External standard. The target is in the calibration bottles at known levels. Samples have an unknown amount. There is no ISTD drop. One curve serves the whole batch.

ESTD initial calibration sequence Blank, five calibration levels, then unknown samples. Target analyte only. One curve for the batch. ESTD  one initial curve, several levels External standard. The curve uses analyte response versus analyte concentration. No ISTD drop. BLK Blank C1 0.5 C2 2.0 C3 10 C4 50 C5 200 UNK-01 unknown UNK-02 unknown QC-01 check Fit one curve from C1C5. Every later bottle uses that curve. Read unknowns and QC from the same curve. Assign Type = Calibration and Dose = C1&C5 on the Samples grid. Method Calibration mode = Initial calibration. Link those dose names on the Quant so the level is used on the curve. Unused doses stay off the fit.
Blank, C1–C5, unknowns, and QC

When to use it

Method Calibration mode = Initial calibration. Quant ISTD column empty.

Set it up

  1. Import or build the method. Create dose names C1, C2, C3, … and type the concentrations on each analyte.
  2. On the Quant, assign those doses (Quant doses). Only assigned doses go on the curve.
  3. Add the sequence. Mark bottles: - Blank → Type Blank - Standards → Type Calibration, Dose C1 … C5 - Checks → Type QC, Dose of the check level - Unknowns → Type Sample
  4. Analyze.

How the curve is read

ESTD multi-level calibration curve Response versus concentration. Five calibration points, a QC, and two unknowns read from the fitted line. ESTD curve  response vs concentration X is the known dose. Y is peak response. Quanto inverts the fit to read unknown and QC bottles. Concentration (dose) Response 0 C1 C2 C3 C4 C5 UNK-01 UNK-02 QC Calibration point (used) Sample read from curve QC check Typical fit: Linear or quadratic, with optional weighting and origin. Clear Use on a cal point to drop it from the fit.
Response versus concentration with unknowns read from the line
Axis Value
X Known dose concentration (÷ sample dilution)
Y Peak response

Quanto fits the Quant’s Curve fit (linear, quadratic, average response factors, power, first- or second-order ln), with the Quant’s weighting and origin. It then inverts the fit for every Sample and QC.

A blank is not a curve point. Use it to confirm the target is gone, not to pin the origin, unless you chose a through-origin fit.

Multi-level rules

After Analyze

Next: add an ISTD (ISTD), split the sequence into brackets (bracketing), or keep this curve and update only some levels later (multi-batch).