全自動移液工作站PP5+1采集LC-MS/MS樣品操作說明

PERSONAL PIPETTOR全自動移液工作站是Apricot Designs研發生產的集多種移液功能于一身的緊湊型全自動移液工作站，具有高效、便捷、精準的特性，能同時兼容1/8/12/16/24/96/384通道的液體處理，開放式的設計可以便于整合疊板機和機械臂等。

特點和優勢

1. 集多種移液功能于一身，僅需單擊運行，即可完成多種不同類型的切換；
2. 5個功能板位+1個洗針槽/廢液槽；
3. 1/8/12/16/24/96/384通道移液；
4. Personal Pipettor+無人值守, 可自動更換吸頭；
5. 簡單直觀的iPad/Microsoft Surface™/PC操作界面；
6. 可選擇使用有線或無線操控儀器；
7. 適用于96/384孔板移液和逐行/逐列梯度稀釋;
8. 移液工作站、鎖頭和EZ-Load吸頭三合一，密封良好，確保實驗精確、可靠和可重復；
9. 可搭載洗針系統/條碼掃描/震蕩器/控溫模塊；
10. 可整合微孔板疊板機和機械手，真正實現自動化無人值守；
11. 體積小巧，最大化節省空間，適合放置于多數標準通風櫥。

以下為有關PERSONAL PIPETTOR全自動移液工作站進行 LC-MS/MS樣品的采集，請參考：

Overview

Purpose
•Show a new and innovative way to collect, store, and prepare plasma samples using microtainers.
•Show that the use of microtainers has saved considerable time and effort in both the Pharmacokinetics (PK) and Analytical groups.
•Show that there are no compound stability or non-specific binding issues with the compounds tested by storing plasma samples in microtainers over long periods of time.
•Show that using microtainers has minimized clotting issues in plasma samples. 

Method
PK
Animal dosing→collection in microtainer→ centrifuge→store in freezer
Analytical 

Receive samples From PK→Arrange in 96-well plate→transfer samples by Apricot (See Figure 1)→Sample prep

Method
Sample preparation
     100 uL plasma sample
           + 10 µL internal standard
           + 400 µL acetonitrile to precipitate the proteins
           - 450 µL of the supernatant
           dry down under nitrogen
           reconstitute with 100 µL of 5% methanol

Column
      Higgins TARGA, C18 (30 x 0.5mm, 5 µm)
      Inject 10 µL
Representative Method
      Initial 0.5 minute hold followed by linear gradient of 25-85% B
      over 1.5 minutes
      100 µL/minute flowrate
     Mobile Phase A: 0.25% Formic Acid in Water
     Mobile Phase B: 0.25% Formic Acid in Methanol
Instrumentation and Hardware
      PE Sciex API 3000 using TIS
      HTS PAL LEAP Autosampler
      Shimadzu Pumps and System Controller
      Data processed with Analyst version 1.3

Figure 1. What is a microtainer?

Microtainers are collection tubes that are used to stabilize and separate whole blood into plasma and blood cell (BC) fractions.

Microtainers can contain a polymer gel separator and the walls can be coated with an anticoagulant (i.e. Heparin or EDTA).

Benefits of using microtainers as a sample collection and storage tube:

• No loss of sample

• Minimal sample transfers

• Minimal labeling

• No decanting

Figure 2. A semi-automated approach using the Apricot Personal Pipettor to transfer samples from microtainers to 96-well plates.

Table 1. Statistical Analysis of THRX-776005 Stability Study in microtainers based on average concentrations.

Figure 3. Stability of THRX-776005 in microtainers over a period of time.

Figure 4. PK profile of THRX-502038 comparing plasma samples collected in microtainers versus samples collected in Eppendorfs.

Results and Conclusion

Using microtainers as a collection, preparation, and storage tube has increased sample processing productivity by 50-fold.

The use of microtainers has negated the problem of plasma clots.

Compound instability in the microtainer is minimal as shown in Figure 3 and defined in Table 1.

Non-specific binding of compounds during routine use and storage has not been seen as shown in Figure 3.

There were no great differences between using Eppendorfs (original method) and using microtainers (current method) to store plasma samples as shown in Figure 4.

To date >20,000 samples have been processed using this method.

The use of microtainers with heparin has shown no evidence of MS signal suppression of our compounds. (Data not shown)