自填式管柱(self-packed columns)之品質控制

填充管柱後,建議在進行樣品純化之前,使用標準樣品進行理論塔板數(N, number of theoretical plates)、理論塔板高度(HETP, height equivalent to a theoretical plate)和峰不對稱因子(As, peak asymmetry factor)測試及計算。藉由定期和重複執行此類標準測試,可以有效地評估膠體(gel, packing material)的品質和耐用性。 如果您想比較各個測試之間的結果,建議在每次測試中在相同條件下使用相同的測量設備。測量結果可能會受到一些因素影響,例如:死體積(dead volume)、流速(flow rate)、溫度(temperature)、沖提液(eluent)和進樣量(sample loading amount)的差異。

下列公式可用於計算理論塔板數(N, number of theoretical plates)。 較大的 N 值對應於層析管柱中較高的填充膠體密度。 此外,在固定滯留時間(retention time),峰形越窄,N 值越高;柱長越長,粒徑越小,N 值越大。 通常,每柱床長(m)的理論塔板數通常用作評估層析管柱純化性能的指標。

N = 5.54 x (tR / W0.5h)2
N/m = N / L
L = Column bed length (m)

理論塔板高度 (HETP, height equivalent to a theoretical plate)有時也用作評估層析管柱性能的指標。 下列公式可用於計算 HETP。 該值越小,填充在柱中的膠體密度越高。

HETP = L / N

洗脫峰形 (peak shape)也是評估層析管柱性能的重要因素。 峰不對稱因子 (As) 的測定是一種相對簡單的估計層析管柱性能的方法。 下列公式可用於計算 As,該值越接近“1”是越理想的,其表示越高的峰形對稱性。

As = b / a

HPLC技術指引:管柱平衡(Column Equilibration)

我應該平衡HPLC管柱多久時間呢?

在執行梯度條件後進行下一次進樣前之HPLC層析管柱平衡時,如果(重新)平衡時間太短,則會出現實驗重複性不佳的風險。如果時間太長,則會浪費時間和溶劑。

在本HPLC技術指引中,您將學習如何通過計算管柱體積來估算最佳平衡體積。此外,還使用了一個實際案例來顯示不同平衡體積對分析結果的影響。有關管柱平衡的詳細說明,您可以在此處下載HPLC技術指引。

Expert Tip: 管柱再生Column regeneration

您的(U)HPLC層析管柱之分離效果是否開始變差?

這份文件我們提供了合適的管柱清洗程序,有助於恢復層析管柱之性能。此外,該程序還可防止污染物在固定相上累積。但是,需考量一些情況,例如:清潔溶劑的類型和有效再生所需的體積。有關層析管柱再生的更多詳細信息,請在此處下載說明文件。

Improving the retention behaviour of hydrophilic compounds on polar stationary phases

Do you analyse hydrophilic compounds? Do you want to improve the retention on polar stationary phases such as PFP or Diol?

In this technical note, two different options for the improved analysis of hydrophilic compounds on YMC-Triart PFP and YMC-Triart Diol-HILIC are shown. The pH value and the amount of organic solvent are examined to learn more about the retention behaviour of basic, acidic and zwitterionic compounds using these phases. Due to their different selectivity, pentafluorophenyl (PFP) or Diol modifications are an ideal choice for the analysis of hydrophilic compounds which often remain unretained or unresolved on conventional C18 phases.

You can download the full method details here.

To get further information about the YMC-Triart series of columns.

Fast UHPLC Analysis of SARS-CoV-2 drug Remdesivir and its active metabolite GS-441524

As the COVID-19 pandemic continues, the worldwide search for an effective treatment is still ongoing. Remdesivir and its active metabolite GS-441524 are both antiviral drugs that have been examined and partially approved for the treatment of COVID-19 due to their capability of inhibiting virus replication.

In this application note, the separation of GS-441524 and Remdesivir in only 3 minutes is shown. A YMC-Triart C18 UHPLC column provides fast and reliable results making it an ideal choice for QC purposes.

You can download the application note with the full method details here.

USP class 管柱分類

USP No Functional group USP Description YMC Product
L01 C18 Octadecyl silane <ODS or C18> chemically bonded to porous silica or ceramic particles – 1.5 to 10 µm in diameter YMC-Triart C18

YMC-Triart C18 ExRS

YMC-Triart Bio C18

Meteoric Core C18

Meteoric Core C18 BIO

Pack Pro C18

Hydrosphere C18

Pack Pro C18 RS

YMC-Pack ODS-A

YMC-Pack ODS-AM

YMC-Pack ODS-AQ

YMC-Pack ODS-AL

J’sphere ODS-H80, ODS-M80, ODS-L80

L03 SIL Porous silica particles – 1.5 to 10 µm in diameter  

YMC-Triart SIL

YMC-Pack SIL, SIL-06

L07 C8 Octyl silane <C8> chemically bonded to porous silica particles – 1.5 to 10 µm in diameter YMC-Triart C8

Meteoric Core C8

YMC-Pack Pro C8

YMC-Pack C8

YMCbasic

L08 NH2 An essentially monomolecular layer of aminopropylsilane <NH2> chemically bonded to totally porous silica gel support – 1.5 to 10 µm in diameter YMC-Pack NH2
L10 CN Nitrile groups <CN> chemically bonded to porous silica particles – 1.5 to 10 µm in diameter YMC-Pack CN
L11 Ph Phenyl groups chemically bonded to porous silica particles – 1.5 to 10 µm in diameter YMC-Triart Phenyl

YMC-Pack Ph

L13 C1 Trimethylsilane <C1> chemically bonded to porous silica particles – 1.5 to 10 µm in diameter YMC-Pack TMS
L20 Diol Dihydroxypropane groups chemically bonded to porous silica particles – 1.5 to 10 µm in diameter YMC-Triart Diol-HILIC

YMC-SEC MAB

YMC-Pack Diol

YMC-Pack Diol-NP

L24 Polyvinyl alcohol Polyvinyl alcohol chemically bonded to porous silica particles – 5 µm in diameter YMC-Pack PVA-Sil
L26 C4 Butyl silane <C4> chemically bonded to porous silica particles – 1.5 to 10 µm in diameter YMC-Triart Bio C4

YMC-Pack Pro C4

YMC-Pack C4

YMC-Pack PROTEIN-RP

L27 SIL Porous silica particles – 30 to 50 µm in diameter YMC-Pack SIL-HG
L33 Diol Packing having the capacity to separate dextrans by molecular size over a range of 4,000 to 500,000 Da. It is spherical, silica-based, and processed to provide pH stability YMC-Pack Diol (SEC)
L40 Cellulose
derivatives
Cellulose tris-3,5-dimethylphenylcarbamate coated porous silica particles, 3 to 20 µm in diameter CHIRAL ART Cellulose-C
L43 PFP Pentafluorophenyl groups chemically bonded to silica particles by a propyl spacer, 1.5 to 10 µm in diameter YMC-Triart PFP
L51 Amylose
derivatives
Amylose tris-3,5-dimethylphenylcarbamate-coated, porous, spherical, silica particles, 3 to 20 µm in diameter CHIRAL ART Amylose-C
L59 Diol Packing for the size exclusion separations of proteins (separation by molecular weight) over the range of 5 to 7000 kDa. It is spherical (1.5 to 10 µm), silica or hybrid packing with a hydrophilic coating YMC-SEC-MAB

YMC-Pack Diol (SEC)

L62 C30 C30 silane bonded phase on a fully porous spherical silica, 3 to 15 µm in diameter YMC Carotenoid
L99 Amylose
derivatives
Amylose tris-(3,5-dimethylphenylcarbamate), immobilized on porous, spherical, silica particles, 3 to 5 µm in diameter CHIRAL ART Amylose-SA
L111 Polyamine Polyamine chemically bonded to porous spherical silica particles, 5 µm in diameter YMC-Pack Polyamine II
L119 Cellulose
derivatives
Cellulose tris-(3,5-dichlorophenylcarbamate), immobilized on porous, spherical, silica particles, 3 to 5 µm in diameter. CHIRAL ART Cellulose-SC

YMC玻璃管柱選擇指引

YMC玻璃管柱可針對客戶實際應用進行最佳化選擇,對於藥物與蛋白質純化日益複雜且高純度的需求,除有多種內徑、可調式長度選擇之外,亦可針對您所需之耐壓、溫控、耐溶劑等實驗需求提供相對應的產品。並且可供應從實驗室一路放大到生產端之完整的產品線供應。以下為簡單選擇指引,詳細規格請參考各產品連結或來電與我們諮詢。

ECO Columns玻璃管柱(適用LPLC、蛋白質樣品)

-低~中壓應用
-管柱內徑: 10, 15, 25, 50, 70, 80 mm
-管柱長度: 120, 200, 450, 750, 1000 mm(管柱長度可自由調整)
-耐壓: 5 ~ 30 bar(依內徑大小耐壓規格略有差異)
溫控夾套(選配): Heating/cooling jacket
-操作快速: Easy screw-lock 設計操作便利

ECOPLUS Columns(適用MPLC)

-中~高壓應用
-管柱內徑: 5, 10, 15, 25, 35, 50 mm
-管柱長度: 125, 250, 500 mm(管柱長度可自由調整)
-耐壓: 15 – 80 bar(依內徑大小耐壓規格略有差異)
-再現性佳
-操作快速:  Quick-Lock 設計操作便利

YMC Pilot

-應用廣泛: 適用於BioLC或逆相(RP)/正相(NP)等應用
-內徑: 100, 140, 200, 300 mm
-可訂製不繡鋼材質
-可選配溫控夾套(Heating/Cooling jacket)
-操作簡單、密封(Seal)技術佳
(更大內徑需求請來信詢問)

YMC PilotPLUS

-Bio液相層析之生物相容性
-內徑: 70, 100, 140, 200 mm
-特殊排液設計使填充管柱簡單且無氣泡產生
-可拆卸式玻璃管柱設計可快速且乾淨的卸除玻璃管柱

如何選擇C18層析管柱-Guides for selecting C18 packing materials

小分子化合物

Low molecular weight compounds

 一般有機化合物

General organic compounds

General organic compounds YMC-Triart C18
Meteoric Core C18
Chemical structures differ slightly among solutes YMC-Triart C18 ExRS
Hydrophobicity differs greatly among solutes YMC-Triart Bio C18
J’sphere ODS-L80
Difficult to separate using standard ODS YMC-Triart C18 ExRS
Hydrosphere C18
YMC-Pack Pro C18 RS
YMC-Pack ODS-AL
J’sphere ODS-H80
J’sphere ODS-L80
YMC-Pack PolymerC18
Basic compounds YMC-Triart C18
YMC-Triart C18 ExRS
Coordination compounds YMC-Triart C18
YMC-Triart C18 ExRS
Sugars and glycosides YMC-Triart C18
Hydrosphere C18
High molecular weight compounds

大分子化合物

High molecular weight peptides and oligonucleic acids YMC-Triart C18
YMC-Triart Bio C18
Meteoric Core C18 BIO
Alkaline mobile phase

鹼性移動相條件

YMC-Triart C18
YMC-Triart C18 ExRS
YMC-Triart Bio C18
YMC-Pack PolymerC18