Chapter 6 · Comparing groups beyond the t-test

How do you allow for the baseline value?

Analysis of covariance, change scores, adjusted means and regression to the mean.

Read first: Clinical trials, Correlation and regression

The problem first

You are a second-year Psychiatry resident. Your thesis randomised 20 patients with depression to a new drug or placebo and recorded the HAM-D score at baseline and at 8 weeks. By chance, the treated arm started 3.1 points more depressed.

A t-test on the 8-week scores gives p = 0.19. A t-test on the change from baseline gives p < 0.001. Your guide asks which analysis the protocol specified.

“You have two p-values for one trial. Which one will you report, and why?”

Start here

Why does the baseline value matter in a randomised trial?

Randomisation makes the groups comparable on average, not identical. In any one trial the arms can start at different mean scores by chance, and a patient's score at 8 weeks depends strongly on where they started.

12162024mean HAM-Dcontrol 21.6treated 24.715.613.3final −2.3baseline +3.1change: −6.0 against −11.4, difference −5.4baseline8 weeks
Made-up teaching data used throughout this chapter. The treated arm started 3.1 points higher. Its final mean is only 2.3 points lower than the control arm, but it fell 5.4 points more. Neither number is the treatment effect adjusted for where each patient started.

Regression to the mean

A single measurement is the patient's usual level plus variation on that day. Patients chosen because their first reading was high tend to have a lower second reading, even with no treatment, because the chance part of the first reading does not repeat. This is regression to the mean. Barnett, van der Pols and Dobson (2005) describe it as a statistical phenomenon that can make natural variation in repeated data look like real change, and note that its effect is larger with more measurement error and when follow-up is examined only in a subgroup selected on its baseline value.

Regression to the mean also explains why change scores do not fix baseline imbalance. Vickers and Altman (BMJ, 2001) wrote that analysing change does not control for baseline imbalance because of regression to the mean: patients with high baseline scores tend to fall more.

Interactive

Raise the entry cut-off and watch untreated patients appear to improve

No one in this population receives any treatment. The apparent fall comes only from selecting people whose first reading was high.

Try this: raise the correlation between the two readings to 0.95. The apparent fall in BP becomes small. Then lower it to 0.3 and the untreated patients seem to improve by more than 10 mmHg.

Made-up teaching population: systolic BP measured twice, 4 weeks apart, mean 130 and SD 15 both times. Nobody receives any treatment. Only people whose first reading is at or above the cut-off are enrolled.

150 mmHg

A stricter entry criterion selects more extreme first readings

0.60

Lower correlation: more of a single reading is day-to-day variation and measurement error

10.8mmHg average fall with no treatment

On average the enrolled patients have a screening BP of 157.0 mmHg and a follow-up BP of 146.2 mmHg. Their first reading was selected for being high, and part of that height was chance, which does not repeat. 9.1% of the population would be enrolled.

this sample of 400: 54 enrolled · mean 157.4 → 149.1 mmHg

8080100100120120140140160160180180screening BP, mmHgfollow-up BPno changeenrolled →−10.8
enrollednot enrolledexpected mean of the enrolled group

Three analyses

Final score, change score or ANCOVA: what does each one do with the baseline?

All three compare the arms at the end of the trial. They differ in how much of each patient's baseline value they subtract.

AnalysisOutcome comparedBaseline usedTeaching trial
Final score8-week HAM-DNot at all (slope 0)−2.30 (95% CI −5.82 to 1.22), p = 0.19
Change score8-week − baselineAll of it (slope 1)−5.40 (95% CI −8.20 to −2.60), p < 0.001
ANCOVA8-week HAM-D, adjusted for baselineThe slope estimated from the data, 0.674−4.39 (95% CI −7.25 to −1.53), p = 0.005
00.250.50.751-2-3-4-5-6slope used to correct for the baseline differencefinal score: −2.30ANCOVA: −4.39change score: −5.40estimate = −2.3 − slope × 3.1
Each estimate equals the difference in final means minus a slope × the baseline difference (3.1 points). The final score uses slope 0 and the change score slope 1. ANCOVA uses the slope that best predicts the 8-week score from the baseline score within the arms. With a baseline imbalance, slope 0 and slope 1 give biased answers in opposite directions.

Why ANCOVA is usually preferred in randomised trials

  1. No bias from chance imbalance. Vickers and Altman: ANCOVA adjusts each patient's follow-up score for their baseline score and is unaffected by baseline differences.
  2. Smaller standard error. Frison and Pocock (1992) quantified the superiority of analysis of covariance over analysis of post-treatment means or mean changes, in both reduced variance and avoidance of bias.
  3. Fewer patients. Vickers and Altman give an example with a correlation of 0.6 between baseline and follow-up: 85 patients for the final score, 68 for the change score and 54 for ANCOVA.
  4. Regulatory advice. The EMA guideline says the baseline value of a continuous primary outcome should usually be included as a covariate, whether the outcome is the raw score or the change from baseline.
00.511.500.250.50.751correlation between baseline and final scorerelative SEfinal scorechange scoreANCOVAr = 0.5
Standard error of each analysis relative to the final-score analysis, for equal SDs at both times: change score √[2(1 − r)], ANCOVA √(1 − r²). At r = 0.6 the variances are 0.80 and 0.64 of the final-score variance, which is where 85, 68 and 54 patients come from. Below r = 0.5 the change score is less precise than simply analysing the final score.

Interactive

Change the baseline imbalance and the correlation, and watch the three estimates and their standard errors

Each run is a new trial of 60 patients with a true effect of −4. The final score and change score move away from −4 in opposite directions when the arms start unequal. ANCOVA stays centred on −4 with the smallest standard error.

Try this: set the baseline imbalance to 0 and press "Run the trial again" a few times. All three estimates scatter around −4, and ANCOVA has the smallest SE. Then set the imbalance to 5: the final-score estimate moves above −4 and the change-score estimate moves below it.

Made-up teaching trial: HAM-D depression score at baseline and at 8 weeks, 30 patients per arm, true treatment effect −4 points. The imbalance is how much higher the treated arm's mean baseline score is, by chance.

3.0 points

0 means both arms start at the same mean score

0.60

Below 0.5 the change score is less precise than the final score

AnalysisEstimate (95% CI)SEpExpected estimateExpected SE
Final score-2.34 (-4.36 to -0.32)1.010.024-2.201.03
Change score-5.34 (-7.21 to -3.47)0.93< 0.001-5.200.92
ANCOVA-4.05 (-5.82 to -2.27)0.89< 0.001-4.000.83

Expected values are averages over many repeats of a trial with this imbalance and correlation. The final score misses the adjustment for baseline (slope 0), the change score subtracts the whole baseline (slope 1), and ANCOVA subtracts the part the data support: here the fitted slope is 0.57.

-12-10-8-6-4-2024true effect −4Final score-2.3Change score-5.3ANCOVA-4.0

observed baseline difference 3.00 points · HAM-D points on the horizontal axis

The method

How does ANCOVA adjust for the baseline?

ANCOVA fits one regression line per arm with a common slope. The treatment effect is the vertical gap between the two parallel lines, which is the difference expected between two patients with the same baseline score.

1015202515202530baseline HAM-D8-week HAM-Dmean baseline 23.1516.6412.26gap −4.39 at every baseline
The teaching data with the fitted ANCOVA model: 8-week HAM-D = 1.04 + 0.674 × baseline − 4.39 × treated. Adjusted means are read off both lines at the same baseline, usually the overall mean. Software calls them estimated marginal means or least-squares means.

final score = a + b × baseline + c × group + error

group
0 for control, 1 for treated
b
common within-arm slope: how much the final score rises per point of baseline
c
treatment effect adjusted for baseline, the number to report with its 95% CI

adjusted mean = arm mean final − b × (arm mean baseline − overall mean baseline)

Control: 15.6 − 0.674 × (21.6 − 23.15) = 16.64. Treated: 13.3 − 0.674 × (24.7 − 23.15) = 12.26. The difference, −4.39, is c. Vickers and Altman note that ANCOVA, despite its name, is a regression method. With the change score as the outcome and baseline as the covariate, the estimate of c is identical, as the EMA guideline states: once baseline is in the model, choosing the raw score or the change is only a matter of interpretation.

ANCOVA as a test of significance

The F test for group asks whether the adjusted means differ. In the teaching data, after baseline is in the model, the group sum of squares is 81.54 on 1 df and the residual mean square is 7.78 on 17 df: F = 10.48, p = 0.005. With one grouping factor of two levels, F = t² (−3.24² = 10.48). One-way ANOVA on the final scores, which ignores baseline, gives F = 1.89, p = 0.19.

Assumptions

What must be true for ANCOVA to be valid?

The assumptions of linear regression, plus one that is specific to ANCOVA: the slope for baseline is the same in both arms.

  1. Linearity: the final score rises in a straight line with the baseline score in each arm. Check a scatter plot. The EMA guideline advises a simple functional form, usually linear, in the absence of prior knowledge.
  2. Homogeneity of regression slopes: no group × baseline interaction. Teaching data: slopes 0.62 (control) and 0.72 (treated), interaction p = 0.78.
  3. Normal residuals with similar spread in both arms: teaching data residual SD 2.79, Shapiro-Wilk p = 0.86, Levene p = 0.19.
  4. Independent patients, and a covariate measured without being affected by the treatment (measured before randomisation).

If the slopes differ, the treatment effect depends on the baseline value, and a single adjusted difference describes the effect only at the baseline where it is calculated. The EMA guideline says the primary model should not include treatment by covariate interactions, and that a substantial interaction expected in advance should be handled in the trial design, with separate estimates in the relevant subgroups.

Interactive

Make the slopes unequal and watch the treatment effect change with baseline severity

With parallel lines the gap is the same at every baseline. With unequal slopes the gap widens or narrows, and the interaction test tells you whether the data support that.

Try this: move the slope difference to −0.5. The two lines stop being parallel, the interaction test becomes significant, and the treatment effect is no longer one number: it depends on how depressed the patient was at baseline.

Made-up teaching data: HAM-D at baseline and at 8 weeks in 20 control and 20 treated patients. With the slider at 0 the true effect is −4 points at every baseline score.

0.00

Control slope is 0.7. Negative values: the drug helps severely depressed patients more

0.581p for group × baseline interaction
Baseline HAM-DTreatment effect (95% CI)p
16-4.8 (-7.9 to -1.6)0.004
23.5 (mean)-4.0 (-5.3 to -2.6)< 0.001
30-3.3 (-6.1 to -0.5)0.022

Fitted slopes: control 0.73, treated 0.84 (difference 0.10). The ordinary ANCOVA assumes one common slope and reports -4.0 (-5.3 to -2.6). When the interaction is real, that single number is the effect at the average baseline only.

0102030101520253035baseline HAM-D8-week HAM-D-4.8-4.0-3.3
controltreatedtreatment effect at baseline 16, the mean, and 30

Choosing covariates

Which covariates, decided when, and which ones never?

Covariates for the primary analysis are chosen in the protocol, before any outcome data are seen, from variables measured before randomisation that are expected to influence the outcome.

  • Prespecify: the EMA guideline requires covariates in the primary analysis to be pre-specified in the protocol, and advises only a few.
  • Stratification factors: factors used to stratify the randomisation should normally be included (EMA). ICH E9 says the same: if factors stratify the design, account for them in the analysis.
  • Not because the groups look unbalanced: baseline imbalance observed post hoc is not an appropriate reason to add a covariate (EMA). Senn (1994) called significance tests of baseline homogeneity philosophically unsound, of no practical value and potentially misleading, and recommended fitting prognostic variables named in the trial plan regardless of their baseline distribution.
  • Never after randomisation: ICH E9 advises against adjusting the main analysis for covariates measured after randomisation, because they may be affected by the treatments. The EMA guideline gives duration of treatment, compliance and rescue medication as examples, and says such adjustment may hide or exaggerate the treatment effect.
drugweight changeafter randomisationHbA1c change−3.82 kgdirect pathUnadjusted effect on HbA1c−0.80 (95% CI −0.89 to −0.71)Adjusted for weight change−0.24 (95% CI −0.32 to −0.15)
Made-up illustration, 500 patients per arm. The drug lowers HbA1c directly and also through weight loss. Weight change is measured after randomisation and is caused by the drug. Adjusting for it removes the part of the effect that works through weight, and the drug appears to do less than it does.

Non-randomised groups

In 1967 Frederic Lord called attention to a basic source of confusion in interpreting comparisons of pre-existing groups, now known as Lord's paradox: change scores and ANCOVA can give different answers. Senn (2006) related the debate to Lord's paradox, and showed that equality of groups at baseline is not a necessary condition for ANCOVA to give unbiased estimates. He added that although many situations can be imagined in which ANCOVA is biased, it is very difficult to imagine one in which the change-score analysis would then be unbiased. In observational studies neither model removes confounding on its own.

Decide

Which adjustment fits your study?

Start from what you want to adjust for.

What do you want to adjust for?

Which adjustment: all possible answers
  • ANCOVA with baseline as covariate: Final score (or change) = group + baseline. Report the adjusted mean difference with its 95% CI. Include stratification factors too.
  • Prespecified interaction or subgroup analysis: The EMA guideline says the primary model should not include treatment by covariate interactions. If a substantial interaction is expected, design the trial to estimate the effect in the subgroups.
  • Caution: no analysis fixes non-randomised groups by itself: Change scores and ANCOVA can give different answers for pre-existing groups (Lord’s paradox). Think about confounding and the causal question before choosing a model.
  • Adjust for it in the primary analysis: Only a few covariates, each justified in the protocol. Randomisation stratification factors should usually be included.
  • Not in the primary analysis: Baseline imbalance observed after the trial is not a reason to add a covariate. A sensitivity analysis with it can check the robustness of the primary result.
  • Do not adjust the primary analysis for it: A variable affected by the treatment can hide or exaggerate the treatment effect (ICH E9, EMA). Use it only in exploratory analyses.

Comparison

Two pairs residents confuse

Change score or ANCOVA?

FeatureChange score analysisANCOVA with baseline
What it subtractsThe whole baseline value (slope 1)b × baseline, with b estimated from the data
Chance baseline imbalanceBiased: overcorrects, because of regression to the meanUnbiased in a randomised trial
PrecisionWorse than the final score when r < 0.5At least as precise as either alternative
Patients needed at r = 0.6 (Vickers and Altman)6854
Easy to explainYes: mean fall in each armYes, if reported as adjusted mean difference
When acceptableHigh correlation (r > 0.8), restricted randomisation (Vickers and Altman)The preferred general approach

ANOVA or ANCOVA?

FeatureOne-way ANOVAANCOVA
PredictorsCategorical factor(s) onlyCategorical factor(s) plus continuous covariate(s)
ComparesRaw group meansAdjusted (estimated marginal) means
Error termAll variation within groupsVariation within groups not explained by the covariate
Teaching trialF = 1.89, p = 0.19F = 10.48, p = 0.005
Extra assumptionsNone beyond ANOVALinear covariate effect, equal slopes, covariate not affected by treatment
As a regressionfinal ~ groupfinal ~ group + baseline

The formula

Worked example: ANCOVA by hand

Made-up teaching data: HAM-D in 10 control and 10 treated patients. Computed with Python (statsmodels 0.14.5).

ArmBaseline HAM-D8-week HAM-DMean baseline (SD)Mean 8-week (SD)
Control17, 29, 22, 20, 20, 19, 23, 26, 20, 2015, 25, 14, 20, 12, 14, 14, 14, 13, 1521.6 (3.57)15.6 (3.92)
Treated27, 25, 30, 19, 29, 25, 23, 17, 26, 2618, 12, 19, 11, 17, 13, 9, 9, 13, 1224.7 (4.08)13.3 (3.56)
  1. Within each arm, work out deviations from the arm's own means. Add over both arms: Sxx (baseline) = 264.5, Sxy (cross-products) = 178.3, Syy (8-week) = 252.5.
  2. Common slope b = Sxy ÷ Sxx = 178.3 ÷ 264.5 = 0.674.
  3. Adjusted difference c = (13.3 − 15.6) − 0.674 × (24.7 − 21.6) = −2.3 − 2.09 = −4.39.
  4. Residual SS = Syy − Sxy² ÷ Sxx = 252.5 − 120.19 = 132.31 on 20 − 3 = 17 df. Residual mean square = 7.78.
  5. SE of c = √[7.78 × (1/10 + 1/10 + 3.1² ÷ 264.5)] = 1.356.
  6. t = −4.39 ÷ 1.356 = −3.24, 17 df, p = 0.005. 95% CI = −4.39 ± 2.110 × 1.356 = −7.25 to −1.53.
Sxx, Sxy, Syy
within-arm sums of squares and cross-products
3.1
difference in mean baseline, treated − control
2.110
critical t for 17 df at 5%, two-sided
Source (adjusted)SSdfMean squareFp
Group (adjusted for baseline)81.54181.5410.480.005
Baseline (adjusted for group)120.191120.1915.440.001
Residual132.31177.78

adjusted difference = −4.39 (95% CI −7.25 to −1.53)

Critical F(1, 17) at 5% = 4.45. The model explains 53% of the variance in 8-week scores (R² = 0.526). The baseline slope is 0.674 (95% CI 0.31 to 1.04).

In a real paper

Baseline adjustment in two published trials

Open-access paper · CC BY

Hamilton DF, Beard DJ, Barker KL, Macfarlane GJ, Tuck CE, Stoddart A, et al. Targeting rehabilitation to improve outcomes after total knee arthroplasty in patients at risk of poor outcomes: randomised controlled trial. BMJ. 2020;371:m3576.

The study: A multicentre UK trial in 334 patients judged at risk of a poor outcome from their Oxford knee score six weeks after knee replacement, randomised to therapist-led outpatient rehabilitation (163) or a single physiotherapy review with home exercise (171). The primary outcome was the Oxford knee score at 52 weeks. The six-week score, which was used to select the patients, was the baseline.

What the authors wrote
“The primary outcome was evaluated using analysis of covariance, with baseline Oxford knee score and study centre as covariates in a fixed effects model.”
In plain words

Analysis of covariance: the 52-week scores were compared between arms after adjusting for each patient's starting score. Baseline Oxford knee score: the prognostic covariate, the score at six weeks. Study centre: patients came from 13 centres, and centre was added as a covariate. Fixed effects model: centre entered as ordinary categories, not as a random effect.

ReportedValueWhat it means
Randomised334163 therapist-led, 171 home exercise
Adjusted difference at 52 weeks1.91 (95% CI −0.18 to 3.99) points, P=0.07The CI includes 0 and excludes the 4-point clinically meaningful difference
Clinically important difference3 points (sample size) and 4 points (abstract)Two different thresholds are stated in the same paper
Check these when you read it
  • Were the covariates (baseline score, centre) prespecified in the protocol or statistical analysis plan?
  • Patients were selected for low early scores, so both arms will improve partly through regression to the mean. Only the comparison between arms is valid.
  • The minimum clinically important difference is given as 3 points in the methods and 4 points in the abstract. Check which one the conclusion uses.
  • Is the adjusted mean difference reported with its 95% CI, and are unadjusted means shown as well?
Open-access paper · CC BY-NC

Bartholdy C, Døssing A, Stisen ZR, Nielsen SM, Christensen R, Danneskiold-Samsøe B, et al. Effect of heated mittens on physical hand function in people with hand osteoarthritis: randomised controlled trial. BMJ. 2024;387:e078222.

The study: 200 people with hand osteoarthritis randomised to electrically heated mittens or control mittens with the heating disconnected, worn daily for six weeks. The primary outcome was the change in the AUSCAN hand function score (0 to 100) at six weeks, measured repeatedly. This variant keeps the change score as the outcome and still adjusts for baseline.

What the authors wrote
“Continuous outcomes with repeated measures (ie, AUSCAN pain, function, and stiffness subscales) were analysed using repeated measures mixed linear models, including participants as random effects, with fixed effect factors for group and week and the corresponding interaction, while adjusting for baseline values.”
In plain words

Repeated measures mixed linear models: every weekly score was used, with a random effect for each participant (see the repeated measures chapter). Group and week and the corresponding interaction: the difference between arms is allowed to change from week to week. Adjusting for baseline values: baseline is a covariate, so reporting the result as change from baseline gives the same between-group difference as analysing the scores themselves.

ReportedValueWhat it means
Participants200 (100 per arm)91 and 95 completed
Least squares mean change in function−6.3 (SE 1.2) heated, −3.3 (SE 1.2) controlAdjusted (least squares) means, not raw means
Between-group difference3.0 points (95% CI −0.4 to 6.3), P=0.09No clear effect on hand function
Check these when you read it
  • Are least squares means labelled as adjusted, and are the raw baseline means shown (68.9 and 67.6 here)?
  • Is the sign of the difference explained? Here it is reported as 3.0 in favour of heated mittens, which equals the control change minus the heated change.
  • Were only baseline values and design factors used as covariates, with nothing measured after randomisation?

Both articles: no retraction, correction or expression of concern found in PubMed, Europe PMC or scite (checked 15 September 2026).

Software

How to run it

One row per patient, with columns for group, baseline score and final score.

SPSS

  • ANCOVA: Analyze > General Linear Model > Univariate. Final score in Dependent Variable, group in Fixed Factor(s), baseline score in Covariate(s).
  • Adjusted means: the Estimated Marginal Means table gives the model-estimated mean and standard error for each group.
  • Homogeneity of slopes: in Model, choose Build terms and add group, baseline and the group × baseline interaction. The full factorial model does not contain covariate interactions. Remove the interaction again for the final ANCOVA.

R

# d: one row per patient with group, baseline, final
fit <- lm(final ~ group + baseline, data = d)   # ANCOVA
summary(fit); confint(fit)

library(emmeans)
emmeans(fit, "group")                           # adjusted (estimated marginal) means

anova(lm(final ~ group * baseline, data = d))   # homogeneity of slopes

jamovi

  • ANCOVA (jmv function ancova): final score as the dependent variable, group as a fixed factor, baseline as a covariate. The estimated marginal means option (emMeans) gives the adjusted means.
  • Homogeneity of slopes: add the group × baseline term to the model terms (modelTerms) to test it, then remove it.

Reporting

How to report it

Say which covariates were prespecified, report unadjusted means by arm and the adjusted difference with its 95% CI, and state that assumptions were checked.

"The primary outcome, HAM-D at 8 weeks, was analysed by analysis of covariance with baseline HAM-D as a prespecified covariate. Mean baseline scores were 21.6 (SD 3.57) in the control arm and 24.7 (SD 4.08) in the treated arm. Unadjusted 8-week means were 15.6 and 13.3. The adjusted means were 16.6 and 12.3, an adjusted difference of −4.4 points (95% CI −7.3 to −1.5; F(1, 17) = 10.48, p = 0.005). There was no evidence that the effect of baseline differed between arms (interaction p = 0.78)."

ICH E9 (1998), section 5.5

"A description should be given of any intentions to use baseline data to improve precision or to adjust estimates for potential baseline differences, for example by means of analysis of covariance."

EMA guideline on adjustment for baseline covariates (2015), summary

"Whenever adjusted analyses are presented, results of the treatment effect in subgroups formed by the covariates (appropriately categorised, if relevant) should be presented to enable an assessment of the model assumptions."

EMA guideline (2015), section 7.2

"Statistical testing for baseline imbalance has no role in a trial where the handling of the randomisation and blinding has been fully satisfactory."

By specialty

Baseline adjustment problems by specialty

HAM-D at baseline and 8 weeks in a trial of a new antidepressant. The treated arm starts 3 points higher by chance.

The final score understates the effect and the change score overstates it. ANCOVA with baseline HAM-D gives the unbiased adjusted difference.

Common myths

Common mistaken beliefs

  • Myth: Analysing change from baseline removes any baseline imbalance.

    Vickers and Altman (2001): analysing change does not control for baseline imbalance because of regression to the mean. In the teaching trial it gave −5.4 against −4.39 from ANCOVA.

  • Myth: Test the baseline table with p-values and adjust only for what is significant.

    Senn (1994) called such tests philosophically unsound and of no practical value. The EMA guideline says statistical testing for baseline imbalance has no role in a properly randomised trial.

  • Myth: ANCOVA is needed only when the arms are unbalanced.

    Adjusting for a covariate strongly or moderately associated with the outcome generally improves the efficiency of the analysis (EMA), even with perfect balance.

  • Myth: The more covariates, the better the adjustment.

    The EMA guideline advises only a few covariates, each justified and prespecified. Many covariates make the analysis less convincing.

  • Myth: Adjust for adherence or weight change to see the drug effect more clearly.

    Variables measured after randomisation may be affected by the treatment. Adjusting for them may hide or exaggerate the effect (ICH E9, EMA). In the illustration, −0.80 became −0.24.

  • Myth: Percentage change from baseline is a fairer outcome.

    Vickers (2001) found that percentage change from baseline had the lowest statistical power of four methods and was highly sensitive to changes in variance.

Why this matters to you

Five consequences of getting baseline adjustment wrong

  1. Two p-values, one trial

    Without a prespecified analysis you can choose p = 0.19 or p < 0.001, and any choice made after seeing both is suspect.

  2. A biased effect size

    With a chance imbalance, the final score understated and the change score overstated the adjusted effect of −4.39 points.

  3. A larger, longer study than needed

    At a baseline-final correlation of 0.6, a final-score analysis needed 85 patients where ANCOVA needed 54 in Vickers and Altman’s example.

  4. Questions at the viva

    "Why did you adjust for compliance?" and "Were your covariates in the protocol?" have only one acceptable answer each.

  5. Misreading papers

    Within-arm improvement in patients selected for high scores is often regression to the mean. Only the adjusted difference between arms estimates the treatment effect.

History

History of baseline adjustment

  1. 1886

    Francis Galton

    Published "Regression towards mediocrity in hereditary stature" in the Journal of the Anthropological Institute of Great Britain and Ireland.

  2. 1967

    Frederic M. Lord

    In "A paradox in the interpretation of group comparisons" (Psychological Bulletin), called attention to a basic source of confusion in interpreting certain group comparison data: Lord’s paradox.

  3. 1992

    L. Frison and S. J. Pocock

    Showed that analysis of covariance is the method of choice for pre- and post-treatment means, and quantified its advantage over post-treatment means and mean changes in variance and bias.

  4. 1994

    Stephen Senn

    Argued that tests of baseline homogeneity are philosophically unsound, of no practical value and potentially misleading, and that prognostic variables should be named in the trial plan and fitted in an analysis of covariance.

  5. 2001

    A. J. Vickers and D. G. Altman

    A BMJ Statistics Note recommended analysis of covariance as the preferred general approach for trials with baseline and follow-up measurements. The same year Vickers showed in a simulation study that percentage change from baseline is statistically inefficient.

  6. 2002

    S. J. Pocock, S. E. Assmann, L. E. Enos and L. E. Kasten

    Surveyed 50 trial reports and found inconsistent covariate adjustment, no clear guidelines on covariate selection, and misuse of significance tests for baseline comparability.

  7. 2005

    Adrian G. Barnett, Jolieke C. van der Pols and Annette J. Dobson

    Described regression to the mean and how to deal with it at the design and analysis stages (International Journal of Epidemiology).

  8. 2006

    Stephen Senn

    In "Change from baseline and analysis of covariance revisited", showed that equal groups at baseline are not a necessary condition for ANCOVA to give unbiased estimates.

  9. 2019

    Lei Clifton and David A. Clifton

    Showed that the change score is always correlated with the baseline score, that change scores do not address regression to the mean or baseline imbalance, and that one should adjust for baseline with ANCOVA.

Guidelines

Baseline covariates in guidelines

ICH E9 (1998), section 5.7

Pre-trial deliberations should identify covariates expected to influence the primary variable and consider how to account for them "in order to improve precision and to compensate for any lack of balance between treatment groups". "If one or more factors are used to stratify the design, it is appropriate to account for those factors in the analysis." "It is not advisable to adjust the main analyses for covariates measured after randomisation because they may be affected by the treatments."

EMA Guideline on adjustment for baseline covariates in clinical trials (EMA/CHMP/295050/2013; adopted 26 February 2015, in effect 1 September 2015)

"Covariates to be included in the primary analysis must be pre-specified in the protocol." "If a baseline value of a continuous primary outcome measure is available, then this should usually be included as a covariate. This applies whether the primary outcome variable is defined as the ‘raw outcome’ or as the ‘change from baseline’." "Variables measured after randomisation and so potentially affected by the treatment should not be included as covariates in the primary analysis."

EMA guideline (2015), section 5.6

"Note that when the baseline is included as a covariate in a standard linear model, the estimated treatment effects are identical for both ‘change from baseline’ (on an additive scale) and the ‘raw outcome’ analysis."

EMA guideline (2015), summary

"Baseline imbalance observed post hoc should not be considered an appropriate reason for including a variable as a covariate in the primary analysis." "The primary model should not include treatment by covariate interactions."

Reading a paper

Five checks on baseline adjustment in any trial

  • Was the primary analysis (final score, change score or ANCOVA) and its covariates stated in the protocol or registry before the results?
  • Is the baseline value of a continuous outcome included as a covariate?
  • Are any covariates measured after randomisation (adherence, weight change, rescue medication, length of stay)?
  • Are unadjusted means by arm reported alongside the adjusted difference and its 95% CI?
  • Do the authors test the baseline table with p-values, or interpret within-arm improvement as a treatment effect?

Why is it called that?

Word surgery

“Regression”

In everyday speech

Going backwards, getting worse.

In research

Regression to the mean: extreme first values tend to be followed by values closer to the average. Also the fitted line of one variable on another.

“Adjusted”

In everyday speech

Altered to look better.

In research

Estimated as if both groups had the same value of the covariate, such as the same mean baseline score.

“Covariate”

In everyday speech

Not an everyday word.

In research

A variable, measured before randomisation, that is related to the outcome and entered in the model alongside the treatment.

Memory trick

Final score: slope 0. Change score: slope 1. ANCOVA: the slope the data give. Choose covariates before randomisation, and never adjust for anything the treatment can change.

Regression

Latin regressio, "a going back, a return", from regredi, "to go back"

In regression to the mean, later values go back towards the average after an extreme first value.

Variance

Latin variantia, from variare, "to change"

ANCOVA analyses the variance of the outcome that remains after the covariate is taken into account.

Baseline

base, from Latin basis, "foundation" + line, from Latin linea, "linen thread, string"

The measurement taken before treatment, the starting point for every comparison.

Covariance

everyday meaning: varying together

How two variables change together. Analysis of covariance uses the covariance of baseline and final scores to estimate the slope b.

Exam answers

Model answer outlines

ANCOVA as a test of significance

MD Community Medicine 2024
  1. Definition: analysis of covariance compares the means of an outcome between groups after adjusting for one or more continuous covariates. It combines ANOVA with linear regression.
  2. Model: outcome = a + b × covariate + c × group + error. H₀: the adjusted group means are equal (c = 0).
  3. Principle: the covariate explains part of the variation in the outcome. Removing it reduces the error variance and corrects the group comparison for differences in the covariate.
  4. Steps: check linearity and equal slopes (group × covariate interaction), fit the model, calculate adjusted means at the overall covariate mean, then F = adjusted mean square for group ÷ residual mean square on (k − 1, N − k − 1) df.
  5. Worked example: HAM-D at 8 weeks, 10 per arm, baseline as covariate. Slope 0.674. Adjusted means 16.64 and 12.26. F(1, 17) = 10.48, p = 0.005. Adjusted difference −4.39 (95% CI −7.25 to −1.53). One-way ANOVA ignoring baseline: F = 1.89, p = 0.19.
  6. Assumptions: linear relation between covariate and outcome, homogeneity of regression slopes, normal residuals with equal variance, independent observations, covariate not affected by the treatment.
  7. Uses: adjusting a trial's final score for its baseline value, improving precision (fewer patients: 54 against 85 at r = 0.6 in Vickers and Altman's example), correcting for chance imbalance, pre-post designs, adjusting for prespecified prognostic factors.
  8. ANCOVA against change scores: change scores do not control for baseline imbalance because of regression to the mean. ANCOVA is the preferred general approach (Vickers and Altman, 2001).
  9. Cautions: prespecify covariates (EMA, 2015), do not adjust for variables measured after randomisation (ICH E9), do not select covariates from baseline significance tests (Senn, 1994), and interpret carefully in non-randomised groups (Lord's paradox).
  10. Software: SPSS Analyze > General Linear Model > Univariate with a covariate. R lm(final ~ group + baseline). jamovi ANCOVA.

Short note: regression to the mean and its handling in a clinical trial

  • Definition: unusually high or low measurements tend to be followed by values closer to the mean, because the chance part of a measurement does not repeat.
  • When it is largest: more measurement error, lower correlation between repeated measurements, and patients selected for extreme baseline values (Barnett et al., 2005).
  • Example: untreated people enrolled for screening BP ≥ 150 mmHg (mean 130, SD 15, correlation 0.6) fall on average from 157.0 to 146.2 mmHg.
  • Handling in design: a randomised control group, more than one baseline measurement, entry criteria and baseline measured on separate occasions.
  • Handling in analysis: compare arms, not within-arm change, and adjust for baseline with ANCOVA. Change scores do not remove it (Vickers and Altman, 2001; Clifton and Clifton, 2019).

The one thing to remember

In a randomised trial with a baseline measurement, analyse the final score with the baseline as a prespecified covariate, and never adjust for anything measured after randomisation.

All worked examples and interactive elements use made-up numbers for teaching. Statistics were computed in Python (SciPy 1.15.3, statsmodels 0.14.5). Software steps were checked against the IBM SPSS Statistics documentation, the CRAN emmeans reference manual, the R stats documentation and the jamovi documentation on 15 September 2026.

Sources: Hamilton DF et al. BMJ 2020;371:m3576 (doi:10.1136/bmj.m3576, PMID 33051212). Bartholdy C et al. BMJ 2024;387:e078222 (doi:10.1136/bmj-2023-078222, PMID 39689944). Vickers AJ, Altman DG. Statistics notes: Analysing controlled trials with baseline and follow up measurements. BMJ 2001;323(7321):1123–4 (doi:10.1136/bmj.323.7321.1123, PMID 11701584). Vickers AJ. BMC Med Res Methodol 2001;1:6 (doi:10.1186/1471-2288-1-6, PMID 11459516). Frison L, Pocock SJ. Stat Med 1992;11(13):1685–704 (doi:10.1002/sim.4780111304, PMID 1485053). Senn S. Stat Med 1994;13(17):1715–26 (doi:10.1002/sim.4780131703, PMID 7997705). Senn S. Stat Med 2006;25(24):4334–44 (doi:10.1002/sim.2682, PMID 16921578). Pocock SJ, Assmann SE, Enos LE, Kasten LE. Stat Med 2002;21(19):2917–30 (doi:10.1002/sim.1296, PMID 12325108). Barnett AG, van der Pols JC, Dobson AJ. Int J Epidemiol 2005;34(1):215–20 (doi:10.1093/ije/dyh299, PMID 15333621). Clifton L, Clifton DA. Trials 2019;20(1):43 (doi:10.1186/s13063-018-3108-3, PMID 30635021). Galton F. J Anthropol Inst G B Irel 1886;15:246–63 (doi:10.2307/2841583). Lord FM. Psychol Bull 1967;68(5):304–5 (doi:10.1037/h0025105). ICH E9 Statistical Principles for Clinical Trials, 1998. EMA Guideline on adjustment for baseline covariates in clinical trials, EMA/CHMP/295050/2013, 26 February 2015.

Past papers

TNMGRMU questions on ancova and baseline adjustment

Questions from MD, MS, DM, MCh and Ph.D. papers that this chapter helps you answer. Tap a sitting to open the university's own question paper.

  1. ×1ANCOVA test of significanceMD Community Medicine · Oct 2024