remote contact closure)的接線圖。 圖4爲樣品129735A未經再猶環分析的層析圖譜。 圖5爲樣品129735A經一次再循環分析的層析圖譜。 I 圖號簡覃説明 C ,代表對掌異構物層析管柱； C 2代表一般yf析管柱； D代表偵測器裝置； E代表溶劑容器； I代表樣品輸入裝置； p代表液壓泵； R代表積分器； t 〇代表樣品注入時間； 11代表六閥式自動切換閥控制器第一次切換之時間； t2代表六閥式自動切換閥控制器第二次切換之時間； t3代表六閥式自動切換間控制器第三次切換之時間； v代表六閥式自動切換閥控制器； w代表廢液收集瓶； 1代表六閥式自動切換閥控制器的第1個接點； 2代表六閥式自動切換閥控制器的第2個接點 3代表六閥式自動切換閥控制器的第3個接點 4代表六閾式自動切換閥控制器的第4個接點 5代表六閥式自動切換閥控制器的第5個接點；及42171 R Printed by Zhengong Consumer Cooperative, Central Bureau of Standards and Quarantine, Ministry of Economic Affairs. 5. Description of the invention (2 results. The present invention is designed to solve the problem that samples with good separation results cannot be obtained in common palm isomer chromatography columns. In high-efficiency liquid chromatography, an automatic switching valve controller is used to switch the chromatography column 'to achieve a special purpose. It has been used as early as 1975. In the previous literature, the switching of chromatography tubes The application of columns in high-performance liquid chromatography mainly focuses on the quantification of general compounds, and mainly uses the properties of different chromatography columns or mobile phases to achieve the separation effect. There has never been any literature report on the use of switching layers. The analytical column is based on the principle of cyclic analysis on a high performance liquid chromatograph to achieve the method of separating optical isomers. Summary of the Invention The purpose of the present invention is to propose a novel recycled high performance liquid phase for the analysis of optical isomers. Chromatography device and method can overcome the problem that good optical isomer separation effect cannot be obtained through common isomer chromatography columns. The invention is based on the time event even t) Hydraulic pump with output function, sample MR input device, six-valve automatic switching valve controller, palm isomer chromatography column, general chromatography column, detector device and integrator High-performance liquid chromatography with automatic repeat cycle analysis. Use the hydraulic pump to output the signal of time event to control the six-valve automatic switching valve controller to perform the switching action at the appropriate timing, and cooperate with the special pipeline between the isomer chromatography column and the general chromatography column Connection method to achieve the effect of recycling analysis samples. Brief description of the drawings Figure 1 is a structural diagram of a recirculating high performance liquid chromatography device. Figure 2 shows the signal timing of the six-valve automatic switching valve controller for hydraulic pump control. The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X29? Mm) (Please read the precautions on the back before filling this page). Ο Install -Order _-line 42171 8 A7 B7 Printed by the Central Standards Bureau of the Ministry of Economic Affairs, Shellfish Consumer Cooperative, V. Description of Invention (3). Figure 3 is a wiring diagram of a remote contact closure on the back of a six-valve automatic switching valve controller. Figure 4 shows the chromatogram of sample 129735A without further analysis. Figure 5 is a chromatogram of sample 129735A after one-cycle analysis. I Brief description of the drawing number C, C stands for palm isomer chromatography column; C 2 stands for general yf analytical column; D stands for detector device; E stands for solvent container; I stands for sample input device; p stands for hydraulic pump ; R represents the integrator; t 〇 represents the sample injection time; 11 represents the first switching time of the six-valve automatic switching valve controller; t2 represents the second switching time of the six-valve automatic switching valve controller; t3 represents six The time of the third switch of the valve type automatic switching controller; v represents the six-valve automatic switching valve controller; w represents the waste liquid collection bottle; 1 represents the first contact of the six-valve automatic switching valve controller; 2 Represents the second contact of the six-valve automatic switching valve controller 3 Represents the third contact of the six-valve automatic switching valve controller 4 Represents the fourth contact of the six-threshold automatic switching valve controller 5 represents six The fifth contact of the valve-type automatic switching valve controller; and
4727B,I 6 - 21GX瓣费 (請先閎讀背面之注$項再填寫本頁) 裝_ 〇 .IT. 421718 A7 B7 五、發明説明（4 ) 6代表7T.閥式自動切換閥控制器的第6個接黯。 曼盟詳細説明 本發明係關於一種分析光學異構物之再循環高效液相層 析裝置，其包括： ⑴溶劑容器（E ); ⑵樣品輸入裝置（I ); ⑶使溶劑容器（E )内之溶劑推動樣品輸入裝置（j)内之樣品 前進之具時間事件輸出功能之液壓泵（p); ⑷接受樣品之六閥式自動切換閥控制器（v)，其可形成會 互相轉換之第一轉換路徑及第二轉換路徑，其中第一轉 換路径係由連接接受樣品之接點2與接點3、連接接點1 與接點6、及連接接點4與接點5所形成者，而第二轉換 路徑係由連接接受樣品之接點2與接點1、連接接點3與 接點4、及連接接點5與接點6所形成者； ⑸接受來自六閥式自動切換閥控制器接點3之樣品的對掌 異構物層析管柱d); ⑹接受來自六閥式自動切換閥控制器接點1之樣品的一般 層析管柱（C 2 );及 ⑺接受來自六閥式自動切換閥控制器接點5之樣品的偵測 器裝置（D)， 其中當樣品排出對掌異構物層析管柱（Cl)，而尚未達到 良好的分離效果時’弟一轉換路徑及第二轉換路徑會接受 液壓系（P)送出之訊號而依次切換，直至達到良好的分離 效果。4727B, I 6-21GX flap fee (please read the note on the back before filling in this page) _ _.IT. 421718 A7 B7 V. Description of the invention (4) 6 represents 7T. Valve type automatic switching valve controller The 6th picks up. MAN League detailed description The present invention relates to a recirculating high-performance liquid chromatography device for analyzing optical isomers, which comprises: ⑴ a solvent container (E); ⑵ a sample input device (I); ⑶ the solvent container (E) Solvent pushes the sample in the sample input device (j), and the hydraulic pump with time event output function (p); 六 Six-valve automatic switching valve controller (v) that accepts samples, which can form the first A conversion path and a second conversion path, wherein the first conversion path is formed by connecting the contact 2 and the contact 3, the connection 1 and the contact 6, and the connection 4 and the contact 5 of the receiving sample, The second conversion path is formed by connecting the contact 2 and the contact 1, the connection 3 and the contact 4, and the connection 5 and the contact 6 of the receiving sample; ⑸ accepting a six-valve automatic switching valve Chromatographic isomer chromatography column of the sample at the controller contact 3 d); ⑹ General chromatography column (C 2) which accepts the sample from the six-valve automatic switching valve controller contact 1; and ⑺ Accept Detector device for sample from contact 5 of six-valve automatic switching valve controller (D) Among them, when the sample is discharged from the palm isomer chromatography column (Cl), but the good separation effect has not been achieved, the first conversion path and the second conversion path will receive the signal from the hydraulic system (P) and switch sequentially until To achieve a good separation effect.
4727B.t>0C 一 Ί 一 本紙張尺度適用中國國家標準（CNS ) A4規格（210X 297公釐） 經濟部中央標準局員工消費合作社印製 421718. A7 B7 五、發明説明（5) 本發明另關於一種分析光學異構物之再循環高效液相層 析方法，其包括下列步驟： ⑴利用具時間事件輸出功能之液壓泵（P)，將溶劑容器（E) 中之溶劑推出，以帶動樣品輸入裝置（Ϊ)所輸入之待分析 樣品向前流動至六閥式自動切換閥控制器（v); ⑵經六閥式自動切換閥控制器（V)之樣品依次供至對掌異 構物層析管柱（C !)及一般層析管柱（C 2)，以分析該樣品 ;及 ⑶利用偵測器裝置（D )價測分離過之樣品， 遠方法之特徵在於該六閥式自動切換閥控制器（V )，於 適當的時機，會自動切換，使樣品可經再循環而達到良好 的分離效果。 再循環尚效液相層析裝置的連接方式如圖1所示。參考 圖1(a)，分析樣品由樣品輸入裝置（1)注入後，由液壓系 C P )將溶劑容器（E )所推出之移動相溶劑，將樣品輸入裝置 (Ϊ )注入的分析樣品向前帶動，依序流經第—轉換路徑之 自動切換閥控制器接點2與接點3，而進入對掌熹構物層析 管柱（C 1 )，進行第一次樣品分離；當樣品排出對掌異構物 層析管柱，依序流經第一轉換路徑之自動切換閥控 制器接點6與接點1，而進入一般層析管柱（CJ ;當樣品 即將要，但尚未排出一般層析管柱（CO時，由液壓泵（p) 运出訊號給六閥式自動切換閥控制器（V)，在切換時間爲 1時（參考圖2)，轉換切換閥管路爲第二轉換路徨（如圖4727B.t > 0C One paper one paper size applies Chinese National Standard (CNS) A4 specification (210X 297 mm) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 421718. A7 B7 V. Description of the invention A method of recycling high-performance liquid chromatography for analyzing optical isomers includes the following steps: ⑴ Use a hydraulic pump (P) with a time event output function to push out the solvent in the solvent container (E) to drive the sample The sample to be analyzed input by the input device (Ϊ) flows forward to the six-valve automatic switching valve controller (v); ⑵ The samples passed through the six-valve automatic switching valve controller (V) are sequentially supplied to the palm isomers Chromatography column (C!) And general chromatography column (C2) to analyze the sample; and (3) the separated sample is measured by the detector device (D). The remote method is characterized by the six-valve type The automatic switching valve controller (V) will automatically switch at the appropriate time, so that the sample can be recycled to achieve a good separation effect. The connection of the recirculating liquid chromatography device is shown in Figure 1. Referring to FIG. 1 (a), after the analysis sample is injected from the sample input device (1), the mobile phase solvent pushed out from the solvent container (E) by the hydraulic system CP) advances the analysis sample injected by the sample input device (Ϊ) forward. Drive, sequentially flow through the second switching path of the automatic switching valve controller contact 2 and contact 3, and enter the palmar structure chromatography column (C 1) for the first sample separation; when the sample is discharged For palm isomer chromatography columns, flow through the automatic switching valve controller contact 6 and contact 1 of the first conversion path in sequence, and enter the general chromatography column (CJ; when the sample is about to be, but not yet discharged For general chromatography column (in CO, the hydraulic pump (p) sends a signal to the six-valve automatic switching valve controller (V). When the switching time is 1 (refer to Figure 2), the switching switching valve pipeline is the first Second conversion route
Mb)所示），之後，樣品依序流經第二轉換路徑之自動切 — 本紙張尺度it财咖家標準（CNS )纟4規格（_2mx297j^整） " --—-_____ (請先閲讀背面之注$項#填寫本頁) .裝. -訂 A7 B7 經濟部中央標準局員工消費合作社印製 五、發明説明（6 換閥控制器接點4與接點3，而再次進入對掌異構物層析管 柱（CJ ’進行第二次樣品分離；當樣品排出對掌異構物層 析管柱（C 1 )後，依序流經第二轉換路徑之自動切換閥控制 态控制器接點6與接點5，而流至偵測器裝置（D )，再由積 分器（R)將馈測結果輸出，而樣品則流入廢液收集瓶 (W) 〇 本發明進一步包含在樣品經第二次分離後，當樣品即將 要’但尚未排出對掌異構物層析管柱（Ci)時，由液壓泵（P: 送出訊號給六閥式自動切換閥控制器（V)，在切換時間爲 t2時’轉換切換周管路爲第一轉換路徑（如圖1(a)所示）， 使樣品依序流經第一轉換路徑之自動切換閥控制器接點6 與接點1 ’而進入一般層析管柱（C 2 );當樣品即將要，但 尚未排出一般層析管柱（c2)時’由液壓泵送出訊號給六閥 式自動切換閥控制器，在切換時間爲“時，轉換切换閥管 路馬第二轉換路徑（如圖1(b)所示），使樣品依序流經第二 轉換路徑之自動切換閥控制器接點4與接點3，而再次進入 對掌異構物層析管柱（Ci),進行第三次樣品分離。 本發明可依實際需要進行多次循環，而達到良好的分離 效果。 液壓泵（!>)控制六閥式自動切換閥控制器之訊號時序 如圖2所不。t 〇係指樣品注入時間，丨1係指第一次切換時 間，h係指第二次切換時間，^係指第三次切換時間，依 此類推。 般層析管柱（C 2 )在本發明中係作爲延遲（如丨吵）管柱，Mb)), after that, the sample will automatically cut through the second conversion path in order — this paper standard it financial standard (CNS) 纟 4 specifications (_2mx297j ^ whole) " -----_____ (please first Read the note at the back of the $ Item # Fill this page). Install.-Order A7 B7 Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the Invention (6 Valve Switch Controller Contact 4 and Contact 3, and enter the right again Palm isomer chromatography column (CJ 'for the second sample separation; after the sample is discharged from the palm isomer chromatography column (C 1), it sequentially flows through the second switching path to the automatic switching valve control state The controller contacts 6 and 5 flow to the detector device (D), and the integrator (R) outputs the test result, and the sample flows into the waste collection bottle (W). The present invention further includes After the sample is separated for the second time, when the sample is about to be 'but not yet discharged from the palm isomer chromatography column (Ci), the hydraulic pump (P: sends a signal to the six-valve automatic switching valve controller (V ), When the switching time is t2, the switching cycle is the first switching path (as shown in Figure 1 (a)), so that the samples are sequentially Enter the general chromatography column (C 2) via the automatic switching valve controller contact 6 and contact 1 'in the first conversion path; when the sample is about to be received but has not been discharged from the general chromatography column (c2)' The hydraulic pump sends a signal to the six-valve automatic switching valve controller. When the switching time is ", the switching valve line is the second switching path (as shown in Figure 1 (b)), so that the sample flows through the first The automatic switching valve controller contact 4 and contact 3 of the second conversion path are re-entered into the palm isomer chromatography column (Ci) for the third sample separation. The present invention can perform multiple cycles according to actual needs The signal timing of the hydraulic pump (! ≫) controlling six-valve automatic switching valve controller is shown in Figure 2. t 〇 refers to the sample injection time, 丨 1 refers to the first switching time , H refers to the second switching time, ^ refers to the third switching time, and so on. The general chromatography column (C 2) is used in the present invention as a delay (such as noisy) column,
47278.DOC ' 9 - 祕張又度適用標準（CNS ) μ規格297公瘦） (請先閱讀背面之注意事項#·填寫本頁) -裝. 訂. 4 2 17 18 A7 B7 五、發明説明（7 其作用在於確保切換閥作切換動作時，樣品已完全通過對 掌異構物層析管柱（CD，而不會造成部分樣品流至偵測器 裝置（D)，無法進行再循環分離。 有關液壓泵（P)控制六閥式自動切換閥控制器（V)的連線 方式説明於表1及圖3。 表Ϊ液壓泵後面8針(pin)連接口中各接點的訊號表 針# 訊號 1 備妥(輸出） 2 最大+5 VDC 100mA 3 接地 4 壓力 0.1 V/1000 PSI 5 停止(輸入） 6 時間事件(輸出） 7 運作(輸入） 8 注射維持(輸出） 請 先 閱 婦 背 之 注 意. 事 項 再· 填 寫 本 頁 裝 訂 --—I 本 經濟部中央橾準局員工消費合咋；iLf获 表1爲液壓泵後面8針（pin)連接口中各接點的訊號，圖3 爲κ閥式自動切換閥控制器後面連接口的接線圖，連接方 式爲將液壓系中的針3 (pin 3)和針6 (pin 6)連接到圖3中所示 終端盒(terminal block)中下方的二個接點上即可。 下列實施例係説明本發明之可實施性，但並非用於限定 本發明之保護範園，而是可以有不達離本發明範圍的各種 變更和修改的。 實施例 -10 經濟部中央橾準局員工消費合作社印製 4 2 17 1 8 - at B7 五、發明説明（8 ) 材料 1.光學異構物樣品： 由“財團法人生物技術開發中心有機合成實驗室”合成而 得之依托多雷克（Etodolac)衍生物，編號爲129735A，其結 構式如下：47278.DOC '9-Secret and Applicable Standard (CNS) μ size 297 male thin) (Please read the precautions on the back # · Fill this page first)-Pack. Order. 4 2 17 18 A7 B7 V. Description of the invention (7 Its role is to ensure that when the switching valve performs a switching action, the sample has completely passed through the isomer chromatography column (CD) without causing part of the sample to flow to the detector device (D), which cannot be recycled and separated. The connection method of the hydraulic pump (P) to control the six-valve automatic switching valve controller (V) is described in Table 1 and Figure 3. Table 的 Signal pin of each contact in the 8-pin connection port on the back of the hydraulic pump # Signal 1 Ready (output) 2 Maximum +5 VDC 100mA 3 Ground 4 Pressure 0.1 V / 1000 PSI 5 Stop (input) 6 Time event (output) 7 Operation (input) 8 Injection maintenance (output) Please read the woman's back first Note: Please fill in this page and bind it --- I Consumption of employees of the Central Bureau of Standards of the Ministry of Economic Affairs; iLf obtained in Table 1 is the signal of each contact in the 8-pin (pin) connection port on the back of the hydraulic pump. Figure 3 shows Wiring diagram of the connection port on the back of the κ valve automatic switching valve controller, connection method In order to connect the pin 3 and pin 6 in the hydraulic system to the two lower contacts in the terminal block shown in Fig. 3, the following examples illustrate the present invention. It can be implemented, but it is not used to limit the scope of protection of the present invention, but various changes and modifications can be made within the scope of the present invention. 2 17 1 8-at B7 V. Description of the invention (8) Materials 1. Samples of optical isomers: Etodolac derivatives derived from the "Organic Synthesis Laboratory of the Biotechnology Development Center of the Consortium" The number is 129735A, and its structural formula is as follows:
兴：手性中心 2 .移動相溶劑 99重量份之己烷與1重量份之異丙醇所组成。 儀器 1. 液壓系：Spectra - Physics 公司 plOOO 型。 2. 樣品注射器；Rheodyne公司7725型。 3. 自動切換間控制器：ANACHEM / UK公司ACP · 3010型， Universal 公司 GS 10c version 型。 4. 六閥式注入器：Rheodyne公司7010型。 5. 層析管柱： ⑴對掌異構物層析管柱：Merck公司之LiChroCART®，長250 mm，内徑4 mm，粒徑大小5 μιη。 ⑵一般層析管柱：Keystone公司之Nucleosil Silica，長50 mm ，内徑4.6 mm，粒徑大小3 μιη。 47278.DOC — 11 _ 本紙張尺度適用中國國家標準（CNS ) Μ规格（210Χ297公釐） (請先間讀背面之注意事項再填寫本頁) _ ____ —裝. 訂 -線_ 4 2 17 18 : A7 B7 經濟部中央標隼局貝工消費合作社印製 五、發明説明（9) 6. 積測器裝置：Shimadzu公司之spo - 6AV型紫外光偵測器。 7. 積分器：Beckman公司427型。 品配置 稱取0,5 mg的樣品，放入10 mL的量瓶中，以移動相容劑 加至10 inL ’授掉直至樣品完全溶解。取2.5 mL的溶液，放^ 入另一個10 mL的量瓶中，以移動相溶劑稀釋至1〇 mL。 HPLC分析條件 1_移動相溶劑：9 9 : 1比例之己烷與異丙醇混合溶劑。 2. 移動相流速：0.8 mL / min 。 3. 樣品注入體積：10 μί。 4. 紫外光偵測器波長：254 ran。 5. 積分器：cs = 0.5，PT = 500，PW = 6，AT = 16 〇 實例1 本實例係關樣品129735A的經一次再循環分析之實驗。首 先，將樣pp 129735A以樣品輸入裝置（I)注人後，.由液壓系 (P )將溶劑容器（E )所推出之移動相溶劑，將樣品輸入裝置 (I )注入的樣品向前夢動至六閥式自動切換閥控制器（V )之 第一轉換路徑’依序流經自動切換閥控制器接點2和接點3 ’進入對掌異構物層析管柱（c ι )，進行第一次之樣品分離 ’當樣品排出對掌異構物層析管柱（c i )後，經自動切換閥 控制器接點6和接點！，進入一般層析管柱（c2);在^爲8 为鐘時’即當樣品即將要但尚未排出一般層析管柱（C 2)時 ，由具時間事件訊號輸出功能之液壓泵（p )送出訊號給六 闕式自動切換閥控制器（V)，將整個系统管路切換至第二 47278.DOC „ 〇 - 丄2 - 本紙張尺度適财關家縣（CNS ) A4規格（加謂了公 (請先聞讀背面之注意事項再填寫本頁) 〇 裝- 訂Xing: Chiral Center 2. Mobile phase solvent consisting of 99 parts by weight of hexane and 1 part by weight of isopropanol. Instrument 1. Hydraulic system: Spectra-Physics plOOO. 2. Sample syringe; Rheodyne Model 7725. 3. Automatic switching controllers: ANACHEM / UK company ACP · 3010 type, Universal company GS 10c version type. 4. Six-valve injector: Model 7010 from Rheodyne. 5. Chromatographic column: ⑴Palm isomer chromatography column: LiChroCART® from Merck, with a length of 250 mm, an inner diameter of 4 mm, and a particle size of 5 μm. ⑵General chromatography column: Keystone's Nucleosil Silica, 50 mm long, 4.6 mm inner diameter, and 3 μm particle size. 47278.DOC — 11 _ This paper size applies to Chinese National Standards (CNS) M specifications (210 × 297 mm) (please read the precautions on the back before filling out this page) _ ____ — Packing. Order-Line_ 4 2 17 18 : A7 B7 Printed by the Central Bureau of Standards, Ministry of Economic Affairs, Shellfish Consumer Cooperative, V. Description of Invention (9) 6. Total detector device: Shimadzu's spo-6AV UV light detector. 7. Integrator: Beckman Company Model 427. Product configuration Weigh a 0.5 mg sample, put it into a 10 mL volumetric flask, and add the mobile compatibilizer to 10 inL ′ and dispense until the sample is completely dissolved. Take 2.5 mL of the solution and place it in another 10 mL volumetric flask and dilute to 10 mL with mobile phase solvent. HPLC analysis conditions 1_ Mobile phase solvent: 9: 1 ratio of hexane and isopropanol mixed solvent. 2. Mobile phase flow rate: 0.8 mL / min. 3. Sample injection volume: 10 μί. 4. UV detector wavelength: 254 ran. 5. Integrator: cs = 0.5, PT = 500, PW = 6, AT = 16 〇 Example 1 This example is an experiment on a sample of 129735A subjected to a recirculation analysis. First, after the sample pp 129735A is injected into the sample input device (I), the mobile phase solvent pushed out by the solvent container (E) is pushed by the hydraulic system (P), and the sample injected by the sample input device (I) is dreamed forward. The first switching path moving to the six-valve automatic switching valve controller (V) 'flows through the automatic switching valve controller's contact 2 and contact 3' in sequence and enters the palm isomer chromatography column (c) , The first sample separation 'When the sample is discharged from the palm isomer chromatography column (ci), the contact 6 and the contact of the valve controller are automatically switched! , Enter the general chromatography column (c2); when ^ is 8 hours, that is, when the sample is about to be discharged but has not been discharged from the general chromatography column (C 2), a hydraulic pump with time event signal output function (p ) Send a signal to the six-way automatic switching valve controller (V) to switch the entire system pipeline to the second 47278.DOC „〇- 丄 2-This paper is suitable for Guancai County (CNS) A4 specifications (plus) The public (please read the notes on the back before filling out this page) 〇 Install-Order
經濟部中央標準局男工消費合作社印I Α7 Β7 五、發明説明（10) 轉換路徑（如圖1 (b )所示），之後，樣品經自動切換閥控制 器接點4和接點3，而再次進入對掌異構物層析管柱（C !) ，進行第二次之樣品分離，結果顯示兩支波峰（分別爲左 旋及右旋）之解析度爲1.3。圖5是樣品129735A經一次再循 環分析的圖譜，表2爲兩支波峰之滯留時間、面積及解析 度之變化。 表2 實例1之結果 滯留時間 面積 波峰解析度 第1波峰 15.17 23804 1.3 第2波峰 16.01 24979 比較實例1 本比較實例係關實例1樣品U9735A未經再循環分析之實 驗。將樣品129735A以樣品輸入裝置（I)注入後，由液壓泵 (P)將溶劑容器（E)所推出之移動相溶劑，將樣品輸入裝置 (I)注入的分析樣品向前帶動，經六閥式自動切換閥控制 器接點2和接點3，進入對掌異構物層析管柱（CJ，進行 樣品之分離，結果顯示兩支波峰（分別爲左旋及右旋）之解 析度爲〇 · 9。圖4是樣品129735A未經再循環分析的圖譜， 表3爲兩支波峰解析度之變化。 表3 比較實例1之結果 滯留時間 面積 波峰解析度 第1波峰 8.61 26308 0.9 第2波峰 8.97 29603Printed by the Male Standards Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs I Α7 Β7 V. Description of the invention (10) Conversion path (as shown in Figure 1 (b)), after that, the sample passes through the automatic switching valve controller contact 4 and contact 3, And re-entered the isomer chromatography column (C!) For the second sample separation, the results showed that the resolution of the two peaks (left-handed and right-handed) was 1.3. Figure 5 shows the spectrum of sample 129735A after a re-cycle analysis. Table 2 shows the change in the retention time, area, and resolution of the two peaks. Table 2 Results of Example 1 Retention time Area Peak resolution 1st peak 15.17 23804 1.3 2nd peak 16.01 24979 Comparative example 1 This comparative example is related to the experiment of sample 1 of U9735A without recycling analysis. After the sample 129735A is injected through the sample input device (I), the mobile phase solvent introduced by the solvent container (E) is pushed by the hydraulic pump (P), and the analysis sample injected by the sample input device (I) is moved forward, and the six valves Type automatic switching valve controller contact 2 and contact 3, enter the palm isomer chromatography column (CJ, the sample is separated, the results show that the two peaks (left-handed and right-handed) have a resolution of 0. · 9. Figure 4 shows the spectrum of sample 129735A without recirculation analysis, and Table 3 shows the changes in the resolution of the two peaks. Table 3 Results of Comparative Example 1 Retention time area Peak resolution First peak 8.61 26308 0.9 Second peak 8.97 29603
4727B.DOC 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） (請先閲讀背面之注意事項再填寫本頁) 裝·4727B.DOC This paper size is applicable to China National Standard (CNS) A4 (210X297 mm) (Please read the precautions on the back before filling this page)
C 訂 線 A7 B7 五、發明説明（u) 由上述實例可知，先前技藝所無法分離之光學異構物， 確實可經由本發明之再循環高效液相層析裝置及方法，而 得到良好之分離。 (請先閲讀背面之注意事項再填寫本頁) -裝- 訂 線 經濟部中央標準局員工消費合作社印製 4 本紙張尺度適用中國國家標準（CNS ) A4規格（2丨Ο X W7公釐）C Order A7 B7 V. Description of the invention (u) From the above examples, it can be seen that the optical isomers that could not be separated by the prior art can indeed be separated well by the recycling high-performance liquid chromatography device and method of the present invention. . (Please read the precautions on the back before filling out this page)-Binding-Binding Line Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economy